Differential proteome analysis of colon carcinoma cell line SW480 after reconstitution of the tumour suppressor Smad4

The tumour suppressor gene Smad4 is frequently inactivated in gastrointestinal carcinomas. Smad4 plays a pivotal role in transducing signals of the transforming growth factor-β (TGF-β) superfamily of proteins. Inactivation of Smad4 seems to occur late during tumour progression when tumours acquire invasive and metastatic properties. Identification of proteins directly or indirectly regulated by Smad4 would, therefore, ease the future design of new diagnostic and therapeutic strategies for gastrointestinal carcinoma. We have used human colon carcinoma cell line SW480 stably transfected with Smad4 as an in-vitro model system to identify Smad4-regulated proteins by applying two-dimensional gel electrophoresis (2DE) then MALDI-PMF/PFF-MS. We identified a total of 47 protein species with a Smad4-dependent expression. From the functions of the candidate proteins we obtained new insights into Smad4’s participation in processes, for example apoptosis, differentiation, and proliferation.     

Synthesis of pyrene and benzo[a]pyrene adducts at the exocyclic amino groups of 2'-deoxyadenosine and 2'-deoxyguanosine by a palladium-mediated C-N bond-formation strategy

Single-electron oxidation of the carcinogenic hydrocarbon benzo[a]pyrene (BaP) is thought to result in a radical cation intermediate and this species has been proposed to cause alkylation at the nitrogens of the purine nucleobases. Although several different nucleoside adducts have been isolated as arising from this mode of metabolic activation, there are no selective, total syntheses of the stable exocyclic amino group adducts formed by the single-electron oxidation of any hydrocarbon with the purine 2'-deoxynucleosides to date. In this paper we disclose the synthesis of the model adducts N(6)-(1-pyrenyl)-2'-deoxyadenosine and N(2)-(1-pyrenyl)-2'-deoxyguanosine as well as the first synthesis of the carcinogen-linked nucleoside derivatives N(6)-(6-benzo[a]pyrenyl)-2'-deoxyadenosine and N(2)-(6-benzo[a]pyrenyl)-2'-deoxyguanosine via a palladium-mediated C-N bond formation. Two different coupling strategies were attempted: coupling of an aryl bromide with a suitably protected nucleoside and the coupling of an arylamine with a suitable halonucleoside. The former had somewhat limited applicability in that only N(6)-(1-pyrenyl)-2'-deoxyadenosine was prepared by this method; on the other hand, the latter was more general. However, there are noteworthy differences in the amination reactions at the C-6 and C-2 positions. Reactions at the C-6 resulted in the competing formation of a 1:2 amine-nucleoside adduct in addition to the desired monoaryl nucleoside. Such a dimer formation was not observed at the C-2. The C-2 adducts, however, displayed an interesting conformational behavior.     

Hydrophobic interaction chromatography of proteins IV. Kinetics of protein spreading

Adsorption of proteins on surfaces of hydrophobic interaction chromatography media is at least a two-stage process. Application of pure protein pulses (bovine serum albumin and beta-lactoglobulin) to hydrophobic interaction chromatography media yielded two chromatographic peaks at low salt concentrations. At these salt concentrations, the adsorption process is affected by a second reaction, which can be interpreted as protein spreading or partial unfolding of the protein. The kinetic constants of the spreading reaction were derived from pulse response experiments at different residence times and varying concentrations by applying a modified adsorption model considering conformational changes. The obtained parameters were used to calculate uptake and breakthrough curves for spreading proteins. Although these parameters were determined at low saturation of the column, predictions of overloaded situations could match the experimental runs satisfactorily. Our findings suggest that proteins which are sensitive to conformational changes should be loaded at high salt concentrations in order to accelerate the adsorption reaction and to obtain steeper breakthrough curves.     

