Detection and quantitative analysis of patient-ventilator interactions in ventilated infants by deep learning networks.

BACKGROUND: The study of patient-ventilator interactions (PVI) in mechanically ventilated neonates is limited by the lack of unified PVI definitions and tools to perform large scale analyses. METHODS: An observational study was conducted in 23 babies randomly selected from 170 neonates who were ventilated with SIPPV-VG, SIMV-VG or PSV-VG mode for at least 12 h. 500 breaths were randomly selected and manually annotated from each recording to train convolutional neural network (CNN) models for PVI classification. RESULTS: The average asynchrony index (AI) over all recordings was 52.5%. The most frequently occurring PVIs included expiratory work (median: 28.4%, interquartile range: 23.2-40.2%), late cycling (7.6%, 2.8-10.2%), failed triggering (4.6%, 1.2-6.2%) and late triggering (4.4%, 2.8-7.4%). Approximately 25% of breaths with a PVI had two or more PVIs occurring simultaneously. Binary CNN classifiers were developed for PVIs affecting ≥1% of all breaths (n = 7) and they achieved F1 scores of >0.9 on the test set except for early triggering where it was 0.809. CONCLUSIONS: PVIs occur frequently in neonates undergoing conventional mechanical ventilation with a significant proportion of breaths containing multiple PVIs. We have developed computational models for seven different PVIs to facilitate automated detection and further evaluation of their clinical significance in neonates. IMPACT: The study of patient-ventilator interactions (PVI) in mechanically ventilated neonates is limited by the lack of unified PVI definitions and tools to perform large scale analyses. By adapting a recent taxonomy of PVI definitions in adults, we have manually annotated neonatal ventilator waveforms to determine prevalence and co-occurrence of neonatal PVIs. We have also developed binary deep learning classifiers for common PVIs to facilitate their automatic detection and quantification.

Computational analysis of neonatal ventilator waveforms and loops.

BACKGROUND: Modern neonatal ventilators allow the downloading of their data with a high sampling rate. We wanted to develop an algorithm that automatically recognises and characterises ventilator inflations from ventilator pressure and flow data. METHODS: We downloaded airway pressure and flow data with 100 Hz sampling rate from Dräger Babylog VN500 ventilators ventilating critically ill infants. We developed an open source Python package, Ventiliser, that includes a rule-based algorithm to automatically discretise ventilator data into a sequence of flow and pressure states and to recognise ventilator inflations and an information gain approach to identify inflation phases (inspiration, expiration) and sub-phases (pressure rise, pressure plateau, inspiratory hold etc.). RESULTS: Ventiliser runs on a personal computer and analyses 24 h of ventilation in 2 min. With longer recordings, the processing time increases linearly. It generates a table reporting indices of each breath and its sub-phases. Ventiliser also allows visualisation of individual inflations as waveforms or loops. Ventiliser identified >97% of ventilator inflations and their sub-phases in an out-of-sample validation of manually annotated data. We also present detailed quantitative analysis and comparison of two 1-hour-long ventilation periods. CONCLUSIONS: Ventiliser can analyse ventilation patterns and ventilator-patient interactions over long periods of mechanical ventilation. IMPACT: We have developed a computational method to recognize and analyse ventilator inflations from raw data downloaded from ventilators of preterm and critically ill infants. There have been no previous reports on the computational analysis of neonatal ventilator data. We have made our program, Ventiliser, freely available. Clinicians and researchers can use Ventiliser to analyse ventilator inflations, waveforms and loops over long periods. Ventiliser can also be used to study ventilator-patient interactions.

Cerebral oxygenation in preterm infants during maternal singing combined with skin-to-skin care.

