Gene expression profiles of adult peripheral and cord blood mononuclear cells altered by lipopolysaccharide.

BACKGROUND: Neonatal Gram-negative sepsis is often characterized by a fulminant clinical course, compared to adults, resulting in higher morbidity and mortality. Genome-wide gene expression analysis can provide insights into the molecular alterations in sepsis. OBJECTIVES: To evaluate in vitro activation of the neonatal and adult immune system, gene expression patterns were compared in mononuclear cells from cord (CBMNC) and adult peripheral blood (APBMNC). METHODS: To better understand the influence of early molecular signals on the effects of sepsis, Affymetrix gene profiling (8,475 genes) was done on RNA isolated from CBMNC and APBMNC without and after incubation with 100 ng/ml lipopolysaccharide (LPS). RESULTS: We demonstrated significant alterations in the expression of 108 CBMNC and APBMNC genes compared with basal levels, 188 significant changes in CBMNC and 97 in APBMNC, including cytokines, chemokines and immunoregulatory genes. Furthermore, we found 5 genes showing a significant interaction effect between cell type and LPS stimulation, including tumor necrosis factor receptor superfamily, member 6 (FAS), absent in melanoma 2, malic enzyme 1, hemoglobin epsilon 1, and trans-prenyltransferase. CONCLUSIONS: These results provide further support for a marked difference in the pathogenesis of neonatal and adult sepsis and may stimulate additional studies to investigate some of the altered genes as potential new targets for diagnostic tools and therapeutic strategies.

Can an EPR support the concept of family-centred, individualized developmental care of premature infants and newborns?

At the University Children's Hospital Heidelberg the concept of 'Developmental, Family-Centred, Individual Care of Premature Infants and Newborns' was introduced to support optimal growth of premature infants. This interdisciplinary concept requires cooperation of different specialists. A well operating communication is a precondition for such cooperation. As a patient's record is not only used for storing information but also for exchanging information, the question was if a complete electronic patient record (EPR), in contrast to the existing patient's record, could sensibly support this new concept of care. To answer this question the whole communication of the staff in the infants ward was analysed using different observation methods. These observations delivered several issues which showed that an EPR could improve communication and workflow. Therefore an EPR for the neonatology at the University Children's Hospital Heidelberg can now be designed on the basis of our communication concept.

Pain management and the effect of guidelines in neonatal units in Austria, Germany and Switzerland.

BACKGROUND: Painful invasive procedures are frequently performed on preterm infants admitted to a neonatal intensive care unit (NICU). The aim of the present study was to investigate current pain management in Austrian, German and Swiss NICU and to identify factors associated with improved pain management in preterm infants. METHODS: A questionnaire was sent to all Austrian, German and Swiss pediatric hospitals with an NICU (n = 370). Pain assessment and documentation, use of analgesics for 13 painful procedures, presence of written guidelines for pain management and the use of 12 analgesics and sedatives were examined. RESULTS: A total of 225 units responded (61%). Pain assessment and documentation and frequent analgesic therapy for painful procedures were performed more often in units using written guidelines for pain management and in those treating >50 preterm infants at <32 weeks of gestation per year. This was also the case for the use of opioid analgesics and sucrose solution. Non-opioid analgesics were used more often in smaller units and in units with written guidelines. There was a broad variation in dosage of analgesics and sedatives within all groups. CONCLUSION: Pain assessment, documentation of pain and analgesic therapy are more frequently performed in NICU with written guidelines for pain management and in larger units with more than 50 preterm infants at <32 weeks of gestation per year.

Effects of erythropoietin on erythrocyte deformability in non-transfused preterm infants.

