[After the pandemic is before the pandemic: And how interdisciplinary research can help here]. / Nach der Pandemie ist vor der Pandemie: Und wie interdisziplinäre Forschung hier helfen kann.

Aviation is among the social sectors most impacted by the COVID-19 pandemic, and at the same time has contributed to the rapid global spread of the SARS-CoV-2 virus. SARS-CoV-2 is one of the coronaviruses that have led to outbreaks such as MERS-CoV in the past. This group of pathogens, as well as others that may be unknown at this time, will continue to challenge our society in the future. In order to be able to react better, a research training group was established at DLR in cooperation with 6 institutes, which will develop interdisciplinary approaches to researching and combating current and future pandemics. Engineers, physicists, software developers, biologists and physicians are working closely together on new concepts and the development of interdisciplinary knowledge in order to better control and contain future pandemics and to be able to react in a more targeted manner. One focus is the reduction of germ contamination in airplanes but also in other means of public transport such as buses and trains. In this review, we provide an overview of the baseline situation and possible approaches to address future pandemic challenges.

Male Fertility After Inguinal Hernia Mesh Repair: A National Register Study.

OBJECTIVE: To determine whether patients who receive an inguinal hernia repair father the same number of children as the background population. BACKGROUND: Although the effect of inguinal hernia repair on male fertility has previously been investigated through indirect measures, no previous studies have evaluated the final measure of male fertility, which is the number of children fathered by patients. METHODS: Prospectively collected data on 32,621 male patients between the ages of 18 and 55 years who received 1 or more inguinal hernia repairs during the years 1998 to 2012 were found in 5 comprehensive Danish linked registers. Patients were matched with 97,805 controls, and the number of fathered children was recorded as the primary outcome. RESULTS: Patients who were operated unilaterally fathered more children than controls (156 vs 147 children per 1000 individuals, P = 0.02), whereas patients who were operated bilaterally fathered the same number of children as controls. Unilateral Lichtenstein operation resulted in an increase in number of children fathered by patients (161 vs 151 children per 1000 patients, P = 0.009). No difference in the number of children fathered was found for any year following operation. Meanwhile, time between operation and first child was longer among controls than patients (log-rank P = 0.003). The youngest (18-30 years of age) bilaterally operated patients fathered the same number of children as controls. CONCLUSIONS: Patients who underwent inguinal hernia repair using Lichtenstein technique or laparoscopic approach did not father fewer children than expected. Thus, inguinal hernia repair using Lichtenstein or laparoscopic approach did not impair male fertility.

Self-directed simulation-based training of emergency cricothyroidotomy: a route to lifesaving skills.

The emergency cricothyroidotomy (EC) is a critical procedure. The high cost of failures increases the demand for evidence-based training methods. The aim of this study was to present and evaluate self-directed video-guided simulation training. Novice doctors were given an individual 1-h simulation training session. One month later, an EC on a cadaver was performed. All EC's were video recorded. An assessment tool was used to rate performance. Performance was compared with a pass/fail level for the EC. We found a high reliability, based on Pearson's r (0.88), and a significant progression of skill during training (p < 0.001). Eleven out of 14 succeeded in creating an airway on the cadaver in 64 s (median, range 39-86 s), but only four achieved a passing score. Our 1-h training protocol successfully raised the competence level of novice doctors; however, the training did not ensure that all participants attained proficiency.

Structural basis for synthesis of inflammatory mediators by human leukotriene C4 synthase.

Cysteinyl leukotrienes are key mediators in inflammation and have an important role in acute and chronic inflammatory diseases of the cardiovascular and respiratory systems, in particular bronchial asthma. In the biosynthesis of cysteinyl leukotrienes, conversion of arachidonic acid forms the unstable epoxide leukotriene A4 (LTA4). This intermediate is conjugated with glutathione (GSH) to produce leukotriene C4 (LTC4) in a reaction catalysed by LTC4 synthase: this reaction is the key step in cysteinyl leukotriene formation. Here we present the crystal structure of the human LTC4 synthase in its apo and GSH-complexed forms to 2.00 and 2.15 A resolution, respectively. The structure reveals a homotrimer, where each monomer is composed of four transmembrane segments. The structure of the enzyme in complex with substrate reveals that the active site enforces a horseshoe-shaped conformation on GSH, and effectively positions the thiol group for activation by a nearby arginine at the membrane-enzyme interface. In addition, the structure provides a model for how the omega-end of the lipophilic co-substrate is pinned at one end of a hydrophobic cleft, providing a molecular 'ruler' to align the reactive epoxide at the thiol of glutathione. This provides new structural insights into the mechanism of LTC4 formation, and also suggests that the observed binding and activation of GSH might be common for a family of homologous proteins important for inflammatory and detoxification responses.

