Mutagenesis of Rat Dopamine β-Hydroxylase: Examination in Cell-Free System

Abstract: Dopamine β-hydroxylase (DBH; EC 1.14.17.1) exists as membrane-bound and soluble forms in neurosecretory vesicles. The features of DBH required for glycosylation and incorporation into membranes were studied in a cell-free system. Translation of full-length DBH with microsomal membranes generated two glycosylated products (GH and GL) depending on the magnesium concentration. Carboxyl-terminal, in contrast to amino-terminal, truncations gave translation products that were glycosylated by microsomal membranes. Site-directed mutants were generated with the second AUG codon and the region of a putative signal sequence cleavage site modified. Translation without membranes indicated that the second AUG is not used to initiate translation. The mutant with Glu⁴¹→ Leu⁴¹ and Ser⁴³→ Thr⁴³ yielded only the GH form with membranes, whereas mutation of Ser⁴³→ Ala⁴³ generated both GH and GL forms. Both glycosylated forms comigrated with the microsomal membranes on sucrose gradients. Endoglycosidase H digestion indicated that the differences between the GH and GL forms are not due to the sugar moiety. The results suggest a role for cleavage of a signal sequence in the formation of different forms of DBH. The possibility that these mutations change the secondary structure near the signal cleavage site, affecting processing, is discussed.     

Comparison of an iterative microcalorimetric method and dialysis equilibrium for calculating thermodynamic parameters of a binding protein which presents a weak affinity for its substrate

An original iterative microcalorimetric method is used to study the interaction of 5-fluorouracil, a cancer chemotherapeutic agent, with human serum albumin. Two equivalent binding sites are demonstrated with an association constant and an enthalpy variation equal to 370 ± 30 m^?1 and ?10 ± 0,5 kcal/mol, respectively. By means of a competitive microcalorimetric method, the partially competitive binding of dipotassium chlorazepate and 5-fluorouracil is shown. This iterative microcalorimetric method can be applied to all techniques involving the measurement of a phenomenon which is proportional to the concentration of the complex.     

Versatile habitat conditioning by damselfish cultivating turf algae on giant clams

In this paper, a monitoring and modelling concept for ecological optimized harbour dredging and fine sediment disposal in large rivers is presented. According to the concept, first a preliminary assessment should be performed previous to the dredging and dumping procedure to derive knowledge about the current status in hydrodynamics, morphology and instream habitat quality. During the performance of the maintenance work, a high-resolution monitoring program has to be organized to measure flow velocities, the suspended sediment concentrations and the extent of the occurring plume. These data can then be compared with natural suspended sediment conditions and serve as input data for numerical sediment transport modelling. Furthermore, bathymetric surveys and biotic sampling enable the detection of possible effects of dredging and disposal in the post-dumping stage. Based on sediment transport modelling approaches, short- to mid-term developments of the sediment plume can be predicted with an additional and final habitat evaluation at the end of the project. This concept was applied and optimized during the maintenance work at the case study winter harbour Linz at the Danube River. The findings of the presented study highlight the necessity of integrated monitoring and modelling approaches for harbour dredging especially in large river systems.     

Repeated Immobilization Stress Increases the Binding of c-Fos-Like Proteins to a Rat Dopamine β-Hydroxylase Promoter Enhancer Sequence

Abstract: The present study addressed the possibility that disinhibition of serotonin (5-HT) autoreceptor-mediated negative feedback might potentiate the elevation of nerve terminal 5-HT output induced by selective 5-HT reuptake blockade. To this end, rats were given citalopram and the 5-HT autoreceptor-blocking agents ( S )-UH-301 (5-HT_1A) and (−)-penbutolol (5-HT_1A/1B), and the effect on extracellular 5-HT in the ventral hippocampus was monitored by means of in vivo microdialysis. Citalopram (5 mg/kg, s.c.) approximately doubled the 5-HT output, a response that was markedly augmented by ( S )-UH-301 (3 mg/kg, s.c.) and (−)-penbutolol (8 mg/kg, s.c.) and by combined treatment with ( S )-UH-301 (3 mg/kg, s.c.) plus (−)-penbutolol (1 μ M ; via the dialysis perfusion medium), but not by (−)-penbutolol (1 μ M ) alone. These findings provide evidence that 5-HT, in particular 5-HT_1A, autoreceptor-mediated negative feedback mechanisms are pivotal in determining the nerve terminal 5-HT output level after 5-HT reuptake inhibition. These findings have important implications for the interplay between different processes controlling 5-HT transmission in vivo and might possibly offer a lead toward novel, therapeutically exploitable principles.     

Recent developments in photochemical and chemical AOPs in water treatment: a mini-review

Photochemical and chemical oxidation methods, belonging to a wide group of advanced oxidation processes (AOPs), are extensively studied but still under constant development. The aim of this study was to review the recent developments mainly in UV/H₂O₂ process and Fenton’s reactions. Ultraviolet (UV) radiation is an essential factor in photochemical oxidation. In the development of UV radiation sources the energy efficiency and the use of toxicant free materials have become major objectives. Also the emitted wavelength plays a significant role. Besides, there are alternatives for UV radiation sources, such as radio frequency (RF) plasmas, which can operate in water without additional chemicals. A remarkable development in the field of chemical oxidation has been novel catalyst materials for Fenton’s reaction enabling the efficient reaction in neutral or almost neutral conditions, instead of pH below 3. In addition, varieties of method integrations, e.g., combinations of AOPs or AOPs integrated to biological processes, and other enhancements such as combining ultrasound (US) with AOP have led to improved water treatment efficiencies.