Studierfenster: an Open Science Cloud-Based Medical Imaging Analysis Platform.

Imaging modalities such as computed tomography (CT) and magnetic resonance imaging (MRI) are widely used in diagnostics, clinical studies, and treatment planning. Automatic algorithms for image analysis have thus become an invaluable tool in medicine. Examples of this are two- and three-dimensional visualizations, image segmentation, and the registration of all anatomical structure and pathology types. In this context, we introduce Studierfenster ( www.studierfenster.at ): a free, non-commercial open science client-server framework for (bio-)medical image analysis. Studierfenster offers a wide range of capabilities, including the visualization of medical data (CT, MRI, etc.) in two-dimensional (2D) and three-dimensional (3D) space in common web browsers, such as Google Chrome, Mozilla Firefox, Safari, or Microsoft Edge. Other functionalities are the calculation of medical metrics (dice score and Hausdorff distance), manual slice-by-slice outlining of structures in medical images, manual placing of (anatomical) landmarks in medical imaging data, visualization of medical data in virtual reality (VR), and a facial reconstruction and registration of medical data for augmented reality (AR). More sophisticated features include the automatic cranial implant design with a convolutional neural network (CNN), the inpainting of aortic dissections with a generative adversarial network, and a CNN for automatic aortic landmark detection in CT angiography images. A user study with medical and non-medical experts in medical image analysis was performed, to evaluate the usability and the manual functionalities of Studierfenster. When participants were asked about their overall impression of Studierfenster in an ISO standard (ISO-Norm) questionnaire, a mean of 6.3 out of 7.0 possible points were achieved. The evaluation also provided insights into the results achievable with Studierfenster in practice, by comparing these with two ground truth segmentations performed by a physician of the Medical University of Graz in Austria. In this contribution, we presented an online environment for (bio-)medical image analysis. In doing so, we established a client-server-based architecture, which is able to process medical data, especially 3D volumes. Our online environment is not limited to medical applications for humans. Rather, its underlying concept could be interesting for researchers from other fields, in applying the already existing functionalities or future additional implementations of further image processing applications. An example could be the processing of medical acquisitions like CT or MRI from animals [Clinical Pharmacology & Therapeutics, 84(4):448-456, 68], which get more and more common, as veterinary clinics and centers get more and more equipped with such imaging devices. Furthermore, applications in entirely non-medical research in which images/volumes need to be processed are also thinkable, such as those in optical measuring techniques, astronomy, or archaeology.

Expression, isolation, and crystallization of the catalytic domain of CopB, a putative copper transporting ATPase from the thermoacidophilic archaeon Sulfolobus solfataricus.

The P-type CPX-ATPases are responsible for the transport of heavy metal ions in archaea, bacteria, and eukaryotes. We have chosen one of the two CPX-ATPases of the thermophile Sulfolobus solfataricus, CopB (= SSO2896) for the investigation of the molecular mechanism of this integral membrane protein. We recombinately expressed three different soluble domains of this protein (named CopB-A, CopB-B, and CopB-C) in Escherichia coli and purified them to homogeneity. 3D crystals of CopB-B, the 29 kDa catalytic ATP binding/phosphorylation domain were produced, which diffracted to a resolution of 2.2 A. CopB-B has heavy metal stimulated phosphatase activity, which was half maximal in the presence of 80 microM Cu2+. The protein forms a phosphorylated intermediate with the substrate gamma-(32P)-ATP. No specific activation of the polypeptide was observed, when CopB-B phosphatase activity was tested in the presence of the purified CopB-C and CopB-A proteins, which provide the cation binding and the phosphatase domains. We conclude that CopB is a putatively copper translocating ATPase, in which structural elements integrally located in the membrane are required for full, coordinated activation of the catalytic ATP binding domain.

Noninvasive auto-photoreduction used as a tool for studying structural changes in heme-copper oxidases by FTIR spectroscopy.

We demonstrate an efficient Fourier transform infrared (FTIR) spectroscopic method, termed "auto-photoreduction," that uses anaerobic photo-induced internal electron transfer to monitor reaction-initiated changes of heme-copper oxidases. It can be applied without the use of either expensive electrochemical equipment, or caged compounds, which cause significant background signals. At high irradiation power, carbon monoxide is released from high-spin heme a of cytochrome c oxidase and heme o from cytochrome bo(3). Photochemistry is initiated at wavelengths <355 nm, and the photochemical action spectrum has a maximum of 290 nm for cytochrome bo(3), which is consistent with the possible intermediate involvement of tyrosinate or an activated state of tyrosine. We propose that the final electron donors are proton channel water molecules. In the pH range of 4-9, the noninvasive auto-photoreduction method yields highly reproducible FTIR redox difference spectra within a broad range, resolving a number of vibrational changes outside the amide I region (1600-1640 cm(-1)). Furthermore, it provides details of redox-induced changes in the spectral region between 1600 and 1100 cm(-1). The auto-photoreduction method should be universally applicable to heme proteins.

Electron transfer at the low-spin heme b of cytochrome bo(3) induces an environmental change of the catalytic enhancer glutamic acid-286.

Intramolecular proton transfer of heme-copper oxidases is performed via the K- and the transmembrane D-channels. A carboxyl group conserved in a subgroup of heme-copper oxidases, located within the D-channel close to the binuclear center (=glutamic acid-286 in cytochrome bo(3) from Escherichia coli) is essential for proton pumping. Upon electron transfer to the fully oxidized (FO) enzyme, this amino acid has been shown to undergo a cyanide-independent environmental change. The redox-induced environmental transition of glutamic acid-286 is preserved in the site-directed mutant Y288F, which has lost its Cu(B) binding capacity. Furthermore, the mixed-valence (MV) redox state of cytochrome bo(3) (in which Cu(B) and high-spin heme are reduced, whereas the low-spin heme stays oxidized) was prepared by anaerobic exposure of the protein to carbon monoxide. This complex was converted (i) to the FO state by reaction with the caged dioxygen donor mu-peroxo) (mu-hydroxo) bis [bis (bipyridyl) cobalt (III)] and (ii) to the fully reduced (FR) state via caged electron donors; the environmental change of glutamic acid-286 could be observed only upon reduction. Taken together, these results from two different lines of evidence clearly show that the redox transition of the low-spin heme b center alone triggers the change in the chemical environment of this acidic side chain. It is suggested that glutamic acid-286 is a kinetic enhancer of proton translocation, which is energetically favoured in mesophilic oxidases.