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Spektroskopische Qualifizierung und Quantifizierung von Hyaluronsäure in Nahrungsergänzungsmitteln
(2023)
Trueness and precision of milled and 3D printed root-analogue implants: A comparative in vitro study
(2023)
Das Interesse an Virtual Reality (VR) für die Hochschullehre steigt aktuell vermehrt durch die Möglichkeit, logistisch schwierige Aufgaben abzubilden sowie aufgrund positiver Ergebnisse aus Wirksamkeitsstudien. Gleichzeitig fehlt es jedoch an Studien, die immersive VR-Umgebungen, nicht-immersive Desktop-Umgebungen und konventionelle Lernmaterialien gegenüberstellen und lehr-lernmethodische Aspekte evaluieren. Aus diesem Grund beschäftigt sich dieser Beitrag mit der Konzeption und Realisierung einer Lernumgebung für die Hochschullehre, die sowohl mit einem Head Mounted Display (HMD) als auch mittels Desktops genutzt werden kann, sowie deren Evaluation anhand eines experimentellen Gruppendesigns. Die Lernumgebung wurde auf Basis einer eigens entwickelten Softwareplattform erstellt und die Wirksamkeit mithilfe von zwei Experimentalgruppen – VR vs. Desktop-Umgebung – und einer Kontrollgruppe evaluiert und verglichen. In einer Pilotstudie konnten sowohl qualitativ als auch quantitativ positive Einschätzungen der Usability der Lernumgebung in beiden Experimentalgruppen herausgestellt werden. Darüber hinaus zeigten sich positive Effekte auf die kognitive und affektive Wirkung der Lernumgebung im Vergleich zu konventionellen Lernmaterialien. Unterschiede zwischen der Nutzung als VR- oder Desktop-Umgebung zeigen sich auf kognitiver und affektiver Ebene jedoch kaum. Die Analyse von Log-Daten deutet allerdings auf Unterschiede im Lern- und Explorationsverhalten hin.
There are several recent works which had proposed an automatic computer-aided diagnosis (CAD) deep learning (DL) model to diagnose coronavirus disease 2019 (COVID-19) using chest x-ray images (CXR) to propose a high-accuracy CAD method to detect COVID-19 automatically. In this study, seven different models including Convolutional Neural Network (CNN) models such as VGG-16 and vision transformer (ViT) models, are proposed. The different proposed models are trained with a three-class balanced dataset consisting of 3,000 CXR images consisting of 1,000 CXR images for each class of COVID-19, Normal, and Lung-Opacity. A publicly available dataset to train and test the models is used from Kaggle-COVID-19-Radiography-Dataset. From the experiments, the accuracy of the VGG16 model is 93.44% and ViT's is 92.33%. Besides, the binary classification between two classes of COVID-19 and Normal CXR with a limited number of just 100 images for each class, using a transfer learning technique, with a validation accuracy of 97.5% is proposed.
Climate change is transforming the risks individuals and households face, with potentially profound socioeconomic consequences such as increased poverty, inequality, and social instability. Social protection is a policy tool that governments use to help individuals and households manage risks linked to income and livelihoods, and to achieve societal outcomes such as reducing poverty and inequality. Despite its potential as a policy response to climate change, the integration of social protection within the climate policy agenda is currently limited. While the concept of risk is key to both sectors, different understandings of the nature and scope of climate change impacts and their implications, as well as of the adequacy of social protection instruments to address them, contribute to the lack of policy and practice integration.
Our goal is to bridge this cognitive gap by highlighting the potential of social protection as a policy response to climate change. Using a comprehensive climate risk lens, we first explore how climate change drives risks that are within the realm of social protection, and their implications, including likely future trends in demand for social protection. Based on this analysis, we critically review existing arguments for what social protection can do and evidence of what it currently does to manage risks arising from climate change. From the analysis, a set of reconceptualised roles emerge for social protection to strategically contribute to climate-resilient development.
Background: Bloodstream infections (BSIs) remain a significant cause of mortality worldwide. Causative pathogens are routinely identified and susceptibility tested but only very rarely investigated for their resistance genes, virulence factors, and clonality. Our aim was to gain insight into the clonality patterns of different species causing BSI and the clinical relevance of distinct virulence genes.
