Development of a rapid clinical flow cytometric assay of neutrophil cell-surface biomarkers

Rationale: Neutrophils are upregulated in infectious processes. They can be measured as surrogates for the presence of an infection. Flow cytometry is a common tool that uses fluorochrome-conjugated antibodies specific to cell-surface biomarkers to identify cells or biomarkers of interest. Many clinical flow cytometry protocols exist that assess lymphocytes, but fewer protocols exist for rapid clinical evaluation of neutrophils. Expression of neutrophil cell-surface markers integrin α9β1 and CD11b has been shown to increase in elderly patients with bacterial pneumonia. A rapid clinical test for these neutrophil cell-surface markers could help to expedite appropriate antibiotic therapy to patients suffering from complex critical illnesses complicated by bacterial pneumonia for which other diagnostic methods are rendered inadequate. Methods: We developed a rapid lyse, no-wash clinical protocol for flow cytometric analysis of neutrophil cell-surface biomarkers integrin α9β1 and CD11b from whole blood samples. Assay development included selecting appropriate neutrophil-specific biomarkers and their corresponding fluorochrome profile, determining the most consistent blood acquisition method (BAM) for obtaining samples for analysis, and evaluating the effect of preservative type and time on neutrophil absolute count (ANC), integrin α9β1 mean fluorescence intensity (MFI), and CD11b MFI. Results: There were significant differences in ANC, CD11b MFI, and integrin α9β1 MFI over time for each BAM and Preservative type. ANC demonstrated the least variability in Citrate, while CD11b MFI and integrin α9β1 MFI were most consistent in Heparin. Therefore, Heparin was selected as the BAM of choice. Streck preservative solution and Streck Blood Collection Tube (BCT) demonstrated the least variability in ANC, integrin α9β1 MFI, and CD11b MFI. Conclusions: Pneumonia is difficult to detect in patients with critical illness, where the usual diagnostic tools are rendered inadequate secondary to multi-system organ failure. More accurate methods of diagnosis for pneumonia can enhance targeted antibiotic therapy and are of paramount public health importance to improve patient care. Development of a rapid clinical flow cytometric protocol is the first step toward precision methods of pneumonia diagnosis; therefore, a rapid lyse/no-wash clinical flow cytometric assay for whole blood specimens was developed for identification of neutrophil cell-surface biomarkers CD11b and integrin α9β1. Heparin was the least variable BAM for assessing CD11b MFI and integrin α9β1 MFI, while Streck preservative solution and Streck BCT demonstrated the least variability over 24 hours from specimen collection. Heparin is a common type of venipuncture tube in hospitals, it is not as cost effective or feasible to purchase a separate blood collection tube (e.g. Streck BCT) for a single laboratory test. Given these real-world concerns, Streck solution added to samples collected in heparin venipuncture tubes is the most reasonable method by which to perform the rapid lyse/no-wash neutrophil flow cytometric assay

A developmentally regulated translational control pathway establishes the meiotic chromosome segregation pattern

Production of haploid gametes from diploid progenitor cells is mediated by a specialized cell division, meiosis, where two divisions, meiosis I and II, follow a single S phase. Errors in progression from meiosis I to meiosis II lead to aneuploid and polyploid gametes, but the regulatory mechanisms controlling this transition are poorly understood. Here, we demonstrate that the conserved kinase Ime2 regulates the timing and order of the meiotic divisions by controlling translation. Ime2 coordinates translational activation of a cluster of genes at the meiosis I–meiosis II transition, including the critical determinant of the meiotic chromosome segregation pattern CLB3. We further show that Ime2 mediates translational control through the meiosis-specific RNA-binding protein Rim4. Rim4 inhibits translation of CLB3 during meiosis I by interacting with the 5′ untranslated region (UTR) of CLB3. At the onset of meiosis II, Ime2 kinase activity rises and triggers a decrease in Rim4 protein levels, thereby alleviating translational repression. Our results elucidate a novel developmentally regulated translational control pathway that establishes the meiotic chromosome segregation pattern.American Cancer Society (Post-doctoral Fellowship)Virginia and D.K. Ludwig Fund for Cancer Research (Post-doctoral Fellowship)National Institutes of Health (U.S.) (Grant GM62207

