Necrotizing enterocolitis after intravitreal bevacizumab in an infant with Incontinentia Pigmenti - a case report.

BACKGROUND: Incontinentia Pigmenti is a rare disease affecting multiple organs. Fifty of patients show affection of the eye with retinopathy and possible amaurosis being the worst outcome. Treatment has commonly been panretinal laser coagulation but intravitreal application of bevacizumab as VEGF-inhibitor has shown to effectively suppress retinal neovascularization. CASE PRESENTATION: A six-week-old female infant with Incontinentia Pigmenti developed a foudroyant necrotizing enterocolitis shortly after intravitreal injection of bevazicumab due to a retinopathy with impending tractional detachment of the left eye. Since the onset of abdominal symptoms occurred immediately after the intravitreal application, a link between the two events seemed likely. Sequential analyses of the VEGF serum concentrations showed a massive suppression of endogenous VEGF with only a very slow recovery over weeks. Such a severe systemic adverse event has not been reported after intravitreal treatment with bevacizumab in an infant. CONCLUSION: This case report shows a relevant systemic uptake of bevacizumab after intravitreal application as suppressed VEGF levels show. There seems to be a connection between suppressed VEGF levels and the onset of necrotizing enterocolitis. Therefore, treatment with bevacizumab should be carefully considered and further research is needed to assess this drug's safety profile.

[Lung Function in Childhood and Adolescence: Influence of Prematurity and Bronchopulmonary Dysplasia]. / Lungenfunktion in Kindheit und Adoleszentenalter: Einfluss von Frühgeburtlichkeit und bronchopulmonaler Dysplasie.

INTRODUCTION: The introduction of prenatal steroids, surfactant replacement therapy and gentle ventilation modes has reduced short term respiratory morbidity and increased survival of very preterm infants. However, there is some evidence that prenatal factors, the extend of prematurity and bronchopulmonary dysplasia (BPD) may affect pulmonary function in childhood and adolescence. METHODS: We have performed a comprehensive review on the outcome of pulmonary function after premature birth before 32 weeks of gestation in the era of surfactant replacement therapy and tried to evaluate the influence of chorioamnionitis, intrauterine growth retardation (IUGR), maternal metabolic syndrome, prematurity and BPD on long term pulmonary function. RESULTS: Some children and adolescents born very preterm may experience significant airflow reduction. The bronchial obstruction in these patients is not entirely reversible by inhalative ß2-mimetics. The degree of pulmonary function impairment is partly correlated with the degree of BPD. Abnormalities in pulmonary diffusion capacity may occur after extreme prematurity, but also in patients with moderate and severe BPD. IUGR may have a negative impact on later pulmonary function in very children. There is insufficient data to assess the preterm impact of chorioamnionitis or maternal metabolic syndrome on later lung function. CONCLUSION: Infants born before 32 weeks of gestational age in the surfactant era still carry an increased risk to suffer an impaired pulmonary function in childhood and adolescence, particularly if they survived with BPD. Long term pulmonary care for these patients should take place in specialized centers.

Outbreak of multidrug-resistant Escherichia coli sequence type 131 in a neonatal intensive care unit: efficient active surveillance prevented fatal outcome.

BACKGROUND: Outbreaks of infections with multidrug-resistant bacteria in neonatal intensive care units (NICUs) pose a major threat, especially to extremely preterm infants. This study describes a 35-day outbreak of multidrug-resistant Escherichia coli (E. coli) in a tertiary-level NICU in Germany. AIM: To underline the importance of surveillance policies in the particularly vulnerable cohort of preterm infants and to describe the efficacy of outbreak control strategies. METHODS: Data were collected retrospectively from medical reports. Infants and environment were tested for E. coli. FINDINGS: The outbreak affected a total of 13 infants between 25(+1) and 35(+0) weeks of gestation with seven infants showing signs of infection. The outbreak strain was identified as E. coli sequence type 131. Environmental screening provided no evidence for an environmental source. Through colonization surveillance and immediate and adequate treatment of potentially infected preterm infants, no fatalities occurred. Outbreak control was achieved by strict contact precautions, enhanced screening and temporary relocation of the NICU. Relocation and reconstruction improved the NICU's structural layout, focusing on isolation capacities. Follow-up indicated carriage for several months in some infants. CONCLUSION: Routine surveillance allowed early detection of the outbreak. The identification of carriers of the outbreak strain was successfully used to direct antibiotic treatment in case of infection. Enhanced hygienic measures and ward relocation were instrumental in controlling the outbreak.

