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Ancient adhesives used in multicomponent tools may be among our best material evidences of cultural evolution and cognitive processes in early humans. African Homo sapiens is known to have made compound adhesives from naturally sticky substances and ochre, a technical behavior proposed to mark the advent of elaborate cognitive processes in our species. Foragers of the European Middle Paleolithic also used glues, but evidence of ochre-based compound adhesives is unknown. Here, we present evidence of this kind. Bitumen was mixed with high loads of goethite ochre to make compound adhesives at the type-site of the Mousterian, Le Moustier (France). Ochre loads were so high that they lowered the adhesive's performance in classical hafting situations where stone implements are glued to handles. However, when used as handheld grips on cutting or scraping tools, a behavior known from Neanderthals, high-ochre adhesives present a real benefit, improving their solidity and rigidity. Our findings help understand the implications of Pleistocene adhesive making.

Life cycle assessment (LCA) and techno-economic analysis (TEA) models are developed for a tertiary wastewater treatment system that employs a biochar-integrated reactive filtration (RF) approach. This innovative system incorporates the utilization of biochar (BC) either in conjunction with or independently of iron-ozone catalytic oxidation (CatOx)-resulting in two configurations: Fe-CatOx-BC-RF and BC-RF. The technology demonstrates 90%-99% total phosphorus removals, adsorption of phosphorus to biochar for recovery, and >90% destructive removal of observed micropollutants. In this work, we conduct an ISO-compliant LCA of a 49.2 m 3 /day (9 gpm) field pilot-scale Fe-CatOx-BC-RF system and a 1130 m 3 /day (0.3 MGD) water resource recovery facility (WRRF)-installed RF system, modeled with BC addition at the same rate of 0.45 g/L to quantify their environmental impacts. LCA results indicated that the Fe-CatOx-BC-RF pilot system is a BC dose-dependent carbon-negative technology at -1.21 kg CO 2 e/m 3 , where biochar addition constitutes a -1.53 kg/m 3 CO 2 e beneficial impact to the process. For the WRRF-installed RF system, modeled with the same rate of BC addition, the overall process changed from 0.02 kg CO 2 e/m 3 to a carbon negative -1.41 kg CO 2 e/m 3 , demonstrating potential as a biochar dose-dependent negative emissions technology. Using the C 100 100-year carbon accounting approach rather than C net reduces these CO 2 e metrics for the process by about 25%. A stochastic TEA for the cost of water treatment using this combinatorial P removal/recovery, micropollutant destructive removal, and disinfection advanced technology shows that at scale, the mean cost for treating 1130 m 3 /day (0.3 MGD) WRRF secondary influent water with Fe-CatOx-BC-RF using the C 100 metric is US$0.18 ± US$0.01/m 3 to achieve overall process carbon neutrality. Using the same BC dose in an estimation of a 3780 m 3 /day (1 MGD) Fe-CatOx-BC-RF facility, the carbon neutral cost of treatment is reduced further to US$0.08 ± $0.01 with added BC accounting for US$0.03/m 3 . Overall, the results demonstrate the potential of carbon negativity to become a water treatment performance standard as important and attainable as pollutant and pathogen removal. PRACTITIONER POINTS: Life cycle assessment (LCA) of a pilot scale tertiary biochar water treatment process with or without catalytic ozonation at a WRRF shows a carbon negative global warming potential of -1.21-kg CO2e/m3 while removing 90%-99% TP and >90% of detected micropollutants. Biochar-integrated reactive filtration use can aid in long-term carbon sequestration by reducing the carbon footprint of advanced water treatment in a dose-dependent manner, allowing an overall carbon-neutral or carbon-negative process. A companion paper to this work (Yu et al., 2023) presents the details related to the process operation and mechanism and evaluates the pollutant removal performance of this Fe-CatOx-BC-RF process in engineering laboratory pilot research and field WRRF pilot-scale water resource recovery trials. Techno-economic analysis (TEA) of this biochar catalytic oxidation reactive filtration process using Monte Carlo stochastic modeling shows a forecasted carbon-neutral process cost with low P and micropollutant removal as US$0.11/m3 ± 0.01 for a 3780-m3/day (1 MGD) scale installation with BC cost at US$0.03/m3 of that total. The results demonstrate the potential of carbon negativity to become a water treatmentperformance standard as important and attainable as pollutant and pathogen removal., ((c) 2023 The Authors. Water Environment Research published by Wiley Periodicals LLC on behalf of Water Environment Federation.)

Biochar (BC) use in water treatment is a promising approach that can simultaneously help address societal needs of clean water, food security, and climate change mitigation. However, novel BC water treatment technology approaches require operational testing in field pilot-scale scenarios to advance their technology readiness assessment. Therefore, the objective of this study is to evaluate the system performance of BC integrated into hydrous ferric oxide reactive filtration (Fe-BC-RF) with and without catalytic ozonation (CatOx) process in laboratory and field pilot-scale scenarios. For this investigation, Fe-BC-RF and Fe-CatOx-BC-RF pilot-scale trials were conducted on synthetic lake water variants and at three municipal water resource recovery facilities (WRRFs) at process flows of 0.05 and 0.6 L/s, respectively. Three native and two iron-modified BCs were used in these studies. The commercially available reactive filtration process (Fe-RF without BC) had 96%-98% total phosphorus (TP) removal from 0.075- and 0.22-mg/L TP, as orthophosphate process influent in these trials. With BC integration, phosphorus removal yielded 94%-98% with the same process-influent conditions. In WRRF field pilot-scale studies, the Fe-CatOx-BC-RF process removed 84%-99% of influent total phosphorus concentrations that varied from 0.12 to 8.1 mg/L. Nutrient analysis on BC showed that the recovered BC used in the pilot-scale studies had an increase in TP from its native concentration, with the Fe-amended BC showing better P recovery at 110% than its unmodified state, which was 16%. Lastly, the field WRRF Fe-CatOx-BC-RF process studies showed successful destructive removals at >90% for more than 20 detected micropollutants, thus addressing a critical human health and environmental water quality concern. The research demonstrated that integration of BC into Fe-CatOx-RF for micropollutant removal, disinfection, and nutrient recovery is an encouraging tertiary water treatment technology that can address sustainable phosphorus recycling needs and the potential for carbon-negative operation. PRACTITIONER POINTS: A pilot-scale hydrous ferric oxide reactive sand filtration process integrating biochar injection typically yields >90% total phosphorus removal to ultralow levels. Biochar, modified with iron, recovers phosphorus from wastewater, creating a P/N nutrient upcycled soil amendment. Addition of ozone to the process stream enables biochar-iron-ozone catalytic oxidation demonstrating typically excellent (>90%) micropollutant destructive removals for the compounds tested. A companion paper to this work explores life cycle assessment (LCA) and techno-economic analysis (TEA) to explore biochar water treatment integrated reactive filtration impacts, costs, and readiness. Biochar use can aid in long-term carbon sequestration by reducing the carbon footprint of advanced water treatment in a dose-dependent manner, including enabling an overall carbon-negative process., (© 2023 The Authors. Water Environment Research published by Wiley Periodicals LLC on behalf of Water Environment Federation.)