Methods: For this study, we whole-genome-sequenced over 400 randomly selected important pathogens isolated from blood cultures in our diagnostic department between 2016 and 2021. Genomic data on virulence factors, resistance genes, and clonality were cross-linked with in-vitro data and demographic and clinical information.
Results: The investigation yielded extensive and informative data on the distribution of genes implicated in BSI as well as on the clonality of isolates across various species.
Conclusion: Associations between survival outcomes and the presence of specific genes must be interpreted with caution, and conducting replication studies with larger sample sizes for each species appears mandatory. Likewise, a deeper knowledge of virulence and host factors will aid in the interpretation of results and might lead to more targeted therapeutic and preventive measures. Monitoring transmission dynamics more efficiently holds promise to serve as a valuable tool in preventing in particular BSI caused by nosocomial pathogens.
Background: the potency of drugs that interfere with glucose metabolism, i.e., glucose transporters (GLUT) and nicotinamide phosphoribosyltransferase (NAMPT) was analyzed in neuroendocrine tumor (NET, BON-1, and QPG-1 cells) and small cell lung cancer (SCLC, GLC-2, and GLC-36 cells) tumor cell lines. (2) Methods: the proliferation and survival rate of tumor cells was significantly affected by the GLUT-inhibitors fasentin and WZB1127, as well as by the NAMPT inhibitors GMX1778 and STF-31. (3) Results: none of the NET cell lines that were treated with NAMPT inhibitors could be rescued with nicotinic acid (usage of the Preiss–Handler salvage pathway), although NAPRT expression could be detected in two NET cell lines. We finally analyzed the specificity of GMX1778 and STF-31 in NET cells in glucose uptake experiments. As previously shown for STF-31 in a panel NET-excluding tumor cell lines, both drugs specifically inhibited glucose uptake at higher (50 μM), but not at lower (5 μM) concentrations. (4) Conclusions: our data suggest that GLUT and especially NAMPT inhibitors are potential candidates for the treatment of NET tumors.
Climate change is increasingly affecting vulnerable groups and resulting in dire social and economic consequences, especially for those in the Global South. Managing current and emerging climate-related risks will require increasing individual’s and communities’ resilience, including enhancing absorptive, adaptive, and transformative capacities. Policymakers are now considering the role that social protection policies and programmes can play in building climate resilience by contributing to these capacities. However, there is a limited understanding of the extent to which social protection instruments can influence these three resilience-related capacities. Lack of assessment tools or frameworks might contribute to limited evidence of social protection’s ability to increase climate resilience. In particular, there appear to be no frameworks or tools that help assess the role of social cash transfers (SCT) in building adaptive capacity. Based on a multi-staged literature review, we develop an adaptive capacity outcomes framework (ACOF) that can help assess SCT’s contribution to building adaptive capacity, and, consequently, resilience. The framework is then tested using impact evaluation and assessment reports from SCT programmes in Indonesia, Zambia, Ethiopia, Bangladesh, and Tanzania. The exercise finds that SCTs alone have a limited contribution to adaptive capacity outcomes, but interventions that combine cash transfers with other components such as nutrition or livelihood training show positive impacts. We find that the ACOF can support assessments of SCT’s contribution towards adaptive capacity. It can help build evidence, evaluate impacts, and through further research, can facilitate learning on SCTs' role in increasing climate resilience.
In the design of robot skills, the focus generally lies on increasing the flexibility and reliability of the robot execution process; however, typical skill representations are not designed for analysing execution failures if they occur or for explicitly learning from failures. In this paper, we describe a learning-based hybrid representation for skill parameterisation called an execution model, which considers execution failures to be a natural part of the execution process. We then (i) demonstrate how execution contexts can be included in execution models, (ii) introduce a technique for generalising models between object categories by combining generalisation attempts performed by a robot with knowledge about object similarities represented in an ontology, and (iii) describe a procedure that uses an execution model for identifying a likely hypothesis of a parameterisation failure. The feasibility of the proposed methods is evaluated in multiple experiments performed with a physical robot in the context of handle grasping, object grasping, and object pulling. The experimental results suggest that execution models contribute towards avoiding execution failures, but also represent a first step towards more introspective robots that are able to analyse some of their execution failures in an explicit manner.