Late Vitamin K Deficient Bleeding in 2 Young Infants--Renaissance of a Preventable Disease.

INTRODUCTION: Late vitamin K deficiency bleeding in young infants is a rare disorder which occurs almost exclusively in breast-fed infants who did not receive proper vitamin K prophylaxis at birth and who might additionally suffer from cholestasis. Its impact on morbidity is high since in 50% of the cases it presents with intracranial hemorrhage with a mortality rate of 20% and life-long neurologic sequelae in 30% of the affected infants. CASE REPORTS: 2 male infants were both admitted to our unit at the age of 5 weeks with subdural hematoma with midline shift due to late vitamin K deficiency bleeding. Both infants did not receive the recommended Vitamin K prophylaxis in Germany. One patient presented with cholestatic jaundice on admission as an additional risk factor. DISCUSSION: Parents who in the apparent best interest for their children refuse the recommended and well established vitamin K prophylaxis at birth leading to the reappearance of late vitamin K deficiency bleeding. These parents also tend to refuse routine immunizations of childhood in later life, which not only have an impact on their own child but might bear a risk for the whole community. CONCLUSION: It is the responsibility of health-care takers to show increased awareness to the growing number of parents refusing vitamin K prophylaxis at birth and educate them properly about the devastating consequences of late vitamin K deficiency bleeding.

Serum GADD45a methylation is a useful biomarker to distinguish benign vs malignant prostate disease.

BACKGROUND: Prostate-specific antigen (PSA) screening for prostate cancer results in a large number of unnecessary prostate biopsies. There is a need for specific molecular markers that can be used in combination with PSA to improve the specificity of PSA screening. We examined GADD45a methylation in blood DNA as a molecular marker for prostate cancer diagnosis. METHODS: The study included 82 men, with PSA levels >4 ng ml(-1) and/or abnormal digital rectal exam, who underwent prostate biopsy. We compared GADD45a methylation in DNA from serum and buffy coat in 44 patients (22 prostate cancer and 22 benign). GADD45a methylation in serum DNA was examined in 82 patients (34 cancer and 48 benign). RESULTS: There was no significant difference in buffy coat GADD45a methylation between cancer and benign patients. Serum GADD45a methylation was significantly higher in cancer than in benign patients. Classification and regression tree predictive model for prostate cancer including risk groups defined by PSA, free circulating DNA (fcDNA) level and GADD45a methylation yielded specificity of 87.5%, sensitivity of 94.1% and receiver operator characteristic curve area of 0.937. CONCLUSIONS: Serum GADD45a methylation in combination with PSA and fcDNA level was useful in distinguishing benign from prostate cancer patients.

An 8-month history of meningitis in an extremely low birth weight infant? - Long-lasting Infection with Ureaplasma parvum.

