Metabolic mapping of the effects of oral alcohol self-administration in rats

A method for the quantitation of protein carbonyls, which have been widely employed as markers of protein oxidative damage, is described. Protein carbonyls were derivatized with tritiated sodium borohydride and the tritiated proteins were separated on SDS-PAGE. Protein bands, visualized by Coomassie blue staining, were then excised and incubated in 30% H2O2 at 60 degrees C for 48 h. Tritium, incorporated into the proteins, was quantitated by liquid scintillation counting after gel solubilization by H2O2. This method can be applied to the measurement of carbonylation of specific proteins as it employs SDS-PAGE and has the advantage that unreacted NaB3H4 in the labeling reaction mixture need not be removed. The present method, when combined with immunochemical detection of protein carbonyls, should be very useful in the quantitation of oxidative damage to individual proteins.     

Über die Verwendung von Glyoxal-bis(tert.-butylnitron) als Spin-Trap

On the Use of Glyoxal Bis (t-Butyl Nitron) as a Spin Trap Nitroxides derived from glyoxal bis(t-butyl nitrone), GBBN , have been studied by e.s.r. spectroscopy. Well resolved e.s.r. spectra exhibiting extra hyperfine splitting by magnetic active nuclei in γ-position have been recorded for the spin adducts of alkyl, as well as ^.N₃ and ^.CH₂Cl radicals. Enhanced stability was found for the spin adducts of halogen atoms. However, a wider application of this spin trap is limited by the strong superposition of the e.s.r. signals of different radical spin adducts which is directly related to almost identical conformations of these nitroxides.     

The fate of nitrogen in agroecosystems: An illustration using Canadian estimates

Agroecosystems rely on inputs of nitrogen (N) to sustain productivity. But added N can leak into adjacent environments, affecting the health of other ecosystems and their inhabitants. Worries about global warming have cast further attention on the N cycle in farmlands because farms are a main source of N₂O, and because carbon sequestration, proposed to help reduce CO₂ loads, requires a build-up of N. Our objective was to estimate, as an illustrative example, the net N balance of Canadian agroecosystems in 1996 and then infer some hypotheses about the routes of N loss, their magnitude, and ways of reducing them. We defined agroecosystems as all agricultural lands in Canada including soil to 1 m depth and all biota, except humans. Only net flows of N across those boundaries were counted in our balance – all others represent internal cycling. Based on our estimates, about 2.35 Tg N entered Canadian agroecosystems from biological fixation, fertilizers, and atmospheric deposition (excluding re-deposited NH₃). In the same year, about 1.03 Tg N were exported in crop products and 0.19 Tg were exported in animals and animal products. Consequently, N inputs exceed exports in products by about 1.13 Tg, a surplus that is either accumulating in agroecosystems or lost to the environment. Because potential soil organic matter gains can account for only a small part of the surplus N, most is probably lost to air or groundwater. Our finding, that N losses amount to almost half of N added, concurs with field experiments that show crop recovery of added N in a given year is often not more than 60%. Better management may reduce the fraction lost somewhat but, because N in ecosystems eventually cycles back to N₂, substantive gains in efficiency may not come easily. As well as trying to reduce losses, research might also focus on steering losses directly to N₂, away from more harmful intermediates. If some of the `missing N' can be assimilated into organic matter, agricultural soils in Canada may need little added N to achieve C sequestration targets.     

Albert Renger—Patzsch's early work: Object and abstraction

Abstract

In photography, one should surely proceed from the essence of the object and attempt to represent it through purely photographic means, regardless of whether it is a human being, a landscape, architecture or something else; whereas today a rape of the object for the sake of formal play is more often the norm. Because the German word Sachlichkeit has taken on practically the opposite meaning today, I have to use a foreign word to properly characterize the position of servitude I maintain before the motif: ‘objectivity’.¹     

Membrane topology of the Rickettsia prowazekii ATP/ADP translocase revealed by novel dual pho-lac reporters

Here, we report the construction and characterization of dual reporters, consisting of both an Escherichia coli alkaline phosphatase (AP) gene and an alpha-fragment of the beta-galactosidase (BG) gene, for studying membrane protein topology by the gene fusion approach. Each of the reporters, when fused to periplasmic domains of polytopic proteins, produces fusions with high AP activity and, when fused to cytoplasmic domains, produces fusions with high BG activity in E. coli strains capable of alpha-complementation. The dual nature of these reporters simplifies interpretation of data obtained with poorly expressed fusions and allows one to evaluate the reliability of topological data. Deleterious effects resulting from the cell's attempt to export the full-length BG are eliminated in this approach. We describe dual indicator plates that allow for discrimination between colonies bearing cytoplasmic fusions, periplasmic fusions, and no fusions. We have generated a set of fusions to the topologically well-studied lactose permease of E. coli and demonstrated that topological information generated by these new reporters is in good agreement with the existing model. We used this new methodology for the determination of membrane topology of the Rickettsia prowazekii ATP/ADP translocase (Tlc). Our results were in agreement with the proposed in silico topological model in which Tlc traverses the cytoplasmic membrane of E. coli 12 times with its N and C termini facing the cytoplasm.     