AIM: Suppression of erythropoiesis due to low plasma erythropoietin levels is an important factor in the development of anaemia of prematurity. Premature infants may therefore be treated with recombinant human erythropoietin (rhEPO). This prospective, randomised and controlled study was designed to find out whether rhEPO treatment improves erythrocyte deformability in preterm infants. METHODS: Sixteen infants were treated with rhEPO (250 IU/kg three times weekly) a total of 15 times beginning on day of life 5 whereas fifteen infants served as controls. Haemoglobin concentration, haematocrit, reticulocyte count, ferritin level and erythrocyte deformability were measured on days 5, 14, 28, 42 and 63. Erythrocyte elongation was determined as an indicator of erythrocyte deformability using a shear stress diffractometer (Rheodyn SSD) at shear forces of 0.3 to 60 Pa. RESULTS: Haemoglobin concentration was significantly higher on days 28 and 42 and reticulocyte percentage on day 28 in the rhEPO group compared to the controls. Serum ferritin was lower in the rhEPO group on day 28. Erythrocyte deformability was significantly increased on days 28 and 42 in the infants receiving rhEPO. We found a strong relationship between erythrocyte elongation and reticulocyte count. CONCLUSION: RhEPO markedly increases the erythropoiesis in preterm infants in the critical first weeks of life and the anaemia of prematurity is obviously reduced. The erythrocyte deformability improved under rhEPO treatment. Erythrocyte deformability was significantly related to the reticulocyte count indicating that the improvement of erythrocyte deformability was due to the formation of well-deformable young erythrocytes.

A haemophagocytic lymphohistiocytosis (HLH)-like picture following breastmilk transmitted cytomegalovirus infection in a preterm infant.

Due to septic illness, neutro- and thrombocytopenia, blocked myelopoiesis, CMV-positive breast milk and CMV-pp65 antigen in bone marrow mononuclear cells, CMV-related HLH was presumed in a breast fed neonate (gestational age 24 weeks). Treatment was successful with foscarnet and methylprednisolone. HLH may be a complication of post-natal CMV-infection acquired from breast milk.

Intracerebellar hemorrhage in premature infants: sonographic detection and outcome.

AIMS: Intracerebellar hemorrhage is a rarely confirmed diagnosis in preterm infants in comparison to peri-/intraventricular hemorrhage. This study evaluates the incidence of intracerebellar hemorrhage and neurological outcome in preterm infants. METHODS: 260 infants with gestational age of 22-32 weeks were studied prospectively by cranial ultrasound. Neurodevelopmental outcome was examined in the first three years of life. RESULTS: 15 infants had intracranial hemorrhage grade II-IV (10 intraventricular, 6 intracerebellar hemorrhage). Neurodevelopmental follow-up showed that one infant with intracerebellar hemorrhage is severely handicapped, two have moderate and two mild impairments and one has no sequelae. CONCLUSION: Cerebellar hemorrhage is not rare if ultrasound examination is specifically focused on cerebellar lesions.

[Interrelation between well-being of the mother and heart rate variability of her preterm infant]. / Zusammenhänge zwischen dem Wohlbefinden der Mutter und der Herzfrequenzvariabilität von Frühgeborenen.

The relationship between mothers' well-being and the heart rate variability of their preterm babies was investigated. In order to study a possible influence of the mother's well-being on the calming quality of her voice and thereby on the heart rate variability of her preterm infant, maternal/paternal stress and competences as well as family functionality were assessed via respective questionnaires. (N = 30) Preterm babies at the postnatal age of approximately 4 weeks were acoustically stimulated with the voice of their own mother. Various heart rate variability measures (NN interval mean value, NN interval median, variance of NN intervals, standard deviation of NN intervals, pnn 6,25, RMSSD, SDSD and RSA) were recorded 15 minutes before, 15 minutes during and 15 minutes after the acoustic stimulation. Non-REM sleep sections of 2 minutes duration were matter of analyses. The correlations between the mothers' well-being and their babies' heart rate variability indicate a strong relationship. The correlations point out that a higher family functionality is associated with a higher heart rate variability of preterm babies. Contradictory to the expectations, higher burden and lower resources as well as lower competences of the mothers were associated with a higher heart rate variability of the preterm babies. Simultaneous real-time investigations of the mothers' and the babies' heart rate variability during a live mother-baby-interaction seems necessary to provide further explanations.

Alpha 2,3-sialyltransferase-IV is essential for L-selectin ligand function in inflammation.