Iron-ozone catalytic oxidation (CatOx) shows promise in addressing challenging wastewater pollutants. This study investigates a CatOx reactive filtration (Fe-CatOx-RF) approach with two 0.4 L/s field pilot studies and an 18-month, 18 L/s full-scale municipal wastewater deployment. We apply ozone to leverage common sand filtration and iron metal salts used in water treatment into a next-generation technology. The process combines micropollutant and pathogen destructive removal with high-efficiency phosphorus removal and recycling as a soil amendment, clean water recovery, and the potential for carbon-negative operation with integrated biochar water treatment. A key process innovation is converting a continuously renewed iron oxide coated, moving bed sand filter into a "sacrificial iron" d-orbital catalyst bed after adding O 3 to the process stream. Results for the Fe-CatOx-RF pilot studies show >95% removal efficiencies for almost all >5 × LoQ detected micropollutants, with removal rates slightly increasing with biochar addition. Phosphorus removal for the pilot site with the most P-impacted discharge was >98% with serial reactive filters. The long-term, full-scale Fe-CatOx-RF optimization trials showed single reactive filter 90% TP removal and high-efficiency micropollutant removals for most of the compounds detected, but slightly less than the pilot site studies. TP removal decreased to a mean of 86% during the 18 L/s, 12-month continuous operation stability trial, and micropollutant removals remained similar to the optimization trial for many detected compounds but less efficient overall. A >4.4 log reduction of fecal coliforms and E. coli in a field pilot sub-study suggests the ability of this CatOx approach to address infectious disease concerns. Life cycle assessment modeling suggests that integrating biochar water treatment into the Fe-CatOx-RF process for P recovery as a soil amendment makes the overall process carbon-negative at -1.21 kg CO 2 e/m 3 . Results indicate positive Fe-CatOx-RF process performance and technology readiness in full-scale extended testing. Further work exploring operational variables is essential to establish site-specific water quality limitations and responsive engineering approaches for process optimization. PRACTITIONERS POINTS: Adding ozone to WRRF secondary influent flows into tertiary ferric/ferrous salt dosed sand filtration amplifies a mature reactive filtration technology into a catalytic oxidation process for micropollutant removal and disinfection. Expensive catalysts are not used. Iron oxide compounds used to remove phosphorus and other pollutants act as sacrificial catalysts with ozone, and these rejected iron compounds can be returned upstream to aid in secondary process TP removal. Biochar addition to the CatOx process improves CO 2 e sustainability and phosphorus removal/recovery for long-term soil and water health. Short duration field pilot scale and 18-month full-scale operation at three WRRFs with good results demonstrate technology readiness., (© 2023 The Authors. Water Environment Research published by Wiley Periodicals LLC on behalf of Water Environment Federation.)

The 17β-hydroxysteroid dehydrogenase type 1 (HSD17B1) has a key role in estrogen biosynthesis as it catalyzes the reduction of estrone to the most potent estrogen, estradiol. Estradiol has a high affinity for estrogen receptors and thus stimulates their transactivation, which leads to cell proliferation and numerous other effects. HSD17B2 catalyzes the oxidation of estradiol to the less potent estrone, thereby decreasing estrogen receptor activation, which results in reduction of estrogen-associated effects. HSD17B1 and HSD17B2 overexpressing E.coli homogenates or recombinant enzymes can be used for screening and development of drugs against various pathologies such as cancer, endometriosis or osteoporosis. Here we describe the preparation of HSD17B1 and HSD17B2 bacterial homogenates and purified recombinant HSD17B1 protein as enzyme sources as well as enzymatic assays based on radiometric and mass-spectrometric detection for enzyme characterization., (Copyright © 2023. Published by Elsevier Inc.)

Decreasing the activation of pathology-activated microglia is crucial to prevent chronic inflammation and tissue scarring. In this study, we used a stab wound injury model in zebrafish and identified an injury-induced microglial state characterized by the accumulation of lipid droplets and TAR DNA-binding protein of 43 kDa (TDP-43) + condensates. Granulin-mediated clearance of both lipid droplets and TDP-43 + condensates was necessary and sufficient to promote the return of microglia back to the basal state and achieve scarless regeneration. Moreover, in postmortem cortical brain tissues from patients with traumatic brain injury, the extent of microglial activation correlated with the accumulation of lipid droplets and TDP-43 + condensates. Together, our results reveal a mechanism required for restoring microglia to a nonactivated state after injury, which has potential for new therapeutic applications in humans., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Dual upflow reactive filtration by a slowly moving sand bed with continuously renewed, hydrous ferric oxide-coated sand is used for removing polluting substances and for meeting the ultralow 0.05 mg/l total phosphorus discharge permit limits at a 1.2 million liters per day (0.32 million gallons per day) water resource recovery facility in Plummer, Idaho, in the United States. A life cycle assessment (LCA) of this reactive filtration installation was carried out to assess the environmental hotspots in the system and analyze alternative system configurations with a focus on CO 2 equivalent (CO 2 e) global warming potential, freshwater and marine eutrophication, and mineral resource scarcity. "What if" scenarios with alternative inputs for the energy, metal salts, and air compressor optimization show trade-offs between the impact categories. Key results that show a comparative reduction of global warming potential include the use of Fe versus Al metal salts, the use of renewable energy, and the energy efficiency benefit of optimizing process inputs, such as compressor air pressure, to match operational demand. The LCA shows a 2 × 10 -2  kg CO 2 e footprint per cubic meter of water, with 47% from housing concrete, and an overall freshwater eutrophication impact reduced by 99% versus no treatment. The use of renewable hydropower energy at this site isolates construction concrete as a target for lowering the CO 2 e footprint. PRACTITIONER POINTS: The main LCA eco-impact hotspots in this dual reactive filtration tertiary treatment are construction concrete and the ferric sulfate used. Iron salts show smaller impact in global warming, freshwater eutrophication, and mineral resource scarcity than "what if scenario" aluminum salts. The energy mix for this site is predominantly hydropower; other energy mix "what if" scenarios show larger impacts. Operational energy efficiency and thermodynamic analysis show that fine tuning the air compressor helps reduce carbon footprint and energy use. LCA shows a favorable 2 x 10-2 kg CO2e/m3 water impact with 99% reduction of freshwater eutrophication potential versus no treatment., (© 2022 The Authors. Water Environment Research published by Wiley Periodicals LLC on behalf of Water Environment Federation.)

