In Silico Investigation of Potential Applications of Gamma Carbonic Anhydrases as Catalysts of CO2 Biomineralization Processes: A Visit to the Thermophilic Bacteria Persephonella hydrogeniphila,Persephonella marina,Thermosulfidibacter takaii,and Thermus thermophilus

Carbonic anhydrases (CAs) have been identified as ideal catalysts for CO₂ sequestration. Here, we report the sequence and structural analyses as well as the molecular dynamics (MD) simulations of four γ-CAs from thermophilic bacteria. Three of these, Persephonella marina, Persephonella hydrogeniphila, and Thermosulfidibacter takaii originate from hydrothermal vents and one, Thermus thermophilus HB8, from hot springs. Protein sequences were retrieved and aligned with previously characterized γ-CAs, revealing differences in the catalytic pocket residues. Further analysis of the structures following homology modeling revealed a hydrophobic patch in the catalytic pocket, presumed important for CO₂ binding. Monitoring of proton shuttling residue His69 (P. marina γ-CA numbering) during MD simulations of P. hydrogeniphila and P. marina’s γ-CAs (γ-PhCA and γ-PmCA), showed a different behavior to that observed in the γ-CA of Escherichia coli, which periodically coordinates Zn²⁺. This work also involved the search for hotspot residues that contribute to interface stability. Some of these residues were further identified as key in protein communication via betweenness centrality metric of dynamic residue network analysis. T. takaii’s γ-CA showed marginally lower thermostability compared to the other three γ-CA proteins with an increase in conformations visited at high temperatures being observed. Hydrogen bond analysis revealed important interactions, some unique and others common in all γ-CAs, which contribute to interface formation and thermostability. The seemingly thermostable γ-CA from T. thermophilus strangely showed increased unsynchronized residue motions at 423 K. γ-PhCA and γ-PmCA were, however, preliminarily considered suitable as prospective thermostable CO₂ sequestration agents.     

Heterogeneity in Youth Symptom Trajectories Following Psychiatric Crisis: Predictors and Placement Outcome.

The authors examined heterogeneity in symptom trajectories among youths following psychiatric crises as well as the psychosocial correlates and placement outcomes associated with identified trajectories. Using semiparametric mixture modeling with 156 youths approved for psychiatric hospitalization, the authors identified 5 trajectories based on symptoms over the 16 months following crisis: high improved, high unimproved, borderline improved, borderline unimproved, and subclinical. Membership in unimproved symptom groups was associated with less suicidality, younger age, more youth hopelessness, and more caregiver empowerment. Improved symptom group membership predicted long-term decreases in days in out-of-home placements. More important, and in contrast with general impressions from the existing literature, findings suggest that a substantive proportion of youths with serious emotional disturbance sustain high levels of symptomatology following intensive mental health services. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The likelihood of violent behaviour: Predictions, postdictions, and hindsight bias.

Studied the effects of variations in base rate (BR) information and temporal context on predictions of violence and hindsight bias (HB). 270 undergraduates estimated the likelihood of violent offenders committing another violent offense based upon fictitious case history information. Ss were given 1 of 2 population BRs of the likelihood of reoffending (30% or 70%) and a recidivism estimation question that was phrased either predictively, predictively with a given outcome of either recidivism or no recidivism, or postdictively. Although different offenders were perceived as differentially likely to reoffend, estimated likelihood of recidivism was affected by neither BR nor temporal phrasing of the estimation task. A 2nd study of 182 undergraduates, using shorter case histories, also found no BR effect. HB effects were weak and inconsistent in both studies. (French abstract) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Stress Superposition in High-Strength Glass

The introduction of surface compression via alkali-ion exchange is frequently used to strengthen alkali-containing glass. However, previous investigators found that when starting with high-strength glass, the strength measured after exchange was significantly less than the sum of the residual compression and the starting fracture strength. Results are presented which suggest that the source of this discrepancy is surface damage produced during the heat treatment accompanying the ion-exchange process. Further, when the surface damage was removed by shallow hydrofluoric acid etching, the resulting fracture strength did approximately equal the sum of the residual stress and the high starting strength.     

