The seeds of lotus japonicus lines transformed with sense, antisense, and sense/antisense galactomannan galactosyltransferase constructs have structually altered galactomannan in their endosperm cell walls

Galactomannan biosynthesis in legume seed endosperms involves two Golgi membrane-bound glycosyltransferases, mannan synthase and galactomannan galactosyltransferase (GMGT). GMGT specificity is an important factor regulating the distribution and amount of (1-->6)-alpha-galactose (Gal) substitution of the (1-->4)-beta-linked mannan backbone. The model legume Lotus japonicus is shown now to have endospermic seeds with endosperm cell walls that contain a high-Gal galactomannan (mannose [Man]/Gal = 1.2-1.3). Galactomannan biosynthesis in developing L. japonicus endosperms has been mapped, and a cDNA encoding a functional GMGT has been obtained from L. japonicus endosperms during galactomannan deposition. L. japonicus has been transformed with sense, antisense, and sense/antisense ("hairpin loop") constructs of the GMGT cDNA. Some of the sense, antisense, and sense/antisense transgenic lines exhibited galactomannans with altered (higher) Man/Gal values in their (T-1 generation) seeds, at frequencies that were consistent with posttranscriptional silencing of GMGT. For T-1 generation individuals, transgene inheritance was correlated with galactomannan composition and amount in the endosperm. All the azygous individuals had unchanged galactomannans, whereas those that had inherited a GMGT transgene exhibited a range of Man/Gal values, up to about 6 in some lines. For Man/Gal values up to 4, the results were consistent with lowered Gal substitution of a constant amount of mannan backbone. Further lowering of Gal substitution was accompanied by a slight decrease in the amount of mannan backbone. Microsomal membranes prepared from the developing T-2 generation endosperms of transgenic lines showed reduced GMGT activity relative to mannan synthase. The results demonstrate structural modification of a plant cell wall polysaccharide by designed regulation of a Golgi-bound glycosyltransferase.

Method for the generation and cultivation of functional three-dimensional mammary constructs without exogenous extracellular matrix

During puberty, pregnancy, lactation and postlactation, breast tissue undergoes extensive remodelling and the disruption of these events can lead to cancer. In vitro studies of mammary tissue and its malignant transformation regularly employ mammary epithelial cells cultivated on matrigel or floating collagen rafts. In these cultures, mammary epithelial cells assemble into three-dimensional structures resembling in vivo acini. We present a novel technique for generating functional mammary constructs without the use of matrix substitutes.

Exploring the usefulness of a consumer activity index in the sponsorship-linked marketing context

Despite the now well developed use of sponsorship-linked marketing, there have been few methodological advances in the measurement of sponsorship constructs and outcomes. This paper offers a preliminary development of an activity index for use in the sponsorship marketing context. The activity index seeks to capture the consumer's extended experience with sport (rugby) and considers the relationship of this overall experience to sponsorship-related outcomes of interest. Initial development of the index, based on a convenience sample of 108 people visiting a sports centre, shows promise.

Using front-end and formative evaluation to design and test persuasive bird feeding warning signs

Wildlife feeding is a wide-spread and controversial practice that can pose serious threats to the safety of both wildlife and visitors. The design and effectiveness of warning signs in recreational areas varies considerably and is rarely the product of theoretical models or scientific research. This study uses front-end and formative evaluation to design and test the perceived effectiveness of warning signs relating to bird feeding. Stage One examined visitors' beliefs, attitudes and bird feeding behaviour and found significant differences between feeders and non-feeders. Stage Two involved designing and evaluating three signs that built on the beliefs, knowledge and mis/conceptions identified in Stage One. Respondents thought the sign that focused on the birds' health and safety would be the most persuasive, however, elements of the other two signs were also positively evaluated. The article concludes with recommendations for the wording of future bird feeding warning signs. (c) 2004 Elsevier Ltd. All rights reserved.