Naturally occurring substances are valuable resources for drug development. In this respect, chalcones are known to be antiproliferative agents against prostate cancer cell lines through various mechanisms or targets. Based on the literature and preliminary results, we aimed to study and optimise the efficiency of a series of chalcones to inhibit androgen-converting AKR1C3, known to promote prostate cancer. A total of 12 chalcones with different substitution patterns were synthesised. Structure-activity relationships associated with these modifications on AKR1C3 inhibition were analysed by performing enzymatic assays and docking simulations. In addition, the selectivity and cytotoxicity of the compounds were assessed. In enzymatic assays, C-6' hydroxylated derivatives were more active than C-6' methoxylated derivatives. In contrast, C-4 methylation increased activity over C-4 hydroxylation. Docking results supported these findings with the most active compounds fitting nicely in the binding site and exhibiting strong interactions with key amino acid residues. The most effective inhibitors were not cytotoxic for HEK293T cells and selective for 17β-hydroxysteroid dehydrogenases not primarily involved in steroid hormone metabolism. Nevertheless, they inhibited several enzymes of the steroid metabolism pathways. Favourable substitutions that enhanced AKR1C3 inhibition of chalcones were identified. This study paves the way to further develop compounds from this series or related flavonoids with improved inhibitory activity against AKR1C3.

Objective: To evaluate and compare the nursing practice environments in Intensive Care Units of a public and private hospital and the prevalence of burnout among nursing professionals., Method: Cross-sectional, descriptive study with a quantitative approach to the data. The data collection took place between October 2018 and March 2019, and it was carried out through a questionnaire with socio-occupational variables and with metrics of intensity. The practice environment was assessed through the Nursing Work Index Revised (NWI-R); and the burnout was assessed by the Maslach Burnout Inventory (IBM). We included professionals with, at least, six months of employment, and those who were on leave or on a fixed-term employment contract were excluded., Results: The sample consisted of 296 professionals. We found Favorable environments in both institutions, however, low results in the subscales autonomy, control and organizational support in the private hospital. The prevalence of burnout among nurses was 2.5% in the public hospital and 9.1% in the private hospital, and among nursing technicians, it was 9.5% and 8.5%, respectively., Conclusion: The control of the environment, autonomy and support were considered critical points, referring to the importance of assessing institutions factors, that can improve the working conditions for the nursing team.

Extreme temperatures and scarce precipitation in deserts have led to abiotic factors often being regarded as more important than biotic ones in shaping desert communities. The presumed low biological activity of deserts is also one reason why deserts are often overlooked by conservation programs. We provide the first quantification of predation intensity from a desert ecosystem using artificial sentinel prey emulating caterpillars, a standardized monitoring tool to quantify relative predation pressure by many invertebrate and vertebrate predators. The study was conducted in a protected natural area affected by oil spills in 1975 and 2014; hence, we assessed the potential effects of oil pollution on predation rates. We found that predation was mostly due to invertebrate rather than vertebrate predators, fluctuated throughout the year, was higher at the ground level than in the tree canopy, and was not negatively affected by the oil spills. The mean predation rate per day (12.9%) was within the range found in other ecosystems, suggesting that biotic interactions in deserts ought not to be neglected and that ecologists should adopt standardized tools to track ecological functions and allow for comparisons among ecosystems., Competing Interests: The authors disclose no conflicts of interest., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Biological aging research is expected to reveal modifiable molecular mechanisms that can be harnessed to slow or possibly reverse unhealthy trajectories. However, there is first an urgent need to define consensus molecular markers of healthy and unhealthy aging. Established aging hallmarks are all linked to metabolism, and a 'rewired' metabolic circuitry has been shown to accelerate or delay biological aging. To identify metabolic signatures distinguishing healthy from unhealthy aging trajectories, we performed nontargeted metabolomics on skeletal muscles from 2-month-old and 21-month-old mice, and after dietary and lifestyle interventions known to impact biological aging. We hypothesized that common metabolic signatures would highlight specific pathways and processes promoting healthy aging, while revealing the molecular underpinnings of unhealthy aging. Here, we report 50 metabolites that commonly distinguished aging trajectories in all cohorts, including 18 commonly reduced under unhealthy aging and 32 increased. We stratified these metabolites according to known relationships with various aging hallmarks and found the greatest associations with oxidative stress and nutrient sensing. Collectively, our data suggest interventions aimed at maintaining skeletal muscle arginine and lysine may be useful therapeutic strategies to minimize biological aging and maintain skeletal muscle health, function, and regenerative capacity in old age.

The African clawed frog, Xenopus laevis, is a versatile model for biomedical research and is largely similar to mammals in terms of organ development, anatomy, physiology, and hormonal signaling mechanisms. Steroid hormones control a variety of processes and their levels are regulated by hydroxysteroid dehydrogenases (HSDs). The subfamily of 20β-HSD type 2 enzymes currently comprises eight members from teleost fish and mammals. Here, we report the identification of three 20β-HSD type 2 genes in X. tropicalis and X. laevis and the functional characterization of the two homeologs from X. laevis. X. laevis Hsd20b2.L and Hsd20b2.S showed high sequence identity with known 20β-HSD type 2 enzymes and mapped to the two subgenomes of the allotetraploid frog genome. Both homeologs are expressed during embryonic development and in adult tissues, with strongest signals in liver, kidney, intestine, and skin. After recombinant expression in human cell lines, both enzymes co-localized with the endoplasmic reticulum and catalyzed the conversion of cortisone to 20β-dihydrocortisone. Both Hsd20b2.L and Hsd20b2.S catalyzed the 20β-reduction of further C 21 steroids (17α-hydroxyprogesterone, progesterone, 11-deoxycortisol, 11-deoxycorticosterone), while only Hsd20b2.S was able to convert corticosterone and cortisol to their 20β-reduced metabolites. Estrone was only a poor and androstenedione no substrate for both enzymes. Our results demonstrate multispecificity of 20β-HSD type 2 enzymes from X. laevis similar to other teleost 20β-HSD type 2 enzymes. X. laevis 20β-HSD type 2 enzymes are probably involved in steroid catabolism and in the generation of pheromones for intraspecies communication. A role in oocyte maturation is unlikely., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

