Highly cited researchers: a moving target

Scientometrics - Highly cited researchers are a category of researchers defined by scientometric rules relating to counts of citations to their scholarly articles. The designation often refers to...     

Expression of genes encoding the E2 and E3 components of the Bacillus stearothermophilus pyruvate dehydrogenase complex and the stoichiometry of subunit interaction in assembly in vitro

Genes encoding the dihydrolipoyl acetyltransferase (E2) and dihydrolipoyl dehydrogenase (E3) components of the pyruvate dehydrogenase (PDH) multienzyme complex from Bacillus stearothermophilus were overexpressed in Escherichia coli. The E2 component was purified as a large soluble aggregate (molecular mass > 1 x 10(6) Da) with the characteristic 532 symmetry of an icosahedral (60-mer) structure, and the E3 as a homodimer with a molecular mass of 110 kDa. The recombinant E2 component in vitro was capable of binding either 60 E3(alpha2) dimers or 60 heterotetramers (alpha2beta2) of the pyruvate decarboxylase (E1) component (also the product of B. stearothermophilus genes overexpressed in E. coli). Assembling the E2 polypeptide chain into the icosahedral E2 core did not impose any restriction on the binding of E1 or E3 to the peripheral subunit-binding domain in each E2 chain. This has important consequences for the stoichiometry of the assembled complex in vivo. The lipoyl domain of the recombinant E2 protein was found to be unlipoylated, but it could be correctly post-translationally modified in vitro using a recombinant lipoate protein ligase from E. coli. The lipoylated E2 component was able to bind recombinant E1 and E3 components in vitro to generate a PDH complex with a catalytic activity comparable with that of the wild-type enzyme. Reversible unfolding of the recombinant E2 and E3 components in 6 M guanidine hydrochloride was possible in the absence of chaperonins, with recoveries of enzymic activities of 95% and 85%, respectively. However, only 26% of the E1 enzyme activity was recovered under the same conditions as a result of irreversible denaturation of both E1alpha and E1beta. This represents the first complete post-translational modification and assembly of a fully active PDH complex from recombinant proteins in vitro.     

Reversible or irreversible modification of [3H]PAF binding on rabbit platelet membranes differentiates various PAF receptor antagonists

[³H]Platelet-activating factor (PAF) binding to rabbit platelet membranes was examined before and after 20 min preincubation at 25°C in the presence of PAF, lysoPAF, or of five different PAF receptor antagonists (L 652731, BN 52021, WEB 2086, BN 52111 and BN 52115). When platelet membranes were not washed after preincubation with PAF or PAF antagonists, no significant specific binding of [³H]PAF was observed, which suggests full occupancy of the binding sites. When membranes were extensively washed, full recovery of specific [³H]PAF binding was attained with L 652731 and partial recoveries (60%, 55% and 30%) were reached with BN52021, WEB 2086 and PAF, respectively; no recovery was seen with the dioxolanes BN 52111 and BN 52115. Scatchard analysis of the binding data indicated that no significant change in the dissociation constant (K_d) and maximum number of binding sites (B_max) occurred after preincubation of platelet membrane with L 652731, whereas a reduction of B_max was observed when PAF and BN 52021 were measured. When platelet membranes were preincubated with WEB 2086, B_max and K_d significantly increased. The data suggest differing binding properties for PAF and the PAF antagonists. Some of the PAF antagonists may tightly bind to the PAF receptor site(s) and/or irreversibly modify or downregulate PAF recognition sites. Our results also suggest that the interaction of PAF receptor antagonists with PAF receptor can be divided into at least two components, namely a reversible component and an irreversible one. Based on a paper presented at the Third International Conference on Platelet-Activating Factor and Structurally Related Alkyl Ether Lipids, Tokyo, Japan, May 1989.     

Activity of Fluorine‐Containing Analogues of WC‐9 and Structurally Related Analogues against Two Intracellular Parasites: Trypanosoma cruzi and Toxoplasma gondii

Two obligate intracellular parasites, Trypanosoma cruzi, the agent of Chagas disease, and Toxoplasma gondii, an agent of toxoplasmosis, upregulate the mevalonate pathway of their host cells upon infection, which suggests that this host pathway could be a potential drug target. In this work, a number of compounds structurally related to WC‐9 (4‐phenoxyphenoxyethyl thiocyanate), a known squalene synthase inhibitor, were designed, synthesized, and evaluated for their effect on T. cruzi and T. gondii growth in tissue culture cells. Two fluorine‐containing derivatives, the 3‐(3‐fluorophenoxy)‐ and 3‐(4‐fluorophenoxy)phenoxyethyl thiocyanates, exhibited half‐maximal effective concentration (EC₅₀) values of 1.6 and 4.9 μm , respectively, against tachyzoites of T. gondii, whereas they showed similar potency to WC‐9 against intracellular T. cruzi (EC₅₀ values of 5.4 and 5.7 μm , respectively). In addition, 2‐[3‐ (phenoxy)phenoxyethylthio]ethyl‐1,1‐bisphosphonate, which is a hybrid inhibitor containing 3‐phenoxyphenoxy and bisphosphonate groups, has activity against T. gondii proliferation at sub‐micromolar levels (EC₅₀=0.7 μm ), which suggests a combined inhibitory effect of the two functional groups.     

