Triple A syndrome is caused by mutations in AAAS encoding the protein ALADIN. We investigated the role of ALADIN in the human adrenocortical cell line NCI-H295R1 by either over-expression or down-regulation of ALADIN. Our findings indicate that AAAS knock-down induces a down-regulation of genes coding for type II microsomal cytochrome P450 hydroxylases CYP17A1 and CYP21A2 and their electron donor enzyme cytochrome P450 oxidoreductase, thereby decreasing biosynthesis of precursor metabolites required for glucocorticoid and androgen production. Furthermore we demonstrate that ALADIN deficiency leads to increased susceptibility to oxidative stress and alteration in redox homeostasis after paraquat treatment. Finally, we show significantly impaired nuclear import of DNA ligase 1, aprataxin and ferritin heavy chain 1 in ALADIN knock-down cells. We conclude that down-regulating ALADIN results in decreased oxidative stress response leading to alteration in steroidogenesis, highlighting our knock-down cell model as an important in-vitro tool for studying the adrenal phenotype in triple A syndrome.

Here we review viral and cellular requirements for entry and intracellular trafficking of foamy viruses (FVs) resulting in integration of viral sequences into the host cell genome. The virus encoded glycoprotein harbors all essential viral determinants, which are involved in absorption to the host membrane and triggering the uptake of virus particles. However, only recently light was shed on some details of FV’s interaction with its host cell receptor(s). Latest studies indicate glycosaminoglycans of cellular proteoglycans, particularly heparan sulfate, to be of utmost importance. In a species-specific manner FVs encounter endogenous machineries of the target cell, which are in some cases exploited for fusion and further egress into the cytosol. Mostly triggered by pH-dependent endocytosis, viral and cellular membranes fuse and release naked FV capsids into the cytoplasm. Intact FV capsids are then shuttled along microtubules and are found to accumulate nearby the centrosome where they can remain in a latent state for extended time periods. Depending on the host cell cycle status, FV capsids finally disassemble and, by still poorly characterized mechanisms, the preintegration complex gets access to the host cell chromatin. Host cell mitosis finally allows for viral genome integration, ultimately starting a new round of viral replication.

Karst aquifers are characterized by highly conductive conduit flow paths embedded in a less conductive fissured and fractured matrix resulting in strong permeability contrasts with structured heterogeneity and anisotropy. Groundwater storage occurs predominantly in the fissured matrix. Hence, most karst models assume quasi steady-state flow in conduits neglecting conduit associated drainable storage (CADS). The concept of CADS considers storage volumes, where karst water is not part of the active flow system but rather hydraulically connected to conduits (for example karstic voids and large fractures). The disregard of conduit storage can be inappropriate when direct water abstraction from karst conduits occurs, e.g. large scale pumping. In such cases, CADS may be relevant. Furthermore, the typical fixed head boundary condition at the karst outlet can be inadequate for water abstraction scenarios because unhampered water inflow is possible. The objective of this paper is to analyze the significance of CADS and flow-limited boundary conditions on the hydraulic behavior of karst aquifers in water abstraction scenarios. To this end, the numerical hybrid model MODFLOW-2005 Conduit Flow Process Mode 1 (CFPM1) is enhanced to account for CADS. Additionally, a fixed-head limited-flow (FHLQ) boundary condition is added that limits inflow from constant head boundaries to a user-defined threshold. The affect and proper functioning of these modifications is demonstrated by simplified model studies. Both enhancements, CAD storage and the FHLQ boundary, are shown to be useful for water abstraction scenarios within karst aquifers. An idealized representation of a large-scale pumping test in a karst conduit is used to demonstrate that the enhanced CFPM1 is potentially able to adequately represent water abstraction processes in both the conduits and the matrix of real karst systems.

We present an unusual case of a late recurrent central neurocytoma that was rediagnosed as an ependymoma and neurocytoma in accordance with changes in histological classifications. Case Description. A 56-year-old male teacher presented with incomplete transverse syndrome due to several intradural extramedullary tumors at the level of lumbar vertebrae 1–3. The histological diagnosis at the time was atypical ependymoma. One year later, two additional tumors were removed at the L5-S1 vertebral level. For 12 years, the patient remained tumor free on followup. Fourteen years after the initial diagnosis, the patient presented with thoracic paresthesias due to two new extramedullary tumors in the C7-T1 and the T8-T9 vertebral levels. After complete removal of the tumors, a radiological survey revealed an intracranial lesion in the third ventricle. Five months later, an additional lesion recurrence was removed surgically. The most recent histological diagnosis revealed an atypical central neurocytoma. In retrospect, the previous tumors were reclassified as neurocytoma according to the additional immunohistochemistry evidence. Discussion. There is no standard adjuvant treatment regimen for atypical neurocytoma; therefore, the patient is currently under close followup. Modern histopathological diagnosis is essential in these cases. Potential routes for dissemination of the tumor should be considered upon first recurrence.