BACKGROUND: Our aim was to investigate the effect of music therapy in combination with skin-to-skin care (SSC) on regional cerebral oxygenation (rSO2) measured with near-infrared spectroscopy (NIRS) in premature infants and to study physiological stability during the interventions. METHODS: This was a prospective single-center observational cohort study conducted in a tertiary neonatal intensive care unit. The study consisted of four phases: (1) baseline measurements in an incubator for 30 min; (2) quiet SSC for 30 min (SSC-Pre); (3) SSC with live maternal singing accompanied by live guitar music for 20 min (SSC-Music); (4) final quiet SSC for another 30 min (SSC-Post). RESULTS: The primary outcome measure of mean rSO2 for the 31 preterm infants analyzed showed a significant increase from baseline during SSC-Music (76.87% vs 77.74%, p = 0.04) and SSC-Post (76.87% vs 78.0%, p = 0.03) phases. There were no significant changes observed in heart rate (HR), peripheral oxygen saturation (SpO2), and cerebral fractional tissue oxygen extraction (cFTOE). The coefficient of variation (CV) of rSO2 and SpO2 decreased during each intervention phase. CONCLUSION: Combining music therapy with SSC appears to be safe in preterm neonates. The impact of the small increase in rSO2 and reduced variability of SpO2 and rSO2 warrants further investigation. IMPACT: Music therapy combined with skin-to-skin care (SSC) is safe in clinically stable premature infants and could be encouraged as part of developmental care. This is the first report where near-infrared spectroscopy (NIRS) was used to detect the simultaneous effect of music therapy and SSC on cerebral rSO2 in preterm infants. Music therapy with SSC caused a modest increase in rSO2 and decreased the coefficient of variation of rSO2 and peripheral oxygen saturation (SpO2), which suggest short-term benefits for preterm infants.

Fifteen-minute consultation: How to interpret and manage ventilator alarms in the neonatal intensive care unit.

Modern neonatal intensive care units use a large number of monitoring and therapeutic devices. Most of them have alarms with varying degree of standardisation. Mechanical ventilator alarms alert clinicians about technical problems with equipment, acute deterioration of the patient, changes in his or her clinical condition or in respiratory mechanics. However, frequent ventilator alarms interfere with developmental care and they may lead to alarm fatigue, missed alarms and clinical incidents. In this article, we discuss the most important ventilator alarms and their clinical significance. We also provide advice how to respond to ventilator alarms and how to set alarm limits.

Neonatal encephalopathy therapy optimization for better neuroprotection with inhalation of CO2: the HENRIC feasibility and safety trial.

BACKGROUND: There is an association between hypocapnia and adverse neurodevelopmental outcome in infants with neonatal encephalopathy (NE). Our aim was to test the safety and feasibility of 5% CO2 and 95% air inhalation to correct hypocapnia in mechanically ventilated infants with NE undergoing therapeutic hypothermia. METHODS: Ten infants were assigned to this open-label, single-center trial. The gas mixture of 5% CO2 and 95% air was administered through patient circuits if the temperature-corrected PCO2 ≤40 mm Hg. The CO2 inhalation was continued for 12 h or was stopped earlier if the base deficit (BD) level decreased <5 mmol/L. Follow-up was performed using Bayley Scales of Infant Development II. RESULTS: The patients spent a median 95.1% (range 44.6-98.5%) of time in the desired PCO2 range (40-60 mm Hg) during the inhalation. All PCO2 values were >40 mm Hg, the lower value of the target range. Regression modeling revealed that BD and lactate had a tendency to decrease during the intervention (by 0.61 and 0.55 mmol/L/h, respectively), whereas pH remained stable. The rate of moderate disabilities and normal outcome was 50%. CONCLUSIONS: Our results suggest that inhaled 5% CO2 administration is a feasible and safe intervention for correcting hypocapnia.

Flow cytometry for near-patient testing in premature neonates reveals variation in platelet function: a novel approach to guide platelet transfusion.

BACKGROUND: Neonatal haemorrhaging is often co-observed with thrombocytopenia; however, no evidence of a causal relationship with low platelet count has been reported. Regardless, the administration of a platelet transfusion is often based upon this parameter. Accurate measurement of platelet function in small volumes of adult blood samples by flow cytometry is well established and we propose that the use of the same technology could provide complementary information to guide the administration of platelet transfusions in premature neonates. METHODS: In 28 neonates born at 27-41 weeks gestation, platelet function after stimulation agonists was measured using fibrinogen binding and P-selectin expression (a marker of degranulation). RESULTS: Platelets of neonates with gestation of ≤36 weeks (n = 20) showed reduced fibrinogen binding and degranulation with ADP, and reduced degranulation with CRP-XL. Degranulation Scores of 7837 ± 5548, 22,408 ± 5301 and 53,131 ± 12,102 (mean ± SEM) identified significant differences between three groups: <29, 29-36 and >36 weeks gestation). Fibrinogen binding and degranulation responses to ADP were significantly reduced in suspected septic neonates (n = 6) and the Fibrinogen Binding scores clearly separated the septic and healthy group (88.2 ± 10.3 vs 38.6 ± 12.2, P = 0.03). CONCLUSIONS: Flow cytometric measurement of platelet function identified clinically different neonatal groups and may eventually contribute to assessment of neonates requiring platelet transfusion.