Rapid determination of double bond configuration and position along the hydrocarbon chain in cyclic fatty acid monomers

This study reports the structural elucidation of diunsaturated 5- or 6-membered ring cyclic fatty acid monomers (CFAM) isolated from heated flaxseed oil by complementary gas chromatography (GC)-mass spectrometry (MS) and GC-matrix isolation-Fourier transform infrared spectroscopy (MI-FTIR). Infrared measurements of CFAM were carried out on methyl ester derivatives as well-resolved chromatograms were obtained on a polar 100% cyanopropyl polysiloxane capillary GC column. By contrast, electron ionization MS of methyl ester derivatives was of limited value because of double bond migration during the ionization process in the mass spectrometer. This communication reports definitive MS fragmentation patterns that can confirm ring position and double bond position along the fatty acid chain in 1,2-disubstituted CFAM determined as 2-alkenyl-4,4-dimethyloxazoline derivatives. Double bond configuration (cis, trans, or conjugatedcis,cis) in CFAM was confirmed by GC-MI-FTIR. The presence of CFAM, degradation products found in used frying oils, is a potential source of dietary toxicity. This work was presented in part at the 84th American Oil Chemists' Society Annual Meeting held in Anaheim, CA, in April 1993.     

Over-production of 5-enolpyruvylshikimate-3-phosphate synthase in Escherichia coli: use of the T7 promoter

5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, the productof the Escherichia coli aroA gene, has been overproduced inE.coli BL21(DE3) under the control of the T7 gene 10 promoterand ribosome binding site, to a level of {small tilde}50% oftotal cell protein. EPSP synthase is the primary target of thepost-emergence herbicide, glyphosate, commonly known as Roundup^TM.A simple two step purification is described, which results in99% pure homogeneous protein (as determined by PAGE). The integrityof the protein has been compared with previously characterizedprotein from .E.coli AB2829(pKD501) by determination of itskinetic parameters, N-terminal protein and DNA sequences, aminoacid analysis and ¹³C-NMR spectroscopy. This new overproducingstrain readily provides the gram quantities of highly pure proteinrequired for NMR studies of the active site and the developmentof novel time-resolved solid-state NMR techniques currentlyunderway in this laboratory.     

Defensive steroids from a carrion beetle (Silpha novaboracensis)

Two novel steroids, 3,7-dihydroxy-14-pregn-4-en-15,20-dione (1) and 3,7, 20-trihydroxy-14-pregn-4-en-15-one (4), have been characterized from the rectal gland ofSilpha novaboracensis (Coleoptera: Silphidae). Judging from the function of comparable pregnanes in another species ofSilpha, the compounds may play an antipredator role.Paper No. 81 of the seriesDefense Mechanisms of Arthropods. Paper No. 80 is Eisner, T., Deyrup, M., Jacobs, R., and Meinwald, J.,J. Chem. Ecol. 12:1407–1415.     

Torrefied agro-industrial residue as filler in natural rubber compounds

This study explored the feasibility of using torrefied biomass as a reinforcing filler in natural rubber compounds. Carbon black was then replaced with the torrefied biomass in elastomer formulations for concentrations varying from 0% to 100% (60 parts per hundred rubber or phr total). Their influence on the curing process, dynamic properties, and mechanical properties was investigated. Results were compared with the properties of vulcanizates containing solely carbon black fillers. Time to cure (t₉₀) for compounds with torrefied biomass fillers increased, while filler-filler interactions (ΔG') decreased, compared to carbon black controls. At low strains, the tan δ values of the torrefied fillers vulcanizates were similar to the controls. Incorporation of torrefied biomass into natural rubber decreased compound tensile strength and modulus but increased elongation. Replacement with torrefied fillers resulted in a weaker filler network in the matrix. Still, results showed that moderate substitution concentrations (~20 phr) could be feasible for some natural rubber applications.