A biodegradable blend of PBAT—poly(butylene adipate-co-terephthalate)—and PLA—poly(lactic acid)—for blown film extrusion was modified with four multi-functional chain extending cross-linkers (CECL). The anisotropic morphology introduced during film blowing affects the degradation processes. Given that two CECL increased the melt flow rate (MFR) of tris(2,4-di-tert-butylphenyl)phosphite (V1) and 1,3-phenylenebisoxazoline (V2) and the other two reduced it (aromatic polycarbodiimide (V3) and poly(4,4-dicyclohexylmethanecarbodiimide) (V4)), their compost (bio-)disintegration behavior was investigated. It was significantly altered with respect to the unmodified reference blend (REF). The disintegration behavior at 30 and 60 °C was investigated by determining changes in mass, Young’s moduli, tensile strengths, elongations at break and thermal properties. In order to quantify the disintegration behavior, the hole areas of blown films were evaluated after compost storage at 60 °C to calculate the kinetics of the time dependent degrees of disintegration. The kinetic model of disintegration provides two parameters: initiation time and disintegration time. They quantify the effects of the CECL on the disintegration behavior of the PBAT/PLA compound. Differential scanning calorimetry (DSC) revealed a pronounced annealing effect during storage in compost at 30 °C, as well as the occurrence of an additional step-like increase in the heat flow at 75 °C after storage at 60 °C. The disintegration consists of processes which affect amorphous and crystalline phase of PBAT in different manner that cannot be understood by a hydrolytic chain degradation only. Furthermore, gel permeation chromatography (GPC) revealed molecular degradation only at 60 °C for the REF and V1 after 7 days of compost storage. The observed losses of mass and cross-sectional area seem to be attributed more to mechanical decay than to molecular degradation for the given compost storage times.
Citizen participation is deemed to be crucial for sustainability and resilience planning. However, generational equity has been missing from recent academic discussions regarding sustainability and resilience. Therefore, the purpose of this paper is to reintroduce the topic of the existence or absence of an intergenerational consensus on the example of a rural community and its perceived brand image attributes and development priorities. The research is based on primary data collected through an online survey, with a sample size of N = 808 respondents in Neunkirchen-Seelscheid, Germany. The data were analyzed using the Kruskal–Wallis test for the presence and/or absence of consensus among the five generations regarding brand image attributes and development priorities. The findings point to divergence between what the median values indicate as the most relevant brand image attributes and development priorities among the citizens and the areas where the Kruskal–Wallis test shows that an intergenerational consensus either does or does not exist. The results imply the need for new concepts and applied approaches to citizen participation for sustainability and resilience, where intergenerational dialogue and equity-building take center stage. In addition to the importance of the theory of citizen participation for sustainability and resilience, our results provide ample evidence for how sustainability and resilience planning documents could potentially benefit from deploying the concept of intergenerational equity. The present research provides sustainability and political science with new conceptual and methodological approaches for taking intergenerational equity into account in regional planning processes in rural and other areas.
Rosenbrock–Wanner methods for systems of stiff ordinary differential equations are well known since the seventies. They have been continuously developed and are efficient for differential-algebraic equations of index-1, as well. Their disadvantage that the Jacobian matrix has to be updated in every time step becomes more and more obsolete when automatic differentiation is used. Especially the family of Rodas methods has proven to be a standard in the Julia package DifferentialEquations. However, the fifth-order Rodas5 method undergoes order reduction for certain problem classes. Therefore, the goal of this paper is to compute a new set of coefficients for Rodas5 such that this order reduction is reduced. The procedure is similar to the derivation of the methods Rodas4P and Rodas4P2. In addition, it is possible to provide new dense output formulas for Rodas5 and the new method Rodas5P. Numerical tests show that for higher accuracy requirements Rodas5P always belongs to the best methods within the Rodas family.
The non-filarial and non-communicable disease podoconiosis affects around 4 million people and is characterized by severe leg lymphedema accompanied with painful intermittent acute inflammatory episodes, called acute dermatolymphangioadenitis (ADLA) attacks. Risk factors have been associated with the disease but the mechanisms of pathophysiology remain uncertain. Lymphedema can lead to skin lesions, which can serve as entry points for bacteria that may cause ADLA attacks leading to progression of the lymphedema. However, the microbiome of the skin of affected legs from podoconiosis individuals remains unclear. Thus, we analysed the skin microbiome of podoconiosis legs using next generation sequencing. We revealed a positive correlation between increasing lymphedema severity and non-commensal anaerobic bacteria, especially Anaerococcus provencensis, as well as a negative correlation with the presence of Corynebacterium, a constituent of normal skin flora. Disease symptoms were generally linked to higher microbial diversity and richness, which deviated from the normal composition of the skin. These findings show an association of distinct bacterial taxa with lymphedema stages, highlighting the important role of bacteria for the pathogenesis of podoconiosis and might enable a selection of better treatment regimens to manage ADLA attacks and disease progression.