In recent years, artificial intelligence techniques have proved to be very successful when applied to problems in physical sciences. Here we apply an unsupervised machine learning (ML) algorithm called principal component analysis (PCA) as a tool to analyse the data from muon spectroscopy experiments. Specifically, we apply the ML technique to detect phase transitions in various materials. The measured quantity in muon spectroscopy is an asymmetry function, which may hold information about the distribution of the intrinsic magnetic field in combination with the dynamics of the sample. Sharp changes of shape of asymmetry functions-measured at different temperatures-might indicate a phase transition. Existing methods of processing the muon spectroscopy data are based on regression analysis, but choosing the right fitting function requires knowledge about the underlying physics of the probed material. Conversely, PCA focuses on small differences in the asymmetry curves and works without any prior assumptions about the studied samples. We discovered that the PCA method works well in detecting phase transitions in muon spectroscopy experiments and can serve as an alternative to current analysis, especially if the physics of the studied material are not entirely known. Additionally, we found out that our ML technique seems to work best with large numbers of measurements, regardless of whether the algorithm takes data only for a single material or whether the analysis is performed simultaneously for many materials with different physical properties., (© 2021 IOP Publishing Ltd.)

17β-Hydroxysteroid dehydrogenase type 2 (17β-HSD2) catalyzes the conversion of highly active estrogens and androgens into their less active forms using NAD + as cofactor. Substrate and cofactor specificities of 17β-HSD2 have been reported and potent 17β-HSD2 inhibitors have been discovered in a ligand-based approach. However, the molecular basis and the amino acids involved in the enzymatic functionality are poorly understood, as no crystal structure of the membrane-associated 17β-HSD2 exists. The functional properties of only few amino acids are known. The lack of topological information impedes structure-based drug design studies and limits the design of biochemical experiments. The aim of this work was the determination of the 17β-HSD2 topology. For this, the first homology model of 17β-HSD2 in complex with NAD + and 17β-estradiol was built, using a multi-fragment "patchwork" approach. To confirm the quality of the model, fifteen selected amino acids were exchanged one by one using site directed mutagenesis. The mutants' functional behavior demonstrated that the generated model was of very good quality and allowed the identification of several key amino acids involved in either ligand or internal structure stabilization. The final model is an optimal basis for further experiments like, for example, lead optimization., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Purpose: The long-term effect of low and moderate doses of ionizing radiation on the lens is still a matter of debate and needs to be evaluated in more detail., Material and Methods: We conducted a detailed histological analysis of eyes from B6C3F1 mice cohorts after acute gamma irradiation ( 60 Co source; 0.063 Gy/min) at young adult age of 10 weeks with doses of 0.063, 0.125, and 0.5 Gy. Sham irradiated (0 Gy) mice were used as controls. To test for genetic susceptibility heterozygous Ercc2 mutant mice were used and compared to wild-type mice of the same strain background. Mice of both sexes were included in all cohorts. Eyes were collected 4 h, 12, 18 and 24 months after irradiation. For a better understanding of the underlying mechanisms, metabolomics analyses were performed in lenses and plasma samples of the same mouse cohorts at 4 and 12 h as well as 12, 18 and 24 months after irradiation. For this purpose, a targeted analysis was chosen., Results: This analysis revealed histological changes particularly in the posterior part of the lens that rarely can be observed by using Scheimpflug imaging, as we reported previously. We detected a significant increase of posterior subcapsular cataracts (PSCs) 18 and 24 months after irradiation with 0.5 Gy (odds ratio 9.3; 95% confidence interval 2.1-41.3) independent of sex and genotype. Doses below 0.5 Gy (i.e. 0.063 and 0.125 Gy) did not significantly increase the frequency of PSCs at any time point. In lenses, we observed a clear effect of sex and aging but not of irradiation or genotype. While metabolomics analyses of plasma from the same mice showed only a sex effect., Conclusions: This article demonstrates a significant radiation-induced increase in the incidence of PSCs, which could not be identified using Scheimpflug imaging as the only diagnostic tool.

Natural products comprise a rich reservoir for innovative drug leads and are a constant source of bioactive compounds. To find pharmacological targets for new or already known natural products using modern computer-aided methods is a current endeavor in drug discovery. Nature's treasures, however, could be used more effectively. Yet, reliable pipelines for the large-scale target prediction of natural products are still rare. We developed an in silico workflow consisting of four independent, stand-alone target prediction tools and evaluated its performance on dihydrochalcones (DHCs)-a well-known class of natural products. Thereby, we revealed four previously unreported protein targets for DHCs, namely 5-lipoxygenase, cyclooxygenase-1, 17β-hydroxysteroid dehydrogenase 3, and aldo-keto reductase 1C3. Moreover, we provide a thorough strategy on how to perform computational target predictions and guidance on using the respective tools.

The expanded use of second-generation antiandrogens revolutionized the treatment landscape of progressed prostate cancer. However, resistances to these novel drugs are already the next obstacle to be solved. Various previous studies depicted an involvement of the enzyme AKR1C3 in the process of castration resistance as well as in the resistance to 2nd generation antiandrogens like enzalutamide. In our study, we examined the potential of natural AKR1C3 inhibitors in various prostate cancer cell lines and a three-dimensional co-culture spheroid model consisting of cancer cells and cancer-associated fibroblasts (CAFs) mimicking enzalutamide resistant prostate cancer. One of our compounds, named MF-15, expressed strong antineoplastic effects especially in cell culture models with significant enzalutamide resistance. Furthermore, MF-15 exhibited a strong effect on androgen receptor (AR) signaling, including significant inhibition of AR activity, downregulation of androgen-regulated genes, lower prostate specific antigen (PSA) production, and decreased AR and AKR1C3 expression, indicating a bi-functional effect. Even more important, we demonstrated a persisting inhibition of AR activity in the presence of AR-V7 and further showed that MF-15 non-competitively binds within the DNA binding domain of the AR. The data suggest MF-15 as useful drug to overcome enzalutamide resistance.