L-selectin belongs to the C-type lectin family of glycoproteins and is constitutively expressed on most leukocytes. L-selectin mediates leukocyte rolling in inflamed microvessels and high endothelial venules (HEV) via binding to specific carbohydrate structures on selectin ligands. Previous studies using sialidase treatment suggested a role of sialic acid residues in L-selectin-dependent rolling. To investigate the role of the alpha2,3-sialyltransferase (ST3Gal)-IV on L-selectin ligand activity in vivo, we studied leukocyte rolling in inflamed venules of the cremaster muscle and in Peyer's patch HEV of ST3Gal-IV-deficient mice and littermate control mice. In cremaster muscle venules with or without TNF-alpha treatment, L-selectin-dependent rolling was almost completely abolished in ST3Gal-IV(-/-) mice. In both models, L-selectin interacts with P-selectin glycoprotein ligand-1 (PSGL-1) presented by adherent leukocytes and leukocyte fragments, but not with endothelial L-selectin ligands. In contrast, L-selectin-dependent rolling in Peyer's patch HEV, which is mediated by unknown endothelial L-selectin ligands, was not impaired in the absence of ST3Gal-IV. Our in vivo data show that PSGL-1, the molecule responsible for L-selectin-mediated leukocyte interactions in inflammation, is dependent on ST3Gal-IV, while alpha2,3-sialylation by ST3Gal-IV is not necessary for L-selectin ligand activity on high endothelial cells of Peyer's patch HEV.

Naive regulatory T cells: a novel subpopulation defined by resistance toward CD95L-mediated cell death.

Most CD4(+)CD25(hi)FOXP3(+) regulatory T cells (T(regs)) from adult peripheral blood express high levels of CD45RO and CD95 and are prone to CD95L-mediated apoptosis in contrast to conventional T cells (T(convs)). However, a T(reg) subpopulation remained consistently apoptosis resistant. Gene microarray and 6-color flow cytometry analysis including FOXP3 revealed an increase in naive T-cell markers on the CD95L-resistant T(regs) compared with most T(regs). In contrast to T(regs) found in adult humans, most CD4(+)CD25(+)FOXP3(+) T cells found in cord blood are naive and exhibit low CD95 expression. Furthermore, most of these newborn T(regs) are not sensitive toward CD95L similar to naive T(regs) from adult individuals. After short stimulation with anti-CD3/CD28 monoclonal antibodies (mAbs), cord blood T(regs) strongly up-regulated CD95 and were sensitized toward CD95L. This functional change was paralleled by a rapid up-regulation of memory T-cell markers on cord blood T(regs) that are frequently found on adult memory T(regs). In summary, we show a clear functional difference between naive and memory T(regs) that could result in different survival rates of those 2 cell populations in vivo. This new observation could be crucial for the planning of therapeutic application of T(regs).

Pharmacokinetics of a netilmicin loading dose on the first postnatal day in preterm neonates with very low gestational age.

OBJECTIVE: Pharmacokinetic parameters are important for dose adjustment of aminoglycosides, but they are highly variable in neonates. In this study the pharmacokinetics of a netilmicin loading dose was investigated on the first postnatal day in preterm neonates with very low gestational age (GA). METHODS: In an open prospective study, 20 neonates with GA between 22.9 and 32.0 weeks and suspected postnatal bacterial infection received an intravenous loading dose of 5 mg/kg netilmicin over 1 h during the first postnatal day. Netilmicin serum concentrations were determined by an enzyme multiplied immunoassay. RESULTS: The systemic clearance of netilmicin normalized to body weight (BW) was not significantly different in three GA subgroups (0.59+/- 0.02 ml/min/kg for GA <24 weeks, 0.72+/-0.14 ml/min/kg for GA 24-27 weeks, and 0.62+/-0.19 ml/min/kg for GA 27-32 weeks, P=0.123). Similar results were also obtained for serum elimination half-time and for the distribution volume normalized to BW. Multiple regression analysis showed that systemic clearance and volume of distribution (both not normalized to BW) significantly correlated with BW (P<0.0001) but not with GA. In the entire group, 20% of peak concentrations were below the target of 6 mg/l, and 63% of trough concentrations were above the target of 2 mg/l. CONCLUSION: In neonates with very low GA, the pharmacokinetic parameters of netilmicin determined after an intravenous loading dose were not dependent on GA when normalized to BW. A number of neonates did not reach targeted peak and trough netilmicin serum concentrations, suggesting that a higher loading dose and a prolonged dosing interval might enhance the effectiveness and safety of netilmicin in preterm neonates immediately after birth.

