Lack of effect of serotonin and norepinephrine on CRF release from hypothalami in vitro

A number of investigations utilizing hypothalami from adrenalectomized animals have provided conflicting results regarding the role of serotonin (5HT) and norepinephrine (NE) in CRF regulation. In order to further investigate these neurotransmitters, we performed three sets of experiments with hypothalami obtained from intact rats. In experiment I, freshly obtained rat hypothalami were randomly grouped and incubated in control medium and medium in the presence of 10(-11) M, 10(-10) M, and 10(-9) M serotonin. Aliquots of 200 microliters of these incubates were bioassayed for CRF activity using a dispersed anterior pituitary cell system, and ACTH secretion from the cells was determined by radioimmunoassay. A preliminary experiment had determined that a 200 microliters aliquot from hypothalami incubated in control medium resulted in a significantly (p less than 0.0001) greater ACTH release than obtained from cells alone. No significant effect of serotonin on hypothalamic CRF release was obtained. In experiment III, individual hypothalami were bisected longitudinally, and one half served as control. The contralateral half was incubated in medium containing 10(-11) M, 10(-10) M, and 10(-9) M serotonin. CRF release in this experiment again revealed no significant effect of serotonin. In experiment II, hypothalami were again randomly grouped and incubated with control medium and in the presence of 10(-8) M and 10(-6) M norepinephrine. This experiment resulted in no significant effect of norepinephrine on CRF release. These results suggest that serotonin and norepinephrine at the concentrations studied have no effect on CRF release from hypothalami obtained from intact rats.     

Critical Design Ethnography: Designing for Change

This article describes critical design ethnography, an ethnographic process involving participatory design work aimed at transforming a local context while producing an instructional design that can be used in multiple contexts. Here, we reflect on the opportunities and challenges that emerged as we built local critiques then reified them into a designed artifact that has been implemented in classrooms all over the world.     

Digestive enzymes of the saltmarsh periwinkleLittorina irrorata (Mollusca: Gastropoda)

Summary The saltmarsh periwinkleLittorina irrorata is well adapted for the digestion of a wide range of polysaccharides. Enzyme extracts attacked cellulose, pectin, xylan, bean gum and mannan (common in cell walls of higher plants), as well as starch and laminarin (representative of major polysaccharide classes in fungal, algal, and animal tissues). Activities were generally highes at a ph of 5 or 6. There was no evidence that chitin was digested, but 19 other enzymes, active toward various carbohydrates, lipids and peptides, were demonstrated. Enzymatic activity toward Azocoll, a general substrate for proteinase activity, was weak compared to that of other aquatic detritivores. The maximum was reached at a pH of 8. Enzymatic activities were generally measured with extracts of the entire visceral hump. Separate stomach or intestine extracts also gave strong activities. The stomach was the most acidic section of the digestive system with an average pH of 5.8; the intestine had an average pH of 7.3.     

Simulation and Neutron Diffraction Studies of Small Biomolecules in Water

Modern biophysics has benefited greatly from the use of X-ray and neutron diffraction from ordered single crystals of proteins and other macromolecules to give highly detailed pictures of these molecules in the solid state. However, the most biologically relevant environments for these molecules are liquid solutions, and their liquid state properties are sensitive to details of the liquid structuring. The best experimental method for studying such structuring is also neutron diffraction, but of course, the inherent disorder of the liquid state means that these experiments cannot hope to achieve the level of informational detail available from single crystal diffraction. Nonetheless, recent advances in neutron beam intensity, beam stability, and detector sensitivity mean that it should be possible, at least in principle, to use such measurements to extract information about structuring in much more complex systems than have previously been studied. We describe a series of neutron diffraction studies of isotopically labeled molecules in aqueous solution which, when combined with results from computer simulations, can be used to extract conformational information of the hydration of the molecules themselves, essentially opening up new avenues of investigation in structural biology.