The impact of a care bundle with an emphasis on hemodynamic assessment on the short-term outcomes in neonates with congenital diaphragmatic hernia.

OBJECTIVE: To evaluate the short-term outcomes of implementing a care bundle emphasizing frequent hemodynamic assessments by echocardiography in neonates with congenital diaphragmatic hernia (CDH). STUDY DESIGN: This was a retrospective cohort study of infants with CDH admitted to a quaternary perinatal unit from January 2013 to March 2021. The primary composite outcome was defined as mortality or use of extracorporeal membrane oxygenation or need for respiratory support at discharge. RESULTS: We identified 37 and 20 CDH infants in Epoch I and II, respectively. More patch repairs (50% vs. 21.9%, p = 0.035) and echocardiograms (6[4-8] vs. 1[0-5], p = 0.003) were performed in Epoch II. While there were no differences in the primary outcome, there was a reduction in mortality in Epoch II (0% vs. 27%, p = 0.01). CONCLUSION: With the implementation of a CDH care bundle with an emphasis on hemodynamic assessment, we demonstrated a significant reduction in mortality.

Non-Native Anionic Ligand Binding and Reactivity in Engineered Variants of the Fe(II)- and α-Ketoglutarate-Dependent Oxygenase, SadA.

Mononuclear non-heme Fe(II)- and α-ketoglutarate-dependent oxygenases (FeDOs) catalyze a site-selective C-H hydroxylation. Variants of these enzymes in which a conserved Asp/Glu residue in the Fe(II)-binding facial triad is replaced by Ala/Gly can, in some cases, bind various anionic ligands and catalyze non-native chlorination and bromination reactions. In this study, we explore the binding of different anions to an FeDO facial triad variant, SadX, and the effects of that binding on HO• vs X• rebound. We establish not only that chloride and bromide enable non-native halogenation reactions but also that all anions investigated, including azide, cyanate, formate, and fluoride, significantly accelerate and influence the site selectivity of SadX hydroxylation catalysis. Azide and cyanate also lead to the formation of products resulting from N3•, NCO•, and OCN• rebound. While fluoride rebound is not observed, the rate acceleration provided by this ligand leads us to calculate barriers for HO• and F• rebound from a putative Fe(III)(OH)(F) intermediate. These calculations suggest that the lack of fluorination is due to the relative barriers of the HO• and F• rebound transition states rather than an inaccessible barrier for F• rebound. Together, these results improve our understanding of the FeDO facial triad variant tolerance of different anionic ligands, their ability to promote rebound involving these ligands, and inherent rebound preferences relative to HO• that will aid efforts to develop non-native catalysis using these enzymes.

A Simulation-Based Pilot Study of a Mobile Application (NRP Prompt) as a Cognitive Aid for Neonatal Resuscitation Training.

INTRODUCTION: Despite standardized neonatal resuscitation program (NRP) training, retention and adherence to the NRP algorithm remain a challenge. Cognitive aids can potentially improve acquisition and application of NRP knowledge and skills. The objective of this study was to determine whether an interactive mobile application providing audiovisual prompts, NRP Prompt, can help novice NRP providers learn the NRP algorithm more effectively and therefore improve their NRP performance. METHODS: First- and second-year residents from family medicine and obstetrics and gynecology attending NRP training were randomized into intervention and control groups. Resident pairs used standard visual aids with NRP Prompt (intervention) or visual aids only (control) in two simulated neonatal resuscitation training sessions with each resident taking turns as a team leader. Pairs were then evaluated in a third simulation that was video recorded, where neither group used cognitive aids. The primary outcome was comparing resuscitation performance. Secondary outcomes included the following: times to positive-pressure ventilation, intubation, and chest compressions. RESULTS: Thirty-nine residents participated, of which 18 received the intervention. Neonatal resuscitation program performance scores did not significantly differ (P = 0.69). Wilcoxon rank-sum tests showed no significant differences in secondary outcomes of times to positive-pressure ventilation (P = 0.43), intubation (P = 0.44), or chest compressions (P = 0.35). CONCLUSIONS: Training using NRP Prompt did not improve performance scores in simulated neonatal resuscitations immediately after training. Potential reasons include voice prompts in their current format being distracting and lack of customizability to user preferences. Future development of prompting applications should apply a user-centered design approach to optimize the ability to meet end-user needs.

Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection.

Genomic characterization of circulating tumor cells (CTCs) may prove useful as a surrogate for conventional tissue biopsies. This is particularly important as studies have shown different mutational profiles between CTCs and ctDNA in some tumor subtypes. However, isolating rare CTCs from whole blood has significant hurdles. Very limited DNA quantities often can't meet NGS requirements without whole genome amplification (WGA). Moreover, white blood cells (WBC) germline contamination may confound CTC somatic mutation analyses. Thus, a good CTC enrichment platform with an efficient WGA and NGS workflow are needed. Here, Vortex label-free CTC enrichment platform was used to capture CTCs. DNA extraction was optimized, WGA evaluated and targeted NGS tested. We used metastatic colorectal cancer (CRC) as the clinical target, HCT116 as the corresponding cell line, GenomePlex® and REPLI-g as the WGA methods, GeneRead DNAseq Human CRC Panel as the 38 gene panel. The workflow was further validated on metastatic CRC patient samples, assaying both tumor and CTCs. WBCs from the same patients were included to eliminate germline contaminations. The described workflow performed well on samples with sufficient DNA, but showed bias for rare cells with limited DNA input. REPLI-g provided an unbiased amplification on fresh rare cells, enabling an accurate variant calling using the targeted NGS. Somatic variants were detected in patient CTCs and not found in age matched healthy donors. This demonstrates the feasibility of a simple workflow for clinically relevant monitoring of tumor genetics in real time and over the course of a patient's therapy using CTCs.

T cell receptor sequencing of early-stage breast cancer tumors identifies altered clonal structure of the T cell repertoire.

Tumor-infiltrating T cells play an important role in many cancers, and can improve prognosis and yield therapeutic targets. We characterized T cells infiltrating both breast cancer tumors and the surrounding normal breast tissue to identify T cells specific to each, as well as their abundance in peripheral blood. Using immune profiling of the T cell beta-chain repertoire in 16 patients with early-stage breast cancer, we show that the clonal structure of the tumor is significantly different from adjacent breast tissue, with the tumor containing ∼2.5-fold greater density of T cells and higher clonality compared with normal breast. The clonal structure of T cells in blood and normal breast is more similar than between blood and tumor, and could be used to distinguish tumor from normal breast tissue in 14 of 16 patients. Many T cell sequences overlap between tissue and blood from the same patient, including ∼50% of T cells between tumor and normal breast. Both tumor and normal breast contain high-abundance "enriched" sequences that are absent or of low abundance in the other tissue. Many of these T cells are either not detected or detected with very low frequency in the blood, suggesting the existence of separate compartments of T cells in both tumor and normal breast. Enriched T cell sequences are typically unique to each patient, but a subset is shared between many different patients. We show that many of these are commonly generated sequences, and thus unlikely to play an important role in the tumor microenvironment.

Spectral- and cepstral-based acoustic features of dysphonic, strained voice quality.

OBJECTIVES: We sought to determine whether spectral- and cepstral-based acoustic measures were effective in distinguishing dysphonic-strained voice quality from normal voice quality and whether these measures were related to auditory-perceptual ratings of strain severity. METHODS: Voice samples from 23 speakers with dysphonia characterized predominantly by strained voice quality and 23 speakers with normal voice were acoustically analyzed. Measures related to the prominence of the cepstral peak and the ratio of low- to high-frequency spectral energies, as well as the variation of each, were computed from continuous speech and a sustained vowel. Correlations to perceptually rated strain severity were determined. RESULTS: Measures related to the cepstrum were the strongest discriminators between dysphonic-strained voice and normal voice. Variation in the ratio of low- to high-frequency spectral energies also significantly differentiated the two speaker groups. All measures were significantly correlated with perceptually rated strain severity, including an acoustic severity index that incorporated both cepstral- and spectral-based measures. CONCLUSIONS: Cepstral- and spectral-based measures that have been previously studied in dysphonia characterized by breathiness and roughness are effective in distinguishing strained dysphonia from normal voice quality. The utility of these acoustic measures is supported by their moderate-to-high relationship with perceptually rated strain severity.