Lack of efficacy of mono-mode of action therapeutics in COVID-19 therapy - How the lack of predictive power of preclinical cell and animal studies leads developments astray.

The diverse experiences regarding the failure of tested drugs in the fight against COVID-19 made it clear that one should at least question the requirement to apply classical preclinical development strategies that demand cell and animal efficacy models to be tested before going into clinical trials. Most animals are not susceptible to infection with SARS-CoV-2, and so this led to one-sided virus replication experiments in cells and the use of animal models that have little in common with the complex pathogenesis of COVID-19 in humans. Therefore, non-clinical development strategies were designed to meet regulatory requirements, but they did not truly reflect the situation in the clinic. This has led the search for effective agents astray in many cases. As proof of this statement, we now bring together the results of such required preclinical experiments and compare with the results in clinical trials. Two clear conclusions that can be drawn from the experience to date: The required preclinical models are unsuitable for the development of innovative treatments medical devices in the case of COVID-19 and mono-action strategies (e.g. direct antivirals) are of very little or no benefit to patients under randomized,blinded conditions. Our hypothesis is that the complex situation of COVID-19 may benefit from multi-mode drugs. Here, the molecular class of aptamers could be a solution.

Genome-wide study of a Neolithic Wartberg grave community reveals distinct HLA variation and hunter-gatherer ancestry.

The Wartberg culture (WBC, 3500-2800 BCE) dates to the Late Neolithic period, a time of important demographic and cultural transformations in western Europe. We performed genome-wide analyses of 42 individuals who were interred in a WBC collective burial in Niedertiefenbach, Germany (3300-3200 cal. BCE). The results showed that the farming population of Niedertiefenbach carried a surprisingly large hunter-gatherer ancestry component (34-58%). This component was most likely introduced during the cultural transformation that led to the WBC. In addition, the Niedertiefenbach individuals exhibited a distinct human leukocyte antigen gene pool, possibly reflecting an immune response that was geared towards detecting viral infections.

A new approach to the temporal significance of house orientations in European Early Neolithic settlements.

This paper shows that local differences in house orientation in settlements from the Early Neolithic in Central Europe reflect a regular chronological trajectory based on Bayesian calibration of 14C-series. This can be used to extrapolate the dating of large-scale settlement plans derived from, among other methods, geophysical surveys. In the southwest Slovakian settlement of Vráble, we observed a progressive counter-clockwise rotation in house orientation from roughly 32° to 4° over a 300 year period. A survey of published and dated village plans from other LBK regions confirms that this counter-clockwise rotation per settlement is a wider Central European trend. We explain this observation as an unintentional, unconscious but systematic leftward deviation in the house builders' cardinal orientation, which has been termed "pseudoneglect" in studies of human perception. This means that whenever houses were intended to be oriented towards a specific direction and be parallel to each other, there was an error in perception causing slight counter-clockwise rotation. This observation is used as a basis to reconstruct dynamics of Early Neolithic settlement in the Slovakian Zitava valley, showing a rapid colonization, followed by increased agglomeration into large villages consisting of strongly autonomous farmsteads.

[Radical prostatectomy through the years]. / Die radikale Prostatektomie im Wandel der Zeit.

The operative aspect of radical prostatectomy has changed dramatically in the past 200 years as significant technological advances have been made, particularly during the past 50 years. The work of Dr. Walsh in the late 1970 s and early 1980 s led to a significant reduction in surgical morbidity and is considered an important milestone of radical prostatectomy, as is the introduction of minimally-invasive (robotic-assisted) surgical techniques. Yet there is no absolute gold standard regarding surgical approaches. Innovative tools, e. g. the addition of "augmented reality", are currently under investigation. This review article for the anniversary issue of "Der Urologe" aims to cover the milestones of the evolution of this "signature" surgery in the field of urology.

The Tumor Suppressor SASH1 Interacts With the Signal Adaptor CRKL to Inhibit Epithelial-Mesenchymal Transition and Metastasis in Colorectal Cancer.

