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The introduction of sulfur into the phosphate linkage of chemically synthesized oligonucleotides creates the stereocenters on phosphorus atoms. Researchers have valued the nature of backbone stereochemistry and early on investigated drug properties for the individual stereocenters in dimers or short oligomers. Only very recently, it has become possible to synthesize fully stereodefined antisense oligonucleotides in good yield and purity. Non-bridging phosphorodithioate (PS2) introduces second sulfur into the phosphorothioate linkage to remove the chirality of phosphorus atom. Here, we describe the application of symmetrical non-bridging PS2 linkages in the context of stereodefined locked nucleic acids (LNAs) antisense oligonucleotides with the goal of reducing chiral complexity and, ultimately, resulting in single molecules. In addition, we propose a rather simple strategy to rapidly identify stereodefined gapmers, combining PS2 and a preferred stereochemistry motif (RSSR), which supports RNase-H-mediated target knockdown. Pharmacological efficacy and metabolic stability are investigated systematically using ApoB as a target sequence, where in vivo data correlate well to what is observed in vitro.

Protein design aims to build novel proteins customized for specific purposes, thereby holding the potential to tackle many environmental and biomedical problems. Here the authors apply some of the latest advances in natural language processing, generative Transformers, to train ProtGPT2, a language model that explores unseen regions of the protein space while designing proteins with nature-like properties.

TRIM4Post-Mining is a H2020/RFCS-funded project that brings together a consortium of European experts from industry and academia to develop an integrated information modelling system. This is designed to support decision making and planning during the transition from coal exploitation to a revitalized post-mining landscape, enabling infrastructure development for agricultural and industrial utilization, and contributing to the recovery of energy and materials from coal mining dumps. The smart system will be founded upon a high-resolution spatiotemporal database, utilizing state-of-the-art multi-scale and multi-sensor monitoring technologies that characterize dynamic processes in coal waste dumps related to timely, dependent deformation and geochemical processes. It will integrate efficient methods for operational and post-mining monitoring, comprehensive spatiotemporal data analytics, feature extraction, and predictive modelling; this will allow for the identification of potential contamination areas and the forecasting of geotechnical risks and ground conditions. For the interactive exploration of alternative land-use planning scenarios—in terms of residual risks, technical feasibility, environmental and social impact, and affordability—up-to-date data and models will be embedded in an interactive planning system based on Virtual Reality and Augmented Reality technology, forming a TRIM—a Transition Information Modelling System. This contribution presents the conceptual approach and main constituents, and describes the state-of-the-art and detailed anticipated methodological approach for each of the constituents. This is supported by the presentation of the first results and a discussion of future work. An anticipated second contribution will focus on the main findings, technology readiness and a discussion of future work.

BackgroundWearable technology is a leading fitness trend in the growing commercial industry and an established method for collecting 24-hour physical behavior data in research studies. High-quality free-living validation studies are required to enable both researchers and consumers to make guided decisions on which study to rely on and which device to use. However, reviews focusing on the quality of free-living validation studies in adults are lacking. ObjectiveThis study aimed to raise researchers’ and consumers’ attention to the quality of published validation protocols while aiming to identify and compare specific consistencies or inconsistencies between protocols. We aimed to provide a comprehensive and historical overview of which wearable devices have been validated for which purpose and whether they show promise for use in further studies. MethodsPeer-reviewed validation studies from electronic databases, as well as backward and forward citation searches (1970 to July 2021), with the following, required indicators were included: protocol must include real-life conditions, outcome must belong to one dimension of the 24-hour physical behavior construct (intensity, posture or activity type, and biological state), the protocol must include a criterion measure, and study results must be published in English-language journals. The risk of bias was evaluated using the Quality Assessment of Diagnostic Accuracy Studies-2 tool with 9 questions separated into 4 domains (patient selection or study design, index measure, criterion measure, and flow and time). ResultsOf the 13,285 unique search results, 222 (1.67%) articles were included. Most studies (153/237, 64.6%) validated an intensity measure outcome such as energy expenditure. However, only 19.8% (47/237) validated biological state and 15.6% (37/237) validated posture or activity-type outcomes. Across all studies, 163 different wearables were identified. Of these, 58.9% (96/163) were validated only once. ActiGraph GT3X/GT3X+ (36/163, 22.1%), Fitbit Flex (20/163, 12.3%), and ActivPAL (12/163, 7.4%) were used most often in the included studies. The percentage of participants meeting the quality criteria ranged from 38.8% (92/237) to 92.4% (219/237). On the basis of our classification tree to evaluate the overall study quality, 4.6% (11/237) of studies were classified as low risk. Furthermore, 16% (38/237) of studies were classified as having some concerns, and 72.9% (173/237) of studies were classified as high risk. ConclusionsOverall, free-living validation studies of wearables are characterized by low methodological quality, large variability in design, and focus on intensity. Future research should strongly aim at biological state and posture or activity outcomes and strive for standardized protocols embedded in a validation framework. Standardized protocols for free-living validation embedded in a framework are urgently needed to inform and guide stakeholders (eg, manufacturers, scientists, and consumers) in selecting wearables for self-tracking purposes, applying wearables in health studies, and fostering innovation to achieve improved validity.

