Structural and Functional Characterization of an Electron Transfer Flavoprotein Involved in Toluene Degradation in Strictly Anaerobic Bacteria.

(R)-Benzylsuccinate is the characteristic initial intermediate of anaerobic toluene metabolism, which is formed by a radical-type addition of toluene to fumarate. Its further degradation proceeds by activation to the coenzyme A (CoA)-thioester and ß-oxidation involving a specific (R)-2-benzylsuccinyl-CoA dehydrogenase (BbsG) affiliated with the family of acyl-CoA dehydrogenases. In this report, we present the biochemical properties of electron transfer flavoproteins (ETFs) from the strictly anaerobic toluene-degrading species Geobacter metallireducens and Desulfobacula toluolica and the facultatively anaerobic bacterium Aromatoleum aromaticum We determined the X-ray structure of the ETF paralogue involved in toluene metabolism of G. metallireducens, revealing strong overall similarities to previously characterized ETF variants but significantly different structural properties in the hinge regions mediating conformational changes. We also show that all strictly anaerobic toluene degraders utilize one of multiple genome-encoded related ETF paralogues, which constitute a distinct clade of similar sequences in the ETF family, for ß-oxidation of benzylsuccinate. In contrast, facultatively anaerobic toluene degraders contain only one ETF species, which is utilized in all ß-oxidation pathways. Our phylogenetic analysis of the known sequences of the ETF family suggests that at least 36 different clades can be differentiated, which are defined either by the taxonomic group of the respective host species (e.g., clade P for Proteobacteria) or by functional specialization (e.g., clade T for anaerobic toluene degradation).IMPORTANCE This study documents the involvement of ETF in anaerobic toluene metabolism as the physiological electron acceptor for benzylsuccinyl-CoA dehydrogenase. While toluene-degrading denitrifying proteobacteria use a common ETF species, which is also used for other ß-oxidation pathways, obligately anaerobic sulfate- or ferric-iron-reducing bacteria use specialized ETF paralogues for toluene degradation. Based on the structure and sequence conservation of these ETFs, they form a new clade that is only remotely related to the previously characterized members of the ETF family. An exhaustive analysis of the available sequences indicated that the protein family consists of several closely related clades of proven or potential electron-bifurcating ETF species and many deeply branching nonbifurcating clades, which either follow the host phylogeny or are affiliated according to functional criteria.

Mechanically metastable structures generated by single pulse laser-induced periodic surface structures (LIPSS) in the photoresist SU8.

Laser-induced periodic surface structures (LIPSS) with a periodicity of 351 nm are generated in the negative photoresist SU8 by single nanosecond laser pulse impact. Friction scans indicate the periodic pattern to comprise alternating regions of crosslinked and non-crosslinked SU8. Intriguingly, even minor mechanical stimuli in the order of nanonewtons cause the unfolding or rather the deletion of the characteristic periodic pattern similarly to the release of a pre-loaded spring. This feature combined with high resilience to heat and photon irradiation makes SU8-LIPSS attractive for applications such as mechanical stress monitors, self-destructing memory and passive micro actuators.

Static FET-PET and MR Imaging in Anaplastic Gliomas (WHO III).

OBJECTIVE: O-(2-[18F]-fluoroethyl)-L-tyrosine-positron emission tomography (FET-PET) imaging is an additional tool for tumor grading and surgery planning. Up to now, not much is known about FET-PET imaging in anaplastic gliomas. Our objective was to assess the FET uptake in anaplastic gliomas, compared with magnetic resonance imaging (MRI), histopathologic markers, and its prognostic value. PATIENTS AND METHODS: Forty-six patients (27 males/19 females) with an anaplastic glioma (WHO III) who received MRI and FET-PET imaging before surgery were retrospectively analyzed. Tumor volume was calculated in MRI and FET-PET imaging using a tumor-to-background ratio (TBR), and maximum FET uptake (TBRmax) was calculated. Overall survival (OS) and histopathologic markers (isocitrate-dehydrogenase 1/2-mutation, oligodendrial differentiation, and Ki67 proliferation index) were assessed. Univariate and multivariate analysis was performed for OS. RESULTS: In univariate analysis a significant correlation of TBRmax to OS was observed (P = 0.031). Tumor volume in FET-PET imaging (TBR > 2.0) (P = 0.028) showed a higher correlation to OS than the volume of the contrast-enhancing tumor part (P = 0.031). The highest correlation was observed for intersection of volume TBR > 1.3 and the volume of the contrast-enhancing tumor part (P = 0.005); fluid-attenuated inversion recovery volume showed no significant correlation to OS (P = 0.401) in the univariate analysis. Anaplastic glioma with oligodendrial differentiation showed significantly higher TBRmax values (P = 0.029), while no significant difference was observed for isocitrate hydrogenase 1/2-mutation (P = 0.752). CONCLUSION: Static FET-PET provides significant prognostic information in anaplastic gliomas, which adds to the value of MRI, supporting the use of both modalities preoperatively to assess individual risks and estimate prognosis. Definition of the histopathologic subtype using static FET-PET remains challenging.

18F-fluoro-ethyl-tyrosine positron emission tomography for grading and estimation of prognosis in patients with intracranial gliomas.

INTRODUCTION: Histopathological examination is the standard for grading and determination of diagnosis in intrinsic brain tumors though the possibility of malignization and tumor heterogeneity always bears the possibility of tumor under-grading or misjudgement regarding the estimation of prognosis. The aim of the present study was to evaluate the use of (18)F-FET-PET (FET-PET) for the grading and estimation of prognosis in newly diagnosed patients with intracranial gliomas in a clinical setting. METHODS: Patients who were treated for a newly diagnosed intracranial glioma between January 2007 and May 2012, and had a preoperative FET-PET and MRI scan between were included. The ratio of counts in a tumor VOI (volume of interest) with maximum uptake to the respective counts in a background VOI was calculated to provide the tumor-to-normal (T/N) ratio. The clinical and histopathological data (tumor grading, pre- and postoperative neurological status, Karnofsky Performance Status Scale scores, and overall survival rates) were recorded. RESULTS: One hundred fifty-two patients (39 WHO II, 26 WHO III, 87 WHO IV) were included. The median T/N ratio was 2.81 (1.1-8.1). The median T/N ratio of low-grade glioma patients was 1.65 (1.1-3.7), and 3.14 (1.61-8.1, p<0.001) in high-grade glioma patients. The median survival for patients with WHO III tumors was 22.8 months (95% CI: 15.87%-NA) and 13.23 months (95% CI: 10.83-15.6.%) for patients with WHO IV tumors (p=0.0001). For T/N≤1.6, no deaths were recorded; for 1.63, median survival was 14.0 months (95% CI: 11.7-16.2%, p<0.001). The test of the maximally selected log-rank statistic resulted in a T/N ratio of 1.88 as the cut-off value, with the greatest difference in overall survival between patients with longer and shorter survival. The ROC curve for differentiation of low- vs. high-grade tumors with regard to the T/N ratio showed an area under the curve (AUC) of 0.903. Regarding the prognostic validity for overall survival ROC-curves for 12-month, 24-month and 48-month survival display a higher validity for the WHO-classification than for the imaging modalities though with an AUC of 0.847 for the 48-month survival T/N ratio and MRI contrast-enhancement have a high prognostic value as well. CONCLUSION: Our study suggests that FET-PET can predict prognosis and survival in patients harboring intracranial gliomas and serves as a valuable tool to supplement the established clinical and histopathological parameters.