Background & Aims: The tumor-suppressor sterile α motif- and Src-homology 3-domain containing 1 (SASH1) has clinical relevance in colorectal carcinoma and is associated specifically with metachronous metastasis. We sought to identify the molecular mechanisms linking decreased SASH1 expression with distant metastasis formation. Methods: SASH1-deficient, SASH1-depleted, or SASH1-overexpressing HCT116 colon cancer cells were generated by the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated 9-method, RNA interference, and transient plasmid transfection, respectively. Epithelial-mesenchymal transition (EMT) was analyzed by quantitative reverse-transcription polymerase chain reaction, immunoblotting, immunofluorescence microscopy, migration/invasion assays, and 3-dimensional cell culture. Yeast 2-hybrid assays and co-immunoprecipitation/mass-spectrometry showed V-Crk avian sarcoma virus CT10 oncogene homolog-like (CRKL) as a novel interaction partner of SASH1, further confirmed by domain mapping, site-directed mutagenesis, co-immunoprecipitation, and dynamic mass redistribution assays. CRKL-deficient cells were generated in parental or SASH1-deficient cells. Metastatic capacity was analyzed with an orthotopic mouse model. Expression and significance of SASH1 and CRKL for survival and response to chemotherapy was assessed in patient samples from our department and The Cancer Genome Atlas data set. Results: SASH1 expression is down-regulated during cytokine-induced EMT in cell lines from colorectal, pancreatic, or hepatocellular cancer, mediated by the putative SASH1 promoter. Deficiency or knock-down of SASH1 induces EMT, leading to an aggressive, invasive phenotype with increased chemoresistance. SASH1 counteracts EMT through interaction with the oncoprotein CRKL, inhibiting CRKL-mediated activation of SRC kinase, which is crucially required for EMT. SASH1-deficient cells form significantly more metastases in vivo, depending entirely on CRKL. Patient tumor samples show significantly decreased SASH1 and increased CRKL expression, associated with significantly decreased overall survival. Patients with increased CRKL expression show significantly worse response to adjuvant chemotherapy. Conclusions: We propose SASH1 as an inhibitor of CRKL-mediated SRC signaling, introducing a potentially druggable mechanism counteracting chemoresistance and metastasis formation.

Can inspiratory muscle training improve weaning outcomes in difficult to wean patients? A protocol for a randomised controlled trial (IMweanT study).

INTRODUCTION: Respiratory muscle dysfunction has been associated with failure to wean from mechanical ventilation. It has therefore been hypothesised that these patients might benefit from inspiratory muscle training (IMT). Evidence, however, is thus far limited to data from small, single-centre studies with heterogeneity in inclusion criteria, training modalities and outcomes. The aim of this study is to evaluate the effects of a novel IMT method on weaning outcomes in selected patients with weaning difficulties. METHODS: This study is designed as a double-blind, parallel-group, randomised controlled superiority trial with 1:1 allocation ratio. Patients with weaning difficulties will be randomly allocated into either an IMT group (intervention) or a sham-IMT group (control). Ninetypatients (45 in each group) will be needed to detect a 28% difference in the proportion of weaning success between groups (estimated difference in primary outcome based on previous studies) with a risk for type I error (α) of 5% and statistical power (1-ß) of 80%. Patients will perform four sets of 6-10 breaths daily against an external load using a tapered flow resistive loading device (POWERbreathe KH2, HaB International, UK). Training intensity in the intervention group will be adjusted to the highest tolerable load. The control group will train against a low resistance that will not be modified during the training period. Training will becontinued until patients are successfully weaned or for a maximum duration of 28 days. Pulmonary and respiratory muscle function, weaning duration, duration of mechanical ventilation, ventilator-free days and length of stay in the intensive care unit will be evaluated as secondary outcomes. Χ2 tests and analysis of covariance with adjustments for baseline values of respective outcomesas covariates will be used to compare results after the intervention period between groups. ETHICS AND DISSEMINATION: Ethics approval was obtained from the local ethical committee (Ethische Commissie Onderzoek UZ/KU Leuven protocol ID: S60516). Results from this randomised controlled trial will be presented at scientific meetings as abstracts for poster or oral presentations and published in peerreviewed journals. TRIAL STATUS: Enrolment into the study have started in August 2017. Data collection and data analysis are expected to be completed in September 2021. TRIAL REGISTRATION NUMBER: NCT03240263.