Genome-Wide Occupancy of SREBP1 and Its Partners NFY and SP1 Reveals Novel Functional Roles and Combinatorial Regulation of Distinct Classes of Genes

The sterol regulatory element-binding protein (SREBP) family member SREBP1 is a critical transcriptional regulator of cholesterol and fatty acid metabolism and has been implicated in insulin resistance, diabetes, and other diet-related diseases. We globally identified the promoters occupied by SREBP1 and its binding partners NFY and SP1 in a human hepatocyte cell line using chromatin immunoprecipitation combined with genome tiling arrays (ChIP-chip). We find that SREBP1 occupies the promoters of 1,141 target genes involved in diverse biological pathways, including novel targets with roles in lipid metabolism and insulin signaling. We also identify a conserved SREBP1 DNA-binding motif in SREBP1 target promoters, and we demonstrate that many SREBP1 target genes are transcriptionally activated by treatment with insulin and glucose using gene expression microarrays. Finally, we show that SREBP1 cooperates extensively with NFY and SP1 throughout the genome and that unique combinations of these factors target distinct functional pathways. Our results provide insight into the regulatory circuitry in which SREBP1 and its network partners coordinate a complex transcriptional response in the liver with cues from the diet

Effect of stomach inflation during cardiopulmonary resuscitation on return of spontaneous circulation in out-of-hospital cardiac arrest patients: A retrospective observational study

Background: Gastric inflation caused by excessive ventilation is a common complication of cardiopulmonary resuscitation. Gastric inflation may further compromise ventilation via increases in intrathoracic pressure, leading to decreased venous return and cardiac output, which may impair out-of-hospital cardiac arrest (OHCA) outcomes. The purpose of this study was to measure the gastric volume of OHCA patients using computed tomography (CT) scan images and evaluate the effect of gastric inflation on return of spontaneous circulation (ROSC). Methods: In this single-center, retrospective, observational study, CT scan was conducted after ROSC or immediately after death. Total gastric volume was measured. Primary outcome was ROSC. Achievement of ROSC was compared in the gastric distention group and the no gastric distention group; gastric distension was defined as total gastric volume in the ≥75th percentile. Additionally, factors associated with gastric distention were examined. Results: A total of 446 cases were enrolled in the study; 120 cases (27%) achieved ROSC. The median gastric volume was 400 ml for all OHCA subjects; 1068 ml in gastric distention group vs. 287 ml in no gastric distention group. There was no difference in ROSC between the groups (27/112 [24.1%] vs. 93/334 [27.8%], p = 0.440). Gastric distention did not have a significant impact, even after adjustments (adjusted odds ratio 0.73, 95% confidence interval [0.42–1.29]). Increased gastric volume was associated with longer emergency medical service activity time. Conclusions: We observed a median gastric volume of 400 ml in patients after OHCA resuscitation. In our setting, gastric distention did not prevent ROSC

Contributions of Emotion Regulation and Brain Structure and Function to Adolescent Internalizing Problems and Stress Vulnerability During the COVID-19 Pandemic: A Longitudinal Study

BACKGROUND: Adolescence is a period of increased vulnerability for internalizing problems, particularly following stressful life events. We examined how emotion regulation and brain structure and function were associated with internalizing problems during the COVID-19 pandemic and moderated the association between pandemic-related stressors and internalizing problems. METHODS: Data are from a longitudinal sample (N = 145, age range, 10-15 years) strategically assessed at 3 crucial time points: before the COVID-19 pandemic, early during the stay-at-home order period, and again 6 months later. We examined associations of amygdala and hippocampal volume and amygdala activation during an emotional processing task before the pandemic, examined use of emotion regulation strategies before and during the pandemic, and examined pandemic-related stressors with internalizing problems. RESULTS: Greater exposure to pandemic-related stressors was associated with higher internalizing problems both early and later in the COVID-19 pandemic. Youths who reported more frequent use of rumination before the pandemic and higher use of expressive suppression and lower use of cognitive reappraisal early in the pandemic had higher internalizing problems early in the pandemic. Higher left amygdala activation to neutral relative to fearful faces before the pandemic was associated with greater internalizing problems and a stronger link between pandemic-related stressors and internalizing problems early in the pandemic. CONCLUSIONS: Stressors related to the COVID-19 pandemic are strongly associated with adolescent internalizing problems, as are individual differences in emotional reactivity and regulation and their underlying neural mechanisms. Interventions that reduce pandemic-related stressors and foster adaptive emotion regulation skills may protect against adolescent psychopathology during this period of heightened exposure to stress