"Psychological characteristics of functional respiratory disorders in children and adolescents-Pilot study".

BACKGROUND: Aim of our prospective, multicenter, nonrandomized study was to identify characteristic features and similarities of patients with functional respiratory disorders regarding socio-familial and behavioral aspects, in comparison with controls in a cross-sectional analysis using standardized psychological questionnaires. Furthermore, we investigated the longitudinal outcome of symptoms, effects of primary interventions and the stability of psychological traits 6 months after diagnosis and primary intervention. METHODS: Initially, 106 patients (68 females, 27 males) and 58 controls (33 females, 25 males) were recruited for the study. Mean age was 12.6 years in patients and 11.9 years in controls. RESULTS: The child behavior checklist (CBCL) showed significantly increased scores for anxious/depressed (p = 0.002) and schizoid/obsessive (p = 0.001) behavior in patients. A trend was evident for internalizing behavior (p = 0.009) and for a higher total score (p = 0.008). In the self-assessment youth self-report (YSR), there was a trend towards higher values for anxious/depressed behavior in patients (p = 0.06) and towards more externalizing behavior (p = 0.029) in the control group. After 6 months, 31% of the patients were free of symptoms, 42% had improved. For themselves, parents reported a decreased burden from 56% to 23% (p < 0.001) and decreased impairment from 57% to 30% (p < 0.008). For their children, parents reported a decrease from 45% to 16% (p < 0.0001) and from 74% to 37% (p < 0.0001), respectively. A longitudinal comparison from T1 to T2 showed no statistically significant changes in all three psychological questionnaires (CBCL, YSR, and SOMS-KJ). CONCLUSIONS: In summary, we show that patients with functional respiratory disorders differ from healthy subjects, with internalizing behavior being a characteristic trait. The outcome in terms of symptoms, perceived psycho-familial burden and impairment after 6 months is encouraging. However, we are aware that our preliminary data offer thought-provoking impulses rather than firm findings.

Testing Procedure for Fatigue Characterization of Steel-CFRP Hybrid Laminate Considering Material Dependent Self-Heating.

Combining carbon fiber reinforced polymers (CFRP) with steel offers the potential of utilizing the desired characteristics of both materials, such as specific strength/stiffness and fatigue strength of fiber reinforced polymers (FRP) and impact resistance of metals. Since in such hybrid laminates multiple material layers are combined, a gradual failure is likely that can lead to changes in mechanical properties. A failure of the metal partner leads to an increase in stress on the FRP, which under fatigue load results in increased self-heating of the FRP. Therefore, a suitable testing procedure is required and developed in this study, to enable a reproducible characterization of the mechanical properties under fatigue load. The resulting testing procedure, containing multiple frequency tests as well as load increase and constant amplitude tests, enabled characterization of the fatigue performance while never exceeding a testing induced change in temperature of 4 K. In addition to the development of the testing procedure, an insight into the manufacturing induced residual stresses occurring in such hybrid laminates, which impacts the load-bearing capacity, was established using finite element simulation. The gathered data and knowledge represents a basis for future in-depth investigations in the area of residual stress influence on the performance of hybrid laminates and highlights its importance, since not only the used testing procedure determines the measured fatigue performance.

Prognostic factors after ultrasound-guided percutaneous ablation of colorectal liver metastases: A systematic review.

