Combining independent component analysis and correlation analysis to probe interregional connectivity in fMRI task activation datasets

A new approach in studying interregional functional connectivity using functional magnetic resonance imaging (fMRI) is presented. Functional connectivity may be detected by means of cross correlating time course data from functionally related brain regions. These data exhibit high temporal coherence of low frequency fluctuations due to synchronized blood flow changes. In the past, this fMRI technique for studying functional connectivity has been applied to subjects that performed no prescribed task ("resting" state). This paper presents the results of applying the same method to task-related activation datasets. Functional connectivity analysis is first performed in areas not involved with the task. Then a method is devised to remove the effects of activation from the data using independent component analysis (ICA) and functional connectivity analysis is repeated. Functional connectivity, which is demonstrated in the "resting brain," is not affected by tasks which activate unrelated brain regions. In addition, ICA effectively removes activation from the data and may allow us to study functional connectivity even in the activated regions.     

The mononuclear metal center of type-I dihydroorotase from aquifex aeolicus

Background

Dihydroorotase (DHO) is a zinc metalloenzyme, although the number of active site zinc ions has been controversial. E. coli DHO was initially thought to have a mononuclear metal center, but the subsequent X-ray structure clearly showed two zinc ions, α and β, at the catalytic site. Aquifex aeolicus DHO, is a dodecamer comprised of six DHO and six aspartate transcarbamoylase (ATC) subunits. The isolated DHO monomer, which lacks catalytic activity, has an intact α-site and conserved β-site ligands, but the geometry of the second metal binding site is completely disrupted. However, the putative β-site is restored when the complex with ATC is formed and DHO activity is regained. Nevertheless, the X-ray structure of the complex revealed a single zinc ion at the active site. The structure of DHO from the pathogenic organism, S. aureus showed that it also has a single active site metal ion.

Results

Zinc analysis showed that the enzyme has one zinc/DHO subunit and the addition of excess metal ion did not stimulate catalytic activity, nor alter the kinetic parameters. The metal free apoenzyme was inactive, but the full activity was restored upon the addition of one equivalent of Zn²⁺ or Co²⁺. Moreover, deletion of the β-site by replacing the His180 and His232 with alanine had no effect on catalysis in the presence or absence of excess zinc. The 2.2 Å structure of the double mutant confirmed that the β-site was eliminated but that the active site remained otherwise intact.

Conclusions

Thus, kinetically competent A. aeolicus DHO has a mononuclear metal center. In contrast, elimination of the putative second metal binding site in amidohydrolyases with a binuclear metal center, resulted in the abolition of catalytic activity. The number of active site metal ions may be a consideration in the design of inhibitors that selectively target either the mononuclear or binuclear enzymes.

     

Helices versus zigzag chains: one-dimensional coordination polymers of Ag(I) and Bis(4-pyridyl)amine

Five one-dimensional coordination polymers were prepared by the reaction of a bent bridging ligand, bis(4-pyridyl)amine (bpa), with an extensive series of AgX salts (X = CF3SO3, PF6, ClO4, NO3). The 1D polymer networks formed with AgCF3SO3 (1), AgPF6 (2.MeCN), and AgClO4 (3.2MeCN) all incorporated MeCN and were found to adopt a zigzag arrangement. The networks formed with AgClO4 (4) and AgNO3 (5) did not contain any solvent and adopted a single-stranded helical arrangement. Two-dimensional H-bonding networks were formed for 1 and 3.2MeCN, with network topologies 4.8(2) and (4, 4), respectively, whereas three-dimensional H-bonded networks of helices were formed for 4, showing an (8, 3)-a network topology, and 5, showing the topology of the alpha-polonium net. The three-dimensional networks both exhibited 4-fold interpenetration. The NO3- anion in 5 appeared to be acting as a template for the 3D structure.