Citations

Abstract A double-antibody radioimmunoassay (RIA) was developed to measure the protein comprising the human eosinophil Charcot-Leyden crystal (CLC). Critical requirements in the development of this assay were the production of large quantities of pure CLC, the solubilization of CLC at neutral pH, and an assessment of the purity of the CLC protein. CLC were prepared from eosinophil-rich leukocyte suspensions by homogenization of cells in hypotonic-buffered saline. Centrifuged eosinophil extracts formed CLC spontaneously at 4°C, but the yield of crystals was significantly augmented by slow freezing of extracts to −20°C and thawing. Lyophilization of crystals was required to obtain material soluble at neutral pH. Soluble CLC protein produced 3 dense bands on polyacrylamide gel electrophoresis (PAGE) at pH 8.9, and these bands gave reactions of identity in immunodiffusion analyses with rabbit anti-CLC antibody. PAGE in 1% sodium dodecyl sulfate revealed a single protein band of 13,000 daltons. Isoelectric focusing of soluble CLC protein yielded 2 major and 4 minor bands with pI’s between 5.7 and 5.1, indicating the slightly acidic nature of the CLC protein. In the RIA, CLC levels were measured by inhibition of the binding of 131I-CLC reacted with specific rabbit anti-CLC antibody. Burro anti-rabbit IgG was used as the second antibody to precipitate immune complexes. The maximum precipitability of 1 ng 131I-CLC by antibody was 97%; the RIA was sensitive to 0.25 ng. CLC protein is immunologically distinct from human eosinophil major basic protein as determined by RIA and Ouchterlony analysis. Serum levels of CLC for 10 normal healthy subjects averaged 37 ± 4.7 ng/ml compared with 8507 ± 2494 ng/ml for 10 patients with diseases associated with a peripheral blood eosinophilia greater than 30%. The RIA for CLC protein provides us with a tool to investigate the biological function(s) of this protein and the role of eosinophils in certain disease states.

Abstract Eosinophilia is a common sequela of treatment in patients infected with the parasitic helminth Wuchereria bancrofti. We investigated the relationship between peripheral blood eosinophil levels and serum levels of eosinophil granule major basic protein (MBP) and Charcot-Leyden crystal (CLC) protein in 37 patients with Bancroft’s filariasis who were treated with diethylcarbamazine (DEC). Before treatment with DEC, eosinophil levels and serum levels of CLC protein, but not MBP, were found to be significantly elevated in both filariasis patients and an uninfected endemic control population. After treatment, patients with microfilaremia developed an initial eosinopenia followed by significant increases in their eosinophil levels and serum levels of both MBP and CLC protein, which peaked 1 to 2 wk after initiating treatment. Before treatment, eosinophil levels and serum levels of MBP and CLC protein were unrelated to the number of microfilariae in circulation, but after treatment, the degree of eosinophilia and the elevations in serum MBP and CLC protein were significantly correlated with the patients’ microfilarial worm burdens before treatment. In microfilaremic patients on DEC therapy, there was a statistically significant increase in serum MBP levels concomitant with the initial decrease in eosinophil levels observed on day 1 of therapy in these patients. Patients with filariasis (elephantiasis, hydrocoeles, or filarial fevers) but without microfilaremia, as well as an uninfected endemic control population, showed no change in their eosinophil levels or serum MBP or CLC protein levels during an identical course of DEC therapy. The eosinophilic response resulting from DEC therapy of patients with microfilaremia provides a unique opportunity to study the relationship between eosinophilia, eosinophil function, and possibly eosinophilopoiesis as reflected by the synthesis and/or release of eosinophil proteins in humans exposed to a quantifiable antigenic stimulus.

Background: There are limited data about the stage D heart failure (advanced HF) in adults with congenital heart disease. Our study objectives were (1) to determine the incidence of new-onset advanced HF in patients and the relationship between advanced HF and all-cause mortality and (2) to determine the relationship between therapies for advanced HF and all-cause mortality. Methods: Retrospective cohort study of adults with congenital heart disease at Mayo Clinic (2003–2019). We defined advanced HF using the European Society of Cardiology diagnostic criteria for advanced HF. Therapies received by the patients with advanced HF were classified into 3 mutually exclusive groups (treatment pathways): (1) conventional cardiac intervention, (2) transplant listing, and (3) palliative care. Results: Of 5309 patients without advanced HF at baseline assessment, 432 (8%) developed advanced HF during follow-up (1.1%/y), and the incidence of advanced HF was higher in patients with severe or complex congenital heart disease. Onset of advanced HF was associated with 6-fold increase in the risk of mortality. Conventional cardiac intervention was associated with significantly higher risk of mortality as compared to transplant listing. The longer the interval from the initial onset of advanced HF to transplant evaluation, the lower the odds of being listed for transplant. Conclusions: Based on these data, we postulate that early identification of patients with advanced HF, and a timely referral for transplant evaluation (instead of conventional cardiac intervention) may offer the best chance of survival for these critically ill patients. Further studies are required to validate this postulation.