Steroids regulate many physiological processes. Hydroxysteroid dehydrogenases (HSDs) modulate the levels of steroids in pre- and post-receptor metabolism. The subfamily of 20β-HSD type 2 currently comprises six members from six different species. The zebrafish ortholog converts cortisone to 20β-dihydrocortisone and is involved in the catabolism of the stress hormone cortisol. Here, we elucidated the substrate preferences of all 20β-HSD type 2 enzymes towards a selected panel of steroids. For quantification of the substrates and their respective 20β-reduced products, we first developed and validated a liquid chromatography-mass spectrometry based method. Applying this method to activity assays with recombinantly expressed enzymes, our findings indicate that the 20β-HSD type 2 enzymes catalyze the 20β-reduction of a plethora of steroids of the glucocorticoid biosynthesis pathway. The observed multispecificity among the homologous 20β-HSD type 2 enzymes implies different physiological roles in different species., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to disclose., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Differentiation of preadipocytes into mature adipocytes is a highly complex cellular process. At lipidome level, the adipogenesis remains poorly characterized. To investigate the lipidomic changes during human adipogenesis, we used the Lipidyzer TM assay, which quantified 743 lipid species from 11 classes. The undifferentiated human SGBS cell strain showed a heterogeneous lipid class composition with the most abundant classes, phosphatidylethanolamines (PE), phosphatidylcholines (PC), and sphingomyelins (SM). The differentiation process was accompanied by increased ceramide concentrations. After completion of differentiation around day 4, massive lipid remodeling occurred during maturation, characterized by substantial synthesis of diacylglycerols (DAG), lysophosphatidylethanolamines (LPE), PC, PE, SM, and triacylglycerols (TAG). Lipid species composition became more homogeneous during differentiation to highly concentrated saturated and monounsaturated long-chain fatty acids (LCFA), with the four most abundant being C16:0, C16:1, C18:0, and C18:1. Simultaneously, the amount of polyunsaturated and very long-chain fatty acids (VLCFA) markedly decreased. High negative correlation coefficients between PE and PC species containing VLCFA and TAG species as well as between ceramides and SM imply that PE, PC, and ceramides might have served as additional sources for TAG and SM synthesis, respectively. These results highlight the enormous remodeling at the lipid level over several lipid classes during adipogenesis.

The aldo-keto reductase (AKR) superfamily comprises NAD(P)H-dependent enzymes that catalyze the reduction of a variety of carbonyl compounds. AKRs are classified in families and subfamilies. Humans exhibit three members of the AKR1B subfamily: AKR1B1 (aldose reductase, participates in diabetes complications), AKR1B10 (overexpressed in several cancer types), and the recently described AKR1B15. AKR1B10 and AKR1B15 share 92% sequence identity, as well as the capability of being active towards retinaldehyde. However, AKR1B10 and AKR1B15 exhibit strong differences in substrate specificity and inhibitor selectivity. Remarkably, their substrate-binding sites are the most divergent parts between them. Out of 27 residue substitutions, six are changes to Phe residues in AKR1B15. To investigate the participation of these structural changes, especially the Phe substitutions, in the functional features of each enzyme, we prepared two AKR1B10 mutants. The AKR1B10 m mutant carries a segment of six AKR1B15 residues (299-304, including three Phe residues) in the respective AKR1B10 region. An additional substitution (Val48Phe) was incorporated in the second mutant, AKR1B10mF48. This resulted in structures with smaller and more hydrophobic binding pockets, more similar to that of AKR1B15. In general, the AKR1B10 mutants mirrored well the specific functional features of AKR1B15, i.e., the different preferences towards the retinaldehyde isomers, the much higher activity with steroids and ketones, and the unique behavior with inhibitors. It can be concluded that the Phe residues of loop C (299-304) contouring the substrate-binding site, in addition to Phe at position 48, strongly contribute to a narrower and more hydrophobic site in AKR1B15, which would account for its functional uniqueness. In addition, we have investigated the AKR1B10 and AKR1B15 activity toward steroids. While AKR1B10 only exhibits residual activity, AKR1B15 is an efficient 17-ketosteroid reductase. Finally, the functional role of AKR1B15 in steroid and retinaldehyde metabolism is discussed., (Copyright © 2019. Published by Elsevier B.V.)

Deciphering biodistribution, biokinetics, and biological effects of nanoparticles (NPs) in entire organs with cellular resolution remains largely elusive due to the lack of effective imaging tools. Here, light sheet fluorescence microscopy in combination with optical tissue clearing was validated for concomitant three-dimensional mapping of lung morphology and NP biodistribution with cellular resolution in nondissected ex vivo murine lungs. Tissue autofluorescence allowed for label-free, quantitative morphometry of the entire bronchial tree, acinar structure, and blood vessels. Co-registration of fluorescent NPs with lung morphology revealed significant differences in pulmonary NP distribution depending on the means of application (intratracheal instillation and ventilator-assisted aerosol inhalation under anesthetized conditions). Inhalation exhibited a more homogeneous NP distribution in conducting airways and acini indicated by a central-to-peripheral (C/P) NP deposition ratio of unity (0.98 ± 0.13) as compared to a 2-fold enhanced central deposition (C/P = 1.98 ± 0.37) for instillation. After inhalation most NPs were observed in the proximal part of the acini as predicted by computational fluid dynamics simulations. At cellular resolution patchy NP deposition was visualized in bronchioles and acini, but more pronounced for instillation. Excellent linearity of the fluorescence intensity-dose response curve allowed for accurate NP dosimetry and revealed ca. 5% of the inhaled aerosol was deposited in the lungs. This single-modality imaging technique allows for quantitative co-registration of tissue architecture and NP biodistribution, which could accelerate elucidation of NP biokinetics and bioactivity within intact tissues, facilitating both nanotoxicology studies and the development of nanomedicines.

This study evaluated the ability of hydrous ferric oxide reactive filtration (HFO-RF) to remove mercury (Hg) from municipal secondary effluent at four study sites. Pilot HFO-RF systems (136 m 3 /day) at two sites demonstrated total Hg concentration removal efficiencies of 96% (inflow/outflow mean total Hg: 43.6/1.6  ng/L) and 80% (4.2/0.8 ng/L). A lightly loaded medium-scale HFO-RF system (950 m 3 /day) had a concentration removal efficiency of 53% (0.98/0.46 ng/L) and removed 0.52 mg/day of total Hg and 2.2 μg/day of methyl-Hg. A full-scale HFO-RF system (11,400 m 3 /day) yielded a total Hg concentration removal efficiency of 97% (87/2.7 ng/L) and removed an estimated 0.36 kg/year of Hg. Results suggest that the quality of secondary effluent, including dissolved organic matter content, affects achievable minimum total Hg concentrations in effluent from HFO-RF systems. Low HFO-RF effluent concentrations (<1 ng/L) can be expected when treating secondary effluent from suspended-growth biological treatment systems. PRACTITIONER POINTS: Trace levels of mercury in municipal secondary effluent can negatively impact receiving waters. Hydrous ferric oxide reactive filtration (HFO-RF) can remove mercury from municipal secondary effluent to levels below the Great Lakes Initiative discharge standard of 1.3 ng/L. Mercury removal to low concentrations (< 1 ng/L) using HFO-RF appears to be associated with secondary effluents with low dissolved organic matter content. HFO-RF can also remove total phosphorus and turbidity to low concentrations., (© 2018 Water Environment Federation.)

