Drugs of Abuse: Sympathomimetics.

Sympathomimetic drugs comprise a broad category of substances including both illicit and prescribed drugs that have deleterious effects when ingested or abused. The clinical syndromes that result from overstimulation of the sympathetic nervous system by reuptake inhibition of biogenic amines, such as norepinephrine and dopamine, carry significant morbidity. Recognition and awareness of the appropriate supportive measures are required to mitigate life-threatening complications of multiple organ systems. The sympathomimetic toxidrome is recognized by a constellation of symptoms including agitation, hyperthermia, tachycardia, and hypertension, and the primary treatment involves supportive care, including the liberal use of benzodiazepines.

Effect of an Educational Intervention on Medical Student Scripting and Patient Satisfaction: A Randomized Trial.

INTRODUCTION: Effective communication between clinicians and patients has been shown to improve patient outcomes, reduce malpractice liability, and is now being tied to reimbursement. Use of a communication strategy known as "scripting" has been suggested to improve patient satisfaction in multiple hospital settings, but the frequency with which medical students use this strategy and whether this affects patient perception of medical student care is unknown. Our objective was to measure the use of targeted communication skills after an educational intervention as well as to further clarify the relationship between communication element usage and patient satisfaction. METHODS: Medical students were block randomized into the control or intervention group. Those in the intervention group received refresher training in scripted communication. Those in the control group received no instruction or other intervention related to communication. Use of six explicit communication behaviors were recorded by trained study observers: 1) acknowledging the patient by name, 2) introducing themselves as medical students, 3) explaining their role in the patient's care, 4) explaining the care plan, 5) providing an estimated duration of time to be spent in the emergency department (ED), and 6) notifying the patient that another provider would also be seeing them. Patients then completed a survey regarding their satisfaction with the medical student encounter. RESULTS: We observed 474 medical student-patient encounters in the ED (231 in the control group and 243 in the intervention group). We were unable to detect a statistically significant difference in communication element use between the intervention and control groups. One of the communication elements, explaining steps in the care plan, was positively associated with patient perception of the medical student's overall communication skills. Otherwise, there was no statistically significant association between element use and patient satisfaction. CONCLUSION: We were unable to demonstrate any improvement in student use of communication elements or in patient satisfaction after refresher training in scripted communication. Furthermore, there was little variation in patient satisfaction based on the use of scripted communication elements. Effective communication with patients in the ED is complicated and requires further investigation on how to provide this skill set.

Hyaluronic acid 35 normalizes TLR4 signaling in Kupffer cells from ethanol-fed rats via regulation of microRNA291b and its target Tollip.

TLR4 signaling in hepatic macrophages is increased after chronic ethanol feeding. Treatment of hepatic macrophages after chronic ethanol feeding with small-specific sized hyaluronic acid 35 (HA35) normalizes TLR4 signaling; however, the mechanisms for HA35 action are not completely understood. Here we used Next Generation Sequencing of microRNAs to identify negative regulators of TLR4 signaling reciprocally modulated by ethanol and HA35 in hepatic macrophages. Eleven microRNAs were up-regulated by ethanol; only 4 microRNAs, including miR291b, were decreased by HA35. Bioinformatics analysis identified Tollip, a negative regulator of TLR4, as a target of miR291b. Tollip expression was decreased in hepatic macrophages from ethanol-fed rats, but treatment with HA35 or transfection with a miR291b hairpin inhibitor restored Tollip expression and normalized TLR4-stimulated TNFα expression. In peripheral blood monocytes isolated from patients with alcoholic hepatitis, expression of TNFα mRNA was robustly increased in response to challenge with lipopolysaccharide. Importantly, pre-treatment with HA35 reduced TNFα expression by more than 50%. Taken together, we have identified miR291b as a critical miRNA up-regulated by ethanol. Normalization of the miR291b → Tollip pathway by HA35 ameliorated ethanol-induced sensitization of TLR4 signaling in macrophages/monocytes, suggesting that HA35 may be a novel therapeutic agent in the treatment of ALD.

MicroRNA 181b-3p and its target importin α5 regulate toll-like receptor 4 signaling in Kupffer cells and liver injury in mice in response to ethanol.

