A Semiautomated Proteomics and Phosphoproteomics Protocol for the Identification of Novel Therapeutic Targets and Predictive Biomarkers in In Vivo Xenograft Models of Pediatric Cancers.

Genomic profiling has identified therapeutic targets for precision treatment of certain cancers, but many patients lack actionable mutations. Additional omics approaches, like proteomics and phosphoproteomics, are essential for comprehensive mapping of cancer-associated molecular phenotypes. In vivo models, such as cell line and patient-derived xenografts (PDX), offer valuable insights into cancer biology and treatment strategies.This chapter presents a semiautomated high-throughput workflow for integrated proteomics and phosphoproteomics analysis on the Kingfish platform coupled with MagReSyn® Zr-IMAC HP. It enhances protein extraction from in vivo xenograft samples and provides better insights into cancers with poor prognosis. The approach successfully identified over 11,000 unique phosphosites and ~6000 proteins in SJSA-1 pediatric osteosarcoma xenografts, demonstrating its efficacy. This workflow is a valuable tool for studying tumor biology and developing precision oncology strategies.

Generation and multi-dimensional profiling of a childhood cancer cell line atlas defines new therapeutic opportunities.

Pediatric solid and central nervous system tumors are the leading cause of cancer-related death among children. Identifying new targeted therapies necessitates the use of pediatric cancer models that faithfully recapitulate the patient's disease. However, the generation and characterization of pediatric cancer models has significantly lagged behind adult cancers, underscoring the urgent need to develop pediatric-focused cell line resources. Herein, we establish a single-site collection of 261 cell lines, including 224 pediatric cell lines representing 18 distinct extracranial and brain childhood tumor types. We subjected 182 cell lines to multi-omics analyses (DNA sequencing, RNA sequencing, DNA methylation), and in parallel performed pharmacological and genetic CRISPR-Cas9 loss-of-function screens to identify pediatric-specific treatment opportunities and biomarkers. Our work provides insight into specific pathway vulnerabilities in molecularly defined pediatric tumor classes and uncovers biomarker-linked therapeutic opportunities of clinical relevance. Cell line data and resources are provided in an open access portal.

Emergency department patients' perceptions of the efficacy and safety of opioid analgesics.

INTRODUCTION: Emergency department (ED) providers are on the forefront of the prescription drug crisis and understand-ing patient's perceptions of opioids may allow physicians to better address misconceptions. The aim was to determine the perceptions of ED patients regarding the efficacy and safety of opioid analgesics. METHODS: Cross-sectional study of a convenience sample of adults at a single urban academic ED. Patients completed a tablet-based survey regarding the efficacy and safety of opioid analgesics. RESULTS: Of the 715 subjects, the sample was predominantly black (80.4 percent), female (59.2 percent), and aged 18-59 years (76.8 percent). The majority (70.1 percent) of respondents reported pain as the reason for visit. Seventy-two percent had previously taken an opioid primarily for acute pain, found them effective for pain (88.2 percent), and would be willing to do so again (62.7 percent). Adverse effects made patients less likely to use them again (OR 0.703, [0.659-0.751]). Gender and age did not affect perceptions of efficacy and safety, but certain racial groups did (OR 1.08, [1.02 to 1.14], p < 0.05). Knowing someone who used opioids in a nonmedical manner did not impact willingness to use opioids. Many (54 percent) believed opioids to be as safe as nonopioid alternatives. The majority (78 percent) consid-ered prescription opioid abuse a public health problem, however underestimated the number of annual opioid-related deaths. CONCLUSION: The majority of ED patients had used opioids and considered the prescription drug crisis a public health problem. Through personal use or media coverage, the majority of participants considered opioids safe, even compared to nonopioid alternatives. Better understanding these viewpoints may improve patient-physician communication about analgesic treatment.

A Mixed Presentation of Serotonin Syndrome vs Neuroleptic Malignant Syndrome in a 12-Year-Old Boy.

BACKGROUND: Neuroleptic malignant syndrome (NMS) and serotonin syndrome (SS) are serious medical conditions associated with commonly prescribed psychiatric medications. Although the mechanisms differ, they can be clinically difficult to distinguish. We report a case of a pediatric patient with complicated psychiatric history that developed features of both syndromes in the setting of polypharmacy. CASE: A 12-year-old boy with a history of developmental delay, attention-deficit hyperactivity disorder, and posttraumatic stress disorder presented to the emergency department with behavior changes consisting of delayed reactions, gait instability, drooling, and slowed movements. Ten days before presentation, his outpatient psychiatrist had made multiple medication changes including discontinuation of cyproheptadine (an appetite stimulant) and initiation of aripiprazole. On arrival, the patient was noted to be tachycardia and hypertensive for age. He was disoriented, intermittently agitated, and tremulous with increased tonicity, clonus in the lower extremities, and mydriasis. He was supportively treated with lorazepam and intravenous fluids while discontinuing potential offending agents. His course was complicated by hypertension and agitation managed with dexmedetomidine infusion and benzodiazepines. His mental status, tremors, and laboratory values began to improve over the next 2 days, and eventually transitioned to the inpatient psychiatric unit on hospital day 7. DISCUSSION: Diagnosis of NMS or SS can be difficult when there is overlap between syndromes, particularly in the setting of multiple potential offending agents or underlying developmental delay. In addition, pediatric patients may present atypically as compared with adult patients with the same condition. CONCLUSION: The use of antipsychotic medications for young children with behavioral problems has risen dramatically in the last decade, increasing their risk for developing SS or NMS.

