Histologic chorioamnionitis in extremely preterm births, microbiological findings and infant outcome.

BACKGROUND: Histologic chorioamnionitis (HCA) is most often caused by ascending bacterial infection originating from the cervicovaginal tract. OBJECTIVES: To investigate whether HCA with a fetal inflammatory response (FIR) has a worse clinical outcome than HCA alone. Further, if FIR or a positive maternal microbiologic culture obtained prior to birth were related to adverse neonatal outcomes in a cohort of extremely preterm (EP) neonates. METHODS: Prospective observational cohort study recruiting EP singleton pregnancies (gestational age at birth ≤28 weeks) with confirmed HCA. FIR was defined by fetal neutrophils in the chorionic vessels and/or umbilical vessels. Positive culture was defined as growth of potentially pathogenic bacteria in a sample from the cervicovaginal tract prior to birth, or if a cervicovaginal culture was lacking, a culture result from the placenta was used. Logistic regression was used to estimate odds ratios and 95% confidence intervals for the associations between FIR, a positive culture result and adverse outcomes, defined as bronchopulmonary dysplasia (BPD), brain pathology assessed by magnetic resonance imaging, retinopathy of prematurity, necrotizing enterocolitis, early-onset neonatal sepsis, and perinatal death. A composite outcome variable included one or more adverse outcomes. RESULTS: We included 71 cases with HCA, of which 51 (72%) had FIR. Maternal age, rate of clinical chorioamnionitis (CCA), preterm pre-labor rupture of membranes (PPROM), the number of women receiving antenatal steroids and antibiotics, and the rate of positive maternal cultures of potentially pathogenic bacteria were all significantly higher in the HCA with FIR. Neonates in the FIR group had significantly higher levels of blood leukocytes compared to those without. FIR was associated with a longer interval from PPROM to delivery (log-rank test: p = .022). Microbiological sampling had been performed in 63 (89%) cases, of which 60 (95%) were cervicovaginal samples. No associations were found between a positive culture and adverse neonatal outcomes, in contrast to FIR, that was significantly associated to BPD and brain pathology. CONCLUSIONS: In a cohort of EP pregnancies with confirmed HCA, the presence of FIR was associated with advanced maternal age, CCA, PPROM, antenatal steroids and antibiotics, and a positive maternal culture of potentially pathogenic bacteria. However, the presence of FIR, and not a positive culture, was associated with adverse neonatal outcomes.

Human Organotypic Airway and Lung Organoid Cells of Bronchiolar and Alveolar Differentiation Are Permissive to Infection by Influenza and SARS-CoV-2 Respiratory Virus.

The ongoing coronavirus disease 2019 (COVID-19) pandemic has led to the initiation of unprecedented research efforts to understand the pathogenesis mediated by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). More knowledge is needed regarding the cell type-specific cytopathology and its impact on cellular tropism. Furthermore, the impact of novel SARS-CoV-2 mutations on cellular tropism, alternative routes of entry, the impact of co-infections, and virus replication kinetics along the respiratory tract remains to be explored in improved models. Most applied virology models are not well suited to address the remaining questions, as they do not recapitulate the histoarchitecture and cellular composition of human respiratory tissues. The overall aim of this work was to establish from single biopsy specimens, a human adult stem cell-derived organoid model representing the upper respiratory airways and lungs and explore the applicability of this model to study respiratory virus infection. First, we characterized the organoid model with respect to growth pattern and histoarchitecture, cellular composition, and functional characteristics. Next, in situ expression of viral entry receptors, including influenza virus-relevant sialic acids and SARS-CoV-2 entry receptor ACE2 and TMPRSS2, were confirmed in organoids of bronchiolar and alveolar differentiation. We further showed successful infection by pseudotype influenza A H7N1 and H5N1 virus, and the ability of the model to support viral replication of influenza A H7N1 virus. Finally, successful infection and replication of a clinical isolate of SARS-CoV-2 were confirmed in the organoids by TCID50 assay and immunostaining to detect intracellular SARS-CoV-2 specific nucleocapsid and dsRNA. The prominent syncytia formation in organoid tissues following SARS-CoV-2 infection mimics the findings from infected human tissues in situ. We conclude that the human organotypic model described here may be particularly useful for virology studies to evaluate regional differences in the host response to infection. The model contains the various cell types along the respiratory tract, expresses respiratory virus entry factors, and supports successful infection and replication of influenza virus and SARS-CoV-2. Thus, the model may serve as a relevant and reliable tool in virology and aid in pandemic preparedness, and efficient evaluation of antiviral strategies.

Placental histology predicted adverse outcomes in extremely premature neonates in Norway-population-based study.

