Differential sensitivities of severe acute respiratory syndrome (SARS) coronavirus spike polypeptide enzyme-linked immunosorbent assay (ELISA) and SARS coronavirus nucleocapsid protein ELISA for serodiagnosis of SARS coronavirus pneumonia

The use of recombinant severe acute respiratory syndrome-coronavirus (SARS-CoV) nucleocapsid protein (N) enzyme-linked immunosorbent assay (ELISA)-based antibody and antigen tests for diagnosis of SARS-CoV infections have been widely reported. However, no recombinant SARS-CoV spike protein (S)-based ELISA is currently available. In this article, we describe the problems and solutions of setting up the recombinant SARS-CoV S-based ELISA for antibody detection. The SARS-CoV S-based immunoglobulin M (IgM) and IgG ELISAs were evaluated and compared with the corresponding N-based ELISA for serodiagnosis of SARS-CoV pneumonia, using sera from 148 healthy blood donors who donated blood 3 years ago as controls and 95 SARS-CoV pneumonia patients in Hong Kong. Results obtained by the recombinant S (rS)-based IgG ELISA using the regenerated S prepared by dialysis with decreasing concentrations of urea or direct addition of different coating buffers, followed by addition of different regeneration buffer, identified 4 M urea and 1 M sarcosine for plate coating and no regeneration buffer as the most optimal conditions for antibody detection. The specificities of the S-based ELISA for IgG and IgM detection were 98.6% and 93.9%, with corresponding sensitivities of 58.9% and 74.7%, respectively. The sensitivity of the rN IgG ELISA (94.7%) is significantly higher than that of the rS IgG ELISA (P < 0.001), whereas the sensitivity of the rS IgM ELISA is significantly higher than that of the rN IgM ELISA (55.2%) (P < 0.01). An ELISA for detection of IgM against S and N could be more sensitive than one that detects IgM against N alone for serodiagnosis of SARS-CoV pneumonia.     

ASO Author Reflections: Geographic Disparities in Referral and Oncologic Outcomes in Intrahepatic Cholangiocarcinoma

Annals of Surgical Oncology -     

Development and usability of a feedback tool, “My Personal Brain Health Dashboard”, to improve setting of self-management goals among people living with HIV in Canada

Quality of Life Research - (1) To develop a personalized health outcome profile as a feedback tool to improve self-management in people living with chronic conditions such as HIV and (2) to...     

Synthesis, rhenium-188 labeling and biodistribution studies of a phenolic ester derivative of trisuccin

Our previous results indicated that the trihydroxamate ligand, trisuccin, was a promising bifunctional chelating agent (BCA) for radiometal labeling of monoclonal antibodies with rhenium and technetium. An interest was developed to evaluate structural modifications of this compound from both synthetic and biological points of view. In this report we describe the synthesis of an esterified trisuccin (referred to as trisester), and conjugation of this new derivative to MAb CC49, radiolabeling of this conjugate with rhenium-188 (188Re), and biodistribution of the labeled conjugate in athymic nude mice. Thus, trisuccin (1) was esterified with benzyl 4-hydroxybenzoate in a DCC/DMAP reaction followed by removal of all benzyl protecting groups with catalytic hydrogenation. The resulting product was conjugated to CC49 by the active ester technique, through formation of the 2-nitrophenyl ester 6, and the conjugate was radiolabeled with generator-produced 188Re. The lead molecule trisuccin 1 was also conjugated to CC49 through the active ester 5 and the conjugate was radiolabeled by the same procedure to serve as the control conjugate. Biodistributions of the labeled conjugates were studied in athymic nude mice, transplanted s.c. with LS174T human colon cancer xenografts. Although an increase in the radiolabeling yield was observed for the esterified ligand-CC49 conjugate, as well as some increase in its immunoreactivity, as compared to those for the parent trisuccin molecule, there were no significant differences in their biodistribution. This new compound therefore may be useful in improving the conjugation and radiolabeling chemistries of this trihydroxamate ligand system.     

The Israeli Sexual Behavior Inventory (ISBI): Scale Construction and Preliminary Validation

This study describes the construction and preliminary validation of the Israeli Sexual Behavior Inventory (ISBI). The ISBI was primarily designed to assess the impact of sexual problems, chronic illness and disability on sexual functioning and experience. Scales were designed to measure three areas of healthy sexual functioning and three areas of sexual dysfunction for both males and females. To provide normative data to which clinical samples can be compared, a large randomly selected sample from an adult male and female population was used for scale construction and preliminary validation. Scale reliabilities, intercorrelations between the ISBI scales, comparisons between the above sample and a clinical sample provide evidence of the ISBI's reliability and validity.     

Ultrastructural confirmation of neuronal protection by melatonin against the neurotoxin 6-hydroxydopamine cell damage

6-Hydroxydopamine (6-OHDA) is a neurotoxin used in the induction of experimental Parkinson's disease in both animals and cultured neuronal cells. Biochemical and molecular approaches showed previously that low doses of 6-OHDA induced apoptosis in PC12 cells, while high doses of this neurotoxin induced necrosis. Melatonin has been shown to protect against the neuronal programmed cell death induced by 6-OHDA, although it was not able to prevent the massive necrotic cellular death occurring after the addition of high doses of the neurotoxin. In the present work, we demonstrate by ultrastructural analysis that although low doses of 6-OHDA induced apoptosis in PC12 cells, it also damaged the non-apoptotic cells, morphologically corresponding this damage to incipient and reversible necrotic lesions. When the doses of the neurotoxin increase, there are still apoptotic cells, although most of the cells show necrotic irreversible lesions. We also found that melatonin partially prevents the incipient necrotic lesions caused by low doses of 6-OHDA. The fact that melatonin was shown in previous work to prevent apoptosis caused by low doses of 6-OHDA, but not necrosis induced by high doses of the neurotoxin, seemed to indicate that this agent is only able to protect against apoptosis. However, our present results, melatonin preventing also the incipient necrotic neuronal lesions, suggest that this hormone may provide a general protection against cell death, suggesting that higher doses should be tried in order to prevent the necrotic cell death induced by high doses of the neurotoxin.