Child and adolescent benzodiazepine exposure and overdose in the United States: 16 years of poison center data.

Background: Recently, there has been an increase in prescription drug abuse and related fatalities. Although opioid analgesics are commonly implicated, there have been significant increases in the prevalence of benzodiazepine exposures and overdoses.Objective: To describe national trends in pediatric benzodiazepine exposures from 2000 to 2015.Methods: A retrospective database analysis was conducted. Data regarding benzodiazepine exposures in children ages 0 to <18 years reported to participating United States poison centers from January 2000 through December 2015 were obtained from the National Poison Data System. Population data were obtained from the US Census Bureau to determine annual population estimates. Data were analyzed using chi-square tests.Results: A total of 296,838 pediatric benzodiazepine exposures were identified during the study period. The rate of pediatric benzodiazepine exposure increased 54% between 2000 and 2015. The severity of medical outcomes also increased, as did the prevalence of co-ingestion of multiple drugs, especially in children ages 12 to <18 years. Nearly half of all reported exposures in 2015 were documented as intentional abuse, misuse, or attempted suicide, reflecting a change from prior years. The most commonly identified pediatric benzodiazepines of exposures were alprazolam, clonazepam, and lorazepam.Conclusions: The rate and severity of reported pediatric benzodiazepine exposure is increasing over time. Adolescent exposures are of specific concern, as co-ingestion and intentional abuse were found to be more common in this group. Medical providers and caretakers should be cognizant of this growing epidemic to avoid preventable harm to adolescents, young children, and infants.

Brugada-Type Pattern on Electrocardiogram Associated with High-Dose Loperamide Abuse.

BACKGROUND: Loperamide has been increasing in popularity recently for its effects separate from treatment of diarrhea. In large doses or in combination with other agents, it can lead to desirable effects in the central nervous system. However, cardiotoxicity has been reported with its abuse. CASE REPORT: A 49-year-old male who had been chronically abusing loperamide was found to have Brugada-like changes on his electrocardiogram (ECG). He had no other clinical symptoms associated with Brugada syndrome and did not have similar findings on previous ECGs. After abstaining from further loperamide use during hospitalization, this pattern slowly resolved without clinically significant dysrthymias. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: A patient with a history of loperamide abuse is at risk for cardiotoxicity. While other dysrhythmias are reported more commonly with loperamide abuse, Brugada-like ECG changes can occur and warrant appropriate consultation and prompt follow-up.

Trends in carisoprodol abuse and misuse after regulatory scheduling: a retrospective review of California poison control calls from 2008 to 2015.

BACKGROUND: In January 2012, carisoprodol was classified as a Schedule IV substance under the controlled substances act from a previously non-controlled, non-scheduled classification. Carisoprodol is marketed as a skeletal muscle relaxant and is commonly cited for its abuse potential. OBJECTIVES: We aimed to compare volume of calls involving carisoprodol abuse or misuse to a statewide poison control system before and after the scheduling change. METHODS: Data were extracted from poison control calls coded as "misuse/abuse" involving carisoprodol from four years before (2008 to 2011) and four years after (2012 to 2015) the scheduling change. The volume of calls from pre- and post-scheduling change was compared after adjusting for yearly California census data. RESULTS: The number of calls related to carisoprodol abuse or misuse was significantly decreased in the four years following the change compared to the four years before. CONCLUSION: Scheduling of carisoprodol was temporally related to decreased exposures as reported to California Poison Control Centers. Governmental regulation may impact a drug's potential for abuse.

Trifluoroacetic acid: Three times the fluoride, three times the toxicity?

Trifluoroacetic acid (TFAA) is a carboxylic acid, similar to acetic acid, used industrially and in laboratories. There is a paucity of data regarding exposure and the concern is that toxicity may mimic that of hydrofluoric acid (HF), causing electrolyte abnormalities, dysrhythmia, and cardiac arrest. We report a case of a 27-year-old male that presents with a dermal exposure to TFAA. His exam was remarkable for a 4% body surface area partial thickness burn to the right forearm. There were no initial electrolyte abnormalities or dysrhythmias and the patient was admitted to telemetry monitoring overnight. Serial laboratories were normal, dysrhythmias did not occur, and patient was ultimately discharged with routine burn care of the wound. Previously reported TFAA exposures are uncommon and tend to be very small body surface area chemical burns without clear systemic toxicity. HF as well as sodium and ammonium bifluorides have been shown to cause clinically significant electrolyte disturbances that can lead to dysrhythmias and fatalities. While TFAA may be structurally similar, it does not appear to have similar toxicity. This is an important difference in presentation and treatment that emergency physicians should be aware of as occupational exposures are likely to present to the emergency department.