Volume Guaranteed Ventilation During Neonatal Transport.

OBJECTIVES: To compare tidal volumes, inflating pressures and other ventilator variables of infants receiving synchronized intermitted mandatory ventilation with volume guarantee during emergency neonatal transport with those of infants receiving synchronized intermitted mandatory ventilation without volume guarantee. DESIGN: Retrospective observational study. SETTING: A regional neonatal emergency transport service. PATIENTS: We enrolled 77 infants undergoing emergency neonatal transfer. Forty-five infants were ventilated with synchronized intermittent mandatory ventilation with volume guarantee and 32 with synchronized intermitted mandatory ventilation without volume guarantee. INTERVENTIONS: Infants received synchronized intermitted mandatory ventilation with or without volume guarantee during interhospital emergency neonatal transport using a Fabian + nCPAP evolution neonatal ventilator (Software Version: 4.0.1; Acutronic Medical Instruments, Hirzel, Switzerland). MEASUREMENTS AND MAIN RESULTS: We downloaded detailed ventilator data with 0.5 Hz sampling rate. We analyzed data with the Python computer language and its data science packages. The mean expiratory tidal volume of inflations was lower and less variable in infants ventilated with volume guarantee than in babies ventilated without volume guarantee (group median 4.8 vs 6.0 mL/kg; p = 0.001). Babies ventilated with synchronized intermittent mandatory ventilation with volume guarantee had on average lower and more variable peak inflating pressures than babies ventilated without volume guarantee (group median 15.5 vs 19.5 cm H2O;p = 0.0004). With volume guarantee, a lower proportion of the total minute ventilation was attributed to ventilator inflations rather than to spontaneous breaths between inflations (group median 66% vs 83%; p = 0.02). With volume guarantee, babies had fewer inflations with tidal volumes greater than 6 mL/kg and greater than 8 mL/kg (group medians 3% vs 44% and 0% vs 7%, respectively; p = 0.0001). The larger tidal volumes in the non-volume guarantee group were not associated with significant hypocapnia except in one case. CONCLUSIONS: During neonatal transport, synchronized intermittent mandatory ventilation with volume guarantee ventilation reduced the occurrence of excessive tidal volumes, but it was associated with larger contribution of spontaneous breaths to minute ventilation compared with synchronized intermitted mandatory ventilation without volume guarantee.

Leak Compensation During Volume Guarantee With the Dräger Babylog VN500 Neonatal Ventilator.

OBJECTIVES: To investigate how compensating for endotracheal tube leaks by targeting the leak-compensated tidal volume affects measured physiologic and ventilator variables during neonatal mechanical ventilation. DESIGN: Retrospective observational study. SETTING: A level III Neonatal ICU. PATIENTS: We enrolled 30 neonates who were ventilated using synchronized intermittent positive pressure mode with volume guarantee and had at least 12 hours of continuous detailed recording of ventilation variables. INTERVENTIONS: Infants were treated using the Dräger VN500 ventilator (Dräger, Lübeck, Germany), which uses a proprietary algorithm to measure and compensate for endotracheal tube leaks. Eleven were ventilated without leak compensation and 19 with leak compensation. MEASUREMENTS AND MAIN RESULTS: Detailed ventilation data were collected and analyzed at 1 Hz, with intermittent blood gas values. The percentage of leak was less than 20% in 73% of leak-compensated inflations, and the volume of the leak compensation was less than 1 mL/kg in 97.3% of inflations. Between the two groups, ventilation variables were comparable, except the percentage of leak that was significantly (p = 0.005) higher in the recordings with leak compensation. Without leak compensation, the mean expired tidal volume was maintained very close to the set level up to 50% leak, but with leaks greater than 50%, it declined progressively. With leak compensation, the mean leak-compensated expired tidal volume was well maintained even with leak greater than 90% although with large variability. Without leak compensation, the difference between the maximum allowed inflating pressure and the peak inflating pressure decreased progressively as the leak increased. This did not occur with leak compensation. The median PCO2 was slightly higher with leak compensation. CONCLUSIONS: During volume guarantee ventilation with a Dräger VN500 ventilator, without leak compensation the expired tidal volume declined after 50% leak. With leak compensation, the tidal volume was maintained even with a large leak. With leak compensation, there was a more stable peak inflating pressure, although the PCO2 was slightly higher.