Increased inflammatory signaling by Kupffer cells contributes to alcoholic liver disease (ALD). Here we investigated the impact of small, specific-sized hyaluronic acid of 35 kD (HA35) on ethanol-induced sensitization of Kupffer cells, as well as ethanol-induced liver injury in mice. Unbiased analysis of microRNA (miRNA) expression in Kupffer cells identified miRNAs regulated by both ethanol and HA35. Toll-like receptor 4 (TLR4)-mediated signaling was assessed in primary cultures of Kupffer cells from ethanol- and pair-fed rats after treatment with HA35. Female C57BL6/J mice were fed ethanol or pair-fed control diets and treated or not with HA35. TLR4 signaling was increased in Kupffer cells by ethanol; this sensitization was normalized by ex vivo treatment with HA35. Next generation sequencing of Kupffer cell miRNA identified miRNA 181b-3p (miR181b-3p) as sensitive to both ethanol and HA35. Importin α5, a protein involved in p65 translocation to the nucleus, was identified as a target of miR181b-3p; importin α5 protein was increased in Kupffer cells from ethanol-fed rats, but decreased by HA35 treatment. Overexpression of miR181b-3p decreased importin α5 expression and normalized lipopolysaccharide-stimulated tumor necrosis factor α expression in Kupffer cells from ethanol-fed rats. In a mouse model of ALD, ethanol feeding decreased miR181b-3p in liver and increased expression of importin α5 in nonparenchymal cells. Treatment with HA35 normalized these changes and also protected mice from ethanol-induced liver and intestinal injury. CONCLUSION: miR181b-3p is dynamically regulated in Kupffer cells and mouse liver in response to ethanol and treatment with HA35. miR181b-3p modulates expression of importin α5 and sensitivity of TLR4-mediated signaling. This study identifies a miR181b-3p-importin α5 axis in regulating inflammatory signaling pathways in hepatic macrophages. (Hepatology 2017;66:602-615).

Myeloid-MyD88 Contributes to Ethanol-Induced Liver Injury in Mice Linking Hepatocellular Death to Inflammation.

BACKGROUND: Toll-like receptor 4 (TLR4) is critical for ethanol (EtOH)-induced liver injury. TLR4 signaling is mediated by 2 proximal adaptor molecules: myeloid differentiation primary response protein (MyD88) and TLR-domain-containing adaptor-inducing interferon-ß (TRIF). Studies utilizing global knockouts of MyD88 and TRIF identified a predominant role for TRIF signaling in the progression of EtOH-induced liver injury. In contrast, IL-1 receptor, which signals solely via the MyD88 pathway, is also known to mediate EtOH-induced liver injury. We postulated that a cell-specific role for MyD88 in myeloid cells might explain these apparently discrepant roles of MyD88. Here we made use of myeloid-specific MyD88-deficient (MyD88LysM-KO ) mice generated by crossing LysM-CRE mice with MyD88fl/fl mice to test this hypothesis. METHODS: MyD88LysM-KO and littermate controls were fed a Lieber-DeCarli EtOH-containing diet or pair-fed control diets for 25 days. RESULTS: Littermate control, but not MyD88LysM-KO , mice developed early stages of EtOH-induced liver injury including elevated plasma alanine aminotransferase and increased hepatic triglycerides. Lobular inflammation and expression of pro-inflammatory cytokines/chemokines was increased in control but not MyD88LysM-KO . Further, EtOH-induced inflammasome activation, indicated by the presence of cleaved caspase-1 and mature IL-1ß protein, was also ameliorated in livers of MyD88LysM-KO mice. In contrast, chronic EtOH-induced apoptosis, assessed via TUNEL staining, was independent of myeloid-MyD88 expression. CONCLUSIONS: Collectively, these data demonstrate a cell-specific role for MyD88 in the development of chronic EtOH-induced liver injury. While MyD88LysM-KO still exhibited hepatocellular apoptosis in response to chronic EtOH, the absence of MyD88 on myeloid cells prevented the development of hepatic steatosis and inflammation.

Limited Excessive Voluntary Alcohol Drinking Leads to Liver Dysfunction in Mice.

