Drug testing, commonly used in health care, workplace, and criminal settings, has become widespread during the past decade. Urine drug screens have been the most common method for analysis because of ease of sampling. The simplicity of use and access to rapid results have increased demand for and use of immunoassays; however, these assays are not perfect. False-positive results of immunoassays can lead to serious medical or social consequences if results are not confirmed by secondary analysis, such as gas chromatography-mass spectrometry. The Department of Health and Human Services' guidelines for the workplace require testing for the following 5 substances: amphetamines, cannabinoids, cocaine, opiates, and phencyclidine. This article discusses potential false-positive results and false-negative results that occur with immunoassays of these substances and with alcohol, benzodiazepines, and tricyclic antidepressants. Other pitfalls, such as adulteration, substitution, and dilution of urine samples, are discussed. Pragmatic concepts summarized in this article should minimize the potential risks of misinterpreting urine drug screens.

Lambda-like dsDNA bacteriophage undergo massive conformational changes in their capsid shell during the packaging of their viral genomes. Capsid shells are complex organizations of hundreds of protein subunits that assemble into intricate quaternary complexes that ultimately are able to withstand over 50 atm. of pressure during genome packaging 1 . The extensive integration between subunits in capsids is unlikely to form in a single assembly step, therefore requiring formation of an intermediate complex, termed a procapsid, from which individual subunits can undergo the necessary refolding and structural rearrangements needed to transition to the more stable capsid. Though various mature capsids have been characterized at atomic resolution, no such procapsid structure is available for a dsDNA virus or bacteriophage that undergoes large scale conformational changes. We present a procapsid x-ray structure at 3.65Å resolution, termed Prohead II, of the lambda like bacteriophage HK97, whose mature capsid structure was previously solved to 3.44 Å 2 . A comparison of the two largely different capsid forms has unveiled an unprecedented expansion mechanism that describes the transition. Crystallographic and Hydrogen/Deuterium exchange data presented here demonstrates that the subunit tertiary structures are significantly different between the two states, with twisting and bending motions occurring in both helical and β-sheet regions. We have also discovered conserved subunit interactions at each 3-fold of the virus capsid, from which capsid subunits maintain their integrity during refolding, facilitating the rotational and translational motions of maturation. Calormetric data of a closely related bacteriophage, P22, showed that capsid maturation was an exothermic process that resulted in a release of 90KJ/mol of energy 3 . We propose the major tertiary changes presented in this study reveal a structural basis for an exothermic maturation process likely present in many dsDNA Bacteriophage and possibly viruses such as Herpes which share the HK97 subunit fold 4 . Main TextHK97 is a favorable system for studying capsid maturation because capsid particles can be assembled in E. coli from the expression of just two viral gene products, gp4 (protease) and gp5 (capsid subunit), and maturation can be triggered and analyzed in vitro 2,5-8 . The 384-residue gp5 subunits assemble into hexameric and pentameric oligomers, termed capsomers, that first assemble to form the Prohead I capsid (P-I). The 17.7-Megadalton, T=7 laevo icosahedral particle contains 12 pentamers and 60 hexamers and encapsidates approximately 60 copies of the gp4 protease 9-11 . Prohead I particles can be made with either a defective protease or without protease and can be disassembled in vitro into free capsomers and than reassembled when exposed to specific chemical treatments 12 . When active gp4 is present, the particles spontaneously mature to the Prohead II (P-II) form following digestion of residues 2-103 from all subunits. The proteolytic fragment...

A more complete picture of the molecules that are critical for the organization of membrane compartments is beginning to emerge through the characterization of proteins in the vesicle-associated membrane protein (also called synaptobrevin) family of membrane trafficking proteins. To better understand the mechanisms of membrane trafficking within the endocytic pathway, we generated a series of monoclonal and polyclonal antibodies against the cytoplasmic domain of vesicle-associated membrane protein 7 (VAMP-7). The antibodies recognize a 25-kD membrane-associated protein in multiple tissues and cell lines. Immunohistochemical analysis reveals colocalization with a marker of late endosomes and lysosomes, lysosome-associated membrane protein 1 (LAMP-1), but not with other membrane markers, including p115 and transferrin receptor. Treatment with nocodozole or brefeldin A does not disrupt the colocalization of VAMP-7 and LAMP-1. Immunoelectron microscopy analysis shows that VAMP-7 is most concentrated in the trans-Golgi network region of the cell as well as late endosomes and transport vesicles that do not contain the mannose-6 phosphate receptor. In streptolysin- O–permeabilized cells, antibodies against VAMP-7 inhibit the breakdown of epidermal growth factor but not the recycling of transferrin. These data are consistent with a role for VAMP-7 in the vesicular transport of proteins from the early endosome to the lysosome.

