The homotetrameric enzyme dihydrodipicolinate synthase (DHDPS, E.C. 4.2.1.52) from Escherichia coli was used as a model for probing oligomeric structure in enzymes. Dimeric mutants of this enzyme have been found in previous work to be largely inactive, due to the trapping of a covalent adduct. Partial restoration of catalytic activity has been achieved by incubation in the presence of the substrate pyruvate to displace the adduct. It was hypothesized that the buttressing of dimeric units against one another in the wildtype tetrameric form of DHDPS provides stability in the dimer interface, necessary to maintain optimum catalytic performance and substrate specificity. We hypothesized that buttressing a dimeric DHDPS mutant against a surface would result in restoration of catalytic activity by mimicking the buttressing proposed to occur in the tetrameric structure. To test this hypothesis, dimeric DHDPS mutants were immobilised against an agarose support and the immobilised enzymes characterised. Three DHDPS mutants were prepared, the double mutant DHDPS-C20S/L167C was produced by mutagenesis and a crystal structure obtained in collaboration with Dr Renwick Dobson. Two other mutants, DHDPS-Ll67C and DHDPS-Ll97Y were also over expressed and purified. The quaternary structures of the three mutants were characterised in solution, DHDPS-Ll67C was determined to be tetrameric, DHDPS-C20S-Ll67C was found to equilibrate between tetramer and dimer and DHDPS-Ll97Y was confirmed as a dimer, consistent with previous findings. Modification experiments indicated that the sulfhydryl groups of DHDPS-C20S/L167C were available for immobilisation. Activation experiments indicated that both DHDPS-Ll67C and DHDPS-Ll97Y activated. These results were in accord with those of others in indicating that the displacement of an a-ketoglutarate adduct from the active site was responsible for the activation of mutant DHDPS enzymes. Wild-type DHDPS and the mutants were immobilised through amine and sulfhydryl groups. The free and immobilised enzymes were rigorously characterised, with thermal stability, pH optima, kinetic and lysine inhibition properties determined and compared to wild-type DHDPS. Following immobilisation, substrate affinity was found to decrease for wild-type and mutant enzymes, wild-type KmPyr = 0.26 mM free, 0.8-1.2 mM immobilised, Km(S)-ASA = 0.10 mM free, 1.5-2.5 mM immobilised. Lysine inhibition was determined to be largely unaffected by immobilisation. The largest change in K, was an increase to double that of the free enzyme. Restoration of some catalytic activity was found following the immobilisation of dimeric DHDPS-Ll97Y, the immobilised enzyme was 31 ± 12% more active than free DHDPS-Ll97Y. DHDPS-C20S/L167C was also found to immobilise as a dimer. Comparison ofthe immobilised DHDPS-C20S/L167C dimer with a derivatised free dimeric form ofthis enzyme indicated that an increase from 3% to 9% of wild-type activity had resulted from immobilisation. These results supported the hypothesis that buttressing of a dimeric mutant of DHDPS against a support surface would increase catalytic activity and that buttressing across the dimerdimer interface is essential for optimal catalytic activity in DHDPS enzymes.

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4 pp. / Why healthy meetings? Many choices go into having a successful 4-H club meeting, training, community workshop, or grower meeting. Extension staff, and volunteers should include healthy lifestyle behaviors into their education and "Walk the Talk" for healthy living in Arizona. Learn how your next meeting or event can promote healthy habits like good nutrition, physical activity and safe food.

4 pp. / (Make My Map in Google (Part 1: Overview, Part 2: Detailed) GPS Watches for Measuring Energy Expenditure during Physical Activity / Maps are a valuable tool among professionals including engineers, environmental specialists, and city developers. Online mapping using GPS has gained popularity in vehicles and cell phones. Recently, online mapping can be used to create your own map (Google My Map) by adding personal photos, related website links, and collaborating with others. Online mapping has now become useful and fun for all ages in a variety of settings. The fact sheets provide a basic (Part 1: Overview) and detailed (Part 2: Detailed) how-to guide to Make a Map in Google [My Maps] using a GPS receiver or GPS watch, as well as enhancing those maps into a poster. We also included interesting facts and useful tips for readers. This article can be used as a guide to encourage youth to map their favorite places, as well as share these places with other youth, friends, family, community groups and organizations.