Optimisation of 16S rRNA gut microbiota profiling of extremely low birth weight infants.

BACKGROUND: Infants born prematurely, particularly extremely low birth weight infants (ELBW) have altered gut microbial communities. Factors such as maternal health, gut immaturity, delivery mode, and antibiotic treatments are associated with microbiota disturbances, and are linked to an increased risk of certain diseases such as necrotising enterocolitis. Therefore, there is a requirement to optimally characterise microbial profiles in this at-risk cohort, via standardisation of methods, particularly for studying the influence of microbiota therapies (e.g. probiotic supplementation) on community profiles and health outcomes. Profiling of faecal samples using the 16S rRNA gene is a cost-efficient method for large-scale clinical studies to gain insights into the gut microbiota and additionally allows characterisation of cohorts were sample quantities are compromised (e.g. ELBW infants). However, DNA extraction method, and the 16S rRNA region targeted can significantly change bacterial community profiles obtained, and so confound comparisons between studies. Thus, we sought to optimise a 16S rRNA profiling protocol to allow standardisation for studying ELBW infant faecal samples, with or without probiotic supplementation. METHODS: Using ELBW faecal samples, we compared three different DNA extraction methods, and subsequently PCR amplified and sequenced three hypervariable regions of the 16S rRNA gene (V1 + V2 + V3), (V4 + V5) and (V6 + V7 + V8), and compared two bioinformatics approaches to analyse results (OTU and paired end). Paired shotgun metagenomics was used as a 'gold-standard'. RESULTS: Results indicated a longer bead-beating step was required for optimal bacterial DNA extraction and that sequencing regions (V1 + V2 + V3) and (V6 + V7 + V8) provided the most representative taxonomic profiles, which was confirmed via shotgun analysis. Samples sequenced using the (V4 + V5) region were found to be underrepresented in specific taxa including Bifidobacterium, and had altered diversity profiles. Both bioinformatics 16S rRNA pipelines used in this study (OTU and paired end) presented similar taxonomic profiles at genus level. CONCLUSIONS: We determined that DNA extraction from ELBW faecal samples, particularly those infants receiving probiotic supplementation, should include a prolonged beat-beating step. Furthermore, use of the 16S rRNA (V1 + V2 + V3) and (V6 + V7 + V8) regions provides reliable representation of ELBW microbiota profiles, while inclusion of the (V4 + V5) region may not be appropriate for studies where Bifidobacterium constitutes a resident microbiota member.

Probiotics for the prevention of necrotising enterocolitis in very low-birth-weight infants: a meta-analysis and systematic review.

We performed an updated meta-analysis incorporating the results of recent randomised controlled trials (RCTs) to measure the effectiveness of probiotic supplementation in preventing necrotising enterocolitis (NEC) and death in very low-birth-weight (VLBW) infants, and to investigate any differences in efficacy by probiotic agent. Using meta-regression analysis, we assessed the contribution of other measured variables on the overall effect size and between-study variability. CONCLUSION: Overall, probiotics lead to significant reductions in NEC incidence and mortality in VLBW infants. Differences in probiotic agents and the influence of prenatal steroids and feeding regimens may explain the differences in outcomes between studies.

Periconceptional maternal micronutrient supplementation is associated with widespread gender related changes in the epigenome: a study of a unique resource in the Gambia.

In addition to the genetic constitution inherited by an organism, the developmental trajectory and resulting mature phenotype are also determined by mechanisms acting during critical windows in early life that influence and establish stable patterns of gene expression. This is the crux of the developmental origins of health and disease hypothesis that suggests undernutrition during gestation and infancy predisposes to ill health in later life. The hypothesis that periconceptional maternal micronutrient supplementation might affect fetal genome-wide methylation within gene promoters was explored in cord blood samples from offspring of Gambian women enrolled into a unique randomized, double blind controlled trial. Significant changes in the epigenome in cord blood DNA samples were further explored in a subset of offspring at 9 months. Gender-specific changes related to periconceptional nutritional supplementation were identified in cord blood DNA samples, some of which showed persistent changes in infant blood DNA samples. Significant effects of periconceptional micronutrient supplementation were also observed in postnatal samples which were not evident in cord blood. In this Gambian population, the increased death rate of individuals born in nutritionally poor seasons has been related to infection and it is of interest that we identified differential methylation at genes associated with defence against infection and immune response. Although the sample size was relatively small, these pilot data suggest that periconceptional nutrition in humans is an important determinant of newborn whole genome methylation patterns but may also influence postnatal developmental patterns of gene promoter methylation linking early with disease risk.

