Prevention of in vitro protein thermal aggregation by the Sulfolobus solfataricus chaperonin. Evidence for nonequivalent binding surfaces on the chaperonin molecule

We have studied the effects of the Sulfolobus solfataricus chaperonin on the aggregation and inactivation upon heating of four model enzymes: chicken egg white lysozyme (one 14.4-kDa chain), yeast alpha-glucosidase (one 68.5-kDa chain), chicken liver malic enzyme (four 65-kDa subunits), and yeast alcohol dehydrogenase (four 37.5-kDa subunits). When the proteins were heated in the presence of an equimolar amount of chaperonin, 1) the aggregation was prevented in all solutions; 2) the inactivation profiles of the single-chain enzymes were comparable with those detected in the absence of the chaperonin, and enzyme activities were regained in the solutions heated in the presence of the chaperonin upon ATP hydrolysis (78 and 55% activity regains for lysozyme and alpha-glucosidase, respectively); 3) the inactivation of the tetrameric enzymes was completely prevented, whereas the activities decreased in the absence of the chaperonin. We demonstrate by gel filtration chromatography that the chaperonin interacted with the structures occurring during thermal denaturation of the model proteins and that the interaction with the single-chain proteins (but not that with the tetrameric proteins) was reversed upon ATP hydrolysis. The chaperonin had nonequivalent surfaces for the binding of the model proteins upon heating: the thermal denaturation intermediates of the single-chain proteins share Surfaces I, while the thermal denaturation intermediates of the tetrameric proteins share Surfaces II. ATP binding to the chaperonin induced a conformation that lacked Surfaces I and carried Surfaces II. These data support the concept that chaperonins protect native proteins against thermal aggregation by two mechanistically distinct strategies (an ATP-dependent strategy and an ATP-independent strategy), and provide the first evidence that a chaperonin molecule bears functionally specialized surfaces for the binding of the protein substrates.     

WOx/ZrO2 catalysts: Part 3. Surface coverage as investigated by low temperature CO adsorption: FT-IR and volumetric studies

The adsorption of CO at 77 K on zirconia-supported tungsten oxide (ZW) samples prepared by equilibrium adsorption or by impregnation and with tungsten content up to 12.4 W atoms nm⁻² was studied with the aim of determining the coverage of ZrO₂ surface by the supported tungsten oxospecies. As an essential background for quantitative volumetric measurements, ZW samples were investigated by FT-IR spectroscopy to identify the surface sites responsible for the CO adsorption. FT-IR showed that CO adsorbed selectively and irreversibly on Zr_cus⁴⁺ centers of the support surface, but not on W_cus⁶⁺ sites. By volumetric determinations, the coverage of the zirconia surface by tungsten oxospecies was about 50% at maximum, irrespective of the preparation procedure and of the support crystallographic modification (monoclinic or tetragonal). Zirconia coverage remained significantly unchanged after leaching with a NH₃ solution that removed tungsten oxospecies not directly interacting with the zirconia surface. This finding indicates that ZrO₂ support coverage depends mainly on the tungsten species strongly anchored to the zirconia surface.     

Monitoring of a Best Management Practice Wetland before and after Maintenance

The U.S. Environmental Protection Agency’s (USEPA) Urban Watershed Management Branch monitored a best management practice (BMP) wetland in the Richmond Creek (RC) watershed. This BMP, designated Richmond Creek #5 (RC-5), is owned and operated by the New York City Department of Environmental Protection’s as part of the Bluebelt program. During monitoring performed in 2003 and 2004, RC-5 was monitored for several water quality parameters including suspended solids (SS), chemical oxygen demand (COD), and phosphorous. Calculated SS loads demonstrated a 61% removal which is in line with the anticipated removals of 55%–57%, however, COD was not being removed from RC-5. Due to visible accumulation of sediment in the forebay and leaves throughout the BMP, maintenance activity was performed in the spring of 2005, to remove the sediment and organic matter (leaves) by vactor truck and physical labor. About four truck loads of sediment and leaf litter, estimated at 35?m3 (46 cubic yards), was removed from the site. Monitoring during the fall of 2005 measured the effects of this maintenance activity. The effluent concentrations of COD were significantly less and less variable after maintenance, and calculated influent and effluent COD loading indicated removals. The reduction in COD effluent coincided with drops in effluent phosphorous levels. Effluent SS concentrations were not significantly altered, implying RC-5 still had capacity to remove sediments before maintenance occurred. Indications are that the maintenance activity significantly reduced the COD effluent and may help communities manage phosphorous loadings.     

