Identification of Human Junctional Adhesion Molecule 1 as a Functional Receptor for the Hom-1 Calicivirus on Human Cells.

The Hom-1 vesivirus was reported in 1998 following the inadvertent transmission of the animal calicivirus San Miguel sea lion virus to a human host in a laboratory. We characterized the Hom-1 strain and investigated the mechanism by which human cells could be infected. An expression library of 3,559 human plasma membrane proteins was screened for reactivity with Hom-1 virus-like particles, and a single interacting protein, human junctional adhesion molecule 1 (hJAM1), was identified. Transient expression of hJAM1 conferred susceptibility to Hom-1 infection on nonpermissive Chinese hamster ovary (CHO) cells. Virus infection was markedly inhibited when CHO cells stably expressing hJAM were pretreated with anti-hJAM1 monoclonal antibodies. Cell lines of human origin were tested for growth of Hom-1, and efficient replication was observed in HepG2, HuH7, and SK-CO15 cells. The three cell lines (of hepatic or intestinal origin) were confirmed to express hJAM1 on their surface, and clustered regularly interspaced short palindromic repeats/Cas9-mediated knockout of the hJAM1 gene in each line abolished Hom-1 propagation. Taken together, our data indicate that entry of the Hom-1 vesivirus into these permissive human cell lines is mediated by the plasma membrane protein hJAM1 as a functional receptor.IMPORTANCE Vesiviruses, such as San Miguel sea lion virus and feline calicivirus, are typically associated with infection in animal hosts. Following the accidental infection of a laboratory worker with San Miguel sea lion virus, a related virus was isolated in cell culture and named Hom-1. In this study, we found that Hom-1 could be propagated in a number of human cell lines, making it the first calicivirus to replicate efficiently in cultured human cells. Screening of a library of human cell surface membrane proteins showed that the virus could utilize human junctional adhesion molecule 1 as a receptor to enter cells and initiate replication. The Hom-1 virus presents a new system for the study of calicivirus biology and species specificity.

Screening for suicidal ideation and attempts among emergency department medical patients: instrument and results from the Psychiatric Emergency Research Collaboration.

Joint Commission National Patient Safety Goal 15 calls for organizations "to identify patients at risk for suicide." Overt suicidal behavior accounts for 0.6% of emergency department (ED) visits, but incidental suicidal ideation is found in 3%-11.6%. This is the first multicenter study of suicide screening in EDs. Of 2,243 patients in six diverse emergency settings, 1,068 (47.7%) were screened with a brief instrument. Depression was endorsed by 369 (34.5%); passive suicidal ideation by 79 (7.3%); and active suicidal ideation by 24 (2.3%). One hundred thirty-seven (12.8%) reported prior attempts, including 35 (3.3%) with current suicidal ideation. Almost half of those with current ideation had a prior attempt (43.8%) versus those without current ideation, 10.3%, χ2 (1) = 75.59, p < .001. Twenty cases (25%) were admitted to medical services, but only 10 (12.5%) received mental health assessment; none were admitted directly to a psychiatry service. The prevalence of suicidal ideation here is similar to previous studies but the frequency of prior attempts has not been reported. The 35 cases with current ideation and prior attempt are at risk. As they did not present psychiatrically, they would likely have gone undetected. Despite reporting these cases to clinical staff, few received risk assessment.

Validation of the BRCA1 antibody MS110 and the utility of BRCA1 as a patient selection biomarker in immunohistochemical analysis of breast and ovarian tumours.

BRCA1 protein measurement has previously been evaluated as a potential diagnostic marker without reaching a conclusive recommendation. In this study, we applied current best practice in antibody validation to further characterize MS110, a widely used antibody targeting BRCA1. Antibody specificity was investigated using different biochemical validation techniques. We found that BRCA1 could not be reliably detected using immunoprecipitation and Western blot in endogenously expressing cells. We used immunohistochemistry on formalin-fixed paraffin-embedded cell pellets to establish compatibility with formalin-fixed paraffin-embedded samples. We demonstrated that in transfected cells and cell lines with known genetic BRCA1 status, MS110 successfully detected BRCA1 giving the expected level of staining in immunohistochemistry. Following this, we investigated the use of BRCA1 protein measurement by immunohistochemistry in a cohort of triple negative breast and serous ovarian tumour samples to explore the use of BRCA1 protein measurement by immunohistochemistry for patient stratification. Using MS110 in repeated standardized experiments, on serial sections from a panel of patient samples, results demonstrated considerable run-to-run variability. We concluded that in formalin-fixed tissue samples, MS110 does detect BRCA1; however, using standard methodologies, BRCA1 expression levels in tissue samples is incompatible with the use of this protein as a statistically robust patient selection marker in immunohistochemistry. These results demonstrate the need for further development to deliver BRCA1 protein quantification by immunohistochemistry as a patient stratification marker.

Teaching physicians to assess suicidal youth presenting to the emergency department.

