Volition and conflict in human medial frontal cortex

Controversy surrounds the role of human medial frontal cortex in controlling actions. Although damage to this area leads to severe difficulties in spontaneously initiating actions, the precise mechanisms underlying such "volitional" deficits remain to be established. Previous studies have implicated the medial frontal cortex in conflict monitoring and the control of voluntary action, suggesting that these key processes are functionally related or share neural substrates. Here, we combine a novel behavioral paradigm with functional imaging of the oculomotor system to reveal, for the first time, a functional subdivision of the pre-supplementary motor area (pre-SMA) into anatomically distinct areas that respond exclusively to either volition or conflict. We also demonstrate that activity in the supplementary eye field (SEF) distinguishes between success and failure in changing voluntary action plans during conflict, suggesting a role for the SEF in implementing the resolution of conflicting actions. We propose a functional architecture of human medial frontal cortex that incorporates the generation of action plans and the resolution of conflict.     

Plk3 functionally links DNA damage to cell cycle arrest and apoptosis at least in part via the p53 pathway

Polo-like kinase 3 (Plk3, previously termed Prk) contributes to regulation of M phase of the cell cycle (Ouyang, B., Pan, H., Lu, L., Li, J., Stambrook, P., Li, B., and Dai, W. (1997) J. Biol. Chem. 272, 28646-28651). Plk3 physically interacts with Cdc25C and phosphorylates this protein phosphatase predominantly on serine 216 (Ouyang, B., Li, W., Pan, H., Meadows, J., Hoffmann, I., and Dai, W. (1999) Oncogene 18, 6029-6036), suggesting that the role of Plk3 in mitosis is mediated, at least in part, through direct regulation of Cdc25C. Here we show that ectopic expression of a kinase-active Plk3 (Plk3-A) induced apoptosis. In response to DNA damage, the kinase activity of Plk3 was rapidly increased in an ATM-dependent manner, whereas that of Plk1 was markedly inhibited. Recombinant Plk3 phosphorylated in vitro a glutathione S-transferase fusion protein containing p53, but not glutathione S-transferase alone. Recombinant Plk1 also phosphorylated p53 but on residues that differed from those targeted by Plk3. Co-immunoprecipitation and pull-down assays demonstrated that Plk3 physically interacted with p53 and that this interaction was enhanced upon DNA damage. In vitro kinase assays followed by immunoblotting showed that serine 20 of p53 was a target of Plk3. Furthermore, expression of a kinase-defective Plk3 mutant (Plk3(K52R)) resulted in significant reduction of p53 phosphorylation on serine 20, which was correlated with a decrease in the expression of p21 and with a concomitant increase in cell proliferation. These results strongly suggest that Plk3 functionally links DNA damage to cell cycle arrest and apoptosis via the p53 pathway.     

Redox properties of the quinoprotein methylamine dehydrogenase from paracoccus denitrificans

Paracoccus denitrificans synthesizes a methylamine dehydrogenase that contains a covalently bound form of pyrroloquinoline quinone as a prosthetic group [Husain, M., & Davison, V.L. (1987) J. Bacteriol. 169, 1712-1717]. Anaerobic reductive titration of this enzyme with dithionite proceeded through a semiquinone intermediate with spectral properties quite distinct from those of the oxidized and reduced species. From these data the molar extinction coefficients were calculated at various wavelengths for the three redox states of this enzyme. The semiquinone was slowly reoxidized under aerobic conditions. The fully reduced enzyme was stable in the presence of oxygen and slowly reoxidized by ferricyanide. Reductive titration of methylamine dehydrogenase with methylamine proceeded directly to the fully reduced form of the enzyme without detectable formation of the semiquinone. Electrochemical titrations of the enzyme yielded an overall midpoint potential value for the two-electron couple (fully oxidized/fully reduced) of 100 +/- 4 mV and an n value of 2.15 +/- 0.15.     

Physiological characteristics of four methylotrophic bacteria and their potential use in single-cell protein production

Summary Four methylotrophic bacteria, isolated at the Kuwait Institute for Scientific Research, were able to grow on methanol as the only carbon source at a maximum temperature of 44°C. An optimized medium composition was obtained through intensive chemostat studies varying both macro- and micro-element concentrations. Various batch and chemostat experiments were carried out at different pH, temperature, dilution rate and methanol concentrations. The results showed optimum pH around 6.8, at temperatures of 37 to 40°C, dilution rate 0.2–0.3 h^–1 and methanol was found to be inhibitory at concentrations above 20 g l^–1. The performance of all four bacteria under chemostat conditions was similar. Chemostat fermentation experiments using the optimized medium at 40°C, pH 6.8, dilution 0.2 h^–1 and 10 g methanol/l in the feed gave a biomass yield coefficient of 0.42–0.44 g/g methanol, 78–79% crude protein content, 58–62% total amino acid content and 10–11.5% nucleic acid content. In conclusion all four methylotrophic strains has good potential for use in the production of single-cell protein.

