Decreased Alpha Klotho Expression in Placentas Exposed to Severe Maternal Vascular Malperfusion.

BACKGROUND: Placental maternal vascular malperfusion (MVM) is characterized by accelerated villous maturation and has been associated with a decrease in the antiaging protein, alpha-klotho (AK). Our aim was to characterize AK protein and gene expression in the placenta and fetal organs. METHODS: We utilized 2 cohorts. First, we characterized AK protein expression in an autopsy cohort where cases were defined as MVM as the cause of fetal death compared to a stillborn control population. Second, we characterized placental and umbilical cord blood AK gene expression in a liveborn population with and without MVM. RESULTS: We found decreased protein expression in the villous trophoblastic cells of placentas exposed to severe MVM and decreased AK gene expression in placental tissue exposed to MVM. We did not see any statistically significant differences in fetal organ or umbilical cord blood AK expression based on the presence or absence of MVM. Furthermore, in liveborn infants, we also found increased odds of preterm birth with lower placental AK expression. CONCLUSIONS: Decreased AK gene and protein expression in the placenta in the setting of MVM is consistent with the theory of placental aging in MVM and is associated with increased odds of preterm birth.

Pre-Medications for Non-Emergency Tracheal Intubation in the United States Neonatal Intensive Care Units.

BACKGROUND: Premedication in neonates undergoing elective intubation effectively minimizes the negative physiological events of bradycardia, systemic hypertension, intracranial hypertension, and hypoxia. Premedication decreases procedure-related pain and discomfort. This study aimed to evaluate the current practice of pre-intubation medications for non-emergent intubations in preterm and term neonates in the United States. STUDY DESIGN: A cross-sectional survey (Appendix) was sent via e-mail to all level 3 and 4 Neonatal Intensive Care Units (NICUs) of the Organization of Neonatal Perinatal Medicine Training Program Directors (ONTPD), NICU directors with pediatric residency only, and Baylor Scott and White Health, Mednax, and Envision health services systems. RESULTS: Of 170 responses, 41% (69/168) routinely premedicate, 38% (64/168) premedicate under specific circumstances, and 21% (35/168) do not administer any routine pre-intubation medications. Only 46% (77/168) of units had a written policy. The most frequently used drugs were fentanyl (68%, 116/170), atropine (39%, 66/170), midazolam (38%, 64/170), and morphine (26%, 45/170). 21% (36/170) used a two-drug combination, and 38% (64/170) used a three-drug combination. The most commonly used two-drug combination was atropine and fentanyl, and the most common three-drug combination was atropine, fentanyl, and a paralytic agent. CONCLUSION:  Despite the well-documented benefits of premedication for NICU intubations, as aligned with AAP recommendations, the US lags behind other nations, with stagnant rates since 2006. This disparity persists despite a rise in written policies, which exhibit significant content variations. The authors advocate for the adoption of standardized, AAP-aligned policies across all NICUs in the US. Continued research is vital to monitor the progress of this crucial practice and address any underlying barriers to implementation.

Association of Adverse Childhood Experiences With Accelerated Epigenetic Aging in Midlife.