INTRODUCTION: Ureaplasma spp. have been implicated in the pathogenesis of both preterm labor and neonatal morbidity including pneumonia and sepsis and the development of chronic lung disease of prematurity. Data on Ureaplasma meningitis are limited and partly controversially discussed. PATIENT: We report the unique case of a 9-month-old infant with progressive internal hydrocephalus of unknown origin and developmental delay due to a history of>200 days of inflammation of the central nervous system. The female extremely low birth weight infant had been referred to our hospital for ventriculoperitoneal shunt implantation. She had been born at 26+3 weeks of gestation with a birth weight of 940 g. With the exception of a moderate respiratory distress syndrome, postnatal period had been reported uneventful. However, internal hydrocephalus had become manifest at 4 weeks of postnatal age. Intraventricular hemorrhage had not been documented by cranial ultrasound and magnetic resonance imaging. Cerebrospinal fluid (CSF) analysis had repetitively revealed pronounced inflammation reflected by pleocytosis (50-86 leukocytes/µL, 60% lymphocytes), CSF protein levels of 578-1,026 mg/dL and undetectable CSF glucose. Although suggesting bacterial meningitis, microbial diagnostics had not been indicative, and empirical antibiotics had not affected the CSF findings. On admission to our hospital, CSF analysis still documented significant inflammation (125 leukocytes/µL, CSF protein 565 mg/dL, CSF glucose<2 mg/dL). RESULTS: Due to a prenatal history of cerclage, we initiated microbial diagnostics on Ureaplasma spp. and Mycoplasma hominis. U. parvum was detected in CSF by culture and PCR, no other pathogens were isolated. On intravenous treatment with chloramphenicol, CSF profile continuously normalized, and cultures and PCR became negative. Treatment was continued for 3 weeks, and the infant was discharged after uncomplicated ventriculoperitoneal shunt placement. During a 12-month observation period she has shown encouraging recovery. CONCLUSION: In preterm infants, in particular, internal hydrocephalus of unknown origin and sustained CSF inflammation are highly suggestive of Ureaplasma meningitis. Our case highlights that infection may escape detection if not explicitly considered, since microbial diagnosis requires complex media and PCR.

Optimization of serum EVELISA for milk testing of Johne's disease.

Johne's disease (JD) or paratuberculosis, caused by Mycobacterium avium ssp. paratuberculosis (MAP), is one of the most economically important diseases of dairy cattle. Control of JD could be achieved by good herd management practices, and diagnosis; however, this approach has been hampered by the low sensitivity of currently available enzyme-linked immunosorbent assay (ELISA) tests. In our previous study, we developed a sensitive serum ELISA test, ethanol-vortex enzyme-linked immunosorbent assay (EVELISA), using ethanol extract of MAP. The objective of this study is to demonstrate that the EVELISA can be used for detection of anti-MAP antibodies in milk samples. In this study, we tested and optimized concentrations of antigen, milk, and secondary antibody for better differentiation of milk samples of cattle with MAP infections from those of cattle in JD-free herds. We evaluated five environmental mycobacteria as absorbents of cross-reactive antibodies in milk and found that the mycobacteria had no significant effect on EVELISA results. Using the optimized conditions, a total of 57 milk samples from Holstein dairy cattle (37 animals found positive on the fecal polymerase chain reaction test and 20 animals from JD-free herds) were tested for anti-MAP antibody in milk by using the EVELISA method. The average of ELISA values in the JD-positive milk samples (mean±SD=0.355±0.455) was significantly higher than that in the JD-negative milk samples (mean±SD=0.071±0.011). These results warrant further studies for evaluation and validation of the EVELISA for milk testing of cattle for JD.

[Intrauterine inflammation and its sequelae: does chorioamnionitis really matter for outcome of very low birth weight infants?]. / Intrauterine Inflammation und ihre Folgen: Spielt Chorioamnionitis eine Rolle für Organschäden bei sehr kleinen Frühgeborenen?

Infections in utero and chorioamnionitis are major risk factors for spontaneous, very early premature birth. Thus chorioamnionitis contributes significantly to prematurity-associated morbidity and mortality. Evidence for a gestation-independent effect of chorioamnionitis on the outcome of very low birth weight infants is much more difficult to obtain as most of the studies addressing this issue lack a normal "control group", as prematurity is mostly associated with some kind of prenatal pathology with a potential influence on neonatal outcome. Moreover, major advances in perinatal and neonatal care for this high-risk group have mitigated the impact of chorioamnionitis on morbidity and mortality of very low birth weight infants. Histological chorioamnionitis is associated with a lower incidence and severity of respiratory distress syndrome. However, short-term maturational effects on the lung are associated with a higher susceptibility for postnatal noxious events, such as mechanical ventilation, thus contributing to the risk of bronchopulmonary dysplasia. Data regarding the importance of chorioamnionitis for brain damage of the very premature infant are inconsistent although meta-analyses have shown an increased risk of cystic periventricular leukomalacia and cerebral palsy after exposure to inflammation in utero. Very recent epidemiological studies suggest a role of chorioamnionitis in the aetiology and pathogenesis of retinopathy of prematurity.