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Caractéristiques physiologiques de quatre bactéries méthylotrophes et leur emploi potentiel dans la production de protéine uni-celluliare

Résumé Quatre bactéries méthylotrophes, isolées à l'Institut pour la Recherche Scientifique du Kuweit, se sont révélées capables de croître sur méthanol comme seule source de carbone à une température maximum de 44°C. Des études intensives en chémostat de variation des concentrations tant en macro-qu'en micro-éléments, ont permis d'obtenir une composition optimum du milieu. Diverses expériences en milieu non renouvelé et en chémostat ont été effectuées à différents pH, températures, taux de dilution et concentration en méthanol. Les résultats ont montré un optimum de pH vers 6.8, de température entre 37 et 40°C, de taux de dilution de 0.2 à 0.3 h^–1. Le méthanol s'est révélé inhibiteur aux concentrations supérieures à 20 g l^–1. La performance des quatres bactéries dans les conditions du chémostat se sont révélées très semblables. Les expériences de fermentátion en chémostat, utilisant le milieu optimum à 40°C, pH 6.8, taux de dilution de 0.2 h^–1 et à 10 g de méthanol par litre dans l'alimentation ont produit un coefficient de rendement en biomasse de 0.42 à 0.44 g de cellules par g de méthanol contenant de 78 à 79% de protéines, de 58 à 62% d'acides aminés totaux et de 10 à 11.5% d'acides nucléiques. En conclusion, les quatre bactéries méthylotrophes présentent une bonne potentialité pour être utilisées dans la production de protéine uni-cellulaire.

     

Rapid In Vitro Propagation of Eclipta alba (L) Hassk by Shoot Tip Culture

An efficient and rapid tissue culture system employing shoot tip explants has been developed for Eclipta alba (L) Hassk, an important medicinal plant of the family Asteraceae. The highest shoot regeneration frequency (95%) as well as the maximum number (32.2 ± 0.4) of shoots was recorded on MS medium amended with BA (5 μM) and NAA (0.5 μM). The regenerated shoots rooted best on MS medium supplemented with 0.2 μM IBA. The in vitro developed plantlets were acclimatized successfully with 100 % survival.     

Cooperative role of endogenous leucotrienes and platelet‐activating factor in ischaemia–reperfusion‐mediated tissue injury

Insufficient oxygen delivery to organs leads to tissue dysfunction and cell death. Reperfusion, although vital to organ survival, initiates an inflammatory response that may both aggravate local tissue injury and elicit remote organ damage. Polymorphonuclear neutrophil (PMN) trafficking to remote organs following ischaemia/reperfusion (I/R) is associated with the release of lipid mediators, including leucotriene (LT) B₄, cysteinyl‐LTs (CysLTs) and platelet‐activating factor (PAF). Yet, their potentially cooperative role in regulating I/R‐mediated inflammation has not been thoroughly assessed. The present study aimed to determine the cooperative role of lipid mediators in regulating PMN migration, tissue oedema and injury using selective receptor antagonists in selected models of I/R and dermal inflammation. Our results show that rabbits, pre‐treated orally with BIIL 284 and/or WEB 2086 and MK‐0571, were protected from remote tissue injury following I/R or dermal inflammation in an additive or synergistic manner when the animals were pre‐treated with two drugs concomitantly. The functional selectivity of the antagonists towards their respective agonists was assessed in vitro, showing that neither BIIL 284 nor WEB 2086 prevented the inflammatory response to IL‐8, C5a and zymosan‐activated plasma stimulation. However, these agonists elicited LTB₄ biosynthesis in isolated rabbit PMNs. Similarly, a cardioprotective effect of PAF and LTB₄ receptor antagonists was shown following myocardial I/R in mice. Taken together, these results underscore the intricate involvement of LTB₄ and PAF in each other's responses and provide further evidence that targeting both LTs and PAF receptors provides a much stronger anti‐inflammatory effect, regulating PMN migration and oedema formation.     

Atrial fibrillation pacing decreases intravascular shear stress in a New Zealand white rabbit model: implications in endothelial function

Atrial fibrillation (AF) is characterized by multiple rapid and irregular atrial depolarization, leading to rapid ventricular responses exceeding 100 beats per minute (bpm). We hypothesized that rapid and irregular pacing reduced intravascular shear stress (ISS) with implication to modulating endothelial responses. To simulate AF, we paced the left atrial appendage of New Zealand White rabbits (n = 4) at rapid and irregular intervals. Surface electrical cardiograms were recorded for atrial and ventricular rhythm, and intravascular convective heat transfer was measured by microthermal sensors, from which ISS was inferred. Rapid and irregular pacing decreased arterial systolic and diastolic pressures (baseline, 99/75 mmHg; rapid regular pacing, 92/73; rapid irregular pacing, 90/68; p < 0.001, n = 4), temporal gradients ( ${\partial\tau/\partial t}$ from 1,275 ± 80 to 1,056 ± 180 dyne/cm² s), and reduced ISS (from baseline at 32.0 ± 2.4 to 22.7 ± 3.5 dyne/cm²). Computational fluid dynamics code demonstrated that experimentally inferred ISS provided a close approximation to the computed wall shear stress at a given catheter to vessel diameter ratio, shear stress range, and catheter position. In an in vitro flow system in which time-averaged shear stress was maintained at ${{\tau_{\rm avg}} = 23 \pm 4\, {\rm dyn}\, {\rm cm}^{-2} {\rm s}^{-1}}$ , we further demonstrated that rapid pulse rates at 150 bpm down-regulated endothelial nitric oxide, promoted superoxide (O ₂ ^.? ) production, and increased monocyte binding to endothelial cells. These findings suggest that rapid pacing reduces ISS and ${\partial\tau/\partial t}$ , and rapid pulse rates modulate endothelial responses.