Perspectives on Fracture Liaison Service in Austria: clinical and economic considerations.

Osteoporosis is a widespread disease and affects over 500,000 people in Austria. Fragility fractures are associated with it and represent not only an individual problem for the patients, but also an enormous burden for the healthcare system. While trauma surgery care is well provided in Vienna, there is an enormous treatment gap in secondary prevention after osteoporotic fracture. Systematic approaches such as the Fracture Liaison Service (FLS) aim to identify patients with osteoporosis after fracture, to clarify diagnostically, to initiate specific therapy, and to check therapy adherence. The aim of this article is to describe the practical implementation and operational flow of an already established FLS in Vienna. This includes the identification of potential FLS inpatients, the diagnostic workup, and recommendations for an IT solution for baseline assessment and follow-up of FLS patients. We summarize the concept, benefits, and limitations of FLS and provide prospective as well as clinical and economic considerations for a city-wide FLS, managed from a central location. Future concepts of FLS should include artificial intelligence for vertebral fracture detection and simple IT tools for the implementation of FLS in the outpatient sector.

Feeding climate and biodiversity goals with novel plant-based meat and milk alternatives.

Plant-based animal product alternatives are increasingly promoted to achieve more sustainable diets. Here, we use a global economic land use model to assess the food system-wide impacts of a global dietary shift towards these alternatives. We find a substantial reduction in the global environmental impacts by 2050 if globally 50% of the main animal products (pork, chicken, beef and milk) are substituted-net reduction of forest and natural land is almost fully halted and agriculture and land use GHG emissions decline by 31% in 2050 compared to 2020. If spared agricultural land within forest ecosystems is restored to forest, climate benefits could double, reaching 92% of the previously estimated land sector mitigation potential. Furthermore, the restored area could contribute to 13-25% of the estimated global land restoration needs under target 2 from the Kunming Montreal Global Biodiversity Framework by 2030, and future declines in ecosystem integrity by 2050 would be more than halved. The distribution of these impacts varies across regions-the main impacts on agricultural input use are in China and on environmental outcomes in Sub-Saharan Africa and South America. While beef replacement provides the largest impacts, substituting multiple products is synergistic.

Enhanced food system efficiency is the key to China's 2060 carbon neutrality target.

Bioenergy with carbon capture and storage, among other negative-emission technologies, is required for China to achieve carbon neutrality-yet it may hinder land-based Sustainable Development Goals. Using modelling and scenario analysis, we investigate how to mitigate the potential adverse impacts on the food system of ambitious bioenergy deployment in China and its trading partners. We find that producing bioenergy domestically while sticking to the food self-sufficiency ratio redlines would lower China's daily per capita calorie intake by 8% and increase domestic food prices by 23% by 2060. Removing China's food self-sufficiency ratio restrictions could halve the domestic food dilemma but risks transferring environmental burdens to other countries, whereas halving food loss and waste, shifting to healthier diets and narrowing crop yield gaps could effectively mitigate these external effects. Our results show that simultaneously achieving carbon neutrality, food security and global sustainability requires a careful combination of these measures.

Nanomaterial Characterization in Complex Media-Guidance and Application.

A broad range of inorganic nanoparticles (NPs) and their dissolved ions possess a possible toxicological risk for human health and the environment. Reliable and robust measurements of dissolution effects may be influenced by the sample matrix, which challenges the analytical method of choice. In this study, CuO NPs were investigated in several dissolution experiments. Two analytical techniques (dynamic light scattering (DLS) and inductively-coupled plasma mass spectrometry (ICP-MS)) were used to characterize NPs (size distribution curves) time-dependently in different complex matrices (e.g., artificial lung lining fluids and cell culture media). The advantages and challenges of each analytical approach are evaluated and discussed. Additionally, a direct-injection single particle (DI sp)ICP-MS technique for assessing the size distribution curve of the dissolved particles was developed and evaluated. The DI technique provides a sensitive response even at low concentrations without any dilution of the complex sample matrix. These experiments were further enhanced with an automated data evaluation procedure to objectively distinguish between ionic and NP events. With this approach, a fast and reproducible determination of inorganic NPs and ionic backgrounds can be achieved. This study can serve as guidance when choosing the optimal analytical method for NP characterization and for the determination of the origin of an adverse effect in NP toxicity.