AIM: We evaluated the role of placental pathology in predicting adverse outcomes for neonates born extremely preterm (EPT) before 28 weeks of gestation. METHODS: This was a prospective observational study of 123 extremely preterm singletons born in a hospital in western Norway, and the placentas were classified according to the Amsterdam criteria. The associations between histologic chorioamnionitis (HCA), by the presence or the absence of a foetal inflammatory response (FIR+ or FIR-), maternal vascular malperfusion (MVM) as a whole and adverse neonatal outcomes were evaluated by logistic regression analyses. Adverse outcomes were defined as perinatal death, necrotising enterocolitis (NEC), bronchopulmonary dysplasia (BPD), brain pathology by magnetic resonance imaging at term-equivalent age, retinopathy of prematurity and early-onset neonatal sepsis. The results are reported as odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: HCA was associated with NEC (OR 12.2, 95% CI 1.1 to 137.1). HCA/FIR+ was associated with BPD (OR 14.9, 95% CI 1.8-122.3) and brain pathology (OR 9.8, 95% CI 1.4-71.6), but HCA/FIR- was not. The only neonatal outcome that MVM was associated with was low birthweight. CONCLUSION: Placental histology provided important information when assessing the risk of adverse neonatal outcomes following EPT birth.

Double paternal uniparental isodisomy 7 and 15 presenting with Beckwith-Wiedemann spectrum features.

Here we describe for the first time double paternal uniparental isodisomy (iUPD) 7 and 15 in a baby boy with features in the Beckwith-Wiedemann syndrome spectrum (BWSp) (placentomegaly, hyperinsulinism, enlarged viscera, hemangiomas, and earlobe creases) in addition to conjugated hyperbilirubinemia. His phenotype was also reminiscent of genome-wide paternal uniparental isodisomy. We discuss the most likely origin of the UPDs: a maternal double monosomy 7 and 15 rescued by duplication of the paternal chromosomes after fertilization. So far, paternal UPD7 is not associated with an abnormal phenotype, whereas paternal UPD15 causes Angelman syndrome. Methylation analysis for other clinically relevant imprinting disorders, including BWSp, was normal. Therefore, we hypothesized that the double UPD affected other imprinted genes. To look for such effects, patient fibroblast RNA was isolated and analyzed for differential expression compared to six controls. We did not find apparent transcription differences in imprinted genes outside Chromosomes 7 and 15 in patient fibroblast. PEG10 (7q21.3) was the only paternally imprinted gene on these chromosomes up-regulated beyond double-dose expectation (sixfold). We speculate that a high PEG10 level could have a growth-promoting effect as his phenotype was not related to aberrations in BWS locus on 11p15.5 after DNA, RNA, and methylation testing. However, many genes in gene sets associated with growth were up-regulated. This case broadens the phenotypic spectrum of UPDs but does not show evidence of involvement of an imprinted gene network.

Induction of alveolar and bronchiolar phenotypes in human lung organoids.

Patient-derived organoids have revolutionized biomedical research and therapies by "transferring the patient into the Petri dish". In vitro access to human lung organoids representing distal lung tissue, i.e. alveolar organoids, would facilitate research pertaining to a wide range of medical conditions and might open for a future approach to individualized treatment.We propose a protocol to derive a single human lung biopsy towards both alveolar and bronchiolar organoids. By modulating Wnt pathway, we obtained a differential gene expression of the main markers for both subtypes, such as a higher expression of surfactant protein C in alveolar organoids or a higher expression of mucine 5AC in bronchiolar organoids. Although the specific cell enrichment was not complete, the differentiation was observed as early as passage 1 based on morphology, and confirmed by QPCR and histology at passage 2. These results are consistent with a functional specification of lung epithelium towards both alveoli- and bronchi-enriched organoids from first passages.

Electromagnetic inductance plethysmography to study airflow after nebulized saline in bronchiolitis.

BACKGROUND: Spirometric effects from therapeutic interventions in infants with severe respiratory distress cannot readily be measured, hampering development of better treatment for acute bronchiolitis. Inhaled normal saline is regularly used in these infants, with little knowledge of how this influences lung physiology. OBJECTIVES: Assess feasibility of infant lung function testing using electromagnetic inductance plethysmography (EIP) in a clinical setting in a busy pediatric department, and explore effects from inhaled normal saline on tidal flow-volume loops in infants with acute bronchiolitis. METHODS: Observational study conducted at the Children's Clinic, Haukeland University Hospital, Bergen, Norway during the winters 2016 and 2017, enrolling children with bronchiolitis below six months of age. EIP was performed immediately before and 5 and 20 min after saline inhalation. EIP is a noninvasive method to measure tidal breathing parameters by quantifying volume changes in the chest and abdomen during respiration. The method consists of an electromagnet/antenna and a patient vest. RESULTS: EIP was successfully applied in 36/45 (80%) enrolled infants at mean (standard deviation) age 2.9 (2.5) months, after a hospital stay of 2.2 (1.9) days. After saline inhalation, tidal expiratory to inspiratory time ratio (Te/Ti) had increased significantly, whereas the other relevant flow/volume parameters had changed numerically in a direction compatible with a more obstructive pattern. CONCLUSIONS: EIP could successfully be used to obtain tidal breathing parameters in infants with respiratory distress and appears a promising tool for assessment of therapeutic interventions in bronchiolitis. Saline inhalations should be used with caution as placebo in intervention studies.

Ventilator flow data predict bronchopulmonary dysplasia in extremely premature neonates.