BACKGROUND: Liver damage is a serious and sometimes fatal consequence of long-term alcohol intake, which progresses from early-stage fatty liver (steatosis) to later-stage steatohepatitis with inflammation and fibrosis/necrosis. However, very little is known about earlier stages of liver disruption that may occur in problem drinkers, those who drink excessively but are not dependent on alcohol. METHODS: We examined how repeated binge-like alcohol drinking in C57BL/6 mice altered liver function, as compared with a single binge-intake session and with repeated moderate alcohol consumption. We measured a number of markers associated with early- and later-stage liver disruption, including liver steatosis, measures of liver cytochrome P4502E1 (CYP2E1) and alcohol dehydrogenase (ADH), alcohol metabolism, expression of cytokine mRNA, accumulation of 4-hydroxynonenal (4-HNE) as an indicator of oxidative stress, and alanine transaminase/aspartate transaminase as a measure of hepatocyte injury. RESULTS: Importantly, repeated binge-like alcohol drinking increased triglyceride levels in the liver and plasma, and increased lipid droplets in the liver, indicators of steatosis. In contrast, a single binge-intake session or repeated moderate alcohol consumption did not alter triglyceride levels. In addition, alcohol exposure can increase rates of alcohol metabolism through CYP2E1 and ADH, which can potentially increase oxidative stress and liver dysfunction. Intermittent, excessive alcohol intake increased liver CYP2E1 mRNA, protein, and activity, as well as ADH mRNA and activity. Furthermore, repeated, binge-like drinking, but not a single binge or moderate drinking, increased alcohol metabolism. Finally, repeated, excessive intake transiently elevated mRNA for the proinflammatory cytokine IL-1B and 4-HNE levels, but did not alter markers of later-stage liver hepatocyte injury. CONCLUSIONS: Together, we provide data suggesting that even relatively limited binge-like alcohol drinking can lead to disruptions in liver function, which might facilitate the transition to more severe forms of liver damage.

Receptor interacting protein 3 protects mice from high-fat diet-induced liver injury.

Multiple pathways of programmed cell death are important in liver homeostasis. Hepatocyte death is associated with progression of nonalcoholic fatty liver disease, and inhibition of apoptosis partially protects against liver injury in response to a high-fat diet (HFD). However, the contribution of necroptosis, a caspase-independent pathway of cell death, to HFD-induced liver injury is not known. Wild-type C57BL/6 and receptor interacting protein (RIP) 3-/- mice were randomized to chow or HFD. HFD-fed C57BL/6 mice increased expression of RIP3, the master regulator of necroptosis, as well as phosphorylated mixed lineage kinase domain-like, an effector of necroptotic cell death, in liver. HFD did not increase phosphorylated mixed lineage kinase domain-like in RIP3-/- mice. HFD increased fasting insulin and glucose, as well as glucose intolerance, in C57BL/6 mice. RIP3-/- mice were glucose-intolerant even on the chow diet; HFD further increased fasting glucose and insulin but not glucose intolerance. HFD also increased hepatic steatosis, plasma alanine aminotransferase activity, inflammation, oxidative stress, and hepatocellular apoptosis in wild-type mice; these responses were exacerbated in RIP3-/- mice. Importantly, increased inflammation and injury were associated with early indicators of fibrosis in RIP3-/- compared to C57BL/6 mice. Culture of AML12 hepatocytes with palmitic acid increased cytotoxicity through apoptosis and necrosis. Inhibition of RIP1 with necrostatin-1 or small interfering RNA knockdown of RIP3 reduced palmitic acid-induced cytotoxicity. CONCLUSION: Absence of RIP3, a key mediator of necroptosis, exacerbated HFD-induced liver injury, associated with increased inflammation and hepatocyte apoptosis, as well as early fibrotic responses; these findings indicate that shifts in the mode of hepatocellular death can influence disease progression and have therapeutic implications because manipulation of hepatocyte cell death pathways is being considered as a target for treatment of nonalcoholic fatty liver disease. (Hepatology 2016;64:1518-1533).

Myeloid Mixed Lineage Kinase 3 Contributes to Chronic Ethanol-Induced Inflammation and Hepatocyte Injury in Mice.

