The effect of compressive loading on proteoglycan turnover in cultured fetal tendon

A fibrocartilaginous tissue develops in tendon at the point where the tendon wraps under bone and is subjected to transverse compressive loading in addition to tension. This tissue is characterized by a high level of large proteoglycan (aggrecan), which could accumulate because of increased synthesis, diminished turnover, or both. To examine the effect of loading on proteoglycan turnover segments of fetal tendon in sterile culture were subjected to cyclic, uniaxial compression loading to 30% of initial thickness once every 6 sec. for 72 h, and then allowed to incorporate 35S-sulfate for 12 h. The rate of loss of newly-synthesized 35S-proteoglycans from tissue was determined during subsequent culture for up to 12 days, with or without continued loading. Proteoglycan was lost from fetal tendon segments rapidly during the first 3 days of culture and slowly thereafter. Loss of newly-synthesized proteoglycan from adult tendon fibrocartilage was linear, with a half life of 12 d. Segments of fetal tendon subjected to cyclic compression before labeling synthesized more proteoglycan. These segments lost a greater percent of labeled proteoglycan to medium during a subsequent 12-day culture period than matched segments that had not experienced loading. Analysis of medium and tissue proteoglycans by SDS polyacrylamide gel electrophoresis and sieve chromatography indicated that small proteoglycans (decorin and biglycan) were retained in both loaded and non-loaded tissue whereas large proteoglycans (migrating in the Vo of a Sepharose CL-4B column) were readily lost. It is concluded that the 3-day loading regimen did not diminish turnover of large proteoglycan. To the contrary, although synthesis of large proteoglycan was enhanced by the loading regimen, these proteoglycans were still rapidly lost from the fetal tissue.     

Some potential applications for intercalation compounds of graphite with high electrical conductivity

Intercalation compounds of graphite of the acceptor type have potential engineering applications because of their attractive electrical conductivity properties. Two kinds of applications are considered in this paper. The first concerns a composite, formed by enclosing an intercalation compound synthesized from high quality crystalline graphite in a matrix of copper. With this form of composite it is found that there are both intrinsic and extrinsic advantages pertaining to the use of a material that has a conductivity higher than and a density lower than that of copper. The second form is a composite compound of intercalated graphite fibers contained in a matrix of epoxy. Extraordinary advantages in this case result from the fact that while intercalation of the fibers produces an order of magnitude increase in their electrical conductivity, when these fibers are incorporated into an epoxy matrix, the composite conductivity is increased by two orders of magnitude over its pristine fiber counterpart. It is projected that these desirable electrical conductivity characteristics portend large scale uses for the acceptor compounds of graphite as substitutes for the present standard conductors, and as a way of upgrading the performance of carbon/graphite materials.     

Comparison of pressure and heat resistance of Clostridium botulinum and other endospores in mashed carrots

Inactivation of bacterial endospores in food requires a combination of pressure and moderate heat. Endospore resistance of seven Clostridium botulinum strains was compared with those of Bacillus spp. (B. cereus, B. subtilis, B. licheniformis, B. smithii, B. amyloliquefaciens) and Thermoanaerobacterium thermosaccharolyticum with respect to pressure (600 to 800 MPa) and temperature (80 to 116 degrees C) treatments in mashed carrots. A large variation was observed in the pressure resistance of C. botulinum spores. Their reduction after treatments with 600 MPa at 80 degrees C for 1 s ranged from more than 5.5 log units to no reduction. Spores of the proteolytic C. botulinum TMW 2.357 exhibited a greater resistance to pressure than spores from all other bacteria examined, with the exception of B. amyloliquefaciens. Heat resistance of spores did not correlate with pressure resistance, either within strains of C. botulinum or when C. botulinum spores were compared with spores of T. thermosaccharolyticum. A quantitative release of dipicolinic acid was observed from C. botulinum spores on combined pressure and temperature treatments only after inactivation of more than 99.999% of the spores. Thus, dipicolinic acid is released by a physicochemical rather than a physiological process. The resistance of spores to combined pressure and temperature treatments correlated with their ability to retain dipicolinic acid. B. amyloliquefaciens, a mesophilic organism that forms highly pressure-resistant spores is proposed as a nonpathogenic target organism for high-pressure process development.     