Human Motion Planning Based on Recursive Dynamics and Optimal Control Techniques

This paper presents an efficient optimal control and recursive dynamics-based computer animation system for simulating and controlling themotion of articulated figures. A quasi-Newton nonlinear programmingtechnique (super-linear convergence) is implemented to solve minimumtorque-based human motion-planning problems. The explicit analyticalgradients needed in the dynamics are derived using a matrix exponentialformulation and Lie algebra. Cubic spline functions are used to make thesearch space for an optimal solution finite. Based on our formulations,our method is well conditioned and robust, in addition to beingcomputationally efficient. To better illustrate the efficiency of ourmethod, we present results of natural looking and physically correcthuman motions for a variety of human motion tasks involving open andclosed loop kinematic chains.     

Contribution of phospholipase C-beta3 phosphorylation to the rapid attenuation of opioid-activated phosphoinositide response

Our objective was: (1) to determine the appropriate dose of new ultrasmall superparamagnetic iron oxide particles for magnetic resonance angiography (MRA). This agent comprised of a single iron oxide crystal stabilized with a carbohydrate-polyethylene glycol coat (PEG-Ferron/NC 100150 injection); (2) to determine the proper flip angle for PEG-Ferron-enhanced 3 D time-of-flight (TOF) MRA sequence; and (3) to compare the enhancement of peripheral vessels following PEG-Ferron and GdDTPA-BMA. MRA parameters were: TR/TE = 50/2.1 ms, NEX = 1, FOV = 30 x 30 x 1.8 cm, and matrix = 256 x 128 x 64. In anesthetized beagle dogs (n = 10), the effects of PEG-Ferron and GdDTPA-BMA on regional signal were monitored for 45 min and compared. The lowest dose of PEG-Ferron (0.05 mmol/kg) produced the best enhancement of primary, secondary and tertiary vessels. The flip angle 60 degrees provided better enhancement than 20 degrees on contrast enhanced images. Unlike GdDTPA-BMA, PEG-Ferron allowed prolonged delineation (> 45 min) of the pelvis and lower extremities circulation. PEG-Ferron provided greater Contrast-to-noise ratio CNR (80.2 +/- 6.2, P < 0.05) than GdDTPA-BMA (63.5 +/- 2.5). It may be possible for blood pool contrast-enhanced 3 D TOF MRA to provide valuable information for visualization of vascular tree including guiding interventions.     

Regulation of periodontal ligament cell functions by interleukin-1beta

Periodontal ligament (PDL) cells maintain the attachment of the tooth to alveolar bone. These cells reside at a site in which they are challenged frequently by bacterial products and proinflammatory cytokines, such as interleukin-1beta (IL-1beta), during infections. In our initial studies we observed that IL-1beta down-regulates the osteoblast-like characteristics of PDL cells in vitro. Therefore, we examined the functional significance of the loss of the PDL cell's osteoblast-like characteristics during inflammation. In this report we show that, during inflammation, IL-1beta can modulate the phenotypic characteristics of PDL cells to a more functionally significant lipopolysaccharide (LPS)-responsive phenotype. In a healthy periodontium PDL cells exhibit an osteoblast-like phenotype and are unresponsive to gram-negative bacterial LPS. Treatment of PDL cells with IL-1beta inhibits the expression of their osteoblast-like characteristics, as assessed by the failure to express transforming growth factor beta1 (TGF-beta1) and proteins associated with mineralization, such as alkaline phosphatase and osteocalcin. As a consequence of this IL-1beta-induced phenotypic change, PDL cells become responsive to LPS and synthesize proinflammatory cytokines. The IL-1beta-induced phenotypic changes in PDL cells were transient, as removal of IL-1beta from PDL cell cultures resulted in reacquisition of their osteoblast-like characteristics and lack of LPS responsiveness. The IL-1beta-induced phenotypic changes occurred at concentrations that are frequently observed in tissue exudates during periodontal inflammation (0.05 to 5 ng/ml). The results suggest that, during inflammation in vivo, IL-1beta may modulate PDL cell functions, allowing PDL cells to participate directly in the disease process by assuming LPS responsiveness at the expense of their normal structural properties and functions.