Expression and regulation of 80K/MARCKS, a major substrate of protein kinase C, in the developing rat heart

Objective: Protein kinase C (PKC) plays a pivotal role in modulating the growth and differentiation of many cell types including the cardiac myocyte. However, little is known about molecules that act immediately downstream of PKC in the heart. In this study we have investigated the expression of 80K/MARCKS, a major PKC substrate, in whole ventricles and in cardiac myocytes from developing rat hearts. Methods: Poly A+ RNA was prepared from neonatal (2-day) and adult (42-day) cardiac myocytes and whole ventricular tissue and mRNA expression determined by reverse transcription-polymerase chain reaction (RT-PCR) using primers designed to identify a 420 bp fragment in the 80K/MARCKS gene. Protein extracts were prepared from either 2-day and 42-day cardiac myocytes or from whole ventricular tissue at 2, 5–11, 14, 17, 21, 28 and 42 days of age. Protein expression was determined by immunoblotting with an 80K/MARCKS antipeptide antibody and PKC activity was determined by measuring the amount of γ32P-ATP transferred to a specific peptide substrate. Results: RT-PCR analysis of 80K/MARCKS mRNA in neonatal (2-day) and adult (42-day) cardiac myocytes showed the expression of this gene in both cell types. Immunoblotting revealed maximum 80K/MARCKS protein expression in whole ventricular tissue at 5 days (a 75% increase above values at 2 days), followed by a transient decrease in expression during the 6–8-day period (61% of the protein expressed at 2 days for 8-day tissue) with levels returning to 5 day levels by 11 days of age. 80K/MARCKS protein was present in cardiac myocytes at 2 days of age whereas it was not detectable in adult cells. In addition, PKC activity levels increased to 160% of levels present at 2 days in 8-day-old ventricles with PKC activity levels returning to 5-day levels by 9 days of age. This was then followed by a steady decline in both 80K/MARCKS protein expression and PKC activity through to adulthood. Conclusions: Expression of the PKC substrate, 80K/MARCKS, in cardiac myocytes changes significantly during development and the transient loss of immunoreactive protein during the 6–8-day developmental period may reflect 80K/MARCKS phosphorylation and subsequent down-regulation as a result of the concomitant up-regulation of PKC activity at this time.

Renewable energy in the UK countryside: policy and practice

Of the technologies currently available for producing energy from renewable sources in the British climate all except one depend on a single ingredient, namely land. Therefore other than offshore wind generation, which has been slow and expensive to establish, renewables have had to be derived almost entirely from the land, whether as sites for turbines or areas on which to grow feedstocks for biomass and biofuels. Of these, only wind turbines have been developed in any number while economic conditions have until now been unfavourable for biomass and biofuel. The UK is unlikely to meet its present targets under the Kyoto agreement, due to a mixture of limited funding and problems of policy. Peter Prag examines the present position and the potential outlook.

English teachers in the digital age – a case study of policy and expert practice from England

This article is a case study of how English teachers in England have coped with the paradigm shift from print to digital literacy. It reviews a large scale national initiative that was intended to upskill all teachers, considers its weak impact and explores the author’s involvement in the evaluation of the project’s direct value to English teachers. It explores how this latter evaluation revealed how best practice in English using ICT was developing in a variable manner. It then reports on a recent small scale research project that investigated how very good teachers have adapted ICT successfully into their teaching. It focuses on how the English teachers studied in the project are developing a powerful new pedagogy situated in the life worlds of their students and suggests that this model may be of benefit to many teachers. The issues this article reports on have resonance in all English speaking countries. This article is also a personal story of the author’s close involvement with ICT and English over 20 years, and provides evidence for his conviction that digital technologies will eventually transform English teaching.

Contested H2O: science, policy and politics in water resources management in Chile

This paper critically explores the politics that mediate the use of environmental science assessments as the basis of resource management policy. Drawing on recent literature in the political ecology tradition that has emphasised the politicised nature of the production and use of scientific knowledge in environmental management, the paper analyses a hydrological assessment in a small river basin in Chile, undertaken in response to concerns over the possible overexploitation of groundwater resources. The case study illustrates the limitations of an approach based predominantly on hydrogeological modelling to ascertain the effects of increased groundwater abstraction. In particular, it identifies the subjective ways in which the assessment was interpreted and used by the state water resources agency to underpin water allocation decisions in accordance with its own interests, and the role that a desocialised assessment played in reproducing unequal patterns of resource use and configuring uneven waterscapes. Nevertheless, as Chile’s ‘neoliberal’ political-economic framework privileges the role of science and technocracy, producing other forms of environmental knowledge to complement environmental science is likely to be contentious. In conclusion, the paper considers the potential of mobilising the concept of the hydrosocial cycle to further critically engage with environmental science.