Abstract Background Physical activity (PA) has beneficial effects on health and health-related quality of life (HRQoL), which is a protective factor of illness and mortality. The purpose of this examination was to investigate if self-reported and device-based measures of PA were related to HRQoL in adolescents. Methods Participants (N = 1565; 54.3% female; M age = 14.37 years, SD age = 1.99) were recruited from 167 sample points across Germany. Adolescents self-reported their PA, supplemented by a 1-week examination of device-based PA using accelerometry. Additionally, they completed the multidimensional KIDSCREEN-27 to assess HRQoL. Results Results showed that self-reported PA was correlated with overall HRQoL, Physical Well-Being, Psychological Well-Being, Social Support & Peers, and School Environment, whereas device-based PA was only correlated with Physical as well as Psychological Well-Being. Further, self-reported PA significantly predicted all facets of HRQoL except for Autonomy and Parent Relations, whereas device-based PA solely heightened the amount of explained variance in the Physical Well-Being subscale. Conclusions Findings demonstrate the importance of self-reported PA as it is related to almost all facets of HRQoL. Both measures of PA are not congruent in their relationship with HRQoL and thus implications have to be carefully considered. Future studies should investigate the direct effect of PA on HRQoL and health in a longitudinal approach to account for the causality of effects.

Abstract The prognosis of metastatic melanoma remains poor due to de novo or acquired resistance to immune and targeted therapies. Previous studies have shown that melanoma cells have perturbed metabolism and that cellular metabolic pathways represent potential therapeutic targets. To support the discovery of new drug candidates for melanoma, we examined 180 metabolic modulators, including phytochemicals and anti-diabetic compounds, for their growth-inhibitory activities against melanoma cells, alone and in combination with the BRAF inhibitor vemurafenib. Two positive hits from this screen, 4-methylumbelliferone (4-MU) and ursolic acid (UA), were subjected to validation and further characterization. Metabolic analysis showed that 4-MU affected cellular metabolism through inhibition of glycolysis and enhanced the effect of vemurafenib to reduce the growth of melanoma cells. In contrast, UA reduced mitochondrial respiration, accompanied by an increase in the glycolytic rate. This metabolic switch potentiated the growth-inhibitory effect of the pyruvate dehydrogenase kinase inhibitor dichloroacetate. Both drug combinations led to increased production of reactive oxygen species, suggesting the involvement of oxidative stress in the cellular response. These results support the potential use of metabolic modulators for combination therapies in cancer and may encourage preclinical validation and clinical testing of such treatment strategies in patients with metastatic melanoma.

Childhood and adolescence are important life periods for the development of health status and physical activity (PA) behaviours. This study analyses the stability and potential changes of self-rated health status, overweight and PA behaviour over time, specifically focusing on the age and the socioeconomic status of children and adolescents. We employ representative longitudinal data for German children and adolescents from the Motorik-Modul Study and the German Health Interview and Examination Survey. Using four different dichotomous health status and PA indicators (self-rated health status [SRHS]; overweight; moderate-to-vigorous PA; and leisure sports engagement), we report within-person transition rates across the panel waves when the survey was taken (2003–2006, 2009–2012, and 2014–2017). Additionally, we report results of logistic regressions estimating the impact of children's age, gender, migration background, and their parents' socioeconomic status on these transition rates. The transition rates show mixed results. While children and adolescents from highly problematic states reporting bad SRHS and no leisure sports engagement at an early stage tend to improve later on, overweight children mostly stay overweight. Age and social inequality indicators correlate with some of the chances of improving or worsening the health and PA states. Most clearly, high parental status prevents the health status and PA from worsening over all transitions, particularly becoming overweight, representing a ratchet effect. The results of the present study underline that health policy needs to target specific groups to reduce social inequality in the health status and PA of children and adolescents.

