Production of herpes B virus recombinant glycoproteins and evaluation of their diagnostic potential

B virus (cercopithecine herpesvirus 1) is the only deadly alphaherpesvirus that is zoonotically transmissible from macaques to humans. The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals. We evaluated the diagnostic potential of recombinant B virus glycoproteins for the detection of immunoglobulin G (IgG) antibodies in monkey and human sera. Glycoproteins B, C, and E and secreted (sgG) and membrane-associated (mgG) segments of glycoprotein G (gG) were expressed in the baculovirus expression system, while gD was expressed in CHO cells. We developed recombinant protein-based IgG enzyme-linked immunosorbent assays (ELISAs) and compared their diagnostic efficacies by using B virus antibody-negative (n = 40) and -positive (n = 75) macaque sera identified by a whole antigen-based ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG- and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys cross-reacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)- and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data indicate that gB, gC, gD, and mgG have a high diagnostic potential for B virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B virus and those induced by other, closely related alphaherpesviruses, including HSV-1 and -2.     

Respondeo: method and content in casuistry

James Tallmon has argued that my criticisms of Jonsen and Toulmin are ill founded. Tallmon argues that Jonsen and Toulmin argue for a method of rhetorical reasoning and not for a particular content. He argues that if one distinguishes the content and method of casuistry the Jonsen-Toulmin model can work. But Tallmon, like Jonsen and Toulmin, cannot escape the need for casuistry to have a content. Tallmon's response evidences that need since he assumes that there is a 'Medical Community' which has a moral vision.     

Sampling the N-terminal proteome of human blood

The proteomes of blood plasma and serum represent a potential gold mine of biological and diagnostic information, but challenges such as dynamic range of protein concentration have hampered efforts to unlock this resource. Here we present a method to label and isolate N-terminal peptides from human plasma and serum. This process dramatically reduces the complexity of the sample by eliminating internal peptides. We identify 772 unique N-terminal peptides in 222 proteins, ranging over six orders of magnitude in abundance. This approach is highly suited for studying natural proteolysis in plasma and serum. We find internal cleavages in plasma proteins created by endo- and exopeptidases, providing information about the activities of proteolytic enzymes in blood, which may be correlated with disease states. We also find signatures of signal peptide cleavage, coagulation and complement activation, and other known proteolytic processes, in addition to a large number of cleavages that have not been reported previously, including over 200 cleavages of blood proteins by aminopeptidases. Finally, we can identify substrates from specific proteases by exogenous addition of the protease combined with N-terminal isolation and quantitative mass spectrometry. In this way we identified proteins cleaved in human plasma by membrane-type serine protease 1, an enzyme linked to cancer progression. These studies demonstrate the utility of direct N-terminal labeling by subtiligase to identify and characterize endogenous and exogenous proteolysis in human plasma and serum.     

Pulmonary edema and acute upper airway obstruction

Pulmonary edema following the relief of acute upper airway obstruction is unusual and unpredictable. The cause is usually attributed to pulmonary and hemodynamic changes resulting from high negative intrathoracic pressures during obstructed respiration. The incidence of this problem is not known. Why pulmonary edema develops in only certain patients is also unclear. We reviewed the records of 27 consecutive adult patients admitted and treated for acute upper airway obstruction. Three of 27 (11%) developed pulmonary edema. Age, sex, diagnosis, duration of obstruction, history of cardiopulmonary disease, and perioperative fluid administration were evaluated. No differences were noted between those who did and those who did not develop pulmonary edema. Pulmonary edema following acute upper airway obstruction appears to be more common than is generally appreciated, however, no specific factors seem related to its occurrence.     

Selectivity for speed gradients in human area MT/V5

Cortical area MT/V5 in the human occipito-temporal cortex is activated by visual motion. In this study, we use functional imaging to demonstrate that a subregion of MT/V5 is more strongly activated by unidirectional motion with speed gradients than by other motion patterns. Our results suggest that like the monkey homolog middle temporal area (MT), human MT/V5 contains neurons selective for the processing of speed gradients. Such neurons may constitute an intermediate stage of processing between neurons selective for the average speed of unidirectional motion and neurons selective for different combinations of speed gradient and different motion directions such as expanding optical flow patterns.     

Clinical trials attitudes and practices of Latino physicians

BackgroundEthnic differences in physicians' attitudes and behaviors related to clinical trials might partially account for disparities in clinical trial participation among Latino patients. Literature regarding Latino physicians' clinical trials attitudes and practices, in comparison to White physicians, was lacking.MethodsCross-sectional data from randomly selected physicians (N = 695), stratified by ethnicity, were analyzed to test associations of ethnicity with physicians' participation in and attitudes toward referral of patients to clinical trials.ResultsChi-square analyses showed significant (p < 0.05) associations of physician race/ethnicity and clinical trials involvement, type of trial for which the physician is likely to recommend a patient, belief in scientific value, and factors that would influence recommendation for a patient to participate. Multivariate analyses resulted in several significant (p < 0.05) predictors of clinical trials outcomes, including physician race/ethnicity.ConclusionsLatino physicians were significantly less involved in clinical trials than White physicians and found less scientific value in them, highlighting areas for future education and intervention.