Isolation and profiling of viable tumor cells from human ex vivo glioblastoma cultures through single-cell transcriptomics.

Single-cell RNA-sequencing (scRNA-seq) is becoming a ubiquitous method in profiling the cellular transcriptomes of both malignant and non-malignant cells from the human brain. Here, we present a protocol to isolate viable tumor cells from human ex vivo glioblastoma cultures for single-cell transcriptomic analysis. We describe steps including surgical tissue collection, sectioning, culturing, primary tumor cells inoculation, growth tracking, fluorescence-based cell sorting, and population-enriched scRNA-seq. This comprehensive methodology empowers in-depth understanding of brain tumor biology at the single-cell level. For complete details on the use and execution of this protocol, please refer to Ravi et al.1.

Spontaneous splenic rupture associated with apixaban: a case report.

BACKGROUND: Spontaneous splenic rupture associated with anticoagulant use is a rare but potentially lethal disorder. Lack of prompt recognition can be associated with poor patient outcomes. The use of novel oral anticoagulants is becoming more common and thus consideration of this disorder while evaluating a patient who presents with abdominal pain while using these agents is extremely important. This is the first reported case of spontaneous splenic rupture associated with apixaban. CASE PRESENTATION: We describe the clinical case of an 83-year-old white man who complained of sudden severe abdominal pain 5 days into a hospital stay for acute-on-chronic congestive heart failure and exacerbation of chronic obstructive pulmonary disease. Neither he nor his wife reported any significant trauma for the past 6 months prior to his admission. His medical history included chronic atrial fibrillation treated with medications including apixaban 2.5 mg twice daily. An urgent abdominal computed tomography scan demonstrated a large splenic hematoma and evidence of intraperitoneal bleeding from which he rapidly declined, developing hypovolemic shock. An emergency splenic arteriogram displayed a patent splenic artery and an embolization was successful in stabilizing him. Due to evidence of recurrent bleeding, an exploratory laparotomy and splenectomy was subsequently performed the following day. CONCLUSIONS: The diagnosis of spontaneous splenic rupture is important to consider in a patient using apixaban who presents with abdominal pain and associated signs of hypotension and anemia. For hemodynamically unstable patients, prompt treatment to stop significant bleeding through splenic artery embolization or splenectomy is warranted and may be lifesaving.

ICU Admission, Discharge, and Triage Guidelines: A Framework to Enhance Clinical Operations, Development of Institutional Policies, and Further Research.

OBJECTIVES: To update the Society of Critical Care Medicine's guidelines for ICU admission, discharge, and triage, providing a framework for clinical practice, the development of institutional policies, and further research. DESIGN: An appointed Task Force followed a standard, systematic, and evidence-based approach in reviewing the literature to develop these guidelines. MEASUREMENTS AND MAIN RESULTS: The assessment of the evidence and recommendations was based on the principles of the Grading of Recommendations Assessment, Development and Evaluation system. The general subject was addressed in sections: admission criteria and benefits of different levels of care, triage, discharge timing and strategies, use of outreach programs to supplement ICU care, quality assurance/improvement and metrics, nonbeneficial treatment in the ICU, and rationing considerations. The literature searches yielded 2,404 articles published from January 1998 to October 2013 for review. Following the appraisal of the literature, discussion, and consensus, recommendations were written. CONCLUSION: Although these are administrative guidelines, the subjects addressed encompass complex ethical and medico-legal aspects of patient care that affect daily clinical practice. A limited amount of high-quality evidence made it difficult to answer all the questions asked related to ICU admission, discharge, and triage. Despite these limitations, the members of the Task Force believe that these recommendations provide a comprehensive framework to guide practitioners in making informed decisions during the admission, discharge, and triage process as well as in resolving issues of nonbeneficial treatment and rationing. We need to further develop preventive strategies to reduce the burden of critical illness, educate our noncritical care colleagues about these interventions, and improve our outreach, developing early identification and intervention systems.

Relations of Parenting to Adolescent Externalizing and Internalizing Distress Moderated by Perception of Neighborhood Danger.

