Thermochemical investigations of hydrolysis of p-nitrophenyl acetate in water-acetontrile mixtures

The α-chymotrypsin-catalyzed hydrolysis of the p-nitrophenyl acetate in the solvent mixtures containing from 1.6 up to 10% (v/v) acetonitrile in the presence of aqueous Tris buffer at pH 8.0 was investigated at 298 K by use of an isoperibolic batch calorimeter. A special experimental arrangement of the reaction components for the investigation of the hydrolytically instable substrate was used. Furthermore, the release of p-nitrophenol was recorded with an UV-vis-spectrophotometer under comparable conditions. The calorimetric curves consist of two parts. The first part is strongly rising and finished by a break point in the ΔT (time) curve. This first step is dominated by the enzyme-catalyzed reaction. After the break point a slow non-enzymatic process determines the course of the calorimetric curve. The molar enthalpy changes of overall reaction (ester hydrolysis and buffer protonation) of -100 ± 8, -106 ± 5 and -102 ± 5 kJ/mol were evaluated by a combination of the results from the spectrophotometric and calorimetric data for 1.6, 4.0 and 10.0% acetonitrile mixtures, respectively. The obtained results indicate that the enzyme-catalyzed hydrolysis is suitable for quantitative determination of the hydrophobic ester p-nitrophenyl acetate in water-acetonitrile mixtures using calorimetric detection. © 2004 Elsevier B.V. All rights reserved

Npn-1 Contributes to Axon-Axon Interactions That Differentially Control Sensory and Motor Innervation of the Limb

The initiation, execution, and completion of complex locomotor behaviors are depending on precisely integrated neural circuitries consisting of motor pathways that activate muscles in the extremities and sensory afferents that deliver feedback to motoneurons. These projections form in tight temporal and spatial vicinities during development, yet the molecular mechanisms and cues coordinating these processes are not well understood. Using cell-type specific ablation of the axon guidance receptor Neuropilin-1 (Npn-1) in spinal motoneurons or in sensory neurons in the dorsal root ganglia (DRG), we have explored the contribution of this signaling pathway to correct innervation of the limb. We show that Npn-1 controls the fasciculation of both projections and mediates inter-axonal communication. Removal of Npn-1 from sensory neurons results in defasciculation of sensory axons and, surprisingly, also of motor axons. In addition, the tight coupling between these two heterotypic axonal populations is lifted with sensory fibers now leading the spinal nerve projection. These findings are corroborated by partial genetic elimination of sensory neurons, which causes defasciculation of motor projections to the limb. Deletion of Npn-1 from motoneurons leads to severe defasciculation of motor axons in the distal limb and dorsal-ventral pathfinding errors, while outgrowth and fasciculation of sensory trajectories into the limb remain unaffected. Genetic elimination of motoneurons, however, revealed that sensory axons need only minimal scaffolding by motor axons to establish their projections in the distal limb. Thus, motor and sensory axons are mutually dependent on each other for the generation of their trajectories and interact in part through Npn-1-mediated fasciculation before and within the plexus region of the limbs

Thermochemical investigations of hydrolysis of p-nitrophenyl acetate in water-acetontrile mixtures

The α-chymotrypsin-catalyzed hydrolysis of the p-nitrophenyl acetate in the solvent mixtures containing from 1.6 up to 10% (v/v) acetonitrile in the presence of aqueous Tris buffer at pH 8.0 was investigated at 298 K by use of an isoperibolic batch calorimeter. A special experimental arrangement of the reaction components for the investigation of the hydrolytically instable substrate was used. Furthermore, the release of p-nitrophenol was recorded with an UV-vis-spectrophotometer under comparable conditions. The calorimetric curves consist of two parts. The first part is strongly rising and finished by a break point in the ΔT (time) curve. This first step is dominated by the enzyme-catalyzed reaction. After the break point a slow non-enzymatic process determines the course of the calorimetric curve. The molar enthalpy changes of overall reaction (ester hydrolysis and buffer protonation) of -100 ± 8, -106 ± 5 and -102 ± 5 kJ/mol were evaluated by a combination of the results from the spectrophotometric and calorimetric data for 1.6, 4.0 and 10.0% acetonitrile mixtures, respectively. The obtained results indicate that the enzyme-catalyzed hydrolysis is suitable for quantitative determination of the hydrophobic ester p-nitrophenyl acetate in water-acetonitrile mixtures using calorimetric detection. © 2004 Elsevier B.V. All rights reserved