Importance: Adverse childhood experiences (ACEs) are associated with the risk of poorer health, and identifying molecular mechanisms may lay the foundation for health promotion in people with ACEs. Objective: To investigate the associations of ACEs with changes in epigenetic age acceleration (EAA), a biomarker associated with various health outcomes in middle-aged adults, in a population with balanced race and sex demographics. Design, Setting, and Participants: Data for this cohort study were from the Coronary Artery Risk Development in Young Adults (CARDIA) study. Participants in CARDIA underwent 8 follow-up exams from baseline (year 0 [Y0]; 1985-1986) to Y30 (2015-2016), and participant blood DNA methylation information was obtained at Y15 (2000-2001) and Y20 (2005-2006). Individuals from Y15 and Y20 with available DNA methylation data and complete variables for ACEs and covariates were included. Data were analyzed from September 2021 to August 2022. Exposures: Participant ACEs (general negligence, emotional negligence, physical violence, physical negligence, household substance abuse, verbal and emotional abuse, and household dysfunction) were obtained at Y15. Main Outcomes and Measures: The primary outcome consisted of results from 5 DNA methylation-based EAA measurements known to be associated with biological aging and long-term health: intrinsic EAA (IEAA), extrinsic EAA (EEAA), PhenoAge acceleration (PhenoAA), GrimAge acceleration (GrimAA), and Dunedin Pace of Aging Calculated From the Epigenome (DunedinPACE), measured at Y15 and Y20. Linear regression and generalized estimating equations were used to assess associations of the burden of ACEs (≥4 vs <4 ACEs) with EAA adjusting for demographics, health-related behaviors, and early life and adult socioeconomic status. Results: A total of 895 participants for Y15 (mean [SD] age, 40.4 [3.5] years; 450 males [50.3%] and 445 females [49.7%]; 319 Black [35.6%] and 576 White [64.4%]) and 867 participants for Y20 (mean [SD] age, 45.4 [3.5] years; 432 males [49.8%] and 435 females [50.2%]; 306 Black [35.3%] and 561 White [64.7%]) were included after excluding participants with missing data. There were 185 participants with (20.7%) vs 710 participants without (79.3%) 4 or more ACEs at Y15 and 179 participants with (20.6%) vs 688 participants without (79.4%) 4 or more ACEs at Y20. Having 4 or more ACEs was positively associated with EAA in years at Y15 (EEAA: ß = 0.60 years; 95% CI, 0.18-1.02 years; PhenoAA: ß = 0.62 years; 95% CI = 0.13-1.11 years; GrimAA: ß = 0.71 years; 95% CI, 0.42-1.00 years; DunedinPACE: ß = 0.01; 95% CI, 0.01-0.02) and Y20 (IEAA: ß = 0.41 years; 95% CI, 0.05-0.77 years; EEAA: ß = 1.05 years; 95% CI, 0.66-1.44 years; PhenoAA: ß = 0.57 years; 95% CI, 0.08-1.05 years; GrimAA: ß = 0.57 years; 95% CI, 0.28-0.87 years; DunedinPACE: ß = 0.01; 95% CI, 0.01-0.02) after adjusting for demographics, health-related behaviors, and socioeconomic status. Conclusions and Relevance: In this cohort study, ACEs were associated with EAA among middle-aged adults after controlling for demographics, behavior, and socioeconomic status. These findings of the associations between early life experience and the biological aging process in midlife may contribute to health promotion in a life course perspective.

Clinical utility of a serum biomarker panel in distinguishing prostate cancer from benign prostate hyperplasia.

Prostate-specific antigen (PSA) screening for prostate cancer (PCa) is limited by the lack of specificity but is further complicated in the benign prostatic hyperplasia (BPH) population which also exhibit elevated PSA, representing a clear unmet need to distinguish BPH from PCa. Herein, we evaluated the utility of FLNA IP-MRM, age, and prostate volume to stratify men with BPH from those with PCa. Diagnostic performance of the biomarker panel was better than PSA alone in discriminating patients with negative biopsy from those with PCa, as well as those who have had multiple prior biopsies (AUC 0.75 and 0.87 compared to AUC of PSA alone 0.55 and 0.57 for patients who have had single compared to multiple negative biopsies, respectively). Of interest, in patients with PCa, the panel demonstrated improved performance than PSA alone in those with Gleason scores of 5-7 (AUC 0.76 vs. 0.56) and Gleason scores of 8-10 (AUC 0.74 vs. 0.47). With Gleason scores (8-10), the negative predictive value of the panel is 0.97, indicating potential to limit false negatives in aggressive cancers. Together, these data demonstrate the ability of the biomarker panel to perform better than PSA alone in men with BPH, thus preventing unnecessary biopsies.

Refractory Pulmonary Interstitial Emphysema in Extreme Premature Newborn.

Pulmonary interstitial emphysema (PIE) occurs when air leaks into the pulmonary interstitium due to overdistension of distal airways, it occurs mainly in neonates with respiratory distress syndrome who need positive pressure ventilation but has also been reported in spontaneously breathing infants. Herein, we report on an extremely low birth weight infant with severe persistent PIE, while on invasive mechanical ventilation (high-frequency oscillatory ventilation, high-frequency jet ventilation, and neurally adjust ventilator assist) managed successfully with 2 weeks of selective right lung ventilation after failure of more conservative measures, including shorter periods of right mainstem intubation, before the prolonged trial that was successful.

Cross-Institutional Evaluation of the Abbott ARCHITECT SARS-CoV-2 IgG Immunoassay.

OBJECTIVE: To describe a cross-institutional approach to verify the Abbott ARCHITECT SARS-CoV-2 antibody assay and to document the kinetics of the serological response. METHODS: We conducted analytical performance evaluation studies using the Abbott ARCHITECT SARS-CoV-2 antibody assay on 5 Abbott ARCHITECT i2000 automated analyzers at 2 academic medical centers. RESULTS: Within-run and between-run coefficients of variance (CVs) for the antibody assay did not exceed 5.6% and 8.6%, respectively, for each institution. Quantitative and qualitative results agreed for lithium heparin plasma, EDTA-plasma and serum specimen types. Results for all SARS-CoV-2 IgG-positive and -negative specimens were concordant among analyzers except for 1 specimen at 1 institution. Qualitative and quantitative agreement was observed for specimens exchanged between institutions. All patients had detectable antibodies by day 10 from symptom onset and maintained seropositivity throughout specimen procurement. CONCLUSIONS: The analytical performance characteristics of the Abbott ARCHITECT SARS-CoV-2 antibody assay within and between 2 academic medical center clinical laboratories were acceptable for widespread clinical-laboratory use.