Expression of PKM2 in wound keratinocytes is coupled to angiogenesis during skin repair in vivo and in HaCaT keratinocytes in vitro.

An injured skin is rapidly restored in a manner of wound healing. We have previously shown that intact insulin signaling and glucose uptake are fundamental to proper wound closure. Consequently, under exacerbated inflammation, compromised insulin action and glucose uptake lead to impaired healing. However, in spite of the increased attention to cell metabolism during tissue regeneration, metabolic mediators that govern cellular and physiological processes throughout skin repair remained largely elusive. Through assessment of mRNA using real-time PCR and protein blot analysis, we report that healing of cutaneous wounds comprise a boosted expression of genes involved in glycolysis, oxidative phosphorylation, pentose phosphate shunt, and glutamine anaplerosis. We further focused on the functional role of pyruvate kinase M (PKM) isoenzymes that catalyze the final and rate-limiting step of glycolysis. Whereas the expression of the metabolic constitutively active Pkm1 isozyme remained almost unchanged, Pkm2 is augmented during the inflammatory phase of healing. The immunohistochemistry and RNA in situ hybridization analysis showed a confined Pkm2 expression to keratinocytes of the hyperproliferative epithelium and, to a lesser extent, infiltrating neutrophils and monocytes as well as later on in macrophages. Notably, the expression of Pkm2 in keratinocytes facing the wound bed side colocalized with VEGF expression. The in vitro knockdown of PKM2 in HaCaT keratinocytes using small interfering (si) RNA confirmed an acute role for PKM2 in facilitating the complete induction of VEGF mRNA and protein expression in keratinocytes; this function is mainly HIF-1α independent. KEY MESSAGES: • Wound healing involves activation of glycolysis, oxidative phosphorylation, pentos-phosphate shunt, and replenishment of tri-carboxylic acid (TCA) cycle through glutamine anaplerosis. • The pyruvate kinase M2 (PKM2) isoform is upregulated during the inflammatory phase of cutaneous healing, mainly in keratinocytes of hyperproliferative epithelia. • In vivo, the expression of VEGF in wound keratinocytes is colocalized with PKM2. • PKM2 is required for full induction of VEGF in HaCaT keratinocytes in vitro.

Reciprocal abrogation of PKM isoforms: contradictory outcomes and differing impact of splicing signal on CRISPR/Cas9 mediates gene editing in keratinocytes.

The healing of wounded skin is a highly organized process involving a massive cell in- and outflux, proliferation and tissue remodelling. It is well accepted that metabolic constraints such as diabetes mellitus, overweight or anorexia impairs wound healing. Indeed, wound inflammation involves a boost of overall metabolic changes. As wound healing converges inflammatory processes that are also common to transformation, we investigate the functional role of the pro-neoplastic factor pyruvate kinase (PK) M2 and its metabolic active splice variant PKM1 in keratinocytes. Particularly, we challenge the impact of reciprocal ablation of PKM1 or two expression. Here, CRISPR/Cas9 genome editing of the PKM gene in HaCaT reveals an unexpected mutational bias at the 3'SS of exon 9, whereas no preference for any particular kind of mutation at exon 10 3' splice, despite the close vicinity (400 nucleotides apart) and sequence similarity between the two sites. Furthermore, as opposed to transient silencing of PKM2, exclusion splicing of PKM2 via genome editing mutually increases PKM1 mRNA and protein expression and compensates for the absence of PKM2, whereas the reciprocal elimination of PKM1 splicing reduces PKM2 expression and impedes cell proliferation, thus unveiling an essential role for PKM1 in growth and metabolic balance of HaCaT keratinocytes.

Large Distortion of Fused Aromatics on Dielectric Interlayers Quantified by Photoemission Orbital Tomography.

Polycyclic aromatic compounds with fused benzene rings offer an extraordinary versatility as next-generation organic semiconducting materials for nanoelectronics and optoelectronics due to their tunable characteristics, including charge-carrier mobility and optical absorption. Nonplanarity can be an additional parameter to customize their electronic and optical properties without changing the aromatic core. In this work, we report a combined experimental and theoretical study in which we directly observe large, geometry-induced modifications in the frontier orbitals of a prototypical dye molecule when adsorbed on an atomically thin dielectric interlayer on a metallic substrate. Experimentally, we employ angle-resolved photoemission experiments, interpreted in the framework of the photoemission orbital tomography technique. We demonstrate its sensitivity to detect geometrical bends in adsorbed molecules and highlight the role of the photon energy used in experiment for detecting such geometrical distortions. Theoretically, we conduct density functional calculations to determine the geometric and electronic structure of the adsorbed molecule and simulate the photoemission angular distribution patterns. While we found an overall good agreement between experimental and theoretical data, our results also unveil limitations in current van der Waals corrected density functional approaches for such organic/dielectric interfaces. Hence, photoemission orbital tomography provides a vital experimental benchmark for such systems. By comparison with the state of the same molecule on a metallic substrate, we also offer an explanation why the adsorption on the dielectric induces such large bends in the molecule.