Nosocomial sepsis in neonatal intensive care: inevitable or preventable?

Very low birth weight (VLBW) infants are at high risk to develop a neonatal nosocomial sepsis. The incidence of neonatal nosocomial, late-onset sepsis (LOS) is about 20-30%, but a rate of up to 43% has been reported among neonates with a birth weight of 400-750 g. Preventive and treatment strategies for neonatal sepsis in VLBW infants are aiming to enhance the infant's host defence mechanisms. Neonatal immunodeficiencies include quantitative and qualitative deficits in phagocytes, complement components, and immunoglobulins. A considerable number of immune strategies has been investigated in carefully designed multicentre trials. These include exchange transfusion, neutrophil transfusion, hematopoietic growth factors such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF), intravenous immunoglobulins (IVIG), and others. Since none of these interventions was able to reduce the mortality rate of immature preterm infants, the current evidence does not support the use of any of the immune strategies for prevention or treatment of neonatal sepsis. Decreasing the burden of intensive care and following strict hygiene programs at NICUs may be the most promising current strategies to minimise nosocomial infection.

[Significance of nitric oxide inhalation (NO) in preterm infants< 34 weeks of gestation]. / Stellenwert der Inhalation von Stickstoffmonoxid (NO) bei Frühgeborenen<34 Gestationswochen.

In 2001, NO was approved as a therapeutic agent in Europe for the treatment of persistent pulmonary hypertension in late preterm infants >34 weeks of gestational age and term newborns. Recent observational studies suggest, that preterm infants <34 weeks of gestation with acute hypoxic lung failure could benefit from inhaled NO (iNO) by improved oxygenation. To date, 21 randomised-controlled trials have enrolled 3 336 preterm infants <34 weeks of gestation for iNO treatment. Overall, iNO treatment does not reduce the rate of bronchopulmonary dysplasia (BPD) or death compared to controls. In addition, iNO treatment of preterm infants with hypoxic respiratory failure or increased risk of BPD does not affect the combined incidence of death and BPD. However, early prophylactic use of iNO in preterm infants with respiratory distress seems to improve survival without BPD or severe cerebral damage. Current data of long term neurological outcome of iNO-treated preterm infants do not seem to justify iNO administration. Outside of well designed clinical trials iNO-treatment of preterm infants can currently not be recommended.

The role of surfactant treatment in preterm infants and term newborns with acute respiratory distress syndrome.

Surfactant treatment in preterm infants and term newborns with (acute respiratory distress syndrome) ARDS-like severe respiratory failure has become part of an individualized treatment strategy in many intensive care units around the world. These babies constitute heterogeneous groups of gestational ages, lung maturity, as well as of the underlying disease processes and postnatal interventions. The pathophysiology of respiratory failure in preterm infants is characterized by a combination of primary surfactant deficiency and surfactant inactivation as a result of plasma proteins leaking into the airways from areas of epithelial disruption and injury. Various pre- and postnatal factors, such as exposure to chorioamnionitis, pneumonia, sepsis and asphyxia, induce an injurious inflammatory response in the lungs of preterm infants, which may subsequently affect surfactant function, synthesis and alveolar stability. Surfactant inactivation--and dysfunction--is also a hallmark in newborns with meconium aspiration syndrome (MAS), pneumonia and other disorders affecting the pulmonary function. Although for the majority of suggested indications no data from randomized controlled trials exist, a surfactant replacement that counterbalances surfactant inactivation seems to improve oxygenation and lung function in many babies with ARDS without any apparent negative side effects. Newborns with MAS will definitely benefit from a reduced need for extracorporeal membrane oxygenation (ECMO). Clinical experience seems to justify surfactant treatment in neonates with ARDS.