Many enzymes from the short-chain dehydrogenase/reductase superfamily (SDR) have already been well characterized, particularly those that participate in crucial biochemical reactions in the human body (e.g. 11β-hydroxysteroid dehydrogenase 1, 17β-hydroxysteroid dehydrogenase 1 or carbonyl reductase 1). Several other SDR enzymes are completely or almost completely uncharacterized, such as DHRS1 (also known as SDR19C1). Based on our in silico and experimental approaches, DHRS1 is described as a likely monotopic protein that interacts with the membrane of the endoplasmic reticulum. The highest expression level of DHRS1 protein was observed in human liver and adrenals. The recombinant form of DHRS1 was purified using the detergent n-dodecyl-β-D-maltoside, and DHRS1 was proven to be an NADPH-dependent reductase that is able to catalyse the in vitro reductive conversion of some steroids (estrone, androstene-3,17-dione and cortisone), as well as other endogenous substances and xenobiotics. The expression pattern and enzyme activities fit to a role in steroid and/or xenobiotic metabolism; however, more research is needed to fully clarify the exact biological function of DHRS1., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The human enzyme 17β-hydroxysteroid dehydrogenase 14 (17β-HSD14) oxidizes the hydroxyl group at position 17 of estradiol and 5-androstenediol using NAD + as cofactor. However, the physiological role of the enzyme remains unclear. We recently described the first class of nonsteroidal inhibitors for this enzyme with compound 1 showing a high 17β-HSD14 inhibitory activity. Its crystal structure was used as starting point for a structure-based optimization in this study. The goal was to develop a promising chemical probe to further investigate the enzyme. The newly designed compounds revealed mostly very high inhibition of the enzyme and for seven of them the crystal structures of the corresponding inhibitor-enzyme complexes were resolved. The crystal structures disclosed that a small change in the substitution pattern of the compounds resulted in an alternative binding mode for one inhibitor. The profiling of a set of the most potent inhibitors identified 13 (K i  = 9 nM) with a good selectivity profile toward three 17β-HSDs and the estrogen receptor alpha. This inhibitor displayed no cytotoxicity, good solubility, and auspicious predicted bioavailability. Overall, 13 is a highly interesting 17β-HSD14 inhibitor, which might be used as chemical probe for further investigation of the target enzyme., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Prolonged storage of biospecimen can lead to artificially altered metabolite concentrations and thus bias data analysis in metabolomics experiments. To elucidate the potential impact of long-term storage on the metabolite profile, a pooled human plasma sample was aliquoted and stored at -80 °C. During a time period of five years, 1012 of the aliquots were measured with the Biocrates AbsoluteIDQ p180 targeted-metabolomics assay at 193 time points. Modeling the concentration courses over time revealed that 55 out of 111 metabolites remained stable. The statistically significantly changed metabolites showed on average an increase or decrease of +13.7% or -14.5%, respectively. In detail, increased concentration levels were observed for amino acids (mean: + 15.4%), the sum of hexoses (+7.9%), butyrylcarnitine (+9.4%), and some phospholipids mostly with chain lengths exceeding 40 carbon atoms (mean: +18.0%). Lipids tended to exhibit decreased concentration levels with the following mean concentration changes: acylcarnitines, -12.1%; lysophosphatidylcholines, -15.1%; diacyl-phosphatidylcholines, -17.0%; acyl-alkyl-phosphatidylcholines, -13.3%; sphingomyelins, -14.8%. We conclude that storage of plasma samples at -80 °C for up to five years can lead to altered concentration levels of amino acids, acylcarnitines, glycerophospholipids, sphingomyelins, and the sum of hexoses. These alterations must be considered when analyzing metabolomics data from long-term epidemiological studies.

Introduction: Global metabolomics analyses using body fluids provide valuable results for the understanding and prediction of diseases. However, the mechanism of a disease is often tissue-based and it is advantageous to analyze metabolomic changes directly in the tissue. Metabolomics from tissue samples faces many challenges like tissue collection, homogenization, and metabolite extraction., Objectives: We aimed to establish a metabolite extraction protocol optimized for tissue metabolite quantification by the targeted metabolomics Absolute IDQ ™ p180 Kit (Biocrates). The extraction method should be non-selective, applicable to different kinds and amounts of tissues, monophasic, reproducible, and amenable to high throughput., Methods: We quantified metabolites in samples of eleven murine tissues after extraction with three solvents (methanol, phosphate buffer, ethanol/phosphate buffer mixture) in two tissue to solvent ratios and analyzed the extraction yield, ionization efficiency, and reproducibility., Results: We found methanol and ethanol/phosphate buffer to be superior to phosphate buffer in regard to extraction yield, reproducibility, and ionization efficiency for all metabolites measured. Phosphate buffer, however, outperformed both organic solvents for amino acids and biogenic amines but yielded unsatisfactory results for lipids. The observed matrix effects of tissue extracts were smaller or in a similar range compared to those of human plasma., Conclusion: We provide for each murine tissue type an optimized high-throughput metabolite extraction protocol, which yields the best results for extraction, reproducibility, and quantification of metabolites in the p180 kit. Although the performance of the extraction protocol was monitored by the p180 kit, the protocol can be applicable to other targeted metabolomics assays., Competing Interests: Compliance with ethical standardsThe authors declare that they have no conflict of interest and no conflict of financial interest. This study was not sponsored by Biocrates Life Sciences AG.According to the providers’ statement, the human plasma purchased from Sera Laboratories was collected from consented donors under IRB approved protocols at facilities located in the United States. All applicable international, national, and institutional guidelines for the care and use of animals were followed. Animal experiments were approved by the Upper Bavarian Government (Gz.55.2-1-54-2531-70-07, 55.2-1-2532-153-11).