Mortality and Epidemiological Changes in Proximal Hip Fractures in the Course of a Pandemic.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has had an immense impact on the treatment protocols of orthopedic and trauma departments, yet its specific effect on mortality in patients with hip fractures due to possible surgical delays is still unclear. The purpose of this paper was to investigate whether the COVID-19 pandemic worsened the mortality rate of hip fracture patients. PATIENTS AND METHODS: This study comprised 175 prospectively included patients who (1) suffered from hip fractures, (2) presented during the Austrian state of emergency period from 15 March 2020 to 30 May 2021, and (3) were admitted to a level I trauma center. This cohort was compared with a retrospective control group of 339 patients admitted for hip fractures during the same timeframe in 2017, 2018, and 2019. RESULTS: An admission reduction of 22% in the COVID period compared with the pre-COVID period was evident (p = 0.018). The 30-day mortality rate was 14.67% (pre-COVID) compared with 15.18% (p = 0.381). No differences in surgical complication rates or relationships between comorbidity burden and survival were observed. There were no significant changes in demographic variables, except for admission rate, gender (p = 0.013), and place of accident (p = 0.049). CONCLUSION: Surgeons should be reassured to take COVID-19 precautions, as this study did not show higher perioperative mortality due to COVID-19 measures. Under the current circumstances, with possibly reduced surgical and hospital bed capacities, it is expected that hip fractures may continue to require a high degree of resources.

Unsupervised Text Segmentation Predicts Eye Fixations During Reading.

Words typically form the basis of psycholinguistic and computational linguistic studies about sentence processing. However, recent evidence shows the basic units during reading, i.e., the items in the mental lexicon, are not always words, but could also be sub-word and supra-word units. To recognize these units, human readers require a cognitive mechanism to learn and detect them. In this paper, we assume eye fixations during reading reveal the locations of the cognitive units, and that the cognitive units are analogous with the text units discovered by unsupervised segmentation models. We predict eye fixations by model-segmented units on both English and Dutch text. The results show the model-segmented units predict eye fixations better than word units. This finding suggests that the predictive performance of model-segmented units indicates their plausibility as cognitive units. The Less-is-Better (LiB) model, which finds the units that minimize both long-term and working memory load, offers advantages both in terms of prediction score and efficiency among alternative models. Our results also suggest that modeling the least-effort principle for the management of long-term and working memory can lead to inferring cognitive units. Overall, the study supports the theory that the mental lexicon stores not only words but also smaller and larger units, suggests that fixation locations during reading depend on these units, and shows that unsupervised segmentation models can discover these units.

Land-based climate change mitigation measures can affect agricultural markets and food security.

Earlier studies have noted potential adverse impacts of land-related emissions mitigation strategies on food security, particularly due to food price increases-but without distinguishing these strategies' individual effects under different conditions. Using six global agroeconomic models, we show the extent to which three factors-non-CO2 emissions reduction, bioenergy production and afforestation-may change food security and agricultural market conditions under 2 °C climate-stabilization scenarios. Results show that afforestation (often simulated in the models by imposing carbon prices on land carbon stocks) could have a large impact on food security relative to non-CO2 emissions policies (generally implemented as emissions taxes). Respectively, these measures put an additional 41.9 million and 26.7 million people at risk of hunger in 2050 compared with the current trend scenario baseline. This highlights the need for better coordination in emissions reduction and agricultural market management policies as well as better representation of land use and associated greenhouse gas emissions in modelling.

Split-HaloTag imaging assay for sophisticated microscopy of protein-protein interactions in planta.