Numerical Investigation of the Effect of Impeller Design Parameters on the Performance of a Multiphase Baffle‐Stirred Reactor

The turbulent gas‐liquid flow field in an industrial 100‐m³ stirred tank was calculated by using computational fluid dynamics based on the finite‐volume method. Turbulent effects were modeled with the shear stress transport model, and gas‐liquid bubbly flow was modeled with the Eulerian‐Eulerian approach using the Grace correlation for the drag force interphase momentum transfer. The relative motion between the rotating impeller and the stationary baffled tank was considered by using a multiple frames of reference algorithm. The effects of Rushton and pitched‐blade impeller design parameters such as blade geometry, location, and pumping direction on the mixing performance were investigated. It was found that a combination of Rushton turbines with up‐pumping pitched‐blade turbines provides the best mixing performance in terms of gas holdup and interfacial area density. The approach outlined in this work is useful for performance optimization of biotechnology reactors, as typically found in fermentation processes.     

Why Phenolic Acids Are Unlikely Primary Allelochemicals in Rice

Allelopathy in rice (Oryza sativa, L.) effective against weeds has been found in about 3.5% of tested rice germplasm in both laboratory and field experimentation. However, the allelochemicals responsible for growth inhibition of rice-associated weeds have not yet been identified. In the literature, phenolic acids are often mentioned as putative allelochemicals. If phenolic acids commonly reach growth inhibitory concentrations in rice ecosystems, it must be expected that the degree of tolerance to phenolic acids will vary among traditional rice cultivars or plant species adapted to rice environments having inherently different phenolic acid concentrations. Phenolic acids concentrations are normally greater in submerged than in aerobic soils. A dose–response study, however, showed that seedlings of rice cultivars adapted to submerged anaerobic soils did not have higher level of tolerance against p-hydroxybenzoic acid than did seedlings of varieties adapted to aerobic upland soils. Moreover, traditional rice cultivars had no greater tolerance than did improved cultivars that were recently bred for traits other than tolerance of phenolic acids. Similarly, there were no differences in tolerance of p-hydroxybenzoic acid between two Echinochloa weed species adapted to either anaerobic or aerobic growth conditions. Thus, neither the rice cultivars nor weed species had evolved different tolerance levels against the phenolic acid. However, all rice cultivars had significantly greater tolerance of p-hydroxybenzoic acid than did either weed species. In a second experiment, the rates at which rice plants released phenolic acids into solution cultures were measured for at least one month, the time period of greatest allelopathic activity following planting under field conditions. The maximum release rate of phenolic acids during the first month of growth was approximately 10 g/plant/day. At a conventional plant density, the release rate of phenolic acids would be approximately 1 mg/m²day. This order of release rate cannot provide concentrations remotely close to phytotoxic levels determined for these rice cultivars and weed species. The results presented in this paper do not preclude the possibility that phenolic acids might be one component in a mixture of chemicals that, when present simultaneously, are allelopathic.     

Synthesis of p-cymene from limonene, a renewable feedstock

Highly selective limonene conversions to p-cymene in short reaction times were achieved under “solvent free” conditions over mesoporous silica–alumina supports heated by microwave irradiation. An increase in the silica content of these mixed oxides led to increases in the specific surface area, porosity (pore size and volume) and the surface acidity. The conversion and selectivity of limonene to p-cymene under microwave irradiation was also found to rise as the silica content in these mixed oxides was increased. By careful choice of the solid and reaction parameters the activities for the conversion of limonene and selectivity to p-cymene (used as an intermediate in fine chemical syntheses) could be fine tuned. Results are presented under both dry media and reflux conditions.     

Characterization of precipitates formed in X7R 0603 BME-MLCC during sintering

In the current study two different batches of X7R-0603 BME-MLCCs displayed dissimilar electrical performance, despite having the same chemical composition, tape casting, and sintering conditions; with the only difference between them being the ore deposits where the raw materials were extracted from to synthesize the BaTiO₃. Specifically, they presented different electrical response to highly accelerated life tests (HALT). Although the chemical analysis of each slip showed the same composition, the trace elements of the BaTiO₃ sources could have acted as dopants or produced different secondary phases. A search for precipitates in the two samples was conducted by means of Scanning (SEM) and Transmission Electron Microscopy (TEM) techniques. SEM observations confirmed the presence of precipitates formed within the structure of the MLCCs exhibiting the greatest decrement in their electrical resistance results during the HALT. In order to further characterize the observed precipitates, samples were prepared by Focused Ion Beam (FIB) lift-out method, to make TEM characterization of specific precipitates feasible. TEM studies were performed on the precipitates to obtain electron diffraction patterns and complementary Energy Dispersive X-Ray Spectroscopy (EDXS) chemical analysis. Based on the crystal and chemical data obtained, it can be concluded that the precipitates are a hexagonal anhydrous silicate oxyapatite phase with a stoichiometry of Ca₃Y₁₆Si₁₀O₁₃, and lattice parameters of a = 0.9353 nm and c = 0.6970 nm; this phase was not found in the JCPDS data base. Differences in raw materials coming from different ore deposits can produce undesired precipitates that affect the electrical performance of MLCCs.