Effectiveness and side effects of an escalating, stepwise approach to indomethacin treatment for symptomatic patent ductus arteriosus in premature infants below 33 weeks of gestation.

OBJECTIVE: Symptomatic patent ductus arteriosus (sPDA) is a common problem in premature infants and can be treated effectively with intravenous indomethacin, leading to permanent ductal closure in 70% to 80% of infants. Infants who do not respond to pharmacologic closure of the duct ultimately have to undergo surgical or interventional closure of the PDA. Optimizing the pharmacologic treatment could offer an interesting approach to reduce the number of infants who need surgical closure of the duct. METHODS: We conducted a retrospective analysis in infants who were <33 weeks' gestation, had sPDA, and were treated with high-dose intravenous indomethacin. From 1993 to 2002, 129 infants with sPDA received indomethacin after diagnosis of sPDA was confirmed by echocardiography. Treatment was started in all infants with intravenous indomethacin (0.2 mg/kg given 5 times at 0 hours, 12 hours, 24 hours, 48 hours, and 72 hours). When the ductus was still open at 36 hours, indomethacin every 12 hours was continued and single doses increased up to 1 mg/kg until ductal closure was achieved. RESULTS: In 68 (53%) of 129 infants who were treated with indomethacin, ductal closure occurred during intermediate-dose indomethacin therapy (up to 1.5 mg/kg total dose). In the 61 initial nonresponders, the continuation of indomethacin led to ductal closure in 59 infants. When infants who were treated with an intermediate dose were compared with the initial nonresponders, no differences in the incidences of renal or electrolyte abnormalities, gastrointestinal bleeding, intraventricular hemorrhage, or periventricular leukomalacia were found. CONCLUSIONS: High-dose indomethacin after intermediate-dose therapy resulted in an overall closure rate of 98.5% (127 of 129). Although single indomethacin doses of up to 1 mg/kg were given, high-dose indomethacin was safe.

[Individual developmental care based on the Newborn Individualized Developmental Care and Assessment Program (NIDCAP)]. / Individuelle, entwicklungsfördernde Pflege basierend auf dem Newborn Individualized Developmental Care and Assessment Program (NIDCAP).

The number of preterm deliveries is increasing. Major advances in neonatology now enable even infants with a very low gestational age to survive. These infants have to spend months in the intensive care unit, while they have to deal with a lot of painful medical procedures, nursing care units, excessive noise and light, all differing from the normal uterine environment. The immature preterm infant nervous system is in a vulnerable phase of development. Early exposure to stress, or the mismatch of the preterm infant brain with the unphysiologic intensive care environment, may be linked to long-term developmental impairments reported in these children. The Newborn Individualized Developmental Care and Assessment Program (NIDCAP) is a systematic method of assessing the individual needs of preterm newborns. In an attempt to mimic uterine environment, care according to the NIDCAP principles is reducing infant distress and in the same way supporting the individual aibilities. Thus NIDCAP may have an effect on the preterm infant neurodevelopmental outcome.

Incontinentia pigmenti in combination with decreased IgG subclass concentrations in a female newborn.

Incontinentia pigmenti (IP) is a rare neurocutaneous disorder caused by mutations in the NEMO (NF-kappaB essential modulator) gene. Skin lesions are typically the first manifestation of IP though they may be accompanied by multiple malformations. This report presents the case of a female newborn with early onset of IP lesions within the 1st day of life. After the age of 1 month she developed frequent episodes of severe gastroenteritis. Examination of the immune system revealed low concentrations of IgG subclasses. This study suggests that, contrary to previous belief, IP is associated with immune deficiency.

Buprenorphine and norbuprenorphine concentrations in human breast milk samples determined by liquid chromatography-tandem mass spectrometry.