Large Scale Probabilistic Drought Characterization Over Europe

A reliable assessment of drought return periods is essential to help decision makers in setting effective drought preparedness and mitigation measures. However, often an inferential approach is unsuitable to model the marginal or joint probability distributions of drought characteristics, such as drought duration and accumulated deficit, due to the relatively limited number of drought events that can be observed in the historical records of the hydrological variables of interest. As an alternative, the marginal and multivariate probability cdf’s of drought characteristics can be derived as functions of the parameters of the cdf of the underlying variable (e.g. precipitation), whose sample series is usually long enough to obtain trustworthy estimates in a statistical sense. In this study, the latter methodology is applied to investigate space-time variability of drought occurrences over Europe by using the CRU TS3.10.01 precipitation dataset for the period 1901–2009. In particular, a methodology able to take into account autocorrelation in the underlying precipitation series is adopted. First, a spatial analysis of historical droughts at European level is carried out. Then, the joint probability distributions of drought duration and accumulated deficit are derived for each cell, with reference to both historical and design drought events. Finally, the corresponding bivariate drought return periods are computed, as the expected values of the interarrival time between consecutive critical droughts.Results show that several heavy drought episodes have widely affected the continent. Among the most recent events, drought occurred during the period 1985–1995 was the worst in terms of extent of the regions characterized by return periods greater than 250 years. Besides Euro-Mediterranean regions, North Western and Central Eastern regions appear more drought prone than the rest of Europe, in terms of low values of return periods.     

The ion photon emission microscope on SNL’s nuclear microprobe and in LBNL’s cyclotron facility

Radiation effects microscopy (REM) has evolved into an essential tool for the study, diagnostics and remedy of single event effects (SEE) in microelectronics devices, However, we are entering an era where the ion energies of the current systems are becoming inadequate for diagnosing SEE problems in modern ICs due to the great thickness of interlevel dielectric, metallization and passivation layers found on top of the active radiation-sensitive Si. Our solution is the ion photon emission microscope (IPEM), which eliminates the need to focus several GeV heavy ions. A tabletop IPEM is currently in use at Sandia National Laboratories (SNL), operating with alpha particles, and showing 4 μm resolution. We have recently developed a second system, and installed it on one of the SNL nuclear microprobe lines to demonstrate the principle and prove its potential as a portable radiation effects microscope that can be installed at the LBNL GeV cyclotron facility. The microprobe system is currently operating with ～2 μm resolution. The determined advantages of installing a similar system at the LBNL cyclotron facility will be discussed, in addition to recently measured optical characteristics of the various phosphor materials being investigated.     

Surfactant-Protected Gold Particles: New Challenge for Gold-on-Carbon Catalysts

N-dodecil-N,N-dimethyl-3-amino-1-propan sulphonate was used for stabilizing gold nanoparticles in water that afforded Au/C (SB-Au/C) when immobilized on activated carbon. Characterization by XPS revealed a high atomic percentage of superficial gold (% at. Au(4f)/C(2s)), compared with other Au/C catalysts prepared by the immobilization of different sols. The activity of the SB-Au/C catalyst was found to be superior to that observed in the liquid-phase oxidation of ethylene glycol using other gold-on-carbon catalysts.     

N-n-Propyl-substituted 3-(dimethylphenyl)piperidines display novel discriminative properties between dopamine receptor subtypes: synthesis and receptor binding studies

3-Phenylpiperidines (PPEs) have been thoroughly investigated in view of their interesting dopaminergic activity, and the N-n-propyl substitution has been suggested as the most effective among several PPEs differently substituted on the phenyl ring. In previous studies, we found that the dimethyl substitution on the phenyl ring of N-unsubstituted PPEs provided compounds active toward alpha2-adrenergic receptors (alpha2-ARs), which proved to possess interesting selectivity properties. The high degree of homology between the binding domains of alpha2-ARs and D4-dopaminergic receptors (D4-DARs) prompted us to verify whether this kind of substitution on the aromatic ring might prove to be active against retinal DARs of the D4 subtype. On the basis of these premises, we synthesized the dimethylphenyl-substituted PPEs 4a-f, in which an n-propyl chain is present on the aminic nitrogen. Radioligand binding assays on bovine retina and striatum membranes for D1-like and D2-like DARs indicated that PPEs 4a, 4b, and 4f possess a high affinity and selectivity for the D4-DAR subtype of bovine retina.