Mental health benefits of outdoor adventures: Results from two pilot studies.

This paper investigates potential mental health benefits of outdoor and adventure education programs. It is argued that experiences made in successful programs can increase self-efficacy, mindfulness and subjective well-being. Furthermore, programs may reduce feelings of time pressure and mental stress amongst participants. Evidence comes from two pilot studies: In the school project "Crossing the Alps" (Study 1), 14-year-old participants reported an increase in life satisfaction, mindfulness and a decrease in the PSQ Subscale 'demand' after a successful nine-day hike through the German, Austrian, and Italian Alps. In the university project "Friluftsliv" (Study 2) participants scored higher in life satisfaction, happiness, mindfulness, and self-efficacy and lower in perceived stress after having spent eight days in the wilderness of the Norwegian Hardangervidda region, miles away from the next locality. The findings suggest that outdoor education and wilderness programs can foster mental health in youths and young adults.

Infrared camera based thermometry for quality assurance of superficial hyperthermia applicators.

The purpose of this work was to provide a feasible and easy to apply phantom-based quality assurance (QA) procedure for superficial hyperthermia (SHT) applicators by means of infrared (IR) thermography. The VarioCAM hr head (InfraTec, Dresden, Germany) was used to investigate the SA-812, the SA-510 and the SA-308 applicators (all: Pyrexar Medical, Salt Lake City, UT, USA). Probe referencing and thermal equilibrium procedures were applied to determine the emissivity of the muscle-equivalent agar phantom. Firstly, the disturbing potential of thermal conduction on the temperature distribution inside the phantom was analyzed through measurements after various heating times (5-50 min). Next, the influence of the temperature of the water bolus between the SA-812 applicator and the phantom's surface was evaluated by varying its temperature. The results are presented in terms of characteristic values (extremal temperatures, percentiles and effective field sizes (EFS)) and temperature-area-histograms (TAH). Lastly, spiral antenna applicators were compared by the introduced characteristics. The emissivity of the used phantom was found to be ε = 0.91 ± 0.03, the results of both methods coincided. The influence of thermal conduction with regard to heating time was smaller than expected; the EFS of the SA-812 applicator had a size of (68.6 ± 6.7) cm(2), averaged group variances were ±3.0 cm(2). The TAHs show that the influence of the water bolus is mostly limited to depths of <3 cm, yet it can greatly enhance or reduce heat generation in this regime: at a depth of 1 cm, measured maximal temperature rises were 14.5 °C for T Bolus = 30 °C and 8.6 °C for T Bolus = 21 °C, respectively. The EFS was increased, too. The three spiral antenna applicators generated similar heat distributions. Generally, the procedure proved to yield informative insights into applicator characteristics, thus making the application of an IR camera a very useful tool in SHT technical QA.

Evidence of autoinducer-dependent and -independent heterogeneous gene expression in Sinorhizobium fredii NGR234.

Populations of genetically identical Sinorhizobium fredii NGR234 cells differ significantly in their expression profiles of autoinducer (AI)-dependent and AI-independent genes. Promoter fusions of the NGR234 AI synthase genes traI and ngrI showed high levels of phenotypic heterogeneity during growth in TY medium on a single-cell level. However, adding very high concentrations of N-(3-oxooctanoyl-)-l-homoserine lactone resulted in a more homogeneous expression profile. Similarly, the lack of internally synthesized AIs in the background of the NGR234-ΔtraI or the NGR234-ΔngrI mutant resulted in a highly homogenous expression of the corresponding promoter fusions in the population. Expression studies with reporter fusions of the promoter regions of the quorum-quenching genes dlhR and qsdR1 and the type IV pilus gene cluster located on pNGR234b suggested that factors other than AI molecules affect NGR234 phenotypic heterogeneity. Further studies with root exudates and developing Arabidopsis thaliana seedlings provide the first evidence that plant root exudates have strong effects on the heterogeneity of AI synthase and quorum-quenching genes in NGR234. Therefore, plant-released octopine appears to play a key role in modulation of heterogeneous gene expression.