Early prediction of bronchopulmonary dysplasia (BPD) may facilitate tailored management for neonates at risk. We investigated whether easily accessible flow data from a mechanical ventilator can predict BPD in neonates born extremely premature (EP). In a prospective population-based study of EP-born neonates, flow data were obtained from the ventilator during the first 48 h of life. Data were logged for >10 min and then converted to flow-volume loops using custom-made software. Tidal breathing parameters were calculated and averaged from ≥200 breath cycles, and data were compared between those who later developed moderate/severe and no/mild BPD. Of 33 neonates, 18 developed moderate/severe and 15 no/mild BPD. The groups did not differ in gestational age, surfactant treatment or ventilator settings. The infants who developed moderate/severe BPD had evidence of less airflow obstruction, significantly so for tidal expiratory flow at 50% of tidal expiratory volume (TEF50) expressed as a ratio of peak tidal expiratory flow (PTEF) (p=0.007). A compound model estimated by multiple logistic regression incorporating TEF50/PTEF, birthweight z-score and sex predicted moderate/severe BPD with good accuracy (area under the curve 0.893, 95% CI 0.735-0.973). This study suggests that flow data obtained from ventilators during the first hours of life may predict later BPD in premature neonates. Future and larger studies are needed to validate these findings and to determine their clinical usefulness.

Lung function at term in extremely preterm-born infants: a regional prospective cohort study.

OBJECTIVES: To compare lung function of extremely preterm (EP)-born infants with and without bronchopulmonary dysplasia (BPD) with that of healthy term-born infants, and to determine which perinatal characteristics were associated with lung function at term and how predictive these measurements were for later respiratory health in EP-born infants. METHODS: Perinatal variables were recorded prospectively, and tidal breathing parameters were measured at term-equivalent age using electromagnetic inductance plethysmography. Respiratory morbidity was defined by hospital readmissions and/or treatment with asthma medications during the first year of life. RESULTS: Fifty-two EP-born infants (mean gestational age 261, range 226-276 weeks) and 45 term-born infants were included. There was evidence of significant airway obstruction, higher tidal volumes and increased minute ventilation in the EP-born infants with and without BPD, although generally more pronounced for those with BPD. Male gender, antenatal steroids and number of days on continuous positive airway pressure were associated with lung function outcomes at term. A prediction model incorporating two unrelated tidal breathing parameters, BPD, birth weight z-score and gender, predicted respiratory morbidity in the first year of life with good accuracy (area under the curve 0.818, sensitivity and specificity 81.8% and 75.0%, respectively). CONCLUSION: Lung function measured at term-equivalent age was strikingly abnormal in EP-born infants, irrespective of BPD. Tidal breathing parameters may be of value in predicting future pulmonary health in infants born premature. TRIAL REGISTRATION NUMBER: NCT01150396; Results.

Electromagnetic inductance plethysmography is well suited to measure tidal breathing in infants.

Reliable, accurate and noninvasive methods for measuring lung function in infants are desirable. Electromagnetic inductance plethysmography has been used to perform infant spirometry and VoluSense Pediatrics (VSP) (VoluSense, Bergen, Norway) represents an updated version of this technique. We aimed to examine its accuracy compared to a validated system measuring airflow via a facemask using an ultrasonic flowmeter. We tested 30 infants with postmenstrual ages between 36 to 43 weeks and weights from 2.3 to 4.8 kg, applying both methods simultaneously and applying VSP alone. Agreement between the methods was calculated using Bland-Altman analyses and we also estimated the effect of applying the mask. Mean differences for all breathing parameters were within ±5.5% and limits of agreement between the two methods were acceptable, except perhaps for peak tidal expiratory flow (PTEF). Application of the facemask significantly increased tidal volume, minute ventilation, PTEF, the ratio of inspiratory to expiratory time and the ratio of expiratory flow at 50% of expired volume to PTEF. VSP accurately measured tidal breathing parameters and seems well suited for tidal breathing measurements in infants under treatment with equipment that precludes the use of a facemask.

A new non-invasive method of infant spirometry demonstrates a level of repeatability that is comparable to traditional methods.

AIM: The FloRight system provides novel non-invasive infant spirometry based on electromagnetic inductance plethysmography. We investigated the consistency of repeated measurements carried out in a Norwegian neonatal intensive care unit (NICU) using the system and how well these were tolerated. METHODS: Tidal flow-volume loops were obtained from 10 preterm infants at discharge, 10 stable growing preterm infants weighing about 1500 g and 10 term-born infants. A nurse experienced with the system measured all patients before and after meals, and these measurements were repeated by nurses new to the system. RESULTS: The measurements were well tolerated by the infants. The repeatability for the two parameters 'tidal volume' (Vt) and 'time to peak tidal expiratory flow to total expiratory time' (Tptef/Te) were relatively poor, similar to previous methods. However, the repeatability was good for the new 'flow-volume gravity mid-point' (FVg) parameter. Repeatability was better for term than preterm infants, when measurements were obtained by the experienced nurse and for measurements carried out before meals. CONCLUSION: The FloRight system proved feasible in a NICU setting. The repeatability of the lung function measurements was similar to those reported for traditional infant spirometry. The nurse's experience and the relationship to meals appeared to be important.