Glial development in the Drosophila CNS requires concomitant activation of glial and repression of neuronal differentiation genes

Two classes of glial cells are found in the embryonic Drosophila CNS, midline glial cells and lateral glial cells. Midline glial development is triggered by EGF-receptor signalling, whereas lateral glial development is controlled by the gcm gene. Subsequent glial cell differentiation depends partly on the pointed gene. Here we describe a novel component required for all CNS glia development. The tramtrack gene encodes two zinc-finger proteins, one of which, ttkp69, is expressed in all non-neuronal CNS cells. We show that ttkp69 is downstream of gcm and can repress neuronal differentiation. Double mutant analysis and coexpression experiments indicate that glial cell differentiation may depend on a dual process, requiring the activation of glial differentiation by pointed and the concomitant repression of neuronal development by tramtrack.     

Structures of sialylated oligosaccharides of human erythropoietin expressed in recombinant BHK-21 cells

The native structures of the Asn-linked oligosaccharides and the O-glycans at Ser126 of human erythropoietin expressed from recombinant BHK cells have been elucidated. Enzymatically released N-glycans were studied by methylation analyses, fast-atom-bombardment mass spectrometry as well as one- and two-dimensional 1H-NMR spectrometry at 600 MHz. Many (82.7%) were found to be tetraantennary N-acetyllactosamine-type (22.8% with one, 3.6% with two and 0.4% with three N-acetyllactosamine repeats) being tetrasialylated (41%), trisialylated (29.6%) and disialylated (12.2%). A few (9.7%; 4.1% 2,4-branched, 5.6%, 2,6-branched) of the chains were triantennary (5.4% trisialyl, 4.3% disialyl) and 4.6% were of the disialyl diantennary type. Almost all of the innermost GlcNAc residues were alpha 1-6 fucosylated and NeuAc was exclusively alpha 2-3 linked to Gal beta 1-4GlcNAc-R; 60% of the protein was found to be O-glycosylated at Ser126; structures were monosialylated (70%) or disialylated (30%) forms of the Gal beta 1-3GalNAc core type. Glycosylation patterns at individual Asn-Xaa-Thr/Ser sites were determined by analytical high-pH anion-exchange chromatography with pulsed amperometric detection. Only tetraantennary chains with 0-3 N-acetyllactosamine repeats were detected at Asn38 and Asn83, while almost all of the di- and triantennary oligosaccharides were attached to Asn24. Batch analysis of different preparations of recombinant erythropoietin revealed the high reproducibility of the production procedure. Structures containing terminal GalNAc-GlcNAc were detected in small amounts in a few batches.     

The influence of preparation conditions on the electrochemical behaviour of CuO in a Li/CuO cell

A dozen CuO samples prepared under various conditions and from different starting materials were evaluated as cathode materials for a primary Li/CuO cell. The “thin electrode” method was used for rapid evaluation of the samples. Both coulombic efficiency and discharge voltage depend considerably on the method of synthesis. No correlation was found between the specific surface area and the resistivity of the samples on the one hand and the cathode performance on the other. Best results were obtained from CuO prepared by the oxidation of Cu₂O under controlled temperature and time of oxidation.     

BB_RADABA: a computer program as a tool for breeding and management of laboratory animals.

The use of well characterized laboratory animals in biomedical research is of importance for quality securing and reproducibility of animal experiments. Therefore, breeders should not only have in mind the reproduction of their animal stocks, but they should also be interested to achieve an optimal resemblance of the animals in the main traits characterizing them as a model of human diseases. Specific software applications which helps the breeders in monitoring the main breeding parameters, selecting suitable breeding pairs and data processing are practically not available. This prompted us to develop a computer program for management of breeding and experimental data, which we called BB_RADABA. In this report we want to give a short survey on this computer program.     

The effect of the scale of horizontal subsurface flow constructed wetlands on flow and transport parameters.

Horizontal subsurface flow constructed wetlands have proven their efficiency in treating wastewater and removing the pollutants of concern. Treatment efficiency depends on the wastewater residence time, which is a function of the hydraulic loading and the physical conditions of the constructed filter system, which can be described with effective parameters such as: hydraulic conductivity, porosity, dispersivity etc. Because spatial variability is often scale dependent, these effective parameters may be affected by the scale of the system being studied. In this paper the results of tracer experiments in constructed filters using saturated horizontal flow at three scales (small and medium lab scales and full-scale system) using the same filter media is reported. Light-weight aggregate (filter media termed Filtralite-P) was used at all scales. Increasing the scale was associated with increasing dispersivity, meanwhile hydraulic conductivity experienced dramatic reduction and variation by increasing the examined scale. Observed changes in the hydraulic parameters indicate that heterogeneity at different scales should be taken into account when the performance of LWA filters are evaluated from small-scale experiments.     

Blood-flow sensing by anionic biopolymers

The purpose of this study was to compare local recurrence, distant metastases, and survival rate in 350 patients with cancer of the middle and low rectum who underwent a radical abdominoperineal resection (APER) or a sphincter-saving resection (SSR) in our Institute. There were 257 APER patients and 93 SSR patients, with a median follow-up of 77 months. At 5 years, the estimates in APER and SSR patients were respectively 11% and 30% for the incidence of pelvic recurrence, 18% and 8% for the incidence of distant metastases, and 64% and 73% for overall survival. In the multivariate analysis it was found that Dukes' stage significantly affected pelvic recurrences, distant metastases rate and overall survival; histologic type affected only the pelvic recurrence rate. However, the final outcome of patients following APER or SSR was similar, suggesting that local failure per se does not affect long-term survival.     

CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering**

CYP154C5 from Nocardia farcinica is a P450 monooxygenase able to hydroxylate a range of steroids with high regio- and stereoselectivity at the 16α-position. Using protein engineering and substrate modifications based on the crystal structure of CYP154C5, an altered regioselectivity of the enzyme in steroid hydroxylation had been achieved. Thus, conversion of progesterone by mutant CYP154C5 F92A resulted in formation of the corresponding 21-hydroxylated product 11-deoxycorticosterone in addition to 16α-hydroxylation. Using MD simulation, this altered regioselectivity appeared to result from an alternative binding mode of the steroid in the active site of mutant F92A. MD simulation further suggested that the entrance of water to the active site caused higher uncoupling in this mutant. Moreover, exclusive 15α-hydroxylation was observed for wild-type CYP154C5 in the conversion of 5α-androstan-3-one, lacking an oxy-functional group at C17. Overall, our data give valuable insight into the structure–function relationship of this cytochrome P450 monooxygenase for steroid hydroxylation.     

Changes in the size and docosahexaenoic acid content of adipocytes during chick embryo development

The development of adipose tissue in the chick embryo was investigated using two groups of fertile eggs which differed by 1.7-fold in their initial yolk lipid levels. The triacylglycerol content of the subcutaneous adipose depot in both groups increased dramatically from day 12 of the 21-day embryonic period, attaining a maximal value just prior to hatching. During this period, the amount of triacylglycerol deposited in the adipose tissue was very highly correlated with the amount of lipid transferred from the yolk. The triacylglycerol content of the depot was also dependent on the initial yolk lipid content. During the hatching period, the amount of adipose triacylglycerol remained approximately constant in the group with the higher initial yolk lipid content but, in the case of the group with the lower initial yolk lipid levels, decreased by approximately 25%. The size distribution of adipocytes isolated from the tissue was determined by computerized image analysis microscopy. The mean adipocyte diameter increased from approximately 6 to 35 μm between days 12 and 19, irrespective of the initial yolk content, although development within the eggs with the lower initial yolk content resulted in a decrease in cell size over the hatching period. Both the triacylglycerol and phospholipid fractions of the isolated adipocytes contained substantial proportions (approximately 6%, w/w) of docosahexaenoic acid (DHA) at days 12 and 14, and lower levels of this fatty acid at the later stages. The amount (mg/depot) of DHA in adipose triacylglycerol decreased dramatically over the hatching period. The amount (mg/brain) of DHA in brain phospholipid increased by more than 5-fold between day 12 of development and hatching. A possible explanation for the data may be that DHA is preferentially mobilized from adipose tissue in order to deliver the fatty acid to the developing neural tissues in a form suitable for uptake.     

A scaled thickness conditioning for solid- and solid-shell discretizations of thin-walled structures

The numerical discretization of thin shell structures yields ill-conditioned stiffness matrices due to an inherent large eigenvalue spectrum. Finite element parametrization that depends on shell thickness, like relative displacement shells, solid shells and other solid finite elements even add to the ill-conditioning by introducing high eigenmodes.To overcome this numerical issue we present a scaled thickness conditioning (STC) approach, a mechanically motivated preconditioner for thin-walled structures discretized with continuum based element formulations. The proposed approach is motivated by the scaled director conditioning (SDC) method for relative displacement shell elements. In contrast to SDC, the novel STC approach yields a preconditioner for the effective linear system. It is applicable independently of element technology employed, coupling to other physical fields, boundary conditions applied and additional algebraic constraints and can be easily extended to multilayer shell formulations.The effect of the proposed preconditioner on the conditioning of the effective stiffness matrix and its eigenvalue spectrum is studied. It is shown that the condition number of the modified system becomes almost independent from the aspect ratio of the employed elements. The improved conditioning has a positive influence on the convergence behavior of iterative linear solvers. In particular, in combination with algebraic multigrid preconditioners the number of iterations could be decreased by more than 85% for some examples and the computation time could be reduced by about 60%.     

Animal experimental studies on the thrombolytic effect of streptokinase

The purpose of this investigation was to explore the feasibility of enhancing the electrochemical stability of dental amalgam restorations by a process of "selective interfacial amalgamation." If dental amalgam restorations can be selectively alloyed at the tooth-amalgam interface, to a minor thickness as compared with the dimensions of the bulk amalgam, so as to present a more electrochemically stable phase than psi2 to the oral environment, corrosion will be reduced and desirable qualities of existing amalgams can be utilized. It was found that selective interfacial amalgamation was indeed possible and that it could be readily accomplished using a silver suspension as a cavity liner. By then placing a conventional silver-tin amalgam according to ordinary dental techniques, mercury expressed during hand condensing reacted with the silver in the liner resulting in the formation of an integral amalgam restoration with the more noble psi1 phase adjacent to tooth structure. In vitro corrosion tests of samples so prepared exhibited corrosion resistance far superior to unlined control samples.