Molecular characterization of the enhancing immunoglobulin of mice. Abstr.

The α-chymotrypsin-catalyzed hydrolysis of the p-nitrophenyl acetate in the solvent mixtures containing from 1.6 up to 10% (v/v) acetonitrile in the presence of aqueous Tris buffer at pH 8.0 was investigated at 298 K by use of an isoperibolic batch calorimeter. A special experimental arrangement of the reaction components for the investigation of the hydrolytically instable substrate was used. Furthermore, the release of p-nitrophenol was recorded with an UV-vis-spectrophotometer under comparable conditions. The calorimetric curves consist of two parts. The first part is strongly rising and finished by a break point in the ΔT (time) curve. This first step is dominated by the enzyme-catalyzed reaction. After the break point a slow non-enzymatic process determines the course of the calorimetric curve. The molar enthalpy changes of overall reaction (ester hydrolysis and buffer protonation) of -100 ± 8, -106 ± 5 and -102 ± 5 kJ/mol were evaluated by a combination of the results from the spectrophotometric and calorimetric data for 1.6, 4.0 and 10.0% acetonitrile mixtures, respectively. The obtained results indicate that the enzyme-catalyzed hydrolysis is suitable for quantitative determination of the hydrophobic ester p-nitrophenyl acetate in water-acetonitrile mixtures using calorimetric detection. © 2004 Elsevier B.V. All rights reserved

Bioavailable 87Sr/86Sr in different environmental samples — Effects of anthropogenic contamination and implications for isoscapes in past migration studies

87Sr/86Sr reference maps (isoscapes) are a key tool for investigating past human and animal migrations. However, there is little understanding of which biosphere samples are best proxies for local bioavailable Sr when dealing with movements of past populations. In this study, biological and geological samples (ground vegetation, tree leaves, rock leachates, water, soil extracts, as well as modern and archeological animal teeth and snail shells) were collected in the vicinity of two early medieval cemeteries (“Thuringians”, 5–6th century AD) in central Germany, in order to characterize 87Sr/86Sr of the local biosphere. Animal tooth enamel is not appropriate in this specific context to provide a reliable 87Sr/86Sr baseline for investigating past human migration. Archeological faunal teeth data (pig, sheep/goat, and cattle) indicates a different feeding area compared to that of the human population and modern deer teeth 87Sr/86Sr suggest the influence of chemical fertilizers. Soil leachates do not yield consistent 87Sr/86Sr, and 87Sr/86Sr of snail shells are biased towards values for soil carbonates. In contrast, water and vegetation samples seem to provide the most accurate estimates of bioavailable 87Sr/86Sr to generate Sr isoscapes in the study area. Long-term environmental archives of bioavailable 87Sr/86Sr such as freshwater bivalve shells and tree cores were examined in order to track potential historic anthropogenic contamination of the water and the vegetation. The data obtained from the archeological bivalve shells show that the modern rivers yield 87Sr/86Sr ratios which are similar to those of the past. However, the tree cores registered decreasing 87Sr/86Sr values over time towards present day likely mirroring anthropogenic activities such as forest liming, coal mining and/or soil acidification. The comparison of 87Sr/86Sr of the Thuringian skeletons excavated in the same area also shows that the vegetation samples are very likely anthropogenically influenced to some extent, affecting especially 87Sr/86Sr of the shallow rooted plants