Nasal drug delivery in traditional persian medicine.

BACKGROUND: Over one hundred different pharmaceutical dosage forms have been recorded in literatures of Traditional Persian Medicine among which nasal forms are considerable. OBJECTIVES: This study designed to derive the most often applied nasal dosage forms together with those brief clinical administrations. MATERIALS AND METHODS: In the current study remaining pharmaceutical manuscripts of Persia during 9th to 18th century AD have been studied and different dosage forms related to nasal application of herbal medicines and their therapeutic effects were derived. RESULTS: By searching through pharmaceutical manuscripts of medieval Persia, different nasal dosage forms involving eleven types related to three main groups are found. These types could be derived from powder, solution or liquid and gaseous forms. Gaseous form were classified into fumigation (Bakhoor), vapor bath (Enkebab), inhalation (Lakhlakheh), aroma agents (Ghalieh) and olfaction or smell (Shomoom). Nasal solutions were as drops (Ghatoor), nasal snuffing drops (Saoot) and liquid snuff formulations (Noshoogh). Powders were as nasal insufflation or snorting agents (Nofookh) and errhine or sternutator medicine (Otoos). Nasal forms were not applied only for local purposes. Rather systemic disorders and specially CNS complications were said to be a target for these dosage forms. DISCUSSION: While this novel type of drug delivery is known as a suitable substitute for oral and parenteral administration, it was well accepted and extensively mentioned in Persian medical and pharmaceutical manuscripts and other traditional systems of medicine as well. Accordingly, medieval pharmaceutical standpoints on nasal dosage forms could still be an interesting subject of study. Therefore, the current work can briefly show the pharmaceutical knowledge on nasal formulations in medieval Persia and clarify a part of history of traditional Persian pharmacy.

Emerging genetic patterns of the European Neolithic: perspectives from a late Neolithic Bell Beaker burial site in Germany.

The transition from hunting and gathering to agriculture in Europe is associated with demographic changes that may have shifted the human gene pool of the region as a result of an influx of Neolithic farmers from the Near East. However, the genetic composition of populations after the earliest Neolithic, when a diverse mosaic of societies that had been fully engaged in agriculture for some time appeared in central Europe, is poorly known. At this period during the Late Neolithic (ca. 2,800-2,000 BC), regionally distinctive burial patterns associated with two different cultural groups emerge, Bell Beaker and Corded Ware, and may reflect differences in how these societies were organized. Ancient DNA analyses of human remains from the Late Neolithic Bell Beaker site of Kromsdorf, Germany showed distinct mitochondrial haplotypes for six individuals, which were classified under the haplogroups I1, K1, T1, U2, U5, and W5, and two males were identified as belonging to the Y haplogroup R1b. In contrast to other Late Neolithic societies in Europe emphasizing maintenance of biological relatedness in mortuary contexts, the diversity of maternal haplotypes evident at Kromsdorf suggests that burial practices of Bell Beaker communities operated outside of social norms based on shared maternal lineages. Furthermore, our data, along with those from previous studies, indicate that modern U5-lineages may have received little, if any, contribution from the Mesolithic or Neolithic mitochondrial gene pool.

Dopamine D4 receptor exon III genotype influence on the auditory evoked novelty P3.

The functional implications of the dopamine D4 receptor gene (DRD4) exon III polymorphism and its role in the modulation of temperament and in the pathophysiology of psychiatric disorders are still a matter of debate. Based on evidence from animal studies, we hypothesised that this polymorphism is involved in the modulation of the cortical response to novelty as reflected by the auditory evoked novelty P3 event-related potential. In a sample of 46 healthy volunteers, we observed an interactive effect of DRD4 exon III genotype and the eye-blink rate, a measure of central dopaminergic activity, on the novelty P3. These findings suggest that the DRD4 exon III polymorphism influences the processing of novelty and that this influence depends on tonic dopaminergic activity.