Aldo-keto reductases (AKRs) are distributed in three families and multiple subfamilies in mammals. The mouse Akr1b3 gene is clearly orthologous to human AKR1B1, both coding for aldose reductase, and their gene products show similar tissue distribution, regulation by osmotic stress and kinetic properties. In contrast, no unambiguous orthologs of human AKR1B10 and AKR1B15.1 have been identified in rodents. Although two more AKRs, AKR1B7 and AKR1B8, have been identified and characterized in mouse, none of them seems to exhibit properties similar to the human AKRs. Recently, a novel mouse AKR gene, Akr1b16, was annotated and the respective gene product, AKR1B16 (sharing 83% and 80% amino acid sequence identity with AKR1B10 and AKR1B15.1, respectively), was expressed as insoluble and inactive protein in a bacterial expression system. Here we describe the expression and purification of a soluble and enzymatically active AKR1B16 from E. coli using three chaperone systems. A structural model of AKR1B16 allowed the estimation of its active-site pocket volume, which was much wider (402 Å 3 ) than those of AKR1B10 (279 Å 3 ) and AKR1B15.1 (60 Å 3 ). AKR1B16 reduced aliphatic and aromatic carbonyl compounds, using NADPH as a cofactor, with moderate or low activity (highest k cat values around 5 min -1 ). The best substrate for the enzyme was pyridine-3-aldehyde. AKR1B16 showed poor inhibition with classical AKR inhibitors, tolrestat being the most potent. Kinetics and inhibition properties resemble those of rat AKR1B17 but differ from those of the human enzymes. In addition, AKR1B16 catalyzed the oxidation of 17β-hydroxysteroids in a NADP + -dependent manner. These results, together with a phylogenetic analysis, suggest that mouse AKR1B16 is an ortholog of rat AKR1B17, but not of human AKR1B10 or AKR1B15.1. These human enzymes have no counterpart in the murine species, which is evidenced by forming a separate cluster in the phylogenetic tree and by their unique activity with retinaldehyde., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The metabolome, although very dynamic, is sufficiently stable to provide specific quantitative traits related to health and disease. Metabolomics requires balanced use of state-of-the-art study design, chemical analytics, biostatistics, and bioinformatics to deliver meaningful answers to contemporary questions in human disease research. The technology is now frequently employed for biomarker discovery and for elucidating the mechanisms underlying endocrine-related diseases. Metabolomics has also enriched genome-wide association studies (GWAS) in this area by providing functional data. The contributions of rare genetic variants to metabolome variance and to the human phenotype have been underestimated until now., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

We investigate extrinsic wormholelike twist defects that effectively increase the genus of space in lattice versions of multicomponent fractional quantum Hall systems. Although the original band structure is distorted by these defects, leading to localized midgap states, we find that a new lowest flat band representing a higher genus system can be engineered by tuning local single-particle potentials. Remarkably, once local many-body interactions in this new band are switched on, we identify various Abelian and non-Abelian fractional quantum Hall states, whose ground-state degeneracy increases with the number of defects, i.e, with the genus of space. This sensitivity of topological degeneracy to defects provides a "proof of concept" demonstration that genons, predicted by topological field theory as exotic non-Abelian defects tied to a varying topology of space, do exist in realistic microscopic models. Specifically, our results indicate that genons could be created in the laboratory by combining the physics of artificial gauge fields in cold atom systems with already existing holographic beam shaping methods for creating twist defects.

Background: From the early 1900s to the 1950s, Yakima Valley orchards were commonly treated with lead arsenate (LA) insecticides. Lead (Pb) and arsenic (As) soil contamination has been identified on former orchard lands throughout Central Washington and pose a threat to human health and the environment., Objectives: The levels of Pb and As in soil and interior dust at participating childcare centers in the Upper Yakima Valley (Yakima County), Washington were sampled to explore exposure potential for young children., Methods: Childcare center soils were collected from two soil depths, homogenized, and analyzed in bulk by a field-portable X-ray fluorescence spectrometer (XRF). Interior dust wipes samples were collected from at least two locations in each facility. All soil samples >250 mg/kg Pb and/or >20 As mg/kg were sieved to 250 μm, tested by XRF a second time, and analyzed via acid digestion and inductively coupled plasma mass spectrometry (ICP-MS) analysis., Results: Bulk and sieved XRF results, as well as ICP-MS to XRF results were strongly correlated. Maximum Pb and As XRF results indicated that 4 (21%) and 8 (42%) of the 19 childcare centers surveyed exceeded the regulatory standard for Pb and As, respectively. Historic land use was significantly associated with elevated Pb and As levels. Interior dust loadings were below United States Environmental Protection Agency (EPA) guidelines., Conclusions: Childcare centers are areas of intensive use for children and when coupled with potential residential exposure in their homes, the total daily exposure is a potential hazard to children., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

17β-HSD14 belongs to the SDR family and oxidizes the hydroxyl group at position 17 of estradiol and 5-androstenediol using NAD + as cofactor. The goal of this study was to identify and optimize 17β-HSD14 nonsteroidal inhibitors as well as to disclose their structure-activity relationship. In a first screen, a library of 17β-HSD1 and 17β-HSD2 inhibitors, selected with respect to scaffold diversity, was tested for 17β-HSD14 inhibition. The most interesting hit was taken as starting point for chemical modification applying a ligand-based approach. The designed compounds were synthesized and tested for 17β-HSD14 inhibitory activity. The two best inhibitors identified in this study have a very high affinity to the enzyme with a K i equal to 7 nM. The strong affinity of these inhibitors to the enzyme active site could be explained by crystallographic structure analysis, which highlighted the role of an extended H-bonding network in the stabilization process. The selectivity of the most potent compounds with respect to 17β-HSD1 and 17β-HSD2 is also addressed.

17β-HSD14 is a SDR enzyme able to oxidize estradiol and 5-androstenediol using NAD(+). We determined the crystal structure of this human enzyme as the holo form and as ternary complexes with estrone and with the first potent, nonsteroidal inhibitor. The structures reveal a conical, rather large and lipophilic binding site and are the starting point for structure-based inhibitor design. The two natural variants (S205 and T205) were characterized and adopt a similar structure.

Introduction: Although cultured cells are nowadays regularly analyzed by metabolomics technologies, some issues in study setup and data processing are still not resolved to complete satisfaction: a suitable harvesting method for adherent cells, a fast and robust method for data normalization, and the proof that metabolite levels can be normalized to cell number., Objectives: We intended to develop a fast method for normalization of cell culture metabolomics samples, to analyze how metabolite levels correlate with cell numbers, and to elucidate the impact of the kind of harvesting on measured metabolite profiles., Methods: We cultured four different human cell lines and used them to develop a fluorescence-based method for DNA quantification. Further, we assessed the correlation between metabolite levels and cell numbers and focused on the impact of the harvesting method (scraping or trypsinization) on the metabolite profile., Results: We developed a fast, sensitive and robust fluorescence-based method for DNA quantification showing excellent linear correlation between fluorescence intensities and cell numbers for all cell lines. Furthermore, 82-97 % of the measured intracellular metabolites displayed linear correlation between metabolite concentrations and cell numbers. We observed differences in amino acids, biogenic amines, and lipid levels between trypsinized and scraped cells., Conclusion: We offer a fast, robust, and validated normalization method for cell culture metabolomics samples and demonstrate the eligibility of the normalization of metabolomics data to the cell number. We show a cell line and metabolite-specific impact of the harvesting method on metabolite concentrations., Competing Interests: The authors declare no competing financial interest. Research involving animal and human rights This article does not contain any studies with human participants or animals performed by any of the authors.