Stabilization, respiratory care and survival of extremely low birth weight infants transferred on the first day of life.

OBJECTIVE: To assess stabilization, respiratory care and survival of extremely low birth weight (ELBW, <1000 g at birth) infants requiring emergency transfer to tertiary NICUs on the first day of life. STUDY DESIGN: Retrospective cohort study of 55 ELBW infants transported by a dedicated neonatal transport service over a 65-month period. Ventilator data were downloaded computationally. RESULTS: 95% of infants were intubated and received surfactant prior to transfer. Median expired tidal volume was 5.0 mL/kg (interquartile range: 4.6-6.2 mL/kg). Infants ventilated with SIPPV had significantly higher mean airway pressure and minute ventilation, but similar FiO2 compared to babies on SIMV. Blood gases showed significant improvement during transport. 55% of infants survived to discharge from NICU. CONCLUSION: Most ELBW infants transferred on the first day of life require mechanical ventilation and can be ventilated with 5 mL/kg tidal volume.

DNA methylation profiling at imprinted loci after periconceptional micronutrient supplementation in humans: results of a pilot randomized controlled trial.

Intrauterine exposures mediated by maternal diet may affect risk of cardiovascular disease, obesity, and type 2 diabetes. Recent evidence, primarily from animal studies and observational data in humans, suggests that the epigenome can be altered by maternal diet during the periconceptional period and that these programming events may underlie later disease risk. A randomized controlled trial of periconceptional micronutrient supplementation in The Gambia, where seasonal nutritional variations affect fetal growth and postnatal outcomes, provided a unique opportunity to test this hypothesis. Specifically, we targeted imprinted genes, which play important roles in allocation of maternal resources while being epigenetically regulated. DNA methylation at 12 differentially methylated regions (DMRs) was analyzed in cord blood samples from 58 offspring of women participating in a double-blind randomized-controlled trial of pre- and periconceptional micronutrient supplementation (including folate, zinc, and vitamins A, B, C, and D). We observed sex-specific effects of micronutrient supplementation, reducing methylation levels at two of the DMRs analyzed, IGF2R in girls and GTL2-2 in boys. This pilot study is the first to analyze DNA methylation in the context of a randomized controlled trial, and it provides suggestive evidence that periconceptional maternal nutrition alters offspring methylation at imprinted loci.

Low inflating pressures during neonatal tidal volume targeted ventilation: occurrence and significance.

OBJECTIVES: We investigated the inflating pressures (Pinfl, the difference between peak inspiratory pressure and positive end-expiratory pressure) in infants receiving volume targeted ventilation. METHODS: Data were collected and analysed from 195 infants. Median Pinfl was determined before each blood gas (n = 3425). Ventilator parameters and blood gases were compared between periods when Pinfl was <5 mbar and periods when it was higher. RESULTS: 1-hour periods when median Pinfl was <5 mbar occurred in 30% of the babies and were associated with similar tidal volumes and minutes ventilation as periods with higher Pinfl. Babies triggered more ventilator inflations, had more spontaneous breaths and lower oxygen requirement when Pinfl was low. There was no difference in blood gases when Pinfl was <5 mbar or when it was higher. CONCLUSIONS: Episodes of low inflating pressure occur frequently in babies receiving volume targeted ventilation, but they do not lead to changes in blood gases.

Synchronized intermittent mandatory ventilation with volume guarantee and pressure support in neonates: Detailed analysis of ventilator parameters.

