Altered white matter connectivity associated with visual hallucinations following occipital stroke

Introduction: Visual hallucinations that arise following vision loss stem from aberrant functional activity in visual cortices and an imbalance of activity across associated cortical and subcortical networks subsequent to visual pathway damage. We sought to determine if structural changes in white matter connectivity play a role in cases of chronic visual hallucinations associated with visual cortical damage. Methods: We performed diffusion tensor imaging (DTI) and probabilistic fiber tractography to assess white matter connectivity in a patient suffering from continuous and disruptive phosphene (simple) visual hallucinations for more than 2 years following right occipital stroke. We compared these data to that of healthy age-matched controls. Results: Probabilistic tractography to reconstruct white matter tracts suggests regeneration of terminal fibers of the ipsilesional optic radiations in the patient. However, arrangement of the converse reconstruction of these tracts, which were seeded from the ipsilesional visual cortex to the intrahemispheric lateral geniculate body, remained disrupted. We further observed compromised structural characteristics, and changes in diffusion (measured using diffusion tensor indices) of white matter tracts in the patient connecting the visual cortex with frontal and temporal regions, and also in interhemispheric connectivity between visual cortices. Conclusions: Cortical remapping and the disruption of communication between visual cortices and remote regions are consistent with our previous functional magnetic resonance imaging (fMRI) data showing imbalanced functional activity of the same regions in this patient (Rafique et al, 2016, Neurology, 87, 1493–1500). Long-term adaptive and disruptive changes in white matter connectivity may account for the rare nature of cases presenting with chronic and continuous visual hallucinations.York University Librarie

Altering Enzymatic Activity: Recruitment of Carboxypeptidase Activity into an RTEM β-Lactamase/Penicillin-Binding Protein 5 Chimera

The D-Ala-D-Ala carboxypeptidases/transpeptidases (penicillin-binding proteins, PBPs) share considerable structural homology with class A β-lactamases (EC 3.5.2.6), although these β-lactamases have no observable D-Ala-D-Ala carboxypeptidase activity. With the objective of recruiting such activity into a β-lactamase background, we have prepared a chimeric protein by inserting a 28-amino acid segment of PBP-5 of Escherichia coli in place of the corresponding region of the RTEM-1 β-lactamase. The segment thus inserted encompasses two residues conserved in both families: Ser-70, which forms the acyl-enzyme intermediate during β-lactam hydrolysis, and Lys-73, whose presence has been shown to be necessary for catalysis. This chimera involves changes of 18 residues and gives a protein that differs at 7% of the residues from the parent. Whereas RTEM β-lactamase has no D-Ala-D-Ala carboxypeptidase activity, that of the chimera is significant and is, in fact, about 1% the activity of PBP-5 on diacetyl-L-Lys-D-Ala-D-Ala; in terms of free energy of activation, the chimera stabilizes the transition state for the reaction to within about 2.7 kcal/mol of the stabilization achieved by PBP-5. Furthermore, the chimera catalyzes hydrolysis exclusively at the carboxyl-terminal amide bond which is the site of cleavage by D-Ala-D-Ala carboxypeptidase. Though containing all those residues that are conserved throughout class A β-Lactamases and are thought to be essential for β-lactamase activity, the chimera has considerably reduced activity ({approx} 10^-5) on penams such as penicillins and ampicillins as substrates. As a catalyst, the chimera shows an induction period of {approx} 30 min, reflecting a slow conformational rearrangement from an inactive precursor to the active enzyme

Ionization behavior of the histidine residue in the catalytic triad of serine proteases

α-Lytic protease is a homologue of the mammalian serine proteases such as trypsin, chymotrypsin, and elastase, and its single histidine residue belongs to the Asp-His-Ser catalytic triad. This single histidine residue has been selectively enriched in the C-2 carbon with 13C. Magnetic resonance studies of the chemical shift and coupling constant (1Jch) behavior of this nucleus as a function of pH suggest that the imidazole ring is neutral above pH 5 and therefore that the group which is known to ionize with pKa near 6.7 must be the aspartic acid residue. Implications of these new pKa assignments for the catalytic mechanism of serine proteases are discussed and include the absence of any need to separate charge during catalysis. The histidine residue plays two roles. (a) It insulates the aspartic acid from an aqueous environment and accordingly raises its pKa. (b) It serves as a bidentate base to accept a proton from the serine at one of its nitrogens and concertedly transfer a proton from its other nitrogen to the buried carboxylate anion during formation of the tetrahedral intermediate

High-Potential C112D/M121X (X = M, E, H, L) Pseudomonas aeruginosa Azurins

Site-directed mutagenesis of Pseudomonas aeruginosa azurin C112D at the M121 position has afforded a series of proteins with elevated Cu^(II/I) reduction potentials relative to the CuII aquo ion. The high potential and low axial hyperfine splitting (Cu^(II) electron paramagnetic resonance A|) of the C112D/M121L protein are remarkably similar to features normally associated with type 1 copper centers

Effect of an agri-environmental measure on nitrate leaching from a beef farming system in Ireland

peer-reviewedAgricultural nitrogen (N) management remains a key environmental challenge. Improving N management is a matter of urgency to reduce the serious ecological consequences of the reactive N. Nitrate (NO3−–N) leaching was measured under suckler beef production systems stocked at two intensities: (1) intensive, 210 kg organic N ha−1 with two cut silage harvests; and (2) rural environmental protection scheme (REPS), 170 kg organic N ha−1 with one cut silage harvest. Three replicate plots of each treatment were instrumented with ceramic cups (8 per plot), randomly placed within each plot at a depth of 1 m to collect soil solution for NO3−–N at 50 kPa suction to collecting vessels one week prior to sampling. Samples were taken on a total of 53 sampling dates over 3 winter drainage periods (2002/03, 2003/04 and 2004/05). Over the course of the experiment the mean annual soil solution NO3−–N concentration exceeded the MAC twice out of 15 means (5 treatments over 3 years). The REPS grazing and silage sub treatments had significantly lower mean annual soil solution total oxidized N (TON) concentrations than the respective intensive treatments in years 2 and 3. Annual total NO3−–N losses over the three years in intensive and REPS systems ranged from 55 to 71 and 15 to 20 kg N ha−1, respectively. Mean N surpluses in intensive and REPS systems were 210 and 95 kg ha−1, respectively with the corresponding mean N inputs of 272 and 124 kg N ha−1. The reduction in N inputs under the REPS system results in lower N leaching losses and contributed to a significant reduction in pressures on water quality