Modern proteins have been shown to share evolutionary relationships via subdomain-sized fragments. The assembly of such fragments through duplication and recombination events led to the complex structures and functions we observe today. We previously implemented a pipeline that identified more than 1,000 of these fragments that are shared by different protein folds and developed a web interface to analyze and search for them. This resource named Fuzzle helps structural and evolutionary biologists to identify and analyze conserved parts of a protein but it also provides protein engineers with building blocks for example to design proteins by fragment combination. Here, we describe a new version of this web resource that was extended to include ligand information. This addition is a significant asset to the database since now protein fragments that bind specific ligands can be identified and analyzed. Often the mode of ligand binding is conserved in proteins thereby supporting a common evolutionary origin. The same can now be explored for subdomain-sized fragments within this database. This ligand binding information can also be used in protein engineering to graft binding pockets into other protein scaffolds or to transfer functional sites via recombination of a specific fragment. Fuzzle 2.0 is freely available at https://fuzzle.uni-bayreuth.de/2.0.

Strong evidence exists that fitness is a physical health resource, which serves to protect one’s health. There is still uncertainty about which fitness level provides the best health outcome and which measurements can be used for analyzing this question. This cross-sectional study analyzed 462 (64.07% female) German middle-aged adults regarding their fitness status, physical activity (Non-Exercise test), body composition (Body Mass Index) and heart-related health status. Motor tests were used to measure the health-related fitness status. The heart-related health status was surveyed by questionnaire and diagnosis was done in part by a physician. Relationships between risk factors and fitness factors are visible during the correlative analysis. They are substantially more visible in the differentiation of people with and without risks. People with low fitness show noticeable risks in activity, Body Mass Index and heart-related health. People with high fitness show health resources for activity, Body Mass Index and heart-related health. This study points out that all fitness dimensions influence one’s heart-related health in a positive way. Fitness is measured objectively and includes all health-related fitness dimensions such as endurance, strength, coordination and flexibility. Apart from this standardization, we ask for more longitudinal studies and more objective health measurements.

The physical activity enjoyment scale (PACES) is a measurement instrument that is commonly used in monitoring and intervention research to assess how much people enjoy being physically active, as this has been related to physical activity adherence. However, while the measurement properties of PACES are well-researched in the English language, there is a gap of research in the German language, especially when looking at adults. Thus, the purpose of this work was to examine reliability, factorial validity, criterion-related validity, and measurement invariance across sex, age groups and time of the PACES for German-speaking adults. Data was obtained from the Motorik-Modul-Study (MoMo) in which 863 adults (53.5% female; mean age = 20.9 years) were examined. To investigate measurement invariance across age groups, data from 2,274 adolescents (50.5% female; mean age = 14.4 years) was obtained additionally. The study provided a nationwide representative sample for Germany. Results showed high internal consistency of PACES in adults (Cronbach's α = .94). Confirmatory factor analyses confirmed the invariance of the measure across age groups, time, and sex. Criterion-related validity could be shown as the global factor significantly correlated with overall physical activity, physical activity in sports clubs, and leisure-time physical activity. The analyses of factorial structure indicated a method effect for positively and negatively worded items. Correlated uniqueness, latent method factor and a hybrid model were applied to analyze the method effect and results indicated that the method effect of positively worded items was predictive of physical activity independently of the global factor. Overall, it can be concluded that PACES is reliable, valid and invariant measure of physical activity enjoyment to be used in German-speaking adults. Further studies are warranted to examine the factorial structure of the PACES and the consequences of the method effect.

