Glucosamine inhibits the synthesis of glycosaminoglycan chains on vascular smooth muscle cell proteoglycans by depletion of cellular ATP

Glucosamine via GlcNAc is a precursor for the synthesis of glycosaminoglycan (GAG) chains on proteoglycans. We previously found that proteoglycans synthesized and secreted by vascular smooth muscle cells (VSMC) in the presence of supplementary glucosamine had GAG of decreased not increased size. We investigated the possibility that the inhibition of GAG chains synthesis on proteoglycans might be related to cellular ATP depletion. Confluent primate VSMCs were exposed to glucosamine, azide, or 2-deoxyglucose (2-DG). Each of these agents depleted cell ATP content by 25-30%. All agents decreased 35S-SO4 incorporation and reduced the size of the proteoglycans, decorin and biglycan as assessed by SDS-PAGE. On withdrawal of the glucosamine, azide or 2-DG ATP levels and proteoglycan synthesis returned towards baseline values. Glucosamine decreased glucose uptake and consumption suggesting that ATP depletion was due preferential phosphorylation of glucosamine over glucose. Thus, glucosamine inhibition of proteoglycan synthesis is due, at least in part, to depletion of cellular ATP content

Increased growth hormone (GH), growth hormone receptor (GHR), and insulin-like growth factor I (IGF-I) gene transcription after hyperosmotic stress in the Brazilian flounder Paralichthys orbignyanus

Growth hormone (GH) action is the result of an intracellular cascade initiated just after its interaction with the growth hormone receptor (GHR)located on the surface of target cells. This cascade culminates with the transcription of target genes, such as the insulin-like growth factors (IGFs), which are responsible for most GH biological effects. In addition to its central role in growth, fish GH is also involved with osmoregulatory control. Within this context, the objective of the present work was to isolate GH, GHR, and IGF-I cDNAs from the Brazilian flounder Paralichthys orbignyanus and evaluate whether these genes are induced by hyperosmotic stress. The obtained results indicated that GH mRNA had a significant peak only 24 h after yperosmotic stress. In gills, GHR mRNA was significantly increased after 7 days. In liver, GHR and IGF-I mRNAs were significantly increased in 72 h and both reached even higher levels after 7 days. These results indicate that hyperosmotic stress can increase GH sensitivity in the gills and liver of P. orbignyanus and, consequently, improve IGF-I production. The management of this parameter could be useful in achieving better growth performance for this and other commercially important species in which GH has a direct correlation with osmoregulatory mechanisms

Assessing the Influence of Field- and GIS-based Inquiry on Student Attitude and Conceptual Knowledge in an Undergraduate Ecology Lab

Combining field experience with use of information technology has the potential to create a problem-based learning environment that engages learners in authentic scientific inquiry. This study, conducted over a 2-yr period, determined differences in attitudes and conceptual knowledge between students in a field lab and students with combined field and geographic information systems (GIS) experience. All students used radio-telemetry equipment to locate fox squirrels, while one group of students was provided an additional data set in a GIS to visualize and quantify squirrel locations. Pre/postsurveys and tests revealed that attitudes improved in year 1 for both groups of students, but differences were minimal between groups. Attitudes generally declined in year 2 due to a change in the authenticity of the field experience; however, attitudes for students that used GIS declined less than those with field experience only. Conceptual knowledge also increased for both groups in both years. The field-based nature of this lab likely had a greater influence on student attitude and conceptual knowledge than did the use of GIS. Although significant differences were limited, GIS did not negatively impact student attitude or conceptual knowledge but potentially provided other benefits to learners