The use of reporter gene fusions to assess cellular processes such as protein targeting and regulation of transcription or translation is established technology in archaeal, bacterial, and eukaryal genetics. Fluorescent proteins or enzymes resulting in chromogenic substrate turnover, like β-galactosidase, have been particularly useful for microscopic and screening purposes. However, application of such methodology is of limited use for strictly anaerobic organisms due to the requirement of molecular oxygen for chromophore formation or color development. We have developed β-lactamase from Escherichia coli (encoded by bla) in conjunction with the chromogenic substrate nitrocefin into a reporter system usable under anaerobic conditions for the methanogenic archaeon Methanosarcina acetivorans. By using a signal peptide of a putative flagellin from M. acetivorans and different catabolic promoters, we could demonstrate growth substrate-dependent secretion of β-lactamase, facilitating its use in colony screening on agar plates. Furthermore, a series of fusions comprised of a constitutive promoter and sequences encoding variants of the synthetic tetracycline-responsive riboswitch (tc-RS) was created to characterize its influence on translation initiation in M. acetivorans. One tc-RS variant resulted in more than 11-fold tetracycline-dependent regulation of bla expression, which is in the range of regulation by naturally occurring riboswitches. Thus, tc-RS fusions represent the first solely cis-active, that is, factor-independent system for controlled gene expression in Archaea.

Habituation is a filter that optimizes the processing of information by our brain in all sensory modalities. It results in an unconscious reduced responsiveness to continuous or repetitive stimulation. In olfaction, the main question is whether habituation works the same way for any odorant or whether we habituate differently to each odorant? In particular, whether chemical, physical or perceptual cues can limit or increase habituation. To test this, the odour intensity of 32 odorants differing in physicochemical characteristics was rated by 58 participants continuously during 120s. Each odorant was delivered at a constant concentration. Results showed odorants differed significantly in habituation, highlighting the multifactoriality of habituation. Additionally habituation was predicted from 15 physico-chemical and perceptual characteristics of the odorants. The analysis highlighted the importance of trigeminality which is highly correlated to intensity and pleasantness. The vapour pressure, the molecular weight, the Odor Activity Value (OAV) and the number of double bonds mostly contributed to the modulation of habituation. Moreover, length of the carbon chain, number of conformers and hydrophobicity contributed to a lesser extent to the modulation of habituation. These results highlight new principles involved in the fundamental process of habituation, notably trigeminality and the physicochemical characteristics associated.

The clinical diagnostics of olfactory dysfunction in children turns out to be challenging due to low attention span, insufficient linguistic development and lack of odor experiences. Several smell tests have been developed for adults. Most of these examinations take a relatively long time and require a high level of concentration. Therefore, the aim of the current study was to evaluate an odor discrimination and olfactory threshold test using the frequently used “Sniffin’ Sticks” in children and adolescents in a simplified two-alternative-forced-choice version (2AFC) and compare it to the original three-alternative-forced-choice test (3AFC). One-hundred-twenty-one healthy participants aged between 5 and 17 years took part in this study. Within each of the two sessions participants underwent olfactory testing using the modified 2AFC as well as the standard 3AFC method. A better test-retest reliability was achieved using the original 3AFC method compared to the modified 2AFC. This was true for the odor discrimination as well as the olfactory threshold. Age had a significant influence on both tests, which should be considered when testing young children. We discuss these findings with relation to the existing norms and recommend using the 3AFC version due to a better test-retest reliability to measure olfactory function in children.

There have recently been considerable advances in our understanding of the neuronal mechanisms underlying multitasking, but the role of multimodal integration for this faculty has remained rather unclear. We examined this issue by comparing different modality combinations in a multitasking (stop-change) paradigm. In-depth neurophysiological analyses of event-related potentials (ERPs) were conducted to complement the obtained behavioral data. Specifically, we applied signal decomposition using second order blind identification (SOBI) to the multi-subject ERP data and source localization. We found that both general multimodal information integration and modality-specific aspects (potentially related to task difficulty) modulate behavioral performance and associated neurophysiological correlates. Simultaneous multimodal input generally increased early attentional processing of visual stimuli (i.e. P1 and N1 amplitudes) as well as measures of cognitive effort and conflict (i.e. central P3 amplitudes). Yet, tactile-visual input caused larger impairments in multitasking than audio-visual input. General aspects of multimodal information integration modulated the activity in the premotor cortex (BA 6) as well as different visual association areas concerned with the integration of visual information with input from other modalities (BA 19, BA 21, BA 37). On top of this, differences in the specific combination of modalities also affected performance and measures of conflict/effort originating in prefrontal regions (BA 6).

Polymer brush patterns were prepared by a combination of electron beam induced damage in self-assembled monolayers (SAMs), creating a stable carbonaceous deposit, and consecutive self-initiated photografting and photopolymerization (SIPGP). This newly applied technique, reactive writing (RW), is investigated with 1H,1H,2H,2H-perfluorooctyltriethoxysilane SAM (PF-SAM) on silicon oxide, which, when modified by RW, can be selectively functionalized by SIPGP. With the monomer N,N-dimethylaminoethyl methacrylate (DMAEMA), we demonstrate the straightforward formation of polymer brush gradients and single polymer lines of sub-100 nm lateral dimensions, with high contrast to the PF-SAM background. The lithography parameters acceleration voltage, irradiation dose, beam current and dwell time were systematically varied to identify the optimal conditions for the maximum conversion of the SAM into a carbonaceous deposit. The results of this approach were compared to patterns prepared by carbon templating (CT) under analogous conditions, revealing a dwell time dependency, which differs from earlier reports. This new technique expands the range of CT by giving the opportunity to not only vary the chemistry of the created polymer patterns with monomer choice but also vary the chemistry of the surrounding substrate.

This review aims to highlight and comprehensively summarize recent developments in the field of mixed-coordinate phosphorus chemistry. Particular attention is focused on the synthetic approaches to compounds containing at least two directly bonded phosphorus atoms in different coordination environments and their unexpected properties that are derived from spectroscopic and crystallographic data. Novel substance classes are discussed in order to supplement previous reviews about mixed-coordinate phosphorus compounds.