Sequential switch of biomineral crystal morphology using trivalent ions

Many biominerals are laminated such that crystal shape or habit changes from layer to layer thus yielding exquisitely designed composite materials with tightly controlled properties. Although lamination in biominerals is usually performed using peptides and proteins, here we introduce a new strategy by which sequential addition or depletion of inorganic trivalent ions in a supersaturated solution can be used to switch the surface morphology of calcium oxalate monohydrate (COM) back and forth, resulting in either the growth of flat crystalline sheets or of nanostructures oriented perpendicular to the surface. We propose that the occupation of a Ca(2+) site by Eu(3+) ion switches the orientation of the COM unit cell. The need to compensate the third charge forces coordination of Eu(3+) to an additional oxalate ion ((-)OOC-COO(-)) in an orientation that is not compatible with the initial unit cell. This mechanism of switching the orientation of the unit cell is unique, as it does not involve the use of expensive and thermally labile biomolecules. Suggestions of how to extend this strategy to engineer non-biological nanocomposites are given.     

Traffic sense--which factors influence the skill to predict the development of traffic scenes?

A study was conducted to evaluate the skill to predict the development of traffic situations. A stop-controlled intersection was filmed over several days, and 12 scenes with varying traffic complexity were selected. In half of the scenes, the traffic rules were violated, in half of the scenes, the rules were observed. A total of 36 participants were asked to watch the scenes and predict how the scene would most likely develop in the 2s after the film was paused. Additionally, the participants rated how certain they were about their prediction, and how complex and dangerous they assessed the scenes to be. With the method used here, experienced drivers were not found to make more correct predictions of situational development, and no difference in skill to predict could be found between genders. Nevertheless, more experienced drivers were more certain in their judgements and evaluated the situations on average as less complex and dangerous than did less experienced drivers. Scenes in which the traffic rules were violated were more difficult to predict correctly. The scenes in which the participants predicted violations were rated as more complex and dangerous. It is concluded that the low-cost method used here is more useful for examining which scenes are generally easy or difficult to predict and how they are experienced subjectively than to investigate differences in performance for different driver categories.     

A comparison of headway and time to collision as safety indicators

The two safety indicators "headway" and "time to collision (TTC)" are discussed and compared with respect to their usefulness in determining the safety of different traffic situations, like different locations in a junction. Over a 6-day-period traffic flow measures were taken in a four-way junction with stop signs on the minor road. It was found that for vehicles in a car following situation headway and TTC are independent of each other. The percentage of small headways is relatively constant across different locations in the junction, while the percentage of small TTC values varies between different locations. It is recommended to use headway for enforcement purposes, because small headways generate potentially dangerous situations. TTC, on the other hand, should be used when a certain traffic environment is to be evaluated in terms of safety, because it indicates the actual occurrences of dangerous situations.     

In vitro and in vivo ligand binding to the 5HT(3) serotonin receptor characterised by time-resolved fluorescence spectroscopy

The binding of the fluorescein-labelled antagonist GR-flu ([1,2,3,9-tetrahydro-3-[(5-methyl-1H-imidazol-4-yl)methyl]-9-(3-amino-(N-fluoresceinthiocarbamoyl)propyl)-4H-carbazol-4-one]) to a purified, detergent-solubilised ligand-gated ion channel, the type-3 serotonin (5-hydroxytryptamine, 5HT) receptor (5HT(3)R), was characterised by frequency-domain time-resolved fluorescence spectroscopy (TRFS). Detailed understanding of how ligands interact with the homopentameric receptor was obtained. While a 1:1 stoichiometry was observed for the GR-flu-receptor complex, the agonist quipazine bound cooperatively to the receptor, suggesting multiple binding sites for this ligand. The GR-flu-binding site of the receptor was proven to provide an acidic environment as shown by determining the fraction of bound GR-flu in the protonated state. Fluorescence anisotropy relaxation experiments indicated a hindered but still high mobility for the receptor-bound GR-flu. Hence, the binding site is expected to present a wide opening to the ligand. Finally, we succeeded in measuring the binding of GR-flu to 5HT(3) receptors in live cells. These results show that the purified and the native receptor behave identically and demonstrate that time-resolved fluorescence measurements are suited to selectively investigate biomolecular interactions in live cells.