Proinflammatory activity of hepatic macrophages plays a key role during progression of alcoholic liver disease (ALD). Since mixed lineage kinase 3 (MLK3)-dependent phosphorylation of JNK is involved in the activation of macrophages, we tested the hypothesis that myeloid MLK3 contributes to chronic ethanol-induced inflammatory responses in liver, leading to hepatocyte injury and cell death. Primary cultures of Kupffer cells, as well in vivo chronic ethanol feeding, were used to interrogate the role of MLK3 in the progression of liver injury. Phosphorylation of MLK3 was increased in primary cultures of Kupffer cells isolated from ethanol-fed rats compared to cells from pair-fed rats. Kupffer cells from ethanol-fed rats were more sensitive to LPS-stimulated cytokine production; this sensitization was normalized by pharmacological inhibition of MLK3. Chronic ethanol feeding to mice increased MLK3 phosphorylation robustly in F4/80(+) Kupffer cells, as well as in isolated nonparenchymal cells. MLK3(-/-) mice were protected from chronic ethanol-induced phosphorylation of MLK3 and JNK, as well as multiple indicators of liver injury, including increased ALT/AST, inflammatory cytokines, and induction of RIP3. However, ethanol-induced steatosis and hepatocyte apoptosis were not affected by MLK3. Finally, chimeric mice lacking MLK3 only in myeloid cells were also protected from chronic ethanol-induced phosphorylation of JNK, expression of inflammatory cytokines, and increased ALT/AST. MLK3 expression in myeloid cells contributes to phosphorylation of JNK, increased cytokine production, and hepatocyte injury in response to chronic ethanol. Our data suggest that myeloid MLK3 could be targeted for developing potential therapeutic strategies to suppress liver injury in ALD patients.

Development of a disaster preparedness curriculum for medical students: a pilot study of incorporating local events into training opportunities.

OBJECTIVE: Contemporary disasters, like the outbreak of Ebola in West Africa, have piqued the interest of medical students in disaster preparedness. The topic is also a requirement of undergraduate medical education.(1) Yet current literature suggests that disaster preparedness education is lacking. Our objective was to pilot a curriculum to augment medical students' disaster preparedness education by marshalling local resources to provide practical hands-on experiences. DESIGN: This pilot curriculum consisted of lectures; simulations; asynchronous learning materials; a large-scale, regional disaster exercise; and preparation for and participation in a real-time mass gathering. Outcomes were measured by student performance on written tests and evaluations of each activity. SETTING: Academic Health Center with associated medical school. PARTICIPANTS: Fifty-two medical students participated in at least one of the six activities during this voluntary pilot program. Premedical students and residents (n=57) participated in some activities. RESULTS: Forty-one medical students took either the pretest or the post-test over the curriculum. Only eight students took both. A paired t test comparing pretest to post-test scores using imputed missing data (t=-11.72, df=40, p≤0.001) was consistent with an analysis using only complete data (t=-2.35, df=7, p=0.05), implying that student scores improved significantly over time. Evaluations indicated a student preference for hands-on over didactic or independent learning activities. CONCLUSIONS: This pilot curriculum was designed to capitalize on practical hands-on training opportunities for our medical students, including participation in a disaster exercise and a mass-gathering event. These opportunities provided effective and engaging disaster preparedness education.

HDL from apoA1 transgenic mice expressing the 4WF isoform is resistant to oxidative loss of function.

HDL functions are impaired by myeloperoxidase (MPO), which selectively targets and oxidizes human apoA1. We previously found that the 4WF isoform of human apoA1, in which the four tryptophan residues are substituted with phenylalanine, is resistant to MPO-mediated loss of function. The purpose of this study was to generate 4WF apoA1 transgenic mice and compare functional properties of the 4WF and wild-type human apoA1 isoforms in vivo. Male mice had significantly higher plasma apoA1 levels than females for both isoforms of human apoA1, attributed to different production rates. With matched plasma apoA1 levels, 4WF transgenics had a trend for slightly less HDL-cholesterol versus human apoA1 transgenics. While 4WF transgenics had 31% less reverse cholesterol transport (RCT) to the plasma compartment, equivalent RCT to the liver and feces was observed. Plasma from both strains had similar ability to accept cholesterol and facilitate ex vivo cholesterol efflux from macrophages. Furthermore, we observed that 4WF transgenic HDL was partially (∼50%) protected from MPO-mediated loss of function while human apoA1 transgenic HDL lost all ABCA1-dependent cholesterol acceptor activity. In conclusion, the structure and function of HDL from 4WF transgenic mice was not different than HDL derived from human apoA1 transgenic mice.

Swimming injuries treated in US EDs: 1990 to 2008.