An ever-increasing number of intracellular multi-protein networks have been identified in plant cells. Split-GFP-based protein-protein interaction assays combine the advantages of in vivo interaction studies in a native environment with additional visualization of protein complex localization. Because of their simple protocols, they have become some of the most frequently used methods. However, standard fluorescent proteins present several drawbacks for sophisticated microscopy. With the HaloTag system, these drawbacks can be overcome, as this reporter forms covalent irreversible bonds with synthetic photostable fluorescent ligands. Dyes can be used in adjustable concentrations and are suitable for advanced microscopy methods. Therefore, we have established the Split-HaloTag imaging assay in plants, which is based on the reconstitution of a functional HaloTag protein upon protein-protein interaction and the subsequent covalent binding of an added fluorescent ligand. Its suitability and robustness were demonstrated using a well-characterized interaction as an example of protein-protein interaction at cellular structures: the anchoring of the molybdenum cofactor biosynthesis complex to filamentous actin. In addition, a specific interaction was visualized in a more distinctive manner with subdiffractional polarization microscopy, Airyscan, and structured illumination microscopy to provide examples of sophisticated imaging. Split-GFP and Split-HaloTag can complement one another, as Split-HaloTag represents an alternative option and an addition to the large toolbox of in vivo methods. Therefore, this promising new Split-HaloTag imaging assay provides a unique and sensitive approach for more detailed characterization of protein-protein interactions using specific microscopy techniques, such as 3D imaging, single-molecule tracking, and super-resolution microscopy.

Overarching control of autophagy and DNA damage response by CHD6 revealed by modeling a rare human pathology.

Members of the chromodomain-helicase-DNA binding (CHD) protein family are chromatin remodelers implicated in human pathologies, with CHD6 being one of its least studied members. We discovered a de novo CHD6 missense mutation in a patient clinically presenting the rare Hallermann-Streiff syndrome (HSS). We used genome editing to generate isogenic iPSC lines and model HSS in relevant cell types. By combining genomics with functional in vivo and in vitro assays, we show that CHD6 binds a cohort of autophagy and stress response genes across cell types. The HSS mutation affects CHD6 protein folding and impairs its ability to recruit co-remodelers in response to DNA damage or autophagy stimulation. This leads to accumulation of DNA damage burden and senescence-like phenotypes. We therefore uncovered a molecular mechanism explaining HSS onset via chromatin control of autophagic flux and genotoxic stress surveillance.

The missing-VP effect in readers of English as a second language.

English sentences with double center-embedded clauses are read faster when they are made ungrammatical by removing one of the required verb phrases. This phenomenon is known as the missing-VP effect. German and Dutch speakers do not experience the missing-VP effect when reading their native language, but they do when reading English as a second language (L2). We investigate whether the missing-VP effect when reading L2 English occurs in native Dutch speakers because their knowledge of English is similar to that of native English speakers (the high exposure account), or because of the difficulty of L2 reading (the low proficiency account). In an eye-tracking study, we compare the size of the missing-VP effect between native Dutch and native English participants, and across native Dutch participants with varying L2 English proficiency and exposure. Results provide evidence for both accounts, suggesting that both native-like knowledge of English and L2 reading difficulty play a role.

The male bias of a generically-intended masculine pronoun: Evidence from eye-tracking and sentence evaluation.

Two experiments tested whether the Dutch possessive pronoun zijn 'his' gives rise to a gender inference and thus causes a male bias when used generically in sentences such as Everyone was putting on his shoes. Experiment 1 (N = 120, 48 male) was a conceptual replication of a previous eye-tracking study that had not found evidence of a male bias. The results of the current eye-tracking experiment showed the generically-intended masculine pronoun to trigger a gender inference and cause a male bias, but for male participants and in stereotypically neutral stereotype contexts only. No evidence for a male bias was thus found in stereotypically female and male context nor for female participants altogether. Experiment 2 (N = 80, 40 male) used the same stimuli as Experiment 1, but employed the sentence evaluation paradigm. No evidence of a male bias was found in Experiment 2. Taken together, the results suggest that the generically-intended masculine pronoun zijn 'his' can cause a male bias for male participants even when the referents are previously introduced by inclusive and grammatically gender-unmarked iedereen 'everyone'. This male bias surfaces with eye-tracking, which taps directly into early language processing, but not in offline sentence evaluations. Furthermore, the results suggest that the intended generic reading of the masculine possessive pronoun zijn 'his' is more readily available for women than for men.

Short- and long-term warming effects of methane may affect the cost-effectiveness of mitigation policies and benefits of low-meat diets.

