Reasoning with graph constraints

Graph constraints were introduced in the area of graph transformation, in connection with the notion of (negative) application conditions, as a form to limit the applicability of transformation rules. However, we believe that graph constraints may also play a significant role in the area of visual software modelling or in the specification and verification of semi-structured documents or websites (i.e. HTML or XML sets of documents). In this sense, after some discussion on these application areas, we concentrate on the problem of how to prove the consistency of specifications based on this kind of constraints. In particular, we present proof rules for two classes of graph constraints and show that our proof rules are sound and (refutationally) complete for each class. In addition, we study clause subsumption in this context as a form to speed up refutation.     

Gadolinium-enhanced magnetic resonance angiography of the pelvis in patients with erectile impotence

This study evaluted the potential of contrast-enhanced digital-subtraction magnetic resonance angiography (CE-DS-MRA) for noninvasive angiographic delineation of the arterial supply of the penis in patients with erectile dysfunction. After induction of an erection with prostaglandin E, a three-dimensional fast imaging with steady-state precision (FISP) sequence with TE of 1.8–2 milliseconds, TR of 4.4–5 milliseconds, and flip angle of 40°–60° was used to obtain high-resolution angiograms of the pelvis and penis during the injection of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) 0.3 mmol/kg body weight, within 30–50 seconds. DS maximum intensity projections (MIPs) and multiplanar reconstructions (MPRs) were compared with clinical work-up and directional Doppler ultrasound in 11 patients. In all 11 patients (100%), the arterial supply of the penis could be delineated from the aortic bifurcation via the iliac and internal pudendal arteries to the dorsal and deep penile arteries. Of the 22 internal pudendal arteries, 6 (27%) were occluded on CE-DS-MRA and 5 (23%) had stenoses, of which 4 (18%) were greater than 50%. In 7 patients (64%) good correlation between CE-DS-MRA and clinical findings and/or Doppler ultrasound was found; in 2 patients (18%), the correlation was moderate, and in 2 patients (18%) results were discrepant. In 6 patients (55%), MRA provided additional information to the clinical and Doppler ultrasound work-up. CE-DS-MRA can delineate small sessels such as the internal pudendal and penile arteries and thus has the potential to become a noninvasive angiography method in the work-up of erectile impotence. Supported by Siemens Medical Systems, Erlangen; Schering AG, Berlin; and Agfa-Gevaert AG, Leverkusen.     

The Oncolytic Adenovirus XVir-N-31, in Combination with the Blockade of the PD-1/PD-L1 Axis,Conveys Abscopal Effects in a Humanized Glioblastoma Mouse Model

Glioblastoma (GBM) is an obligatory lethal brain tumor with a median survival, even with the best standard of care therapy, of less than 20 months. In light of this fact, the evaluation of new GBM treatment approaches such as oncolytic virotherapy (OVT) is urgently needed. Based on our preliminary preclinical data, the YB-1 dependent oncolytic adenovirus (OAV) XVir-N-31 represents a promising therapeutic agent to treat, in particular, therapy resistant GBM. Preclinical studies have shown that XVir-N-31 prolonged the survival of GBM bearing mice. Now using an immunohumanized mouse model, we examined the immunostimulatory effects of XVir-N-31 in comparison to the wildtype adenovirus (Ad-WT). Additionally, we combined OVT with the inhibition of immune checkpoint proteins by using XVir-N-31 in combination with nivolumab, or by using a derivate of XVir-N-31 that expresses a PD-L1 neutralizing antibody. Although in vitro cell killing was higher for Ad-WT, XVir-N-31 induced a much stronger immunogenic cell death that was further elevated by blocking PD-1 or PD-L1. In vivo, an intratumoral injection of XVir-N-31 increased tumor infiltrating lymphocytes (TILs) and NK cells significantly more than Ad-WT not only in the virus-injected tumors, but also in the untreated tumors growing in the contralateral hemisphere. This suggests that for an effective treatment of GBM, immune activating properties by OAVs seem to be of greater importance than their oncolytic capacity. Furthermore, the addition of immune checkpoint inhibition (ICI) to OVT further induced lymphocyte infiltration. Consequently, a significant reduction in contralateral non-virus-injected tumors was only visible if OVT was combined with ICI. This strongly indicates that for an effective eradication of GBM cells that cannot be directly targeted by an intratumoral OV injection, additional ICI therapy is required.     