INTRODUCTION: Liver metastases from colorectal cancer are common and ablation therapy is a favourable treatment option for selected patients not suited for surgical resection. This study aimed to systematically review the literature and present prognostic factors associated with survival and local recurrence after percutaneous ultrasound-guided ablation treatment. MATERIALS AND METHODS: This review is reported according to the PRISMA. PubMed, Embase and Scopus were searched and records were independently screened by two authors, initially on title and abstract and subsequently on full-text basis. The quality of the studies was assessed using the Newcastle-Ottawa quality assessment scale. RESULTS: Of 2.882 records screened, 18 studies were included. The median survival was 23 months. One-year survival was median 95% and 3-year survival was median 58%. Complete ablation response and adjuvant chemotherapy produce considerably improved survival and low local recurrence rate outcomes. CONCLUSION: Percutaneous ultrasound-guided ablation technique for colorectal liver metastases provides impressive survival rates for patients not suited for surgical resection. However, there are some factors related to poorer prognosis, which may be considered when selecting patients.

High-affinity binders selected from designed ankyrin repeat protein libraries.

We report here the evolution of ankyrin repeat (AR) proteins in vitro for specific, high-affinity target binding. Using a consensus design strategy, we generated combinatorial libraries of AR proteins of varying repeat numbers with diversified binding surfaces. Libraries of two and three repeats, flanked by 'capping repeats,' were used in ribosome-display selections against maltose binding protein (MBP) and two eukaryotic kinases. We rapidly enriched target-specific binders with affinities in the low nanomolar range and determined the crystal structure of one of the selected AR proteins in complex with MBP at 2.3 A resolution. The interaction relies on the randomized positions of the designed AR protein and is comparable to natural, heterodimeric protein-protein interactions. Thus, our AR protein libraries are valuable sources for binding molecules and, because of the very favorable biophysical properties of the designed AR proteins, an attractive alternative to antibody libraries.

Allosteric inhibition of aminoglycoside phosphotransferase by a designed ankyrin repeat protein.

Aminoglycoside phosphotransferase (3')-IIIa (APH) is a bacterial kinase that confers antibiotic resistance to many pathogenic bacteria and shares structural homology with eukaryotic protein kinases. We report here the crystal structure of APH, trapped in an inactive conformation by a tailor-made inhibitory ankyrin repeat (AR) protein, at 2.15 A resolution. The inhibitor was selected from a combinatorial library of designed AR proteins. The AR protein binds the C-terminal lobe of APH and thereby stabilizes three alpha helices, which are necessary for substrate binding, in a significantly displaced conformation. BIAcore analysis and kinetic enzyme inhibition experiments are consistent with the proposed allosteric inhibition mechanism. In contrast to most small-molecule kinase inhibitors, the AR proteins are not restricted to active site binding, allowing for higher specificity. Inactive conformations of pharmaceutically relevant enzymes, as can be elucidated with the approach presented here, represent powerful starting points for rational drug design.

Crystal structure of a consensus-designed ankyrin repeat protein: implications for stability.

Consensus-designed ankyrin repeat (AR) proteins are thermodynamically very stable. The structural analysis of the designed AR protein E3_5 revealed that this stability is due to a regular fold with highly conserved structural motifs and H-bonding networks. However, the designed AR protein E3_19 exhibits a significantly lower stability than E3_5 (9.6 vs. 14.8 kcal/mol), despite 88% sequence identity. To investigate the structural correlations of this stability difference between E3_5 and E3_19, we determined the crystal structure of E3_19 at 1.9 A resolution. E3_19 as well has a regular AR domain fold with the characteristic H-bonding patterns. All structural features of the E3_5 and E3_19 molecules appear to be virtually identical (RMSD(Calpha) approximately 0.7 A). However, clear differences are observed in the surface charge distribution of the two AR proteins. E3_19 features clusters of charged residues and more exposed hydrophobic residues than E3_5. The atomic coordinates of E3_19 have been deposited in the Protein Data Bank. PDB ID: 2BKG.

Molecular dynamics study of the stabilities of consensus designed ankyrin repeat proteins.