A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots.

OBJECTIVES: Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low-grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the measurement of IL6, IL8, IL10, and TNFα in finger stick dried blood spot (DBS) samples. METHODS: The protocol uses a multiplex electrochemiluminescent immunoassay platform. The following measures of assay performance were evaluated: reliability (inter-assay percent coefficient of variation; %CV), precision (intra-assay %CV), lower limit of detection (LLD), linearity of dilution, and agreement with results from matched plasma samples. RESULTS: Analysis of three control samples across the assay range indicated an acceptable level of precision and reliability for each cytokine. Linearity of dilution returned average values that ranged from 104.1 to 127.6% of expected. Lower limits of detection for IL6, IL8, and IL10 were <0.5, and <1.0 pg/ml for TNFα. Level of agreement in results between matched DBS and plasma samples was high for all cytokines except for IL8. CONCLUSIONS: Finger stick DBS sampling provides a viable alternative to venipuncture for the quantification of IL6, IL10, and TNFα at low concentrations associated with chronic inflammation. The presence of red blood cells may interfere with the quantification of IL8 in DBS. In facilitating blood collection in nonclinical settings this method can advance scientific understandings of how social and ecological contexts shape immune function and health over the life course.

Development of a Mobile Hospital for Disaster Relief.

Mobile supplemental hospitals were an important asset to community response in preparing for the recent pandemic. MED-1 is a Mobile Emergency Department that has adapted and evolved to the changing needs of communities in times of disaster and nondisaster. An overview of the asset (MED-1), the operations, and use is provided to demonstrate how mobile supplemental hospitals can effectively meet a range of healthcare needs. Innovative utilization of MED-1 has secured its future as an effective resource averaging 100 days of deployment per year.

Profiles of gene expression in maternal blood predict offspring birth weight in normal pregnancy.

The association between lower birth weight and increased disease risk in adulthood has drawn attention to the physiological processes that shape the gestational environment. We implement genome-wide transcriptional profiling of maternal blood samples to identify subsets of genes and associated transcription control pathways that predict offspring birth weight. Female participants (N = 178, mean = 27.0 years) in a prospective observational birth cohort study were contacted between 2009 and 2014 to identify new pregnancies. An in-home interview was scheduled for early in the third trimester (mean = 30.3 weeks) to collect pregnancy-related information and a blood sample, and birth weight was measured shortly after delivery. Transcriptional activity in white blood cells was determined with a whole-genome gene expression direct hybridization assay. Fifty transcripts were differentially expressed in association with offspring birth weight, with 18 up-regulated in relation to lower birth weight, and 32 down-regulated. Examination of transcription control pathways identified increased activity of NF-κB, AP-1, EGR1, EGR4, and Gfi families, and reduced the activity of CEBP, in association with lower birth weight. Transcript origin analyses identified non-classical CD16+ monocytes, CD1c+ myeloid dendritic cells, and neutrophils as the primary cellular mediators of differential gene expression. These results point toward a systematic regulatory shift in maternal white blood cell activity in association with lower offspring birth weight, and they suggest that analyses of gene expression during gestation may provide insight into regulatory and cellular mechanisms that influence birth outcomes.

Combinatorial screening using orthotopic patient derived xenograft-expanded early phase cultures of osteosarcoma identify novel therapeutic drug combinations.

Lead discovery in osteosarcoma has been hampered by the lack of new agents, limited representative clinical samples and paucity of accurate preclinical models. We developed orthotopic patient-derived xenografts (PDXs) that recapitulated the molecular, cellular and histologic features of primary tumors, and screened PDX-expanded short-term cultures and commercial cell lines of osteosarcoma against focused drug libraries. Osteosarcoma cells were most sensitive to HDAC, proteasome, and combination PI3K/MEK and PI3K/mTOR inhibitors, and least sensitive to PARP, RAF, ERK and MEK inhibitors. Correspondingly, PI3K signaling pathway genes were up-regulated in metastatic tumors compared to primary tumors. In combinatorial screens, as a class, HDAC inhibitors showed additive effects when combined with standard-of-care agents gemcitabine and doxorubicin. This lead discovery strategy afforded a means to perform high-throughput drug screens of tumor cells that accurately recapitulated those from original human tumors, and identified classes of novel and repurposed drugs with activity against osteosarcoma.

Is the World Health Organization predicted exposure assessment model for space spraying of insecticides applicable to agricultural farmers?