OBJECTIVE: To analyse the relationship between peak inflating pressure, expired tidal volume, respiratory rate, and inspiratory time of volume-guaranteed ventilator inflations and pressure-supported spontaneous breaths during synchronized intermittent positive pressure mode with volume guarantee and pressure support (SIMV-VG-PS) in neonates. METHODS: Ventilator parameters were downloaded every second from 16 babies ventilated with SIMV-VG-PS mode using Dräger Babylog VN500 ventilators over 137 days. Transcutaneous carbon dioxide (tcCO2 ) data were also collected. Data were computationally analysed using Python. The average of each ventilator parameter was determined during each minute separately for ventilator inflations and for spontaneous breaths. These values were compared and their effect on tcCO2 levels was also analysed. RESULTS: The relationship between the peak inflating pressure of the volume guaranteed inflations (PIPVG ) and pressure-supported spontaneous breaths (PIPPS ) was highly variable. The PIPPS /PIPVG ratio differed significantly from the value (0.66) targeted by clinicians (group median: 0.80, range: 0.50-1.00). PIPPS frequently exceeded PIPVG . When PIPPS /PIPVG was >0.66, the expired tidal volume and the rate of the pressure-supported spontaneous breaths were also significantly (p < 0.0001) higher, but there was no difference in tcCO2 levels. The flow-cycled spontaneous breaths had significantly shorter inspiratory times than ventilator inflations. CONCLUSIONS: During SIMV-VG-PS it is difficult to ensure a pressure support level proportionate to the inflating pressure of ventilator inflations and to achieve the stability of tidal volumes.

Acceleration during neonatal transport and its impact on mechanical ventilation.

OBJECTIVE: During interhospital transfer, critically ill neonates frequently require mechanical ventilation and are exposed to physical forces related to movement of the ambulance. In an observational study, we investigated acceleration during emergency transfers and if they result from changes in ambulance speed and direction or from vibration due to road conditions. We also studied how these forces impact on performance of the fabian+nCPAP evolution neonatal ventilator and on patient-ventilator interactions. METHODS: We downloaded ventilator parameters at 125 Hz and acceleration data at 100 Hz sampling rates, respectively, during the emergency transfer of 109 infants. Study subjects included term, preterm and extremely preterm infants. We computationally analysed the magnitude, direction and frequency of ambulance acceleration. We also analysed maintenance and variability of ventilator parameters and the shape of pressure-volume loops. RESULTS: While acceleration was <1 m/s2 most of the time, most babies were occasionally exposed to accelerations>5 m/s2. Vibration was responsible for most of the acceleration, rather than speed change or vehicle turning. There was no significant difference between periods of high or low vibration in ventilation parameters, their variability and how well targeted parameters were kept close to their target. Speed change or vehicle turning did not affect ventilator parameters or performance. However, during periods of intense vibration, pressure-volume ventilator loops became significantly more irregular. CONCLUSIONS: Infants are exposed to significant acceleration and vibration during emergency transport. While these forces do not interfere with overall maintenance of ventilator parameters, they make the pressure-volume loops more irregular.

Comparison of flexible nasogastric tube and semi-rigid catheter during less invasive surfactant administration.

BACKGROUND: Various flexible and semi-rigid catheter techniques have been reported for surfactant delivery during less invasive surfactant administration (LISA) in preterm infants. Data on the effect of catheter selection on procedural success rates and adverse events are limited. Our objective was to compare the rates of success and adverse events of LISA performed with nasogastric tube and semi-rigid catheter. METHODS: This was a post-hoc analysis of data from a quality improvement project. LISA was performed according to the standardized local protocol. Baseline characteristics, data on performance of LISA, degree of difficulty in laryngoscopy and vital parameters after the initiation of LISA were collected and outcomes were compared between groups. RESULTS: Fifty-six infants were included (21 with nasogastric tube, and 35 with semi-rigid catheter). Procedure success rate (defined as a single LISA attempt resulting in intratracheal administration of the planned dose of surfactant), incidence of adverse events, heart rate and oxygen saturation values and outcomes did not differ significantly between the two groups. When using a nasogastric tube for LISA, a significantly higher fraction of inspired oxygen was needed in the 3rd (0.62 vs. 0.48, P=0.024), 4th (0.61 vs. 0.37, P<0.001) and 5th minute (0.48 vs. 0.37, P=0.001) to maintain normal oxygen saturations. CONCLUSIONS: Use of the semi-rigid catheter was associated with better oxygenation during and shortly after the procedure. Our results may help neonatal units to develop local guidelines.

Particular genomic and virulence traits associated with preterm infant-derived toxigenic Clostridium perfringens strains.

