The Comprehensive Assessment of Prospective Memory (CAPM) is a questionnaire designed to evaluate frequency of prospective memory (PM) failures in people with brain injury. The aims of this study were to investigate the psychometric properties of the CAPM, including test-retest reliability and internal consistency, and to establish normative data by comparing CAPM scores between groups on the basis of sex, age, and education. Data were collected on 95 people aged 15-60 years living in the community, with no history of brain injury, using the CAPM. The results showed that the test-retest reliability and internal consistency for the CAPM were within acceptable ranges, indicating that the CAPM provides a stable and homogenous measure of an individual's self-report of PM failures. Normative data are presented in two age groups based on the significant difference found between the age groups 15-30 years and 31-60 years. These established norms can be used to describe perceived or observed behaviours indicative of PM failure in patients with brain injury by comparing CAPM ratings from significant others with the norms. The CAPM questionnaire provides researchers or clinicians with a stable and reliable assessment option that specifically focuses on PM for individuals with brain injury.

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Chromatin Immunoprecipitation (ChIP) is an invaluable method to characterize interactions between proteins and genomic DNA, such as the genomic localization of transcription factors and post-translational modification of histones. DNA and proteins are reversibly and covalently crosslinked using formaldehyde. Then the cells are lysed to release the chromatin. The chromatin is fragmented into smaller sizes either by micrococcal nuclease (MNase) or sonication and then purified from other cellular components. The protein-DNA complexes are enriched by immunoprecipitation (IP) with antibodies that target the epitope of interest. The DNA is released from the proteins by heat and protease treatment, followed by degradation of contaminating RNAs with RNase. The resulting DNA is analyzed using various methods, including PCR, qPCR, or sequencing. This protocol outlines each of these steps for both yeast and human cells.

Chemical tools to control the activities and interactions of chromatin components have broad impact on our understanding of cellular and disease processes. It is important to accurately identify their molecular effects to inform clinical efforts and interpretations of scientific studies. Chaetocin is a widely used chemical that decreases H3K9 methylation in cells. It is frequently attributed as a specific inhibitor of the histone methyltransferase activities of SUV39H1/SU(VAR)3-9, although prior observations showed chaetocin likely inhibits methyltransferase activity through covalent mechanisms involving its epipolythiodixopiperazine disulfide ‘warhead’ functionality. The continued use of chaetocin in scientific studies may derive from the net effect of reduced H3K9 methylation, irrespective of a direct or indirect mechanism. However, there may be other molecular impacts of chaetocin on SUV39H1 besides inhibition of H3K9 methylation levels that could confound the interpretation of past and future experimental studies. Here we test a new hypothesis that chaetocin may have an additional downstream impact aside from inhibition of methyltransferase activity. Using a combination of truncation mutants, a yeast two-hybrid system, and direct in vitro binding assays, we show that the human SUV39H1 chromodomain (CD) and HP1 chromoshadow domain (CSD) directly interact. Chaetocin inhibits this binding interaction through its disulfide functionality with some specificity by covalently binding with the CD of SUV39H1, whereas the histone H3-HP1 interaction is not inhibited. Given the key role of HP1 dimers in driving a feedback cascade to recruit SUV39H1 and to establish and stabilize constitutive heterochromatin, this additional molecular consequence of chaetocin should be broadly considered.

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