Trypanosome brucei, the causative agent of African sleeping sickness, harbours a highly ordered, subpellicular microtubule cytoskeleton that defines many aspects of morphology, motility and virulence. This array of microtubules is associated with a large number of proteins involved in its regulation. Employing proximity-dependent biotinylation assay (BioID) using the well characterised cytoskeleton-associated protein CAP5.5 as a probe, we identified CAP50 (Tb927.11.2610). This protein colocalises with the subpellicular cytoskeleton microtubules but not with the flagellum. Depletion by RNAi results in defects in cytokinesis, morphology and partial disorganisation of microtubule arrays. Published proteomics data indicate a possible association of CAP50 with two other, yet uncharacterised, cytoskeletal proteins, CAP52 (Tb927.6.5070) and CAP42 (Tb927.4.1300), which were therefore included in our analysis. We show that their depletion causes phenotypes similar to those described for CAP50 and that they are essential for cellular integrity.

An increasing number of brands rely on ecolabelling as associative branding strategy for communicating their sustainability performance. Specifically, associative branding aims to link a brand to an ecolabel to embed sustainability into brand knowledge to provide an added value for consumers. In this regard, the present study applies a combined measurement approach that examines both implicit and explicit information processing to assess consumers’ brand associations related to sustainability. Specifically, a pre/post-testing is conducted to evaluate the framing effect of ecolabel exposure in a brand advertisement on the enhancement of brand knowledge. The results of the current study provide evidence that in particular implicit brand sustainability is a favorable brand association that positively affects consumers’ decision-making and preferences. Furthermore, the results reveal a relevant framing effect of ecolabelling on the enhancement of brand knowledge, on both implicit and explicit levels. In addition, the study results indicate the existence of three efficacy parameters that influence brand knowledge transfer: adequate ecolabel-brand fit, early ecolabel recognition time, and distinct implicit-explicit knowledge acquisition.

Correction After publication of the article [1], it has been brought to our attention that the values cited under “indices t1-t0” have been reversed and should be displayed in the opposite order. The corrected Table 1 would appear as follows

Developmental programs have the fidelity to form neural circuits with the same structure and function among individuals of the same species. It is less well understood, however, to what extent entire neural circuits of different individuals are similar. Previously, we reported the neuronal connectome of the visual eye circuit from the head of a Platynereis dumerilii larva (Randel et al., 2014). We now report a full-body serial section transmission electron microscopy (ssTEM) dataset of another larva of the same age, for which we describe the connectome of the visual eyes and the larval eyespots. Anatomical comparisons and quantitative analyses of the two circuits reveal a high inter-individual stereotypy of the cell complement, neuronal projections, and synaptic connectivity, including the left-right asymmetry in the connectivity of some neurons. Our work shows the extent to which the eye circuitry in Platynereis larvae is hard-wired.

The feasibility of exploiting secretory phospholipase A2 (sPLA2) enzymes, which are overexpressed in tumors, to activate drug release from liposomes precisely at the tumor site has been demonstrated before. Although the efficacy of the developed formulations was evaluated using in vitro and in vivo models, the pattern of sPLA2-assisted drug release is unknown due to the lack of a suitable bio-relevant model. We report here on the development of a novel bioluminescence living-cell-based luciferase assay for the monitoring of sPLA2-triggered release of luciferin from liposomes. To this end, we engineered breast cancer cells to produce both luciferase and sPLA2 enzymes, where the latter is secreted to the extracellular medium. We report on setting up a robust and reproducible bioassay for testing sPLA2-sensitive, luciferin remote-loaded liposomal formulations, using 1,2-distearoyl-sn-glycero-3-phosphatidylcholine/1,2-distearoyl-sn-glycero-3-phosphatidylglycerol (DSPC/DSPG) 7:3 and DSPC/DSPG/cholesterol 4:3:3 as initial test systems. Upon their addition to the cells, the liposomes were degraded almost instantaneously by sPLA2 releasing the encapsulated luciferin, which provided readout from the luciferase-expressing cells. Cholesterol enhanced the integrity of the formulation without affecting its susceptibility to sPLA2. PEGylation of the liposomes only moderately broadened the release profile of luciferin. The provided bioassay represents a useful tool for monitoring active drug release in situ in real time as well as for testing and optimizing of sPLA2-sensitive lipid formulations. In addition, the bioassay will pave the way for future in-depth in vitro and in vivo studies.