Buprenorphine (BUP) is considered to be safe during pregnancy. However, the extent of BUP transfer into breast milk has not been investigated thoroughly. Because the drug concentration in the milk is 1 of the determinants in the assessment of the exposure risk, a rapid and sensitive LC-MS/MS method has been developed and evaluated to measure BUP and norbuprenorphine (norBUP) concentrations in milk. A solid-phase and 2 liquid-liquid extraction procedures have been compared. The lower limits of detection and quantification were 0.05 ng/mL and 0.18 ng/mL for BUP and 0.05 ng/mL and 0.20 ng/mL for norBUP, respectively, using a sample volume of 0.5 mL milk. BUP and norBUP concentrations determined from 10 random breast milk samples collected over 4 successive days from a lactating woman during buprenorphine maintenance therapy ranged from 1.0 to 14.7 and 0.6 to 6.3 ng/mL, respectively. Drug exposure of the infant may be considered to be low. Further investigations may seek to extend these preliminary findings to evaluate an infant's level of BUP exposure through breast milk.

Drotrecogin alpha (activated) in neonatal septic shock.

A 12-d-old neonate suffering from group B streptococcal septic shock was treated with 24 microg/kg/h recombinant human activated protein C [rhAPC, drotrecogin alpha (activated)] for 96 h. The protein C activity increased from 5% to 53% after rhAPC infusion. The patient recovered within 14 d without any adverse effects.

Cleavage of L1 in exosomes and apoptotic membrane vesicles released from ovarian carcinoma cells.

PURPOSE: The L1 adhesion molecule (CD171) is overexpressed in human ovarian and endometrial carcinomas and is associated with bad prognosis. Although expressed as a transmembrane molecule, L1 is released from carcinoma cells in a soluble form. Soluble L1 is present in serum and ascites of ovarian carcinoma patients. We investigated the mode of L1 cleavage and the function of soluble L1. EXPERIMENTAL DESIGN: We used ovarian carcinoma cell lines and ascites from ovarian carcinoma patients to analyze soluble L1 and L1 cleavage by Western blot analysis and ELISA. RESULTS: We find that in ovarian carcinoma cells the constitutive cleavage of L1 proceeds in secretory vesicles. We show that apoptotic stimuli like C2-ceramide, staurosporine, UV irradiation, and hypoxic conditions enhance L1-vesicle release resulting in elevated levels of soluble L1. Constitutive cleavage of L1 is mediated by a disintegrin and metalloproteinase 10, but under apoptotic conditions multiple metalloproteinases are involved. L1 cleavage occurs in two types of vesicles with distinct density features: constitutively released vesicles with similarity to exosomes and apoptotic vesicles. Both types of L1-containing vesicles are present in the ascites fluids of ovarian carcinoma patients. Soluble L1 from ascites is a potent inducer of cell migration and can trigger extracellular signal-regulated kinase phosphorylation. CONCLUSIONS: We suggest that tumor-derived vesicles may be an important source for soluble L1 that could regulate tumor cell function in an autocrine/paracrine fashion.

Effects of Intralipid infusion on hemorheology and peripheral resistance in neonates and children.

Deleterious microcirculatory effects of Intralipid (IL) infusion may be caused by hemorheological or vascular effects. The aim of this investigation was to study vascular and hemorheological effects of IL in preterm and fullterm neonates and children. Ten preterm newborns, 10 fullterm neonates, and 10 children received an initial infusion of IL (0.6 g/kg) over 4 h. Calf blood flow (venous occlusion plethysmography), blood pressure (Dinamap), whole blood and plasma viscosity (capillary viscometer), red blood cell deformability (rheoscope), and erythrocyte aggregation (aggregometer) were measured before and after administration of IL. Plasma triglyceride levels showed the greatest increase in preterm infants. Whole blood viscosity decreased by about 10% in all three groups because of a similar reduction in hematocrit. Red blood cell aggregation decreased by about 20% after IL infusion. Blood pressure rose by 10%, and peripheral blood flow declined by about 10% in the three groups. Vascular hindrance, a calculation of blood pressure divided by blood flow and viscosity, was raised by about 20%, suggesting marked vasoconstriction of peripheral arteries. Vasoconstriction rather than hemorheological changes during infusion of IL may play a crucial role in the pathogenesis of circulatory alterations in parenterally-fed neonates.