Abstract Objective To identify potential diagnostic and prognostic biomarkers for clinical management and clinical trials in amyotrophic lateral sclerosis. Methods We analysed proteomics data of ALS patient‐induced pluripotent stem cell‐derived motor neurons available through the AnswerALS consortium. After stratifying patients using clinical ALSFRS‐R and ALS‐CBS scales, we identified differentially expressed proteins indicative of ALS disease severity and progression rate as candidate ALS‐related and prognostic biomarkers. Pathway analysis for identified proteins was performed using STITCH. Protein sets were correlated with the effects of drugs using the Connectivity Map tool to identify compounds likely to affect similar pathways. RNAi screening was performed in a Drosophila TDP‐43 ALS model to validate pathological relevance. A statistical classification machine learning model was constructed using ridge regression that uses proteomics data to differentiate ALS patients from controls. Results We identified 76, 21, 71 and 1 candidate ALS‐related biomarkers and 22, 41, 27 and 64 candidate prognostic biomarkers from patients stratified by ALSFRS‐R baseline, ALSFRS‐R progression slope, ALS‐CBS baseline and ALS‐CBS progression slope, respectively. Nineteen proteins enhanced or suppressed pathogenic eye phenotypes in the ALS fly model. Nutraceuticals, dopamine pathway modulators, statins, anti‐inflammatories and antimicrobials were predicted starting points for drug repurposing using the connectivity map tool. Ten diagnostic biomarker proteins were predicted by machine learning to identify ALS patients with high accuracy and sensitivity. Interpretation This study showcases the powerful approach of iPSC‐motor neuron proteomics combined with machine learning and biological confirmation in the prediction of novel mechanisms and diagnostic and predictive biomarkers in ALS.

ObjectiveTo determine the association between race/ethnicity and COVID‐19 outcomes in individuals with systemic lupus erythematosus (SLE).MethodsIndividuals with SLE from the US with data entered into the COVID‐19 Global Rheumatology Alliance registry between March 24, 2020 and August 27, 2021 were included. Variables included age, sex, race, and ethnicity (White, Black, Hispanic, other), comorbidities, disease activity, pandemic time period, glucocorticoid dose, antimalarials, and immunosuppressive drug use. The ordinal outcome categories were: not hospitalized, hospitalized with no oxygenation, hospitalized with any ventilation or oxygenation, and death. We constructed ordinal logistic regression models evaluating the relationship between race/ethnicity and COVID‐19 severity, adjusting for possible confounders.ResultsWe included 523 patients; 473 (90.4%) were female and the mean ± SD age was 46.6 ± 14.0 years. A total of 358 patients (74.6%) were not hospitalized; 40 patients (8.3%) were hospitalized without oxygen, 64 patients (13.3%) were hospitalized with any oxygenation, and 18 (3.8%) died. In a multivariable model, Black (odds ratio [OR] 2.73 [95% confidence interval (95% CI) 1.36–5.53]) and Hispanic (OR 2.76 [95% CI 1.34–5.69]) individuals had higher odds of more severe outcomes than White individuals.ConclusionBlack and Hispanic individuals with SLE experienced more severe COVID‐19 outcomes, which is consistent with findings in the US general population. These results likely reflect socioeconomic and health disparities and suggest that more aggressive efforts are needed to prevent and treat infection in this population.