The purpose of this study is to extend previous research on brand innovation by uncovering the process of family winery branding in relation to the new product launch in the VUCA market on the case of three Serbian wineries. The study deploys qualitative oriented and empirical approach in presenting a multi-case study. Three semi-structured telephone interviews were conducted with owners and/or managers in these three wineries. The results demonstrate that all three family wineries are offering high-end product for the domestic market with smaller one still experimenting with strategic direction of innovating for high-end market while the two larger ones putting focus either on autochthonous grape varieties with eye-cathicng labels or authentic brand identity with strong storytelling. Another important aspect identified is the frugal nature of product launch in the family wineries due to limited resources. The paper presents is among only few studies on new product development in wine business literature.
The perceptual upright results from the multisensory integration of the directions indicated by vision and gravity as well as a prior assumption that upright is towards the head. The direction of gravity is signalled by multiple cues, the predominant of which are the otoliths of the vestibular system and somatosensory information from contact with the support surface. Here, we used neutral buoyancy to remove somatosensory information while retaining vestibular cues, thus "splitting the gravity vector" leaving only the vestibular component. In this way, neutral buoyancy can be used as a microgravity analogue. We assessed spatial orientation using the oriented character recognition test (OChaRT, which yields the perceptual upright, PU) under both neutrally buoyant and terrestrial conditions. The effect of visual cues to upright (the visual effect) was reduced under neutral buoyancy compared to on land but the influence of gravity was unaffected. We found no significant change in the relative weighting of vision, gravity, or body cues, in contrast to results found both in long-duration microgravity and during head-down bed rest. These results indicate a relatively minor role for somatosensation in determining the perceptual upright in the presence of vestibular cues. Short-duration neutral buoyancy is a weak analogue for microgravity exposure in terms of its perceptual consequences compared to long-duration head-down bed rest.
Solar photovoltaic power output is modulated by atmospheric aerosols and clouds and thus contains valuable information on the optical properties of the atmosphere. As a ground-based data source with high spatiotemporal resolution it has great potential to complement other ground-based solar irradiance measurements as well as those of weather models and satellites, thus leading to an improved characterisation of global horizontal irradiance. In this work several algorithms are presented that can retrieve global tilted and horizontal irradiance and atmospheric optical properties from solar photovoltaic data and/or pyranometer measurements. The method is tested on data from two measurement campaigns that took place in the Allgäu region in Germany in autumn 2018 and summer 2019, and the results are compared with local pyranometer measurements as well as satellite and weather model data. Using power data measured at 1 Hz and averaged to 1 min resolution along with a non-linear photovoltaic module temperature model, global horizontal irradiance is extracted with a mean bias error compared to concurrent pyranometer measurements of 5.79 W m−2 (7.35 W m−2) under clear (cloudy) skies, averaged over the two campaigns, whereas for the retrieval using coarser 15 min power data with a linear temperature model the mean bias error is 5.88 and 41.87 W m−2 under clear and cloudy skies, respectively.
During completely overcast periods the cloud optical depth is extracted from photovoltaic power using a lookup table method based on a 1D radiative transfer simulation, and the results are compared to both satellite retrievals and data from the Consortium for Small-scale Modelling (COSMO) weather model. Potential applications of this approach for extracting cloud optical properties are discussed, as well as certain limitations, such as the representation of 3D radiative effects that occur under broken-cloud conditions. In principle this method could provide an unprecedented amount of ground-based data on both irradiance and optical properties of the atmosphere, as long as the required photovoltaic power data are available and properly pre-screened to remove unwanted artefacts in the signal. Possible solutions to this problem are discussed in the context of future work.