[Preterm and term infants with acute respiratory distress syndrome (ARDS): what is the role of surfactant substitution?]. / Früh- und Neugeborene mit einem Acute Respiratory Distress Syndrome (ARDS): welche Rolle spielt die Surfactantsubstitution?

BACKGROUND: Surfactant treatment in preterm infants and term newborns with ARDS-like severe respiratory failure has become part of an individualised treatment strategy in many intensive care units around the world. DISCUSSION: These babies constitute heterogeneous groups with regards to gestational age, lung maturity, underlying disease processes and postnatal interventions. The pathophysiology of respiratory failure in preterm infants is characterised by a combination of primary surfactant deficiency and surfactant inactivation as a result of plasma proteins leaking into the airways from areas of epithelial disruption and injury. Various pre- and postnatal factors, - such as exposure to chorioamnionitis, pneumonia, fluid lung, sepsis and asphyxia - can induce an injurious inflammatory response in the lung which may subsequently affect surfactant function, synthesis and alveolar stability. CONCLUSION: Surfactant inactivation and dysfunction is also a hallmark in newborns with meconium aspiration syndrome (MAS), for which a beneficial effect of exogenous surfactant replacement. i.e., reduction of need for ECMO, could be shown. Although for the majority of the above-mentioned diseases process data from randomised, controlled trials are lacking, it is evident from clinical experience that surfactant replacement which counterbalances surfactant inactivation seems to improve oxygenation and lung function in many babies with ARDS without apparent negative side effects. Thus surfactant treatment seems to be justified in many neonates with ARDS.

[Leukemoid reaction in extremely immature preterm infants]. / Leukämoide Reaktion bei extrem unreifen Frühgeborenen.

Extremely immature preterm infants rarely present with a leukocytosis exceeding 30,000/microL. The pathogenetic sequence leading to leukemoid reactions in non-malignant diseases remains to be elucidated. Potential triggers for leukemoid reactions in premature infants include prenatal corticosteroids, chorioamnionitis and funisitis or systemic infection. In the two-year period from 2006 to 2007 all infants with a gestational age of less than 26 weeks were screened for leukocytosis. Among our cases, one preterm infant presented with a leukocyte count of 229,300/microL at the age of 48 hours, lasting throughout the first three weeks of life. Impairment of microcirculation and resulting organ dysfunction were not observed. Thus, invasive therapeutic procedures, which are routinely initiated in hyperleukocytosis in accompanying malignant diseases, may not have the same significance in extremely immature preterm infants and should be executed in these patients on an individual basis and with extreme caution.

All-trans retinoic acid and intra-amniotic endotoxin-mediated effects on fetal sheep lung.

All-trans retinoic acid (RA) is a potent modulator of lung development. Chorioamnionitis, which is frequently associated with preterm birth, causes fetal lung inflammation and improves lung function but also results in alveolar simplification and microvascular injury. Endotoxin-mediated chorioamnionitis reduces RA concentration in the fetal lung to 16% of control values. We hypothesized that administration of RA to the fetus before induction of chorioamnionitis would preserve septation of the distal airspaces. Time-mated ewes with singletons were assigned to receive a fetal intramuscular treatment with 20,000 IU of RA in olive oil (or olive oil only) 3 hr prior to intra-amniotic injection of endotoxin (20 mg, E. coli 055:B5) or saline, at 124-day gestational age and 7 days after the fetal treatment. The right cranial lung lobe was processed for morphometric analysis. RA treatment did not affect chorioamnionitis-induced fetal and systemic inflammation or interleukin-8 concentrations in lung tissue. RA administration alone did not alter lung structure. Relative to control lungs (5 +/- 3 mL/kg), lung volume increased similarly with endotoxin (22 +/- 4 mL/kg) or RA plus endotoxin (20 +/- 3 mL/kg; P < 0.05). Alveolar wall thickness was 4.2 +/- 0.3 mum after endotoxin-induced chorioamnionitis, 6.0 +/- 0.4 mum in controls (P < 0.05 versus endotoxin) and 5.5 +/- 0.2 mum after RA and endotoxin (P < 0.05 versus control, n.s. versus endotoxin). The ratio of airspace versus tissue was 4.6 +/- 0.3 in endotoxin-induced chorioamnionitis, 2.1 +/- 0.3 in controls and 4.1 +/- 0.5 after RA and endotoxin. We conclude that fetal treatment with RA did not prevent inflammation-induced alveolar simplification.