Trends in Opioid Analgesic Use in Encounters Involving Physician Trainees in U.S. Emergency Departments.

BACKGROUND: Opioid analgesic use has increased dramatically in emergency departments (EDs), but the relative contribution of physician trainees has not been explored. We assessed trends in opioid utilization focusing on ED encounters where a physician trainee was involved. METHODS: We studied ED visits from the National Hospital Ambulatory Medical Care Survey, 2001-2011. Adult ED visits in which an opioid was administered in the ED or prescribed at discharge were stratified by whether or not there was trainee involvement. Trends in use over time for five common opioids (codeine, hydrocodone, hydromorphone, morphine, oxycodone) were tested using survey-weighted logistic regression. RESULTS: From 2001-02 to 2009-11, the proportion of ED visits where an opioid analgesic was used increased 31.5% from 21.9% (95% CI: 20.3-23.6) of visits to 28.8% (95% CI: 27.5-30.1). Trainee involvement in ED visits was stable, with 9.3% (95% CI: 7.7-11.3) seen by a trainee in 2001-02 vs. 10.2% (95% CI: 8.1-12.7) in 2010-11. Opioid use in visits with trainee involvement did not change significantly over time relative to visits without a trainee (increase of 36.8% compared to 31.2% without trainees, P = 0.652). Trends in opioid utilization for trainee visits paralleled non-trainee visits. Hydromorphone had the greatest relative increase in use for all providers. Adjusted for patient- and hospital-level factors, the probability of receiving opioids when a trainee was involved increased to a greater extent than among non-trainee visits (30.9% vs. 24.0%). CONCLUSION: Opioid utilization patterns for visits involving trainees reflect similar trends in attending practice, and highlights the more liberal opioid prescribing climate over time.

Accuracy of rapid sequence intubation medication dosing in obese patients intubated in the ED.

OBJECTIVE: There are limited data regarding appropriateness of sedative and paralytic dosing of obese patients undergoing rapid sequence intubation (RSI) in the emergency department. The goal of this study was to compare rates of appropriate succinylcholine and etomidate doses in obese and nonobese patients. METHODS: Retrospective review using a database of endotracheally intubated patients using RSI in an urban, tertiary care academic emergency department, from November 2009 to June 2011. Dosing for succinylcholine and etomidate was calculated as milligrams per kilogram of total body weight (TBW) for each patient, defining appropriate dosing as succinylcholine 1-1.5 mg/kg TBW and etomidate 0.2-0.4 mg/kg TBW. Logistic regression analysis was used to estimate the association between appropriate dosing and World Health Organization body mass index classification. RESULTS: A total of 440 patients were included in the study, 311 (70.7%) classified as nonobese and 129 (29.3%) as obese. two hundred thirty-three (56%) received an inappropriate succinylcholine dose and 107 (24%) received an inappropriate etomidate dose. Obese patients were more likely to be underdosed with succinylcholine (odds ratio [OR], 63.7; 95% confidence interval [CI], 17.8-228.1) and etomidate (OR, 178.3; 95% CI, 37.6-844.7). Nonobese patients were more likely to be overdosed with succinylcholine (OR, 62.5; 95% CI, 17.9-250) and etomidate (OR, 166.7; 95% CI, 37.0-1000). CONCLUSION: Obese patients were more likely to be underdosed during RSI compared with nonobese patients, whereas nonobese patients were more likely to be overdosed with RSI medications. Most obese and nonobese patients were inappropriately dosed with RSI medications, suggesting that physicians are not dosing these medications based on weight.

Trends in Emergency Department Resource Utilization for Poisoning-Related Visits, 2003-2011.

In recent years, there has been an increase in poisoning-related emergency department (ED) visits. This study examines trends in ED resource utilization for poisoning-related visits over time. A retrospective review of data from the National Hospital Ambulatory Medical Care Survey, 2003-2011, was conducted. All ED visits with a reason for visit or ICD-9 code related to poisoning were included. We examined the number of ED visits and resources used including diagnostic studies and procedures performed, medications provided, admission rates, and length of stay. The proportion of visits involving resource use was tabulated and trends analyzed using survey-weighted logistic regression, grouping into 2-year periods to ensure adequate sample size. Of an estimated 843 million ED visits between 2003 and 2011, 8 million (0.9 %) were related to poisoning. Visits increased from 1.8 million (0.8 %) visits in 2003-2004 to 2.9 million (1.1 %) visits in 2010-2011, p = 0.001. Use of laboratory studies, EKGs, plain radiographs, and procedures remained stable across the study period. CT use was more than doubled, increasing from 5.2 to 13.7 % of visits, p = 0.001. ED length of stay increased by 35.5 % from 254 to 344 min, p = 0.001. Admission rates increased by 45.3 %, from 15.0 to 21.8 %, p = 0.046. Over the entire study period, 52.0 % of poisoned patients arrived via ambulance, and 3.0 % of patients had been discharged from the hospital within the previous 7 days. Poisoning-related ED visits increased over the 8-year study period; poisonings are resource-intensive visits and require increasingly longer lengths of ED stay or hospital admission.