Trust your guts: fostering embodied knowledge and sustainable practices through voice interaction
(2023)
Despite various attempts to prevent food waste and motivate conscious food handling, household members find it difficult to correctly assess the edibility of food. With the rise of ambient voice assistants, we did a design case study to support households’ in situ decision-making process in collaboration with our voice agent prototype, Fischer Fritz. Therefore, we conducted 15 contextual inquiries to understand food practices at home. Furthermore, we interviewed six fish experts to inform the design of our voice agent on how to guide consumers and teach food literacy. Finally, we created a prototype and discussed with 15 consumers its impact and capability to convey embodied knowledge to the human that is engaged as sensor. Our design research goes beyond current Human-Food Interaction automation approaches by emphasizing the human-food relationship in technology design and demonstrating future complementary human-agent collaboration with the aim to increase humans’ competence to sense, think, and act.
The cystic fibrosis transmembrane conductance regulator (CFTR) anion channel and the epithelial Na+ channel (ENaC) play essential roles in transepithelial ion and fluid transport in numerous epithelial tissues. Inhibitors of both channels have been important tools for defining their physiological role in vitro. However, two commonly used CFTR inhibitors, CFTRinh-172 and GlyH-101, also inhibit non-CFTR anion channels, indicating they are not CFTR specific. However, the potential off-target effects of these inhibitors on epithelial cation channels has to date not been addressed. Here, we show that both CFTR blockers, at concentrations routinely employed by many researchers, caused a significant inhibition of store-operated calcium entry (SOCE) that was time-dependent, poorly reversible and independent of CFTR. Patch clamp experiments showed that both CFTRinh-172 and GlyH-101 caused a significant block of Orai1-mediated whole cell currents, establishing that they likely reduce SOCE via modulation of this Ca2+ release-activated Ca2+ (CRAC) channel. In addition to off-target effects on calcium channels, both inhibitors significantly reduced human αβγ-ENaC-mediated currents after heterologous expression in Xenopus oocytes, but had differential effects on δβγ-ENaC function. Molecular docking identified two putative binding sites in the extracellular domain of ENaC for both CFTR blockers. Together, our results indicate that caution is needed when using these two CFTR inhibitors to dissect the role of CFTR, and potentially ENaC, in physiological processes.
There has been a growing interest in taste research in the HCI and CSCW communities. However, the focus is more on stimulating the senses, while the socio-cultural aspects have received less attention. However, individual taste perception is mediated through social interaction and collective negotiation and is not only dependent on physical stimulation. Therefore, we study the digital mediation of taste by drawing on ethnographic research of four online wine tastings and one self-organized event. Hence, we investigated the materials, associated meanings, competences, procedures, and engagements that shaped the performative character of tasting practices. We illustrate how the tastings are built around the taste-making process and how online contexts differ in providing a more diverse and distributed environment. We then explore the implications of our findings for the further mediation of taste as a social and democratized phenomenon through online interaction.
Background
Consumers rely heavily on online user reviews when shopping online and cybercriminals produce fake reviews to manipulate consumer opinion. Much prior research focuses on the automated detection of these fake reviews, which are far from perfect. Therefore, consumers must be able to detect fake reviews on their own. In this study we survey the research examining how consumers detect fake reviews online.
Methods
We conducted a systematic literature review over the research on fake review detection from the consumer-perspective. We included academic literature giving new empirical data. We provide a narrative synthesis comparing the theories, methods and outcomes used across studies to identify how consumers detect fake reviews online.
Results
We found only 15 articles that met our inclusion criteria. We classify the most often used cues identified into five categories which were (1) review characteristics (2) textual characteristics (3) reviewer characteristics (4) seller characteristics and (5) characteristics of the platform where the review is displayed.
Discussion
We find that theory is applied inconsistently across studies and that cues to deception are often identified in isolation without any unifying theoretical framework. Consequently, we discuss how such a theoretical framework could be developed.
