Restoration of glutathione levels in vascular smooth muscle cells exposed to high glucose conditions

Hyperglycaemia-induced oxidative stress may play a key role in the pathogenesis of diabetic vascular disease. The purpose of the present study was to determine the effects of glucose on levels of glutathione (a major intracellular antioxidant), the expression of gamma-glutamylcysteine synthetase (the rate-limiting enzyme in glutathione de novo synthesis) and DNA damage in human vascular smooth muscle cells in vitro. High glucose conditions and buthionine sulphoximine, an inhibitor of gamma-glutamylcysteine synthetase, reduced intracellular glutathione levels in vascular smooth muscle cells. This reduction was accompanied by a decrease in the mRNA expression of both subunits of gamma-glutamylcysteine synthetase as well as an increase in DNA damage. In high glucose conditions incubation of the vascular smooth muscle cells with alpha-lipoic acid and L-cystine restored glutathione levels. We suggest that the decrease in GSH levels seen in high glucose conditions is mediated by the availability of cysteine (rate-limiting substrate in de novo glutathione synthesis) and the gene expression of the gamma- glutamylcysteine synthetase enzyme. Glutathione depletion is associated with an increase in DNA damage, which can be reduced when glutathione levels are restored.

The 'Belfast' timber roof truss - is it sustainable and relevant today?

In the early 19th century the requirement for clear span industrial buildings brought about the development of a variety of timber truss types. The Belfast truss was introduced circa 1860 to meet the demand for efficient wide span industrial buildings. It has essentially a bow-string configuration with a curved top chord, straight horizontal bottom chord and close-spaced lattice web. Several thousand still exist in Ireland, many in buildings of historic significance. This paper sets out to demonstrate the efficiency of the Belfast truss and to show that, by modern structural design criteria, the concept, member sizes and joint details were well chosen. Trusses in historic buildings can be replicated almost exactly as originally fabricated. Results of a theoretical study are compared with the experimental behaviour of two full-scale trusses: one a replacement truss, tested in the laboratory; the other an 80-year-old truss tested on site. In addition, experimental results from a manufacturers archive material of full-scale truss tests carried out about 100 years ago are compared with theoretical models. As well as considering their significance in building conservation the paper proposes that Belfast trusses are an attractive sustainable alternative to other roof structures. The analysis, design, fabrication and testing of trusses have resulted in a better understanding of their behaviour which is not only of historic interest and fundamental to the repair/restoration of existing trusses, but also relevant to the design of modern timber trusses and the promotion of a sustainable form of roof construction.

ZAP-70 restoration in mice by in vivo thymic electroporation.

Viral and non-viral vectors have been developed for gene therapy, but their use is associated with unresolved problems of efficacy and safety. Efficient and safe methods of DNA delivery need to be found for medical application. Here we report a new monopolar system of non-viral electro-gene transfer into the thymus in vivo that consists of the local application of electrical pulses after the introduction of the DNA. We assessed the proof of concept of this approach by correcting ZAP-70 deficient severe combined immunodeficiency (SCID) in mice. The thymic electro-gene transfer of the pCMV-ZAP-70-IRES-EGFP vector in these mice resulted in rapid T cell differentiation in the thymus with mature lymphocytes detected by three weeks in secondary lymphoid organs. Moreover, this system resulted in the generation of long-term functional T lymphocytes. Peripheral reconstituted T cells displayed a diversified T cell receptor (TCR) repertoire, and were responsive to alloantigens in vivo. This process applied to the thymus could represent a simplified and effective alternative for gene therapy of T cell immunodeficiencies.