A non-conserved sequence in the 5'region of the CYH2 intron from Saccharomyces cerevisiae controls splicing efficiency of the pre-mRNA

The CYH2 gene from Saccharomyces cerevisiae containing one 510 bp intron is spliced inefficiently. We have shown previously that a non-conserved sequence within the intron is responsible for this low splicing efficiency. Using synthetic oligonucleotides comprising the identified region we show in this report that a very short region contains the specificity to act negatively on the splicing efficiency of the CYH2 gene. Furthermore, this sequence influences the splicing efficiency only when it is placed close to the 5' splice site of the gene. Investigations with chimeric CYH2/beta-actin genes show that this sequence acts independent from its natural surroundings. We propose that this sequence might interact with splicing factor(s).     

Adaptation of Oxidative Phosphorylation Machinery Compensates for Hepatic Lipotoxicity in Early Stages of MAFLD

Alterations in mitochondrial function are an important control variable in the progression of metabolic dysfunction-associated fatty liver disease (MAFLD), while also noted by increased de novo lipogenesis (DNL) and hepatic insulin resistance. We hypothesized that the organization and function of a mitochondrial electron transport chain (ETC) in this pathologic condition is a consequence of shifted substrate availability. We addressed this question using a transgenic mouse model with increased hepatic insulin resistance and DNL due to constitutively active human SREBP-1c. The abundance of ETC complex subunits and components of key metabolic pathways are regulated in the liver of these animals. Further omics approaches combined with functional assays in isolated liver mitochondria and primary hepatocytes revealed that the SREBP-1c-forced fatty liver induced a substrate limitation for oxidative phosphorylation, inducing enhanced complex II activity. The observed increased expression of mitochondrial genes may have indicated a counteraction. In conclusion, a shift of available substrates directed toward activated DNL results in increased electron flows, mainly through complex II, to compensate for the increased energy demand of the cell. The reorganization of key compounds in energy metabolism observed in the SREBP-1c animal model might explain the initial increase in mitochondrial function observed in the early stages of human MAFLD.     

Analysis of free amino acids in green coffee beans

To investigate amino acid changes in green coffee beans in the post-harvest period, amino acid concentrations were determined in green beans and after modelled drying, fermentation and storage. After the drying at alternating temperatures up to maximally 40°C, considerable changes in the concentrations of individual amino acids were identified. At the beginning of the storage period, significant changes in concentration were found to a minor extent. Under the condition of drying, it was mainly the concentration of glutamic acid that changed considerably. There was an increase in all the samples by 500 mg/kg dry matter on average, which corresponds to an increase of about 50% of the original value. In contrast, the concentration of aspartic acid in most of the samples decreased clearly due to drying. For the predominant part of the coffee samples, there was a significant increase in the hydrophobic amino acids Val, Phe, Ile and Leu. Changes of the quantities of other amino acids were non-uniform and only insignificant. Constant drying at 80°C for most of the amino acids brought about only minor concentration changes compared to those values obtained at 40°C. Modelled fermentation had no significant effect on the concentrations of the individual amino acids. After a 4-week storage of dried beans, amino acid concentrations did not change further. It is very possible that different post-harvest treatment parameters may influence the amount of aroma precursor compounds in the coffee beans.     

How platinum oxide affects the degradation analysis of PEM fuel cell cathodes

In this work, proton exchange membrane fuel cell cathodes are degraded with accelerated-stress-tests.These PtCo containing cathodes are analyzed at begin-of-life and end-of-test with a dedicated diagnostic procedure. For every individual load point, the oxygen transport resistance and voltage losses due to the formation of platinum oxides were obtained in addition to commonly measured electrochemical surface area, high frequency resistance, as well as cathode ionomer resistance. These data were used to break down the voltage losses into six different contributors. With this break down, performance gains and performance losses were determined at end-of-test. At low current densities, it was found that voltage losses due to degradation are dominated by the loss of specific activity and catalyst surface area - in line with the state-of-the-art knowledge. But by quantifying the losses from platinum oxide formation explicitly, we show that end-of-test an unassigned voltage loss is not only present at highest current densities, but already at low current density. More precisely, the unassigned voltage loss shows a linear increase with decreasing half cell voltage and is independent from the chosen accelerated stress test. As this unassigned loss depends on half cell voltage, it might arise from ionomer adsorption.