ESKAPEE Pathogen Biofilm Control on Surfaces with Probiotic Lactobacillaceae and Bacillus species
(2023)
Combatting the rapidly growing threat of antimicrobial resistance and reducing prevalence and transmission of ESKAPEE pathogens in healthcare settings requires innovative strategies, one of which is displacing these pathogens using beneficial microorganisms. Our review comprehensively examines the evidence of probiotic bacteria displacing ESKAPEE pathogens, with a focus on inanimate surfaces. A systematic search was conducted using the PubMed and Web of Science databases on 21 December 2021, and 143 studies were identified examining the effects of Lactobacillaceae and Bacillus spp. cells and products on the growth, colonization, and survival of ESKAPEE pathogens. While the diversity of study methods limits evidence analysis, results presented by narrative synthesis demonstrate that several species have the potential as cells or their products or supernatants to displace nosocomial infection-causing organisms in a variety of in vitro and in vivo settings. Our review aims to aid the development of new promising approaches to control pathogen biofilms in medical settings by informing researchers and policymakers about the potential of probiotics to combat nosocomial infections. More targeted studies are needed to assess safety and efficacy of different probiotic formulations, followed by large-scale studies to assess utility in infection control and medical practice.
Isovaleric acidemia (IVA), due to isovaleryl-CoA dehydrogenase (IVD) deficiency, results in the accumulation of isovaleryl-CoA, isovaleric acid and secondary metabolites. The increase in these metabolites decreases mitochondrial energy production and increases oxidative stress. This contributes to the neuropathological features of IVA. A general assumption in the literature exists that glycine N-acyltransferase (GLYAT) plays a role in alleviating the symptoms experienced by IVA patients through the formation of N-isovalerylglycine. GLYAT forms part of the phase II glycine conjugation pathway in the liver and detoxifies excess acyl-CoA’s namely benzoyl-CoA. However, very few studies support GLYAT as the enzyme that conjugates isovaleryl-CoA to glycine. Furthermore, GLYATL1, a paralogue of GLYAT, conjugates phenylacetyl-CoA to glutamine. Therefore, GLYATL1 might also be a candidate for the formation of N-isovalerylglycine. Based on the findings from the literature review, we proposed that GLYAT or GLYATL1 can form N-isovalerylglycine in IVA patients. To test this hypothesis, we performed an in-silico analysis to determine which enzyme is more likely to conjugate isovaleryl-CoA with glycine using AutoDock Vina. Thereafter, we performed in vitro validation using purified enzyme preparations. The in-silico and in vitro findings suggested that both enzymes could form N-isovaleryglycine albeit at lower affinities than their preferred substrates. Furthermore, an increase in glycine concentration does not result in an increase in N-isovalerylglycine formation. The results from the critical literature appraisal, in-silico, and in vitro validation, suggest the importance of further investigating the reaction kinetics and binding behaviors between these substrates and enzymes in understanding the pathophysiology of IVA.
Indoor spaces exhibit microbial compositions that are distinctly dissimilar from one another and from outdoor spaces. Unique in this regard, and a topic that has only recently come into focus, is the microbiome of hospitals. While the benefits of knowing exactly which microorganisms propagate how and where in hospitals are undoubtedly beneficial for preventing hospital-acquired infections, there are, to date, no standardized procedures on how to best study the hospital microbiome. Our study aimed to investigate the microbiome of hospital sanitary facilities, outlining the extent to which hospital microbiome analyses differ according to sample-preparation protocol. For this purpose, fifty samples were collected from two separate hospitals—from three wards and one hospital laboratory—using two different storage media from which DNA was extracted using two different extraction kits and sequenced with two different primer pairs (V1–V2 and V3–V4). There were no observable differences between the sample-preservation media, small differences in detected taxa between the DNA extraction kits (mainly concerning Propionibacteriaceae), and large differences in detected taxa between the two primer pairs V1–V2 and V3–V4. This analysis also showed that microbial occurrences and compositions can vary greatly from toilets to sinks to showers and across wards and hospitals. In surgical wards, patient toilets appeared to be characterized by lower species richness and diversity than staff toilets. Which sampling sites are the best for which assessments should be analyzed in more depth. The fact that the sample processing methods we investigated (apart from the choice of primers) seem to have changed the results only slightly suggests that comparing hospital microbiome studies is a realistic option. The observed differences in species richness and diversity between patient and staff toilets should be further investigated, as these, if confirmed, could be a result of excreted antimicrobials.
