Quantification of mitochondrial DNA mutation load

Mitochondrial DNA (mtDNA) mutations are an important cause of genetic disease and have been proposed to play a role in the ageing process. Quantification of total mtDNA mutation load in ageing tissues is difficult as mutational events are rare in a background of wild-type molecules, and detection of individual mutated molecules is beyond the sensitivity of most sequencing based techniques. The methods currently most commonly used to document the incidence of mtDNA point mutations in ageing include post-PCR cloning, single-molecule PCR and the random mutation capture assay. The mtDNA mutation load obtained by these different techniques varies by orders of magnitude, but direct comparison of the three techniques on the same ageing human tissue has not been performed. We assess the procedures and practicalities involved in each of these three assays and discuss the results obtained by investigation of mutation loads in colonic mucosal biopsies from ten human subjects.

Temporal regulation of expression of immediate early and second phase transcripts by endothelin-1 in cardiomyocytes

Background: Endothelin-1 stimulates Gq protein-coupled receptors to promote proliferation in dividing cells or hypertrophy in terminally differentiated cardiomyocytes. In cardiomyocytes, endothelin-1 rapidly (within minutes) stimulates protein kinase signaling, including extracellular-signal regulated kinases 1/2 (ERK1/2; though not ERK5), with phenotypic/physiological changes developing from approximately 12 h. Hypertrophy is associated with changes in mRNA/protein expression, presumably consequent to protein kinase signaling, but the connections between early, transient signaling events and developed hypertrophy are unknown. Results: Using microarrays, we defined the early transcriptional responses of neonatal rat cardiomyocytes to endothelin-1 over 4 h, differentiating between immediate early gene (IEG) and second phase RNAs with cycloheximide. IEGs exhibited differential temporal and transient regulation, with expression of second phase RNAs within 1 h. Of transcripts upregulated at 30 minutes encoding established proteins, 28 were inhibited >50% by U0126 (which inhibits ERK1/2/5 signaling), with 9 inhibited 25-50%. Expression of only four transcripts was not inhibited. At 1 h, most RNAs (approximately 67%) were equally changed in total and polysomal RNA with approximately 17% of transcripts increased to a greater extent in polysomes. Thus, changes in expression of most protein-coding RNAs should be reflected in protein synthesis. However, approximately 16% of transcripts were essentially excluded from the polysomes, including some protein-coding mRNAs, presumably inefficiently translated. Conclusion: The phasic, temporal regulation of early transcriptional responses induced by endothelin-1 in cardiomyocytes indicates that, even in terminally differentiated cells, signals are propagated beyond the primary signaling pathways through transcriptional networks leading to phenotypic changes (that is, hypertrophy). Furthermore, ERK1/2 signaling plays a major role in this response.

Transforming tradition: performances of Jingju ('Beijing Opera') in the UK

Jingju (‘Beijing opera’) is China's most iconic traditional theatre, marketed as a global signifier of Chinese theatre and national identity. Although troupes from mainland China regularly tour Europe, audiences in the UK have also had access to Jingju via two indigenous organizations: the UK Beijing Opera Society (now defunct) and the London Jing Kun Opera Association (now in its ninth year). These organizations consist of Chinese, overseas Chinese and Western performers performing both Jingju and Kunju (‘Kun opera’). Where there is a mix of ethnicity, can ‘traditional Chinese theatre’ still be conceived of as ‘traditional’? How is Jingju mapped onto non-Chinese bodies? Can Jingju performances by ethnically white performers reflect diasporic identities? Drawing on the theories of Judith Butler and Homi Bhabha, this article argues that by highlighting the performativity of identity, the performance of Jingju by non-Chinese performers challenges the notion of Jingju as a global signifier of ‘authentic traditional Chinese theatre’.

State and society in the English countryside: the rural community movement 1918-1939

This paper assesses the relationship between state and society in interwar rural England, focusing on the hitherto neglected role of the Rural Community Councils (RCCs). The rise of statutory social provision in the early twentieth century created new challenges and opportunities for voluntaryism, and the rural community movement was in part a response. The paper examines the early development of the movement, arguing that a crucial role was played by a close-knit group of academics and local government officials. While largely eschewing party politics, they shared a commitment to citizenship, democracy and the promotion of rural culture; many of them had been close associates of Sir Horace Plunkett. The RCCs engaged in a wide range of activities, including advisory work, adult education, local history, village hall provision, support for rural industries and an ambivalent engagement with parish councils. The paper concludes with an assessment of the achievements of the rural community movement, arguing that it was constrained by its financial dependence on voluntary contributions.