Two designed ankyrin repeat (AR) proteins (E3_5 and E3_19) are high homologous (with about 87% sequence identity) and their crystal structures have a Calpha atom-positional root-mean-square difference of about 0.14 nm. However, it was found that E3_5 is considerably more stable than E3_19 in guanidinium hydrochloride and thermal denaturation experiments. With the goal of providing insights into the various factors contributing to the stabilities of the designed AR proteins and suggesting possible mutations to enhance their stabilities, homology modeling and molecular dynamics (MD) simulations with explicit solvent have been performed. Because the crystal structure of E3_19 was solved later than that of E3_5, a homology model of E3_19 based on the crystal structure of E3_5 was also used in the simulations. E3_5 shows a very stable trajectory in both crystal and solution simulations. In contrast, the C-terminal repeat of E3_19 unfolds in the simulations starting from either the modeled structure or the crystal structure, although it has a sequence identical to that of E3_5. A continuum electrostatic model was used to estimate the effect of single mutations on protein stability and to study the interaction between the internal ARs and the C-terminal capping AR. Mutations involving charged residues were found to have large effects on stability. Due to the difference in charge distribution in the internal ARs of E3_19 and E3_5, their interaction with the C-terminal capping AR is less favorable in E3_19. The simulation trajectories suggest that the stability of the designed AR proteins can be increased by optimizing the electrostatic interactions within and between the different repeats.

Tyrosine phosphatase PTP1B interacts with TRPV6 in vivo and plays a role in TRPV6-mediated calcium influx in HEK293 cells.

This study investigates the role of tyrosine phosphorylation and dephosphorylation in the regulation of the Ca(2+) permeant TRPV6 channel. HEK293 cells co-transfected with TRPV6 and the tyrosine phosphatase PTP1B show a constitutive Ca(2+) entry which was independent of tyrosine phosphorylation under resting conditions. Following depletion of intracellular Ca(2+) stores, TRPV6-mediated Ca(2+) entry could be increased in the presence of a tyrosine phosphatase inhibitor (bis-(N,N-dimethyl-hydroxamido) hydroxo-vanadate; DMHV). Inhibition of Src-kinases completely abolished DMHV-induced increase in TRPV6-mediated Ca(2+) influx. Co-transfection with Src led to tyrosine phosphorylation of TRPV6 which could be dephosphorylated by PTP1B. In vivo interaction of TRPV6 with PTP1B was visualized using the bimolecular fluorescence complementation (BiFC) method and proved by co-immunoprecipitation of both proteins. These data indicate that tyrosine phosphorylation is involved in the regulation of the TRPV6 channel protein.

NMR structure of the apoptosis- and inflammation-related NALP1 pyrin domain.

Signaling in apoptosis and inflammation is often mediated by proteins of the death domain superfamily in the Fas/FADD/Caspase-8 or the Apaf-1/Caspase-9 pathways. This superfamily currently comprises the death domain (DD), death effector domain (DED), caspase recruitment domain (CARD), and pyrin domain (PYD) subfamilies. The PYD subfamily is most abundant, but three-dimensional structures are only available for the subfamilies DD, DED, and CARD, which have an antiparallel arrangement of six alpha helices as common fold. This paper presents the NMR structure of PYD of NALP1, a protein that is involved in the innate immune response and is a component of the inflammasome. The structure of NALP1 PYD differs from all other known death domain superfamily structures in that the third alpha helix is replaced by a flexibly disordered loop. This unique feature appears to relate to the molecular basis of familial Mediterranean fever (FMF), a genetic disease caused by single-point mutations.

[Inguinal hernia operation and fertility in men]. / Operation for ingvinalhernie og fertilitet hos mænd.

It is still unknown whether or not inguinal hernia repair with or without the use of mesh causes infertility in male patients. The objective of this article was to discuss the available literature, to ascertain if the preclinical and clinical findings point towards a causal relationship. The review showed that the question remains difficult to answer due to a limited number of patients included and the discordant conclusions drawn in past studies. The largest recent cohort study came to the conclusion that there was no clinically significant risk of infertility associated with inguinal hernia repair. However, further studies are needed.

Identification of a basic surface area of the FADD death effector domain critical for apoptotic signaling.

Death effector domains (DEDs) are protein-protein interaction domains found in the death inducing signaling complex (DISC). Performing a structure-based alignment of all DED sequences we identified a region of high diversity in alpha-helix 3 and propose a classification of DEDs into class I DEDs typically containing a stretch of basic residues in the alpha-helix 3 region whereas DEDs of class II do not. Functional assays using mutants of Fas-associated death domain revealed that this basic region influences binding and recruitment of caspase-8 and cellular FLICE inhibitor protein to the DISC.