OBJECTIVE: Swimming is one of the most popular recreational activities in the United States. The objective of this study was to investigate the epidemiology of the complete spectrum of injuries associated with swimming and swimming pools treated in US hospital emergency departments. METHODS: Data from the National Electronic Injury Surveillance System from 1990 to 2008 were analyzed. Injury rates were calculated using US census swimming participation data. RESULTS: An estimated 1688924 swimming injuries occurred during the 19-year study, averaging 1 injury every 6 minutes. During the study period, the number of injuries and rate of injury among individuals 7 years or older significantly increased. Within this trend, injuries peaked in 1999 and significantly decreased during the last 10 years but still showed an overall increase of 18.6% in number and 29.3% in rate from 1900 to 2008. Patients 17 years or younger accounted for 60.5% of injuries, and patients 7 to 17 years of age had a greater mean annual swimming injury rate (18.78 per 10000 participants) than patients older than 17 years (9.15). Most injuries occurred in or around a swimming pool (87.0%), and most were soft tissue injuries (54.7%), followed by strains/sprains (16.4%), fractures/dislocations (11.3%), and submersion (4.9%). Injuries to patients younger than 7 years, submersion injuries, and injuries occurring at home were more likely to result in hospital admission or fatality. CONCLUSIONS: The observed increase in injuries among individuals older than 7 years underscores the need for increased prevention efforts, including education about safe swimming practices, supervision, and environmental modifications.

Pediatric eye injuries treated in US emergency departments, 1990-2009.

This study investigates activity- and consumer product-related eye injuries treated in US hospital emergency departments among children <18 years old using National Electronic Injury Surveillance System data from 1990 through 2009. An estimated 1,406,200 (95% confidence interval = 1,223,409-1,588,992) activity- and consumer product-related pediatric eye injuries occurred during the study period, averaging 70,310 annually. The annual number of injuries declined significantly by 17%. Patients ≤ 4 years of age accounted for 32% of all injuries and had the highest mean annual eye injury rate (11.31 per 10,000 population). Eye injuries associated with sports and recreation (24%) and chemicals (17%) occurred most frequently. The majority (69%) of eye injuries occurred at home. Opportunities exist to further decrease these injuries. Pediatricians should educate child caregivers and children about risks for eye injuries in the home and about use of appropriate protective eyewear during sports.

Pediatric volleyball-related injuries treated in US emergency departments, 1990-2009.

This study describes the epidemiology of pediatric volleyball-related injuries treated in US hospital emergency departments. Data for children younger than 18 years obtained from the National Electronic Injury Surveillance System of the US Consumer Product Safety Commission from 1990 through 2009 were analyzed. An estimated 692 024 volleyball-related injuries to children younger than 18 years occurred during the study period. The annual number of injuries declined significantly by 23% during the study period; however, the annual injury rate remained unchanged, and the number of volleyball-related concussions/closed head injuries increased significantly. Upper (48%) and lower (39%) extremity injuries occurred most frequently, as did strains/sprains (54%). Contact with the net/pole was associated with concussions/closed head injury our findings indicate opportunities for making volleyball an even safer sport for children. Protective padding, complying with US volleyball standards, should cover all volleyball poles and protruding hardware to prevent impact-related injuries.

The serum of dysautonomia patients enhances proliferation and signaling in Schwann cells.

Disorders of the autonomic nervous system, or dysautonomias, affect a large segment of the population, especially women, and represent a diagnostic challenge. Identification of biomarkers for autonomic disorders, and the subsequent development of screening methods, would benefit diagnosis and symptom management. We studied the effect of sera from fifteen well-characterized dysautonomia patients (mean age 49+/-16 years, 10 females, 5 males) and ten control subjects (mean age 31+/-14 years, 5 females, 5 males) on the proliferation of cultured Schwann cells and activity of mitogen-activated protein kinases (MAPKs) in these cells. We correlated characteristics of patients with the effects on cell proliferation and signaling. Overall, we observed a significant increase in proliferation when Schwann cells were incubated with sera from female dysautonomia patients when compared to control subjects and male patients. Interestingly, removal of IgGs significantly reduced the proliferative effect of patient sera. We also observed significant activation of p38 MAPK following incubation with both male and female patient sera. These results suggest that patient sera contain factors that contribute to aberrant Schwann cell proliferation and signaling and may ultimately lead to autonomic nerve dysfunction. Our observations represent a promising first step in the identification of dysautonomia biomarkers.