In memoriam Willy Lehnert
(2023)
Pitfalls of using sequence databases for heterologous expression studies - a technical review
(2023)
Synthesis of DNA fragments based on gene sequences available in public resources has become an efficient and affordable method that gradually replaced traditional cloning efforts such as PCR cloning from cDNA. However, database entries based on genome sequencing results are prone to errors which can lead to false sequence information and, ultimately, errors in functional characterization of proteins such as ion channels and transporters in heterologous expression systems. We have identified five common problems that repeatedly appear in public resources: 1) Not every gene has yet been annotated; 2) Not all gene annotations are necessarily correct; 3) Transcripts may contain automated corrections; 4) There are mismatches between gene, mRNA, and protein sequences; and 5) Splicing patterns often lack experimental validation. This technical review highlights and provides a strategy to bypass these issues in order to avoid critical mistakes that could impact future studies of any gene/protein of interest in heterologous expression systems. Abstract figure legend Projects involving heterologous gene expression are often characterised by similar steps. Initially, database research (A) is necessary to retrieve information of full of partial sequences of a gene of interest. A multitude of genome assemblies are annotated and deposited in public databases or that are available for refined search options using individual sequence information. The search results need to be scrutinised and compared to already available information (B). Once the sequence has been determined, DNA synthesis (C) by PCR or commercial synthesis are necessary for further cloning procedures (D). Eventually, the DNA needs to be transfected (E) and expressed in, e.g., eukaryotic cells (F). Finally, the expression of the gene of interest needs to be documented and its function analysed (G). This article is protected by copyright. All rights reserved.
Microbiome analyses are essential for understanding microorganism composition and diversity, but interpretation is often challenging due to biological and technical variables. DNA extraction is a critical step that can significantly bias results, particularly in samples containing a high abundance of challenging-to-lyse microorganisms. Taking into consideration the distinctive microenvironments observed in different bodily locations, our study sought to assess the extent of bias introduced by suboptimal bead-beating during DNA extraction across diverse clinical sample types. The question was whether complex targeted extraction methods are always necessary for reliable taxonomic abundance estimation through amplicon sequencing or if simpler alternatives are effective for some sample types. Hence, for four different clinical sample types (stool, cervical swab, skin swab, and hospital surface swab samples), we compared the results achieved from extracting targeted manual protocols routinely used in our research lab for each sample type with automated protocols specifically not designed for that purpose. Unsurprisingly, we found that for the stool samples, manual extraction protocols with vigorous bead-beating were necessary in order to avoid erroneous taxa proportions on all investigated taxonomic levels and, in particular, false under- or overrepresentation of important genera such as Blautia, Faecalibacterium, and Parabacteroides. However, interestingly, we found that the skin and cervical swab samples had similar results with all tested protocols. Our results suggest that the level of practical automation largely depends on the expected microenvironment, with skin and cervical swabs being much easier to process than stool samples. Prudent consideration is necessary when extending the conclusions of this study to applications beyond rough estimations of taxonomic abundance.
The epithelial sodium channel (ENaC) is a key regulator of sodium homeostasis that contributes to blood pressure control. ENaC open probability is adjusted by extracellular sodium ions, a mechanism referred to as sodium self-inhibition (SSI). With a growing number of identified ENaC gene variants associated with hypertension, there is an increasing demand for medium- to high-throughput assays allowing the detection of alterations in ENaC activity and SSI. We evaluated a commercially available automated two-electrode voltage-clamp (TEVC) system that records transmembrane currents of ENaC-expressing Xenopus oocytes in 96-well microtiter plates. We employed guinea pig, human and Xenopus laevis ENaC orthologs that display specific magnitudes of SSI. While demonstrating some limitations over traditional TEVC systems with customized perfusion chambers, the automated TEVC system was able to detect the established SSI characteristics of the employed ENaC orthologs. We were able to confirm a reduced SSI in a gene variant, leading to C479R substitution in the human α-ENaC subunit that has been reported in Liddle syndrome. In conclusion, automated TEVC in Xenopus oocytes can detect SSI of ENaC orthologs and variants associated with hypertension. For precise mechanistic and kinetic analyses of SSI, optimization for faster solution exchange rates is recommended.
Atomic oxygen is a key species in the mesosphere and thermosphere of Venus. It peaks in the transition region between the two dominant atmospheric circulation patterns, the retrograde super-rotating zonal flow below 70 km and the subsolar to antisolar flow above 120 km altitude. However, past and current detection methods are indirect and based on measurements of other molecules in combination with photochemical models. Here, we show direct detection of atomic oxygen on the dayside as well as on the nightside of Venus by measuring its ground-state transition at 4.74 THz (63.2 µm). The atomic oxygen is concentrated at altitudes around 100 km with a maximum column density on the dayside where it is generated by photolysis of carbon dioxide and carbon monoxide. This method enables detailed investigations of the Venusian atmosphere in the region between the two atmospheric circulation patterns in support of future space missions to Venus.