These data suggest that the neutrophils of newborns are selectively deficient in BPI, a central effector of antibacterial activity against gram-negative bacteria. BPI deficiency correlates with decreased antibacterial activity of newborn neutrophil extracts against serum-resistant E coli and could contribute to the increased incidence of gram-negative sepsis among newborns relative to healthy adults.neonatal sepsis, gram-negative bacteria, endotoxin, neutrophil, polymorphonuclear leukocyte, innate immunity, bactericidal/permeability-increasing protein, defensin, myeloperoxidase.

Microglia and infiltrating macrophages are thought to orchestrate the central nervous system (CNS) response to injury; however, the similarities between these cells make it challenging to distinguish their relative contributions. We genetically labeled microglia and CNS-associated macrophages to distinguish them from infiltrating macrophages. Using single-cell RNA sequencing, we describe multiple microglia activation states, one of which was enriched for interferon associated signaling. Although blood-derived macrophages acutely infiltrated the demyelinated lesion, microglia progressively monopolized the lesion environment where they surrounded infiltrating macrophages. In the microglia-devoid sciatic nerve, the infiltrating macrophage response was sustained. In the CNS, the preferential proliferation of microglia and sparse microglia death contributed to microglia dominating the lesion. Microglia ablation reversed the spatial restriction of macrophages with the demyelinated spinal cord, highlighting an unrealized macrophages-microglia interaction. The restriction of peripheral inflammation by microglia may be a previously unidentified mechanism by which the CNS maintains its “immune privileged” status.

Background Previous studies have demonstrated nurses are at risk of suicide. This is the first national longitudinal study of U.S. nurse suicide. Aims To identify the longitudinal incidence, method, and risks of nurse suicide in the United States. Methods 2005 to 2016 Centers for Disease Control and Prevention National Violent Death Reporting System retrospective analysis of suicide incident rate ratios (IRR). Results A total of 1,824 nurse and 152,495 non‐nurse suicides were evaluated. Nurses were at greater risk of suicide than the general population (female IRR 1.395, 95% confidence intervals [CI] 1.323, 1.470, p < .001; male IRR 1.205, 95% CI 1.083, 1.338, p < .001). Female nurses who completed suicide did so most frequently by pharmacologic poisoning (n = 399, 27.2% vs. n = 8,843, 26.9%), whereby male nurses and the general public used firearms (n = 148, 41.7% vs. n = 57,887, 48.4%). Job problems were more likely in nurses (female odds ratio [OR] 1.989, 95% CI 1.695, 2.325, p < .001; male OR 1.814, 95% CI 1.380, 2.359, p < .001), as well as mental health history (female OR 1.126, 95% CI 1.013, 1.253, p < .027; male OR 1.302, 95% CI 1.048, 1.614, p = .016) and leaving a suicide note (female OR 1.221, 95% CI 1.096, 1.360, p < .001; male OR 1.756 [1.412, 2.181], p < .001). Linking Evidence to Action The increased risk of suicide in nurses is congruent with previous reports. The consistency in results increases confidence that findings are generalizable and warrant action. The use of pharmacologic poisoning as a method of suicide, most often by opioids and benzodiazepines, indicates a need for improved identification and treatment of nurses with substance use. Workplace wellness programs need to focus on reducing workplace stressors. Further research is indicated to determine best prevention methods. Policy indications include the need to accurately track gender in nursing, enhance substance use disorder programs, and mandate suicide prevention activities.

Objectives. To measure the level of burnout among pharmacy practice faculty members at US colleges and schools of pharmacy and to identify factors associated with burnout. Methods. Using a cross-sectional, electronic, anonymous survey-design, we measured faculty burnout (n52318) at US colleges and schools of pharmacy using the Maslach Burnout Inventory-Educators Survey (MBI-ES), which measures burnout dimensions: emotional exhaustion, depersonalization, and personal accomplishment. We assessed MBI-ES scores, demographics and possible predictors of burnout. Results. The response rate was 32.7% (n5758). Emotional exhaustion was identified in 41.3% and was higher in women, assistant professors, and those without a hobby. Participants without a mentor had higher scores of depersonalization. Those with children ages 1-12 years had higher emotional exhaustion and depersonalization compared to those with older children. Conclusion. Pharmacy practice faculty members at US colleges and schools of pharmacy are suffering from burnout, exhibited mainly through emotional exhaustion.

This list serves as a starting point to enhance coping with stressors as a healthcare student or professional in order to help mitigate burnout, depression, and suicidality. The next steps include adapting digital health strategies to specifically fit the needs of healthcare providers, with the ultimate goal of facilitating in-person care when warranted.