Contributions of Emotion Regulation and Brain Structure and Function to Adolescent Internalizing Problems and Stress Vulnerability During the COVID-19 Pandemic: A Longitudinal Study

Background: Adolescence is a period of increased vulnerability for internalizing problems, particularly following stressful life events. We examined how emotion regulation and brain structure and function were associated with internalizing problems during the COVID-19 pandemic and moderated the association between pandemic-related stressors and internalizing problems. Methods: Data are from a longitudinal sample (N = 145, age range, 10–15 years) strategically assessed at 3 crucial time points: before the COVID-19 pandemic, early during the stay-at-home order period, and again 6 months later. We examined associations of amygdala and hippocampal volume and amygdala activation during an emotional processing task before the pandemic, examined use of emotion regulation strategies before and during the pandemic, and examined pandemic-related stressors with internalizing problems. Results: Greater exposure to pandemic-related stressors was associated with higher internalizing problems both early and later in the COVID-19 pandemic. Youths who reported more frequent use of rumination before the pandemic and higher use of expressive suppression and lower use of cognitive reappraisal early in the pandemic had higher internalizing problems early in the pandemic. Higher left amygdala activation to neutral relative to fearful faces before the pandemic was associated with greater internalizing problems and a stronger link between pandemic-related stressors and internalizing problems early in the pandemic. Conclusions: Stressors related to the COVID-19 pandemic are strongly associated with adolescent internalizing problems, as are individual differences in emotional reactivity and regulation and their underlying neural mechanisms. Interventions that reduce pandemic-related stressors and foster adaptive emotion regulation skills may protect against adolescent psychopathology during this period of heightened exposure to stress

2002-2003 Florida\u27s Young Musician Showcase

https://spiral.lynn.edu/conservatory_otherseasonalconcerts/1090/thumbnail.jp

A Rapid Host-protein Test for Differentiating Bacterial From Viral Infection: Apollo Diagnostic Accuracy Study

OBJECTIVES: To determine the diagnostic accuracy of a rapid host-protein test for differentiating bacterial from viral infections in patients who presented to the emergency department (ED) or urgent care center (UCC). METHODS: This was a prospective multicenter, blinded study. MeMed BV (MMBV), a test based on tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), interferon gamma-inducible protein-10 (IP-10), and C-reactive protein (CRP), was measured using a rapid measurement platform. Patients were enrolled from 9 EDs and 3 UCCs in the United States and Israel. Patients \u3e3 months of age presenting with fever and clinical suspicion of acute infection were considered eligible. MMBV results were not provided to the treating clinician. MMBV results (bacterial/viral/equivocal) were compared against a reference standard method for classification of infection etiology determined by expert panel adjudication. Experts were blinded to MMBV results. They were provided with comprehensive patient data, including laboratory, microbiological, radiological and follow-up. RESULTS: Of 563 adults and children enrolled, 476 comprised the study population (314 adults, 162 children). The predominant clinical syndrome was respiratory tract infection (60.5% upper, 11.3% lower). MMBV demonstrated sensitivity of 90.0% (95% confidence interval [CI]: 80.3-99.7), specificity of 92.8% (90.0%-95.5%), and negative predictive value of 98.8% (96.8%-99.6%) for bacterial infections. Only 7.2% of cases yielded equivocal MMBV scores. Area under the curve for MMBV was 0.95 (0.90-0.99). CONCLUSIONS: MMBV had a high sensitivity and specificity relative to reference standard for differentiating bacterial from viral infections. Future implementation of MMBV for patients with suspected acute infections could potentially aid with appropriate antibiotic decision-making