Abstract Purpose: In many cancers, nivolumab in combination with ipilimumab improves response rates compared with either agent alone, but the combination has not been evaluated in childhood cancer. We conducted a phase I/II trial of nivolumab plus ipilimumab in children and young adults with recurrent/refractory solid tumors. Patients and Methods: ADVL1412, Part C assessed safety of nivolumab plus ipilimumab at two dose levels (DL): DL1 1 mg/kg of each drug and DL2 3 mg/kg nivolumab plus 1 mg/kg ipilimumab. Part D evaluated response at the recommended phase II dose (RP2D) in Ewing sarcoma, rhabdomyosarcoma, and osteosarcoma. Part E tested DL3 (1 mg/kg nivolumab plus 3 mg/kg ipilimumab) in Ewing sarcoma and rhabdomyosarcoma. Tumor response was measured using RECIST v1.1. Pharmacokinetics and PD-L1 expression on archival tissues were assessed. Results: Fifty-five eligible patients enrolled. Based on safety, tolerability, and similar drug exposure to the same doses administered in adults, DL2 was defined as the pediatric RP2D. Among 41 patients treated at the RP2D, 2 patients experienced dose-limiting toxicities during cycle 1, and 4 patients experienced toxicities beyond that period. Two patients had clinically significant sustained partial responses (1 rhabdomyosarcoma, 1 Ewing sarcoma) and 4 had stable disease. Among 8 patients treated at DL3, 3 dose-limiting toxicities (DLT) occurred, all immune-related adverse events; no objective responses were observed. Conclusions: The RP2D of nivolumab (3 mg/kg) plus ipilimumab (1 mg/kg) is well tolerated in children and young adults with solid tumors and shows some clinical activity. Increased dose of ipilimumab (3 mg/kg) plus nivolumab (1 mg/kg) was associated with increased toxicity without clinical benefit.

The influence of pharmacotherapy regimens on surgical patient outcomes is increasingly appreciated in the era of enhanced recovery protocols and institutional focus on reducing postoperative complications. Specifics related to medication selection, dosing, frequency of administration, and duration of therapy are evolving to optimize pharmacotherapeutic regimens for many enhanced recovery protocolized elements. This review provides a summary of recent pharmacotherapeutic strategies, including those configured within electronic health record (EHR) applications and functionalities, that are associated with the minimization of the frequency and severity of postoperative complications (POCs), shortened hospital length of stay (LOS), reduced readmission rates, and cost or revenue impacts. Further, it will highlight preventive pharmacotherapy regimens that are correlated with improved patient preparation, especially those related to surgical site infection (SSI), venous thromboembolism (VTE), nausea and vomiting (PONV), postoperative ileus (POI), and emergence delirium (PoD) as well as less commonly encountered POCs such as acute kidney injury (AKI) and atrial fibrillation (AF). The importance of interprofessional collaboration in all periprocedural phases, focusing on medication management through shared responsibilities for drug therapy outcomes, will be emphasized. Finally, examples of collaborative care through shared mental models of drug stewardship and non-medical practice agreements to improve operative throughput, reduce operative stress, and increase patient satisfaction are illustrated.

ABSTRACT Model organism (MO) research provides a basic understanding of biology and disease due to the evolutionary conservation of the molecular and cellular language of life. MOs have been used to identify and understand the function of orthologous genes, proteins, cells and tissues involved in biological processes, to develop and evaluate techniques and methods, and to perform whole-organism-based chemical screens to test drug efficacy and toxicity. However, a growing richness of datasets and the rising power of computation raise an important question: How do we maximize the value of MOs? In-depth discussions in over 50 virtual presentations organized by the National Institutes of Health across more than 10 weeks yielded important suggestions for improving the rigor, validation, reproducibility and translatability of MO research. The effort clarified challenges and opportunities for developing and integrating tools and resources. Maintenance of critical existing infrastructure and the implementation of suggested improvements will play important roles in maintaining productivity and facilitating the validation of animal models of human biology and disease.

ABSTRACT Background Evaluation and interpretation of the literature on obstructive sleep apnea (OSA) allows for consolidation and determination of the key factors important for clinical management of the adult OSA patient. Toward this goal, an international collaborative of multidisciplinary experts in sleep apnea evaluation and treatment have produced the International Consensus statement on Obstructive Sleep Apnea (ICS:OSA). Methods Using previously defined methodology, focal topics in OSA were assigned as literature review (LR), evidence‐based review (EBR), or evidence‐based review with recommendations (EBR‐R) formats. Each topic incorporated the available and relevant evidence which was summarized and graded on study quality. Each topic and section underwent iterative review and the ICS:OSA was created and reviewed by all authors for consensus. Results The ICS:OSA addresses OSA syndrome definitions, pathophysiology, epidemiology, risk factors for disease, screening methods, diagnostic testing types, multiple treatment modalities, and effects of OSA treatment on multiple OSA‐associated comorbidities. Specific focus on outcomes with positive airway pressure (PAP) and surgical treatments were evaluated. Conclusion This review of the literature consolidates the available knowledge and identifies the limitations of the current evidence on OSA. This effort aims to create a resource for OSA evidence‐based practice and identify future research needs. Knowledge gaps and research opportunities include improving the metrics of OSA disease, determining the optimal OSA screening paradigms, developing strategies for PAP adherence and longitudinal care, enhancing selection of PAP alternatives and surgery, understanding health risk outcomes, and translating evidence into individualized approaches to therapy.