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.     

Escape trajectories are deflected when fish larvae intercept their own C-start wake

Fish larvae may intercept their own wake during sharp turns, which might affect their escape performance. We analysed C-starts of larval zebrafish (Danio rerio, Hamilton, 1822) using a computational fluid dynamics approach that simulates free swimming (swimming trajectory is determined by fluid forces) by coupling hydrodynamics and body dynamics. The simulations show that fish may intercept their own wake when they turn by 100–180°. During stage 1 of a C-start, the fish generates a strong jet at the tail that is shed into the wake. During stage 2, the fish intercepts this wake. Counterfactual simulations showed that wake interception increased the lateral force on the fish and reduced the fish''s turning angle by more than 5°. Wake interception caused no significant acceleration tangential to the trajectory of the fish and did not affect total power output. While experimental and simulation evidence suggests that fish larvae can either undershoot or intercept but not overshoot their wake, our simulations show that larger fish might be able to avoid intercepting their wake by either under- or overshooting. As intercepting its own wake modifies the fish''s escape trajectory, fish should account for this effect when planning their escape route.     

Ice‐Templated Scaffolds with Microridged Pores Direct DRG Neurite Growth

Successful spinal cord repair is thought to be promoted with hierarchically structured scaffolds. These should combine aligned porosity with additional linear features on the micrometer scale to guide axons across multiple length scales. Such scaffolds are generated through the carefully controlled directional solidification of an aqueous biopolymer solution, followed by lyophilization. Under specific freezing conditions this yields a highly regular and aligned lamellar architecture. This architecture exhibits uniform ridges of controlled height and width on the lamellar surface. These ridges run parallel to the pore axis, serving as secondary guidance features. The ridges are capable of linearly aligning 62.4% of chick dorsal root ganglia neurites to within ±10° of the ridge direction. Notably, neurites sprouting perpendicular to the ridge are guided into alignment with these microridged features.