4 pp. / "(Make My Map in Google (Part 1: Overview, Part 2: Detailed) GPS Watches for Measuring Energy Expenditure during Physical Activity" / Maps are a valuable tool among professionals including engineers, environmental specialists, and city developers. Online mapping using GPS has gained popularity in vehicles and cell phones. Recently, online mapping can be used to create your own map (Google My Map) by adding personal photos, related website links, and collaborating with others. Online mapping has now become useful and fun for all ages in a variety of settings. The fact sheets provide a basic (Part 1: Overview) and detailed (Part 2: Detailed) how-to guide to Make a Map in Google [My Maps] using a GPS receiver or GPS watch, as well as enhancing those maps into a poster. We also included interesting facts and useful tips for readers. This article can be used as a guide to encourage youth to map their favorite places, as well as share these places with other youth, friends, family, community groups and organizations.

220 pp. / A collection of hands-on, interactive activities designed to engage youth in understanding purposes, uses, applications, and designs of rainwater harvesting systems. Together the activities promote a culture of conservation through the development of rainwater harvesting demonstration and use projects, encouragement of community awareness and action, and optimally the reduction of groundwater and surface water use.

This work analyzes data taken in 1979 using a modification of the solar detector at SCLERA (Santa Catalina Laboratory for Experimental Relativity) designed to enhance spatial properties of the previously observed solar oscillations. Unlike previous solar observations taken at SCLERA, where the data consisted of single solar diameter measurements, the 1979 data consisted of six recorded limb profiles. This has important ramifications for the amount of signal present in the data which was generated by the terrestrial atmosphere, for the origin of the observed solar oscillations in fluctuations of the solar limb darkening function, and, most importantly, for the spatial symmetry properties of the observed solar eigenfunctions. The data consisted of 18 days of observations averaging ten hours per day and covering a total of 41 days. A linked Fourier transform of all 18 days was done for signal generated from each limb profile, and combinations of these six Fourier transforms made to increase sensitivity to symmetric or antisymmetric properties of the observed solar eigenmodes. The following results were found: 1. The observed oscillations are manifestations of fluctuations in the solar limb darkening function. 2. Terrestrial atmospheric contributions to the observed signal are negligible; thus, the sun constitutes the only possible source of the signal. 3. Given a resolution element of 1/(41 days) or 0.28 μHz, the solar oscillations observed represent individual solar eigenstates. 4. The spatial properties of the eigenstates are consistent with their interpretation in terms of spherical harmonics defined with respect to the observed solar rotational axis. 5. The eigenstates are temporally coherent for > 2 days and, in selected samples, for > 41 days. 6. The observed spacing of groups of eigenmodes is shown to be indicative of solar rotational effects; this spacing implies that the core of the sun is rotating approximately six times faster than the observed surface rotational velocity.

This work investigates the response of the solar atmosphere to mechanical and thermal driving due to global solar oscillations. It was discovered that the coupling of thermal and mechanical modes was very important in reconciling theoretical predictions of the expected change in the solar limb due to solar oscillations and experimental observations of the variability in the solar limb darkening function undertaken at SCLERA (Santa Catalina Laboratory for Experimental Relativity). The coupling between the thermal and mechanical modes occur mainly due to the nonlocal nature of the radiation field. Previous theoretical calculations that used approximations for the radiative transfer that ignored the nonlocal nature of the radiation field predicted expected temperature perturbations (compared to the fluid displacement) that were much too small to be observed. Much larger ratios were found when the radiative transfer was treated properly. A particular solar oscillation can be influenced by the presence of a large number of other modes, if these modes can change the average properties of the medium. If the basic nonlinear equations are statistically averaged, the influence of the "mean field" can be investigated. This nonlinear effect can become important in the analysis for single modes in the upper photosphere.

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