Clostridium perfringens is an anaerobic toxin-producing bacterium associated with intestinal diseases, particularly in neonatal humans and animals. Infant gut microbiome studies have recently indicated a link between C. perfringens and the preterm infant disease necrotizing enterocolitis (NEC), with specific NEC cases associated with overabundant C. perfringens termed C. perfringens-associated NEC (CPA-NEC). In the present study, we carried out whole-genome sequencing of 272 C. perfringens isolates from 70 infants across 5 hospitals in the United Kingdom. In this retrospective analysis, we performed in-depth genomic analyses (virulence profiling, strain tracking and plasmid analysis) and experimentally characterized pathogenic traits of 31 strains, including 4 from CPA-NEC patients. We found that the gene encoding toxin perfringolysin O, pfoA, was largely deficient in a human-derived hypovirulent lineage, as well as certain colonization factors, in contrast to typical pfoA-encoding virulent lineages. We determined that infant-associated pfoA+ strains caused significantly more cellular damage than pfoA- strains in vitro, and further confirmed this virulence trait in vivo using an oral-challenge C57BL/6 murine model. These findings suggest both the importance of pfoA+ C. perfringens as a gut pathogen in preterm infants and areas for further investigation, including potential intervention and therapeutic strategies.

Disruption of paraoxonase 3 impairs proliferation and antioxidant defenses in human A549 cells and causes embryonic lethality in mice.

We had shown previously that paraoxonase 3 (PON3), a putative circulating antioxidant, was systemically upregulated in late-gestation rat, sheep, and human fetuses. Our overarching hypothesis is that preterm human infants are delivered with low levels of PON3 and that this contributes to a state of oxidative stress. We sought to determine whether absence of Pon3 was associated with reduced neonatal viability in mice and studied the offspring from crosses between Pon3(+/-) mice. The number of Pon3(-/-) animals at E10.5 and E17.5 was significantly lower than the expected 25% (9.3 and 7.9% respectively, P < 0.001). On the first day of postnatal life, this was reduced further (2.4%, significantly less than the proportion in fetal life, P = 0.04). Pon3(+/-) animals had lower body and placental weights than wild-type littermates at E17.5, an effect that was independent of the parent of origin of the mutant allele. We then studied the effect of PON3 knockdown in a human cell line, A549. Stable knockdown of PON3 using short-hairpin RNA reduced cell proliferation in 21% oxygen. We then studied the effect of transient knockdown of PON3 using short interfering RNA (siRNA) in the same cell line in low (2%) or ambient (21%) oxygen. Knockdown of PON3 using siRNA reduced total antioxidant capacity in 21% (P = 0.008) but not 2% oxygen. We conclude that the absence of Pon3 in mice resulted in increased rates of early fetal and neonatal death. Knockdown of PON3 in human cells reduced cell proliferation and total antioxidant capacity.

Incidence, predictors of success and outcome of LISA in very preterm infants.

OBJECTIVES: The aim of this study was to examine the success rate of less invasive surfactant administration (LISA), to identify early predictive factors for the outcome of LISA, and to compare neonatal outcomes between the LISA failure group and the group of infants who were successfully treated with LISA. DESIGN: A retrospective cohort study. PATIENTS: Infants born at less than 33 weeks of gestation (n = 158) and treated with LISA for respiratory distress syndrome. RESULTS: LISA was successful in 86 cases (54.4%); 72 preterm infants (45.6%) needed additional surfactant therapy and/or mechanical ventilation in the first 72 h (LISA failure). In a multivariate logistic regression analysis, six independent predictors of LISA success were identified: core temperature at the time of admission (adjusted odds ratio (OR): 3.56), dose of poractant alfa (<200 mg/kg; adjusted OR: 0.254), elevated C-reactive protein (>10 mg/L) at 24 h of life (adjusted OR: 0.28), highest respiratory severity score (RSS) during the first hour of life or at the time of LISA (adjusted OR: 0.463), maternal age (adjusted OR: 0.923), and birth weight (adjusted OR: 1.003). The receiver operating curve created by using the identified factors indicates good predictive power with an area under the curve of 0.85. LISA failure was associated with a substantially higher risk of complications. CONCLUSION: LISA success can be predicted by variables available before the intervention. Failure of LISA is relatively frequent event in very preterm infants and is associated with adverse outcomes. Prevention of hypothermia during early stabilization and appropriate dosing of surfactant may increase LISA success rates and improve patient outcome.