Essential contribution of caspase 3/CPP32 to apoptosis and its associated nuclear changes

Caspases are fundamental components of the mammalian apoptotic machinery, but the precise contribution of individual caspases is controversial. CPP32 (caspase 3) is a prototypical caspase that becomes activated during apoptosis. In this study, we took a comprehensive approach to examining the role of CPP32 in apoptosis using mice, embryonic stem (ES) cells, and mouse embryonic fibroblasts (MEFs) deficient for CPP32. CPP32(ex3-/-) mice have reduced viability and, consistent with an earlier report, display defective neuronal apoptosis and neurological defects. Inactivation of CPP32 dramatically reduces apoptosis in diverse settings, including activation-induced cell death (AICD) of peripheral T cells, as well as chemotherapy-induced apoptosis of oncogenically transformed CPP32(-/-) MEFs. As well, the requirement for CPP32 can be remarkably stimulus-dependent: In ES cells, CPP32 is necessary for efficient apoptosis following UV- but not gamma-irradiation. Conversely, the same stimulus can show a tissue-specific dependence on CPP32: Hence, TNFalpha treatment induces normal levels of apoptosis in CPP32 deficient thymocytes, but defective apoptosis in oncogenically transformed MEFs. Finally, in some settings, CPP32 is required for certain apoptotic events but not others: Select CPP32(ex3-/-) cell types undergoing cell death are incapable of chromatin condensation and DNA degradation, but display other hallmarks of apoptosis. Together, these results indicate that CPP32 is an essential component in apoptotic events that is remarkably system- and stimulus-dependent. Consequently, drugs that inhibit CPP32 may preferentially disrupt specific forms of cell death.     

Reductions in self-injury produced by transcutaneous electrical nerve stimulation

Transcutaneous electrical nerve stimulation is used to reduce pain but also may be useful for self-injurious behavior (SIB). In the current investigation, a microcurrent electromedical device, classified as a transcutaneous electrical nerve stimulator (TENS), was applied with a man with Down syndrome who displayed SIB that persisted in the absence of social contingencies. Although clinically significant results were not maintained, a clear difference in the rates of SIB during active and inactive TENS was observed.     

Types of social support as predictors of psychiatric morbidity in a cohort of British Civil Servants (Whitehall II Study)

BACKGROUND: Few studies have examined prospectively both the direct and buffering effects of types of social support and social networks on mental health. This paper reports longitudinal associations between types of social support and psychiatric morbidity from the Whitehall II study. METHODS: Social support was measured by the Close Persons Questionnaire and psychiatric morbidity by the General Health Questionnaire at baseline (1985-1988) and at first follow-up (1989) in 7697 male and female London-based civil servants aged 35-55 years at baseline. The cohort was followed up and baseline measures were used to predict psychiatric disorder measured by the General Health Questionnaire at second follow-up (1991-1993). RESULTS: Longitudinal analyses showed that low confiding/emotional support in men and high negative aspects of close relationships in men and women were associated with greater risk of psychiatric morbidity even after adjustment for baseline General Health Questionnaire score. There was no evidence of a buffering effect among men or women who experienced life events or chronic stressors. Controlling for a personality measure of hostility did not affect the observed relations. CONCLUSIONS: The present findings illustrate that different types of support are risk factors for psychological distress and that they operate in different ways for men and women. Direct effects of emotional support are predictive of good mental health in men and negative aspects of close relations predict poor mental health in both men and women. Emotional support is predictive of good mental health in women whereas, confiding alone is not.     

Delivery of multiple CD8 cytotoxic T cell epitopes by DNA vaccination

Development of CD8 alphabeta CTL epitope-based vaccines requires an effective strategy capable of co-delivering large numbers of CTL epitopes. Here we describe a DNA plasmid encoding a polyepitope or "polytope" protein, which contained multiple contiguous minimal murine CTL epitopes. Mice vaccinated with this plasmid made MHC-restricted CTL responses to each of the epitopes, and protective CTL were demonstrated in recombinant vaccinia virus, influenza virus, and tumor challenge models. CTL responses generated by polytope DNA plasmid vaccination lasted for 1 yr, could be enhanced by co-delivering a gene for granulocyte-macrophage CSF, and appeared to be induced in the absence of CD4 T cell-mediated help. The ability to deliver large numbers of CTL epitopes using relatively small polytope constructs and DNA vaccination technology should find application in the design of human epitope-based CTL vaccines, in particular in vaccines against EBV, HIV, and certain cancers.     

Characterization of microdialysis acidification for capillary isoelectric focusing-microelectrospray ionization mass spectrometry

A microdialysis junction, based on a microdialysis membrane connecting a separation capillary and a short, sharply tapered microelectrospray emitter capillary, is demonstrated for on-line combination of capillary isoelectric focusing (CIEF) with electrospray ionization mass spectrometry (ESI-MS). The microdialysis junction provides the necessary electrical connection across the dialysis membrane for defining the electric fields needed for the CIEF separation and the electrospray process. Additionally, postseparation acidification of focused protein zones eluted from the CIEF capillary is achieved using the microdialysis junction while separation efficiency and resolution is maintained. A microelectrospray emitter produces a stable electrospray of protein analytes without the need for a makeup liquid flow and eliminates any subsequent sample dilution and reduction in MS sensitivity. The microdialysis junction is advantageous over the coaxial liquid sheath interface as evidenced by the simplicity in operation procedures, the enhancement in detection sensitivity, and the linear correlation between protein migration time and isoelectric point in CIEF-ESI-MS.     

Hepatobiliary and pancreatic mucinous cystadenocarcinomas with mesenchymal stroma: analysis of estrogen receptors/progesterone receptors and expression of tumor-associated antigens

In an attempt to establish a standardized rating system for CT of the paranasal sinuses, the Committee on Rhinology and Paranasal Sinus Disease of the American Academy of Otolaryngology-Head and Neck Surgery instituted a protocol for the review of sinus CT scans at six international sites. Fifty identical scans were rated by four otolaryngologists at each site according to five established sinus CT staging systems. Twenty of 24 reviewers repeated the rating session at least 1 week later to determine intrarater variability. The number of CT scans that could not be classified by a particular rating system ranged from 1.3% to 5.5%. The range of intrarater agreement (kappa = 0.39 to 0.74) exceeded that of interrater agreement (kappa = 0.18 to 0.49). A skewed distribution of CT scans resulted in a system with high rater agreement but poor ability to differentiate among disease states. The use of a numeric rating system to assign a score to each scan produced a comprehensive and disease-sensitive system, but one with low rater agreement. A precise definition of mucosal thickening in terms of millimeters appeared to enhance the raters' ability to assign stage and improve a system's comprehensiveness and reproducibility. On the basis of these findings, recommendations are made for the use of CT rating systems to study clinical outcomes in patients with chronic sinusitis.