Methane's short atmospheric life has important implications for the design of global climate change mitigation policies in agriculture. Three different agricultural economic models are used to explore how short- and long-term warming effects of methane can affect the cost-effectiveness of mitigation policies and dietary transitions. Results show that the choice of a particular metric for methane's warming potential is key to determine optimal mitigation options, with metrics based on shorter-term impacts leading to greater overall emission reduction. Also, the promotion of low-meat diets is more effective at reducing greenhouse gas emissions compared to carbon pricing when mitigation policies are based on metrics that reflect methane's long-term behaviour. A combination of stringent mitigation measures and dietary changes could achieve substantial emission reduction levels, helping reverse the contribution of agriculture to global warming.

Can N2 O emissions offset the benefits from soil organic carbon storage?

To respect the Paris agreement targeting a limitation of global warming below 2°C by 2100, and possibly below 1.5°C, drastic reductions of greenhouse gas emissions are mandatory but not sufficient. Large-scale deployment of other climate mitigation strategies is also necessary. Among these, increasing soil organic carbon (SOC) stocks is an important lever because carbon in soils can be stored for long periods and land management options to achieve this already exist and have been widely tested. However, agricultural soils are also an important source of nitrous oxide (N2 O), a powerful greenhouse gas, and increasing SOC may influence N2 O emissions, likely causing an increase in many cases, thus tending to offset the climate change benefit from increased SOC storage. Here we review the main agricultural management options for increasing SOC stocks. We evaluate the amount of SOC that can be stored as well as resulting changes in N2 O emissions to better estimate the climate benefits of these management options. Based on quantitative data obtained from published meta-analyses and from our current level of understanding, we conclude that the climate mitigation induced by increased SOC storage is generally overestimated if associated N2 O emissions are not considered but, with the exception of reduced tillage, is never fully offset. Some options (e.g. biochar or non-pyrogenic C amendment application) may even decrease N2 O emissions.

Modeling the Influence of Language Input Statistics on Children's Speech Production.

We trained a computational model (the Chunk-Based Learner; CBL) on a longitudinal corpus of child-caregiver interactions in English to test whether one proposed statistical learning mechanism-backward transitional probability-is able to predict children's speech productions with stable accuracy throughout the first few years of development. We predicted that the model less accurately reconstructs children's speech productions as they grow older because children gradually begin to generate speech using abstracted forms rather than specific "chunks" from their speech environment. To test this idea, we trained the model on both recently encountered and cumulative speech input from a longitudinal child language corpus. We then assessed whether the model could accurately reconstruct children's speech. Controlling for utterance length and the presence of duplicate chunks, we found no evidence that the CBL becomes less accurate in its ability to reconstruct children's speech with age.

Biomass residues as twenty-first century bioenergy feedstock-a comparison of eight integrated assessment models.

In the twenty-first century, modern bioenergy could become one of the largest sources of energy, partially replacing fossil fuels and contributing to climate change mitigation. Agricultural and forestry biomass residues form an inexpensive bioenergy feedstock with low greenhouse gas (GHG) emissions, if harvested sustainably. We analysed quantities of biomass residues supplied for energy and their sensitivities in harmonised bioenergy demand scenarios across eight integrated assessment models (IAMs) and compared them with literature-estimated residue availability. IAM results vary substantially, at both global and regional scales, but suggest that residues could meet 7-50% of bioenergy demand towards 2050, and 2-30% towards 2100, in a scenario with 300 EJ/year of exogenous bioenergy demand towards 2100. When considering mean literature-estimated availability, residues could provide around 55 EJ/year by 2050. Inter-model differences primarily arise from model structure, assumptions, and the representation of agriculture and forestry. Despite these differences, drivers of residues supplied and underlying cost dynamics are largely similar across models. Higher bioenergy demand or biomass prices increase the quantity of residues supplied for energy, though their effects level off as residues become depleted. GHG emission pricing and land protection can increase the costs of using land for lignocellulosic bioenergy crop cultivation, which increases residue use at the expense of lignocellulosic bioenergy crops. In most IAMs and scenarios, supplied residues in 2050 are within literature-estimated residue availability, but outliers and sustainability concerns warrant further exploration. We conclude that residues can cost-competitively play an important role in the twenty-first century bioenergy supply, though uncertainties remain concerning (regional) forestry and agricultural production and resulting residue supply potentials.