Agricultural farmers in developing countries are at high risk of pesticide exposure and adverse effects because of unsafe practices and inappropriate legislation. Biological monitoring is considered a useful tool for pesticide exposure assessment; however, its use is limited in developing countries due to a lack of techniques and resources such as laboratory analysis, trained staff and budgets. This study examines whether the World Health Organization predicted exposure assessment model (WHO-PEAM) is a suitable alternative tool for assessing insecticide exposure among agricultural farmers. WHO-PEAM was used to predict daily doses (PDD) of chlorpyrifos for a group of Vietnamese rice farmers using a set of exposure parameters obtained from a questionnaire survey of participant famers during a field study. These results were compared to absorbed daily doses (ADD) of chlorpyrifos for the farmers measured using a biological monitoring program, in which 24-h urine samples were collected and analysed for the chlorpyrifos metabolite, 3,5,6-trichloro-2-pyridinol (TCP) using LC/MS. Validation of the model results was tested using the Wilcoxon signed-rank test (WSR) and two-way mixed-model intraclass correlation coefficient (ICC). The mean of total ADD was 20 µg/kg/day while that of total PDD was 22 µg/kg/day. The WSR test revealed no statistically significant difference in the average values of ADDT and PDDT. ICC indicated substantial agreement for both single and average measures between ADDT and PDDT (ICC, 0.62 and 0.77, respectively). The results demonstrate that a refined WHO-PEAM model can be readily used as a field method, without biological monitoring, to evaluate chlorpyrifos exposure among agricultural farmers in Vietnam and similar developing countries.

Statewide At-Risk Tracking and Intervention for Nurses: Identifying and Intervening With Nursing Students at Risk of Attrition in Texas.

AIM: The purpose of this study was to identify students at risk of attrition and implement interventions to decrease the risk. BACKGROUND: The ability to identify and intervene with students deemed at risk of attrition can be a valuable tool for increasing the RN workforce. "Statewide At-Risk Tracking and Interventions for Nurses" (SATIN) involved students from 27 initial RN licensure nursing programs in Texas. METHOD: At-risk status of each nursing student was identified, and each nursing program provided interventions based on students' needs. RESULTS: The Weaver Reading Program was the most effective of the six intervention strategies used with SATIN participants. Lack of utilization of the interventions was the number one limitation of the study. CONCLUSION: Use of the Weaver Reading Program as well as personal engagement with students, one-on-one mentoring, and continued time investment are recommended to all programs.

Structure-function characterization of three human antibodies targeting the vaccinia virus adhesion molecule D8.

Vaccinia virus (VACV) envelope protein D8 is one of three glycosaminoglycan adhesion molecules and binds to the linear polysaccharide chondroitin sulfate (CS). D8 is also a target for neutralizing antibody responses that are elicited by the smallpox vaccine, which has enabled the first eradication of a human viral pathogen and is a useful model for studying antibody responses. However, to date, VACV epitopes targeted by human antibodies have not been characterized at atomic resolution. Here, we characterized the binding properties of several human anti-D8 antibodies and determined the crystal structures of three VACV-mAb variants, VACV-66, VACV-138, and VACV-304, separately bound to D8. Although all these antibodies bound D8 with high affinity and were moderately neutralizing in the presence of complement, VACV-138 and VACV-304 also fully blocked D8 binding to CS-A, the low affinity ligand for D8. VACV-138 also abrogated D8 binding to the high-affinity ligand CS-E, but we observed residual CS-E binding was observed in the presence of VACV-304. Analysis of the VACV-138- and VACV-304-binding sites along the CS-binding crevice of D8, combined with different efficiencies of blocking D8 adhesion to CS-A and CS-E allowed us to propose that D8 has a high- and low-affinity CS-binding region within its central crevice. The crevice is amenable to protein engineering to further enhance both specificity and affinity of binding to CS-E. Finally, a wild-type D8 tetramer specifically bound to structures within the developing glomeruli of the kidney, which express CS-E. We propose that through structure-based protein engineering, an improved D8 tetramer could be used as a potential diagnostic tool to detect expression of CS-E, which is a possible biomarker for ovarian cancer.

Cassini finds molecular hydrogen in the Enceladus plume: Evidence for hydrothermal processes.

Saturn's moon Enceladus has an ice-covered ocean; a plume of material erupts from cracks in the ice. The plume contains chemical signatures of water-rock interaction between the ocean and a rocky core. We used the Ion Neutral Mass Spectrometer onboard the Cassini spacecraft to detect molecular hydrogen in the plume. By using the instrument's open-source mode, background processes of hydrogen production in the instrument were minimized and quantified, enabling the identification of a statistically significant signal of hydrogen native to Enceladus. We find that the most plausible source of this hydrogen is ongoing hydrothermal reactions of rock containing reduced minerals and organic materials. The relatively high hydrogen abundance in the plume signals thermodynamic disequilibrium that favors the formation of methane from CO2 in Enceladus' ocean.