Fire and death: the pyrin domain joins the death-domain superfamily.

Apoptosis and inflammation are important cellular processes that are highly regulated through specific protein-protein interactions (PPI). Proteins involved in these signaling cascades often carry PPI domains that belong to the death-domain superfamily. This includes the structurally well-characterized Death Domain (DD), the Death Effector Domain (DED) and the Caspase Recruitment Domain (CARD) subfamilies. Recently, a fourth member of the DD superfamily was identified, the Pyrin Domain (PYD). Based on sequence alignments, homology to other domains occurring in death-signalling pathways, and secondary-structure prediction, the PYD was predicted to have an overall fold similar to other DD superfamily members. Just recently, NMR structures of two PYDs have been determined. The PYD structures not only revealed the DD superfamily fold as previously predicted, but also distinct features that are characteristic exclusively for this subfamily. This review summarizes recent findings and developments regarding structural aspects of the DD superfamily, with a special emphasis on the PPIs of the DD superfamily.

Crystal structure of a divalent metal ion transporter CorA at 2.9 angstrom resolution.

CorA family members are ubiquitously distributed transporters of divalent metal cations and are considered to be the primary Mg2+ transporter of Bacteria and Archaea. We have determined a 2.9 angstrom resolution structure of CorA from Thermotoga maritima that reveals a pentameric cone-shaped protein. Two potential regulatory metal binding sites are found in the N-terminal domain that bind both Mg2+ and Co2+. The structure of CorA supports an efflux system involving dehydration and rehydration of divalent metal ions potentially mediated by a ring of conserved aspartate residues at the cytoplasmic entrance and a carbonyl funnel at the periplasmic side of the pore.

Intracellular kinase inhibitors selected from combinatorial libraries of designed ankyrin repeat proteins.

The specific intracellular inhibition of protein activity at the protein level allows the determination of protein function in the cellular context. We demonstrate here the use of designed ankyrin repeat proteins as tailor-made intracellular kinase inhibitors. The target was aminoglycoside phosphotransferase (3')-IIIa (APH), which mediates resistance to aminoglycoside antibiotics in pathogenic bacteria and shares structural homology with eukaryotic protein kinases. Combining a selection and screening approach, we isolated 198 potential APH inhibitors from highly diverse combinatorial libraries of designed ankyrin repeat proteins. A detailed analysis of several inhibitors revealed that they bind APH with high specificity and with affinities down to the subnanomolar range. In vitro, the most potent inhibitors showed complete enzyme inhibition, and in vivo, a phenotype comparable with the gene knockout was observed, fully restoring antibiotic sensitivity in resistant bacteria. These results underline the great potential of designed ankyrin repeat proteins for modulation of intracellular protein function.

Designed to be stable: crystal structure of a consensus ankyrin repeat protein.

Ankyrin repeat (AR) proteins mediate innumerable protein-protein interactions in virtually all phyla. This finding suggested the use of AR proteins as designed binding molecules. Based on sequence and structural analyses, we designed a consensus AR with fixed framework and randomized interacting residues. We generated several combinatorial libraries of AR proteins consisting of defined numbers of this repeat. Randomly chosen library members are expressed in soluble form in the cytoplasm of Escherichia coli constituting up to 30% of total cellular protein and show high thermodynamic stability. We determined the crystal structure of one of those library members to 2.0-A resolution, providing insight into the consensus AR fold. Besides the highly complementary hydrophobic repeat-repeat interfaces and the absence of structural irregularities in the consensus AR protein, the regular and extended hydrogen bond networks in the beta-turn and loop regions are noteworthy. Furthermore, all residues found in the turn region of the Ramachandran plot are glycines. Many of these features also occur in natural AR proteins, but not in this rigorous and standardized fashion. We conclude that the AR domain fold is an intrinsically very stable and well-expressed scaffold, able to display randomized interacting residues. This scaffold represents an excellent basis for the design of novel binding molecules.