Cognitive flexibility and performance in children and adolescents with threshold and sub-threshold bipolar disorder

Greater understanding of cognitive function in children and adolescents with bipolar disorder (BD) is of critical importance to improve our ability to design targeted treatments to help with real-world impairment, including academic performance. We sought to evaluate cognitive performance among children with either BD type I, II, or “not otherwise specified” (NOS) participating in multi-site Course and Outcome of Bipolar Youth study compared to typically developing controls (TDC) without psycho-pathology. In particular, we sought to test the hypothesis that BD-I and BD-II youths with full threshold episodes of mania or hypomania would have cognitive deficits, including in reversal learning, vs. those BD-NOS participants with sub-threshold episodes and TDCs. N = 175 participants (BD-I = 81, BD-II = 11, BD-NOS = 28, TDC = 55) completed Cambridge Neuropsychological Automated Testing Battery (CANTAB) tasks. A priori analyses of the simple reversal stage of the CANTAB intra-/extra-dimensional shift task showed that aggregated BD-I/II participants required significantly more trials to complete the task than either BD-NOS participants with sub-syndromal manic/hypomanic symptoms or than TDCs. BD participants across sub-types had impairments in sustained attention and information processing for emotionally valenced words. Our results align with prior findings showing that BD-I/II youths with distinct episodes have specific alterations in reversal learning. More broadly, our study suggests that further work is necessary to see the interaction between neurocognitive performance and longitudinal illness course. Additional work is required to identify the neural underpinnings of these differences as targets for potential novel treatments, such as cognitive remediation

Effect of Thromboprophylaxis on Clinical Outcomes After COVID-19 Hospitalization

BACKGROUND: Patients hospitalized with COVID-19 have an increased incidence of thromboembolism. The role of extended thromboprophylaxis after hospital discharge is unclear. OBJECTIVE: To determine whether anticoagulation is superior to placebo in reducing death and thromboembolic complications among patients discharged after COVID-19 hospitalization. DESIGN: Prospective, randomized, double-blind, placebo-controlled clinical trial. (ClinicalTrials.gov: NCT04650087). SETTING: Done during 2021 to 2022 among 127 U.S. hospitals. PARTICIPANTS: Adults aged 18 years or older hospitalized with COVID-19 for 48 hours or more and ready for discharge, excluding those with a requirement for, or contraindication to, anticoagulation. INTERVENTION: 2.5 mg of apixaban versus placebo twice daily for 30 days. MEASUREMENTS: The primary efficacy end point was a 30-day composite of death, arterial thromboembolism, and venous thromboembolism. The primary safety end points were 30-day major bleeding and clinically relevant nonmajor bleeding. RESULTS: Enrollment was terminated early, after 1217 participants were randomly assigned, because of a lower than anticipated event rate and a declining rate of COVID-19 hospitalizations. Median age was 54 years, 50.4% were women, 26.5% were Black, and 16.7% were Hispanic; 30.7% had a World Health Organization severity score of 5 or greater, and 11.0% had an International Medical Prevention Registry on Venous Thromboembolism risk prediction score of greater than 4. Incidence of the primary end point was 2.13% (95% CI, 1.14 to 3.62) in the apixaban group and 2.31% (CI, 1.27 to 3.84) in the placebo group. Major bleeding occurred in 2 (0.4%) and 1 (0.2%) and clinically relevant nonmajor bleeding occurred in 3 (0.6%) and 6 (1.1%) apixaban-treated and placebo-treated participants, respectively. By day 30, thirty-six (3.0%) participants were lost to follow-up, and 8.5% of apixaban and 11.9% of placebo participants permanently discontinued the study drug treatment. LIMITATIONS: The introduction of SARS-CoV-2 vaccines decreased the risk for hospitalization and death. Study enrollment spanned the peaks of the Delta and Omicron variants in the United States, which influenced illness severity. CONCLUSION: The incidence of death or thromboembolism was low in this cohort of patients discharged after hospitalization with COVID-19. Because of early enrollment termination, the results were imprecise and the study was inconclusive. PRIMARY FUNDING SOURCE: National Institutes of Health