ObjectivesTo assess the relationship between proximity to specific sports facilities and participation in the corresponding sports activities for adolescents in Germany.MethodsA sample of 1,768 adolescents aged 11-17 years old and living in 161 German communities was examined. Distances to the nearest sports facilities were calculated as an indicator of proximity to sports facilities using Geographic Information Systems (GIS). Participation in specific leisure-time sports activities in sports clubs was assessed using a self-report questionnaire and individual-level socio-demographic variables were derived from a parent questionnaire. Community-level socio-demographics as covariates were selected from the INKAR database, in particular from indicators and maps on land development. Logistic regression analyses were conducted to examine associations between proximity to the nearest sports facilities and participation in the corresponding sports activities.ResultsThe logistic regression analyses showed that girls residing longer distances from the nearest gym were less likely to engage in indoor sports activities; a significant interaction between distances to gyms and level of urbanization was identified. Decomposition of the interaction term showed that for adolescent girls living in rural areas participation in indoor sports activities was positively associated with gym proximity. Proximity to tennis courts and indoor pools was not associated with participation in tennis or water sports, respectively.ConclusionsImproved proximity to gyms is likely to be more important for female adolescents living in rural areas.

BACKGROUND: The measurement of activity energy expenditure (AEE) via accelerometry is the most commonly used objective method for assessing human daily physical activity and has gained increasing importance in the medical, sports and psychological science research in recent years. OBJECTIVE: The purpose of this study was to determine which of the following procedures is more accurate to determine the energy cost during the most common everyday life activities; a single regression or an activity based approach. For this we used a device that utilizes single regression models (GT3X, ActiGraph Manufacturing Technology Inc., FL., USA) and a device using activity-dependent calculation models (move II, movisens GmbH, Karlsruhe, Germany). MATERIAL AND METHODS: Nineteen adults (11 male, 8 female; 30.4±9.0 years) wore the activity monitors attached to the waist and a portable indirect calorimeter (IC) as reference measure for AEE while performing several typical daily activities. The accuracy of the two devices for estimating AEE was assessed as the mean differences between their output and the reference and evaluated using Bland-Altman analysis. RESULTS: The GT3X overestimated the AEE of walking (GT3X minus reference, 1.26 kcal/min), walking fast (1.72 kcal/min), walking up-/downhill (1.45 kcal/min) and walking upstairs (1.92 kcal/min) and underestimated the AEE of jogging (-1.30 kcal/min) and walking upstairs (-2.46 kcal/min). The errors for move II were smaller than those for GT3X for all activities. The move II overestimated AEE of walking (move II minus reference, 0.21 kcal/min), walking up-/downhill (0.06 kcal/min) and stair walking (upstairs: 0.13 kcal/min; downstairs: 0.29 kcal/min) and underestimated AEE of walking fast (-0.11 kcal/min) and jogging (-0.93 kcal/min). CONCLUSIONS: Our data suggest that the activity monitor using activity-dependent calculation models is more appropriate for predicting AEE in daily life than the activity monitor using a single regression model.

The rehabilitation of patients should not only be limited to the first phases during intense hospital care but also support and therapy should be guaranteed in later stages, especially during daily life activities if the patient’s state requires this. However, aid should only be given to the patient if needed and as much as it is required. To allow this, automatic self-initiated movement support and patient-cooperative control strategies have to be developed and integrated into assistive systems. In this work, we first give an overview of different kinds of neuromuscular diseases, review different forms of therapy, and explain possible fields of rehabilitation and benefits of robotic aided rehabilitation. Next, the mechanical design and control scheme of an upper limb orthosis for rehabilitation are presented. Two control models for the orthosis are explained which compute the triggering function and the level of assistance provided by the device. As input to the model fused sensor data from the orthosis and physiology data in terms of electromyography (EMG) signals are used.