In situ determination of adsorption kinetics of proteins in a finite bath

A method for fast in situ measurement of adsorption kinetics based on a finite bath was developed. We modified the conventional finite bath by replacing the external loop by a dip probe which enables in situ measurement of the concentration change in the contactor. Deposition of adsorbent particles on the reflection surface of the dip probe compromised measurements. Different membranes, a polyamide, a polypropylene and a nylon membrane were tested to protect the internal reflection surface of the dip probe from fouling with adsorbent particles. The nylon membrane provided efficient protection and high mass transfer evaluated by response time experiments. Unspecific adsorption of the model protein on the membrane could also be excluded. To corroborate the measurements of the dip probe the results were compared to a conventional finite bath and to a shallow-bed. The uptake curves for human polyclonal IgG at different concentrationes (0.1-3 g/l) on rProtein A Sepharose FF and MabSelect were used as model system. The effective diffusion coefficients were determined using a pore diffusion model. These values were in good agreement for all methods.     

Low frequency sound scattering from spherical assemblages of bubbles using effective medium theory

The determination of the acoustic field scattered by an underwater assembly of gas bubbles or similar resonant monopole scatterers is of considerable theoretical and practical interest. This problem is addressed from a theoretical point of view within the framework of the effective medium theory for the case of spherically shaped assemblages. Although being valid more generally, the effective medium theory is an ideal instrument to study multiple scattering effects such as low frequency collective resonances, acoustically coupled breathing modes of the entire assembly. Explicit expressions for the scattering amplitude and cross sections are derived, as well as closed form expressions for the resonance frequency and spectral shape of the fundamental collective mode utilizing analytical S-matrix methods. This approach allows, in principle, a simultaneous inversion for the assembly radius and void fraction directly from the scattering cross sections. To demonstrate the validity of the approach, the theory is applied to the example of idealized, spherically shaped schools of swim bladder bearing fish. The analytic results of the theory are compared to numerical first-principle benchmark computations and excellent agreement is found, even for densely packed schools and frequencies across the bladder resonance.     

A-sign und a.trust bündeln Aktivitäten

AktuellDivitec Metal

Aufbereitung von Elektro(nik)-Altger?ten     

Zur Lumineszenz des Zinkoxyds

The temperature dependance of efficiency and decay of the ultraviolet and green luminescence bands at UV- and cathode ray-excitation is investigated. In addition to earlier results the Riehl-effect is only observed when excited by cathode rays. The UV-band decays hyperbolically at all temperatures while the decay of the green band depends on kind and intensity of the excitation as well as on temperature. The results can be interpreted by considering the increasing number of electrons in the conduction band under different kinds of excitation and the variation of conductivity with temperature. Furthermore, the glow curves of various ZnO-samples (untreated and annealed in reducing atmosphere) show one or two peaks respectively, thus revealing two trap depths. As the light sum increases with the efficiency it is assumed that by reduction both activators and traps are created. The occurrence of thermoluminescence with many conduction electrons being present leads to the assumption of hole traps.     

Radiative and nonradiative lifetimes in excited states of Ar,Kr and Xe atoms in a neon matrix

Synchrotron radiation with its intense continuum and its excellent time structure has been exploited for time resolved luminescence spectroscopy in the solid state. By selective excitation of n = 1, n′ = 2 exciton states of Xe, Kr and Ar atoms in a neon matrix we were able to identify the emitting states involved. Lifetimes within the cascade of radiative and radiationless relaxation between excited states as well as the radiative lifetimes for transitions to the ground state have been derived from the decay curves. Energy positions and radiative lifetimes of the emitting states correspond quite well with those of the free atoms. Radiative and radiationless relaxation processes take place within the manifold of excited states of the guest atoms. The rate constants for radiationless decay confirm an energy gap law. The order of the radiationless processes reaches in some cases extremely high values. Selection rules for spin and angular momentum are essential to understand the observed radiationless transition rates.     

A mathematical model of an anthrax epizoötic in the Kruger National Park

An outbreak of anthrax in a single homogeneous geographical region is modelled. The spread of the disease to neigbouring regions is also simulated and the model is compared with data from the 1970 epizoötic in the Kruger National Park.Two possible driving mechanisms for the epizoötic are proposed and examined, viz. environmental contamination or direct contact between vulnerable animals and fresh carcases. In the latter case, it is shown that the model exhibits a threshold effect, in that an epizoötic will only arise if the vulnerable population exceeds a threshold value which is determined in terms of one of the parameters of the model.