OBJECTIVES: The objective of this study was to determine whether a 5-module self-paced computerized educational program improves residents' skills in assessing and managing youth presenting to the emergency department in acute psychiatric distress. METHODS: The evaluation used a quasi-experimental posttest-only design assessing both knowledge of the educational context of the program and self-rated pretest knowledge of program content with 32 residents recruited from 1 medical center in Cleveland, Ohio. RESULTS: About half of the respondents were female (48%); almost two thirds were white (65%), and few were trained in psychiatric assessment of children/adolescents. On average, residents had significantly higher scores on the posttest compared with the self-rated pretest (6.4 ± 1.1 vs 3.8 ± 2.3; P < 0.001), an effect size of 1.32. Residents responded positively to the modules and rated them highly on educational content (4.2 ± 0.5 on a 5-point scale) and satisfaction with clinical applicability (8.2 ± 1.2 on a 10-point scale) and found the program easy to navigate (8.5 ± 1.9 on a 10-point scale). CONCLUSIONS: A brief, self-administered, Web-based training program shows promise for improving residents' knowledge about suicidal behaviors in youth.

Breathlessness in advanced disease. 2: Patient assessment and management.

This second of a two part unit on breathlessness addresses patient assessment and management, including common treatments. Interventions that can be taught to, and used independently by, patients and carers are explored.

Breathlessness in advanced disease. 1: Definitions, epidemiology and pathophysiology.

This first in a two part unit on breathlessness in advanced disease defines the condition and discusses the epidemiology. It gives an overview of the complex pathophysiology that contributes to breathlessness.

Reproductive decisions for women with HIV: motherhood's role in envisioning a future.

Cultural influences might exert more influence on HIV-positive women's reproductive choices than HIV-related conditions. In this article we report on grounded theory research on how women with HIV made reproductive decisions during a time of transition from HIV as potentially fatal to mothers and newborns to its current status as an often controllable chronic illness. Eighty HIV-positive women of childbearing age in three United States cities were interviewed, and the interviews were analyzed using grounded theory techniques. The core concept of the findings was that women's decisions were based on their judgment of the relative weight of positive aspects of motherhood versus the often negative pressures of social and public opinion. These findings have relevance for future research as well as for program development and policy.

Non-invasive ventilation in COPD. 2: Starting and monitoring NIV.

Part 2 of this two-part unit examines the role of non-invasive ventilation (NIV) in treating hypercapnic respiratory failure and discusses the care of patients on NIV.

Non-invasive ventilation in COPD 1: Managing exacerbations.

This two-part unit discusses the use of non-invasive ventilation (NIV) in patients with respiratory failure, and in particular that associated with COPD. Part 1 examines how an exacerbation of COPD can lead to hypercapnic respiratory failure and how this might be managed.

Letters from a suicide.

A linguistic analysis of a series of letters written by a young man who died by suicide to 1 friend over a period of 2 years prior to his death revealed several trends, including a decrease in negative emotion words and an increase in death words. The implication of these trends was discussed.

Rehydration fluid temperature affects voluntary drinking in horses dehydrated by furosemide administration and endurance exercise.

To determine whether temperature of rehydration fluid influences voluntary rehydration by horses, six 2-3-year-old horses were dehydrated (4-5% body weight loss) by a combination of furosemide administration and 30 km of treadmill exercise. For the initial 5 min following exercise, horses were offered a 0.9% NaCl solution at 10, 20, or 30 degrees C. Subsequently, after washing and cooling out, voluntary intake of water at 10, 20, or 30 degrees C from 20 to 60 min after exercise was measured. Fluid intake (FI) during the first 5 min of recovery was 9.8+/-2.5,12.3+/-2.1 and 9.7+/-2.0L (p>0.05) for saline at 10, 20, and 30 degrees C, respectively. Although not a significant finding, horses offered 0.9% NaCl at 20 degrees C tended to take fewer (p=0.07), longer drinks than when saline at either 10 or 30 degrees C was offered. Between 20 and 60 min of recovery, intake of water at 20 degrees C (7.7+/-0.8L) and 30 degrees C (6.6+/-1.2L) was greater (p<0.05) than that at 10 degrees C (4.9+/-0.5L). Thus, total FI was 14.7+/-2.5,19.9+/-2.5, and 16.3+/-2.4L for rehydration fluids at 10, 20, and 30 degrees C, respectively (p<0.05, value for 20 degrees C water greater than that for 10 degrees C water). Although the amount of metabolic heat transferred to the initial saline drink was correlated with the decrease in core temperature during the initial 5 min of recovery, heat transfer to ingested fluid was most likely responsible for the dissipation of, at most, 5% of the heat generated during endurance exercise. In conclusion, following exercise these dehydrated-normothermic horses voluntary drank the greatest amount of fluid at near ambient (20 degrees C) temperature. Although not determined in this study, greater satiation of thirst by oropharyngeal cooling may have contributed to lesser intake of colder (10 degrees C) fluid.