GAGE proteins are highly similar, primate-specific molecules with unique primary structure and undefined cellular roles. They are restricted to cells of the germ line in adult healthy individuals, but are broadly expressed in a wide range of cancers. In a yeast two-hybrid screen we identified the metazoan transcriptional regulator, Germ cell-less (GCL), as an interaction partner of GAGE12I. GCL directly binds LEM-domain proteins (LAP2β, emerin, MAN1) at the nuclear envelope, and we found that GAGE proteins were recruited to the nuclear envelope inner membrane by GCL. Based on yeast two-hybrid analysis and pull-down experiments of GCL polypeptides, GCL residues 209-320 (which includes the BACK domain) were deduced sufficient for association with GAGE proteins. GAGE mRNAs and GCL mRNA were demonstrated in human testis and most types of cancers, and at the protein level GAGE members and GCL were co-expressed in cancer cell lines. Structural studies of GAGE proteins revealed no distinct secondary or tertiary structure, suggesting they are intrinsically disordered. Interestingly GAGE proteins formed stable complexes with dsDNA in vitro at physiological concentrations, and GAGE12I bound several different dsDNA fragments, suggesting sequence-nonspecific binding. Dual association of GAGE family members with GCL at the nuclear envelope inner membrane in cells, and with dsDNA in vitro, implicate GAGE proteins in chromatin regulation in germ cells and cancer cells.

This corrects the article on p. e1000281 in Vol. 4, PMID: 19043566. Hypermutable Non-Synonymous Sites Are Under Stronger Negative Selection.

Mutation rate varies greatly between nucleotide sites of the human genome and depends both on the global genomic location and the local sequence context of a site. In particular, CpG context elevates the mutation rate by an order of magnitude. Mutations also vary widely in their effect on the molecular function, phenotype, and fitness. Independence of the probability of occurrence of a new mutation's effect has been a fundamental premise in genetics. However, highly mutable contexts may be preserved by negative selection at important sites but destroyed by mutation at sites under no selection. Thus, there may be a positive correlation between the rate of mutations at a nucleotide site and the magnitude of their effect on fitness. We studied the impact of CpG context on the rate of human-chimpanzee divergence and on intrahuman nucleotide diversity at non-synonymous coding sites. We compared nucleotides that occupy identical positions within codons of identical amino acids and only differ by being within versus outside CpG context. Nucleotides within CpG context are under a stronger negative selection, as revealed by their lower, proportionally to the mutation rate, rate of evolution and nucleotide diversity. In particular, the probability of fixation of a non-synonymous transition at a CpG site is two times lower than at a CpG site. Thus, sites with different mutation rates are not necessarily selectively equivalent. This suggests that the mutation rate may complement sequence conservation as a characteristic predictive of functional importance of nucleotide sites.

Quantifying the distribution of fitness effects among newly arising mutations in the human genome is key to resolving important debates in medical and evolutionary genetics. Here, we present a method for inferring this distribution using Single Nucleotide Polymorphism (SNP) data from a population with non-stationary demographic history (such as that of modern humans). Application of our method to 47,576 coding SNPs found by direct resequencing of 11,404 protein coding-genes in 35 individuals (20 European Americans and 15 African Americans) allows us to assess the relative contribution of demographic and selective effects to patterning amino acid variation in the human genome. We find evidence of an ancient population expansion in the sample with African ancestry and a relatively recent bottleneck in the sample with European ancestry. After accounting for these demographic effects, we find strong evidence for great variability in the selective effects of new amino acid replacing mutations. In both populations, the patterns of variation are consistent with a leptokurtic distribution of selection coefficients (e.g., gamma or log-normal) peaked near neutrality. Specifically, we predict 27-29% of amino acid changing (nonsynonymous) mutations are neutral or nearly neutral (|s|1%). Our results are consistent with 10-20% of amino acid differences between humans and chimpanzees having been fixed by positive selection with the remainder of differences being neutral or nearly neutral. Our analysis also predicts that many of the alleles identified via whole-genome association mapping may be selectively neutral or (formerly) positively selected, implying that deleterious genetic variation affecting disease phenotype may be missed by this widely used approach for mapping genes underlying complex traits.

This article reports the results from a national survey directed to the department chairs of political science to assess the current and future state of distance learning in that discipline. The insights of this research are relevant to all social science fields and offer important insights to other academic disciplines as well. Key findings of the study include the low utilization of distance learning courses, a low degree of importance currently attributed to distance learning and modest expectations of future growth, ambivalent acceptance of a future role for distance learning, the common use of Internet-related technologies, low levels of faculty knowledge and interest about distance learning, limited institutional support, and serious doubts about the appropriateness and quality of instruction at a distance. We propose a model of the size and scope of distance learning as a function of three factors: the capacity of distance learning technologies, market demand, and faculty and university interest in distance learning. The article concludes with suggestions of critical areas for future research in this dynamic, fluid post-secondary environment.