Host-derived succinate accumulates in the airways during bacterial infection. Here, we show that luminal succinate activates murine tracheal brush (tuft) cells through a signaling cascade involving the succinate receptor 1 (SUCNR1), phospholipase Cβ2, and the cation channel transient receptor potential channel subfamily M member 5 (TRPM5). Stimulated brush cells then trigger a long-range Ca2+ wave spreading radially over the tracheal epithelium through a sequential signaling process. First, brush cells release acetylcholine, which excites nearby cells via muscarinic acetylcholine receptors. From there, the Ca2+ wave propagates through gap junction signaling, reaching also distant ciliated and secretory cells. These effector cells translate activation into enhanced ciliary activity and Cl- secretion, which are synergistic in boosting mucociliary clearance, the major innate defense mechanism of the airways. Our data establish tracheal brush cells as a central hub in triggering a global epithelial defense program in response to a danger-associated metabolite.
Estimates of global horizontal irradiance (GHI) from reanalysis and satellite-based data are the most important information for the design and monitoring of PV systems in Africa, but their quality is unknown due to the lack of in situ measurements. In this study, we evaluate the performance of hourly GHI from state-of-the-art reanalysis and satellite-based products (ERA5, CAMS, MERRA-2, and SARAH-2) with 37 quality-controlled in situ measurements from novel meteorological networks established in Burkina Faso and Ghana under different weather conditions for the year 2020. The effects of clouds and aerosols are also considered in the analysis by using common performance measures for the main quality attributes and a new overall performance value for the joint assessment. The results show that satellite data performs better than reanalysis data under different atmospheric conditions. Nevertheless, both data sources exhibit significant bias of more than 150 W/m2 in terms of RMSE under cloudy skies compared to clear skies. The new measure of overall performance clearly shows that the hourly GHI derived from CAMS and SARAH-2 could serve as viable alternative data for assessing solar energy in the different climatic zones of West Africa.
PURPOSE
Cervical cancer (CC) is caused by a persistent high-risk human papillomavirus (hrHPV) infection. The cervico-vaginal microbiome may influence the development of (pre)cancer lesions. Aim of the study was (i) to evaluate the new CC screening program in Germany for the detection of high-grade CC precursor lesions, and (ii) to elucidate the role of the cervico-vaginal microbiome and its potential impact on cervical dysplasia.
METHODS
The microbiome of 310 patients referred to colposcopy was determined by amplicon sequencing and correlated with clinicopathological parameters.
RESULTS
Most patients were referred for colposcopy due to a positive hrHPV result in two consecutive years combined with a normal PAP smear. In 2.1% of these cases, a CIN III lesion was detected. There was a significant positive association between the PAP stage and Lactobacillus vaginalis colonization and between the severity of CC precursor lesions and Ureaplasma parvum.
CONCLUSION
In our cohort, the new cervical cancer screening program resulted in a low rate of additional CIN III detected. It is questionable whether these cases were only identified earlier with additional HPV testing before the appearance of cytological abnormalities, or the new screening program will truly increase the detection rate of CIN III in the long run. Colonization with U. parvum was associated with histological dysplastic lesions. Whether targeted therapy of this pathogen or optimization of the microbiome prevents dysplasia remains speculative.
Stably stratified Taylor–Green vortex simulations are performed by lattice Boltzmann methods (LBM) and compared to other recent works using Navier–Stokes solvers. The density variation is modeled with a separate distribution function in addition to the particle distribution function modeling the flow physics. Different stencils, forcing schemes, and collision models are tested and assessed. The overall agreement of the lattice Boltzmann solutions with reference solutions from other works is very good, even when no explicit subgrid model is used, but the quality depends on the LBM setup. Although the LBM forcing scheme is not decisive for the quality of the solution, the choice of the collision model and of the stencil are crucial for adequate solutions in underresolved conditions. The LBM simulations confirm the suppression of vertical flow motion for decreasing initial Froude numbers. To gain further insight into buoyancy effects, energy decay, dissipation rates, and flux coefficients are evaluated using the LBM model for various Froude numbers.