Parental monitoring and warmth have traditionally been studied in the context of White, middle-class families. This article explores optimal levels of these parenting behaviors in preventing adolescent psychopathology in impoverished, urban high-crime areas while accounting for child perceptions of neighborhood danger. In this study, data were collected longitudinally at 2 time points 1 year apart from a sample of 254 African American young adolescents (T1: M age = 12.6 years, 41% male) and their parents. Parental monitoring and warmth, child perception of neighborhood danger, and child internalizing and externalizing behaviors were measured using questionnaires. Child internalizing behaviors were also measured using a time sampling technique capturing in vivo accounts of daily distress. Findings indicated associations between parental monitoring and children's externalizing behaviors along with linear and quadratic associations between parental monitoring and internalizing behaviors. Monitoring and warmth were differentially related to symptoms depending on neighborhood danger level. When children perceived less danger, more monitoring related to less externalizing. When children perceived more danger, more warmth related to less internalizing. In addition, adolescents' perceptions of neighborhood danger emerged as equally strong as monitoring and warmth in predicting symptoms. This study underscores the influence of carefully considering parenting approaches and which techniques optimally prevent adolescents' externalizing, as well as prevent internalizing difficulties. It also highlights how context affects mental health, specifically how perceptions of danger negatively influence adolescents' psychopathology, emphasizing the importance of initiatives to reduce violence in communities.

The Relation of Severity and Type of Community Violence Exposure to Emotional Distress and Problem Behaviors Among Urban African American Adolescents.

Severity level and type of exposure to community violence were examined to determine their effect on emotional distress and problem behaviors among 234 low-income urban African American early adolescents. There were 4 violence exposure scales developed from a principal component analysis of the Richters and Martinez (1993) exposure to violence scale: moderate and severe witnessing and moderate and severe victimization. Regression analyses indicated that moderate victimization was the most consistent predictor of emotional distress and behavioral problems, whereas moderate witnessing did not relate to any of the dependent variables. Severe victimization predicted depression and delinquency, whereas severe witnessing predicted posttraumatic stress disorder symptoms and delinquency. Witnessing and victimization scales based on severity of exposure better represented the experience than combining all data into a single exposure or simply witnessing and victimization scales.

Metoprolol-induced visual hallucinations: a case series.

INTRODUCTION: Metoprolol is a widely used beta-adrenergic blocker that is commonly prescribed for a variety of cardiovascular syndromes and conditions. While central nervous system adverse effects have been well-described with most beta-blockers (especially lipophilic agents such as propranolol), visual hallucinations have been only rarely described with metoprolol. CASE PRESENTATIONS: Case 1 was an 84-year-old Caucasian woman with a history of hypertension and osteoarthritis, who suffered from visual hallucinations which she described as people in her bedroom at night. They would be standing in front of the bed or sitting on chairs watching her when she slept. Numerous medications were stopped before her physician realized the metoprolol was the causative agent. The hallucinations resolved only after discontinuation of this medication.Case 2 was a 62-year-old Caucasian man with an inferior wall myocardial infarction complicated by cardiac arrest, who was successfully resuscitated and discharged from the hospital on metoprolol. About 18 months after discharge, he related to his physician that he had been seeing dead people at night. He related his belief that since he 'had died and was brought back to life', he was now seeing people from the after-life. Upon discontinuation of the metoprolol the visual disturbances resolved within several days.Case 3 was a 68 year-old Caucasian woman with a history of severe hypertension and depression, who reported visual hallucinations at night for years while taking metoprolol. These included awakening during the night with people in her bedroom and seeing objects in her room turn into animals. After a new physician switched her from metoprolol to atenolol, the visual hallucinations ceased within four days. CONCLUSION: We suspect that metoprolol-induced visual hallucinations may be under-recognized and under-reported. Patients may frequently fail to acknowledge this adverse effect believing that they are just dreaming, or may be embarrassed to report visions that they feel will not be perceived by others to be real. Similarly, healthcare providers can also fail to recognize this visual toxicity or attribute visual hallucinations to concurrent illness or other medications. Clinicians must maintain diligent surveillance when managing patients receiving this drug.

Exposure to community violence and protective and risky contexts among low income urban African American adolescents: a prospective study.

This study examined protective and risky companionship and locations for exposure to community violence among African American young adolescents living in high crime, urban areas. The Experience Sampling Method (ESM), an in vivo data collection method, was employed to gather information from 233 students (62% female) over 3 years, beginning in the 6th grade. Questionnaire variables of exposure to community violence were regressed onto ESM companionship and location variables, cross-sectionally and longitudinally, separately for boys and girls. At different points, time spent with parents, in school, and outside in private space was associated with less exposure to violence for boys and girls, while time spent with girls was protective for boys. In addition, time spent outside in public and with older peers was associated with increased risk for boys and girls. These findings are discussed in relation to previous and potential future research, and to strategies to prevent exposure to community violence.