Analysis of Detergent-Insoluble and Whole Cell Lysate Fractions of Resting Neutrophils Using High-Resolution Mass Spectrometry

Neutrophilic granulocytes play a major role in the initiation and resolution of the inflammatory response, and demonstrate significant transcriptional and translational activity. Although much was known about neutrophils prior to the introduction of proteomics, the use of MS-based methodologies has provided an unprecedented tool to confirm and extend previous findings. In the present study, we performed a Gel-LC-MS/MS analysis of neutrophil detergent insoluble and whole cell lysate fractions of resting neutrophils. We achieved a set of identifications through the use of high-resolution mass spectrometry and validation of its data. We identified a total of 1249 proteins with a wide range of intensities from both detergent-insoluble and whole cell lysate fractions, allowing a mapping of proteins such as those involved in intracellular transport (Rab and Sec family proteins) and cell signaling (S100 proteins). These results represent the most comprehensive proteomic characterization of resting human neutrophils to date, and provide important information relevant for further studies of the immune system in health and disease. The methods applied here can be employed to help us understand how neutrophils respond to various physiologic and pathophysiologic conditions and could be extended to protein quantitation after cell activation.

Ethical issues in neuroscience research on substance dependence treatment and prevention

Previous chapters have presented the latest findings in neuroscience research, and have pointed to potential treatment and prevention strategies. However, there are many ethical implications of the research itself, as well as the treatment and prevention strategies, that must be considered. The rapid pace of change in the field of neuroscience brings with it a host of new ethical issues, which need to be addressed. This chapter considers the important ethical and human rights issues that are raised by neuroscience research on psychoactive substance dependence.

Ethical considerations for drug abuse epidemiological research

The mortality and morbidity caused by alcohol, tobacco and illicit drug misuse represents a significant public health burden (Ezzati et al., 2002). A key part of the public health response is the collection of epidemiological and social science data to define at-risk populations to identify opportunities for intervention and to evaluate the effectiveness of policies in preventing or treating drug misuse and drug-related harm. The systematic use of epidemiological and social science research methods to study illicit drug use is barely 40 years old in the United States and United Kingdom, which have pioneered this approach. Because of the sensitive nature of epidemiological research on illicit drug use a unique set of ethical challenges need to be explicitly addressed by the field. Although ethics guidelines have been proposed (Council for International Organizations of Medical Sciences, 1991), scholarship on the ethics of epidemiology is scant, and consensus on core values not yet achieved (Coughlin, 2000).