Nonventilatory strategies for prevention and treatment of bronchopulmonary dysplasia--what is the evidence?

This review briefly summarizes the evidence for a number of mainly drug-related strategies to prevent or treat bronchopulmonary dysplasia (BPD). Oxygen supplementation is frequently used in neonatal units and oxygen toxicity plays an important role in the pathogenesis of BPD. However, current evidence for an optimal oxygen saturation for extremely premature infants is scarce. This gap in knowledge will hopefully be closed by a number of ongoing or prospective trials addressing this issue. The role of inhalational nitric oxide in the prevention of BPD is still unclear despite existing data from a number of large randomized trials. Early administration of caffeine seems to confer a benefit with regard to BPD. Prophylactic or early application of surfactant may also be beneficial. High intramuscular doses of vitamin A slightly reduce the incidence of the disease. There is currently no evidence supporting other nutritional interventions to prevent BPD. Anti-inflammatory drugs, like alpha(1)-proteinase inhibitor, pentoxifylline and azithromycin, and antioxidants, like N-acetylcysteine and superoxide dismutase, have not been proven effective yet. Diuretics can ameliorate lung function, but there is no evidence supporting their long-term use. Ureaplasma urealyticum colonization of airways is associated with an increased risk of BPD. However, there is no proof for an effect of erythromycin on BPD. The potential roles for therapies like bombesin-like peptide-blocking antibodies or Clara cell 10-kDa protein have yet to be defined.

Connective tissue growth factor does not affect transforming growth factor-beta 1-induced Smad3 phosphorylation and T lymphocyte proliferation inhibition.

Transforming growth factor-beta 1 (TGF-beta(1)) is a key regulator of immune tolerance. TGF-beta(1) controls T lymphocyte activation and is involved in the immunosuppressive function and generation of regulatory T lymphocytes. Connective tissue growth factor (CTGF) has an essential role in the formation of connective tissue and blood vessels. CTGF expression is induced by TGF-beta(1) in several cell types and CTGF mediates several of the downstream actions of TGF-beta(1). Since little is known about the potential synergy between CTGF and TGF-beta(1) in T lymphocyte biology, the purpose of the present study was to determine whether CTGF can modulate TGF-beta(1)-mediated effects on human CD4+ T lymphocytes. Human recombinant CTGF was expressed in HEK293 cells. rCTGF was biologically active demonstrated by induction of proliferation in the endothelial cell line EA hy 926. rCTGF alone did not potentiate or diminish anti-CD3-induced CD4+ T lymphocyte proliferation and did not activate the Smad signaling pathway in CD4+ T lymphocytes. Furthermore, rCTGF did not attenuate TGF-beta(1)-mediated inhibition of CD4+ T lymphocyte proliferation and TGF-beta(1)-induced Smad signaling in CD4+ T lymphocytes. These results indicate that rCTGF had no detectable effects of its own on human CD4+ T lymphocytes and did not potentiate the effects of low amounts of TGF-beta(1) on human CD4+ T lymphocytes. Overall, these data support the hypothesis that CTGF does not act on CD4+ T lymphocytes.