Fat cell metabolism has an impact on body homeostasis and its proper function. Nevertheless, the knowledge about simultaneous metabolic processes, which occur during adipogenesis and in mature adipocytes, is limited. Identification of key metabolic events associated with fat cell metabolism could be beneficial in the field of novel drug development, drug repurposing, as well as for the discovery of patterns predicting obesity risk. The main objective of our work was to provide comprehensive characterization of metabolic processes occurring during adipogenesis and in mature adipocytes. In order to globally determine crucial metabolic pathways involved in fat cell metabolism, metabolomics and transcriptomics approaches were applied. We observed significantly regulated metabolites correlating with significantly regulated genes at different stages of adipogenesis. We identified the synthesis of phosphatidylcholines, the metabolism of even and odd chain fatty acids, as well as the catabolism of branched chain amino acids (BCAA; leucine, isoleucine and valine) as key regulated pathways. Our further analysis led to identification of an enzymatic switch comprising the enzymes Hmgcs2 (3-hydroxy-3-methylglutaryl-CoA synthase) and Auh (AU RNA binding protein/enoyl-CoA hydratase) which connects leucine degradation with cholesterol synthesis. In addition, propionyl-CoA, a product of isoleucine degradation, was identified as a putative substrate for odd chain fatty acid synthesis. The uncovered crosstalks between BCAA and lipid metabolism during adipogenesis might contribute to the understanding of molecular mechanisms of obesity and have potential implications in obesity prediction., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The fractional quantum Hall (FQH) effect illustrates the range of novel phenomena which can arise in a topologically ordered state in the presence of strong interactions. The possibility of realizing FQH-like phases in models with strong lattice effects has attracted intense interest as a more experimentally accessible venue for FQH phenomena which calls for more theoretical attention. Here we investigate the physical relevance of previously derived geometric conditions which quantify deviations from the Landau level physics of the FQHE. We conduct extensive numerical many-body simulations on several lattice models, obtaining new theoretical results in the process, and find remarkable correlation between these conditions and the many-body gap. These results indicate which physical factors are most relevant for the stability of FQH-like phases, a paradigm we refer to as the geometric stability hypothesis, and provide easily implementable guidelines for obtaining robust FQH-like phases in numerical or real-world experiments.

Steroid hormones are involved in the regulation of a variety of processes like embryonic development, sex differentiation, metabolism, immune responses, circadian rhythms, stress response, and reproduction in vertebrates. Teleost fishes and humans show a remarkable conservation in many developmental and physiological aspects, including the endocrine system in general and the steroid hormone related processes in particular. This review provides an overview of the current knowledge about steroid hormone biosynthesis and the steroid hormone receptors in teleost fishes and compares the findings to the human system. The impact of the duplicated genome in teleost fishes on steroid hormone biosynthesis and perception is addressed. Additionally, important processes in fish physiology regulated by steroid hormones, which are most dissimilar to humans, are described. We also give a short overview on the influence of anthropogenic endocrine disrupting compounds on steroid hormone signaling and the resulting adverse physiological effects for teleost fishes. By this approach, we show that the steroidogenesis, hormone receptors, and function of the steroid hormones are reasonably well understood when summarizing the available data of all teleost species analyzed to date. However, on the level of a single species or a certain fish-specific aspect of physiology, further research is needed., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Unlabelled: Persistence with and adherence to osteoporosis therapy are critical for fracture reduction. This non-interventional study is evaluating medication-taking behavior of women with postmenopausal osteoporosis (PMO) receiving denosumab in Germany, Austria, Greece, and Belgium. Patients were representative of the PMO population and highly persistent with and adherent to denosumab at 12 months., Introduction: Persistence with and adherence to osteoporosis therapy are important for optimal treatment efficacy, namely fracture reduction. This ongoing, non-interventional study will evaluate medication-taking behavior of women with postmenopausal osteoporosis (PMO) receiving denosumab in routine practice in four European countries., Methods: The study enrolled women who had been prescribed subcutaneous denosumab (60 mg every 6 months) in accordance with prescribing information and local guidelines. Persistence was defined as receiving the subsequent injection within 6 months + 8 weeks of the previous injection. Adherence was defined as receiving two consecutive injections within 6 months ± 4 weeks of each other. Medication coverage ratio (MCR) was calculated using the time a patient was covered with denosumab, as assessed from prescription records. Treatment was assigned prior to and independently of enrollment; outcomes are recorded during routine practice., Results: These planned 12-month interim analyses included data from 1500 patients from 141 sites. Mean age was 66.4-72.4 years, mean baseline total hip T-scores ranged from -2.0 to -2.1 and femoral neck T-scores from -2.2 to -2.6, and 30.7-62.1% of patients had prior osteoporotic fracture. Persistence was 87.0-95.3%, adherence 82.7-89.3%, and MCR 91.3-95.4%. In a univariate analysis, increased age, decreased mobility, and increased distance to the clinic were associated with significantly decreased persistence; parental history of hip fracture was associated with significantly increased persistence., Conclusions: These data extend the real-world evidence regarding persistence with and adherence to denosumab, both of which are critical for favorable clinical outcomes, including fracture risk reduction.

The Harper-Hofstadter model provides a fractal spectrum containing topological bands of any integer Chern number C. We study the many-body physics that is realized by interacting particles occupying Harper-Hofstadter bands with |C|>1. We formulate the predictions of Chern-Simons or composite fermion theory in terms of the filling factor ν, defined as the ratio of particle density to the number of single-particle states per unit area. We show that this theory predicts a series of fractional quantum Hall states with filling factors ν=r/(r|C|+1) for bosons, or ν=r/(2r|C|+1) for fermions. This series includes a bosonic integer quantum Hall state in |C|=2 bands. We construct specific cases where a single band of the Harper-Hofstadter model is occupied. For these cases, we provide numerical evidence that several states in this series are realized as incompressible quantum liquids for bosons with contact interactions.

Aldo-keto reductases (AKRs) comprise a superfamily of proteins involved in the reduction and oxidation of biogenic and xenobiotic carbonyls. In humans, at least 15 AKR superfamily members have been identified so far. One of these is a newly identified gene locus, AKR1B15, which clusters on chromosome 7 with the other human AKR1B subfamily members (i.e. AKR1B1 and AKR1B10). We show that alternative splicing of the AKR1B15 gene transcript gives rise to two protein isoforms with different N termini: AKR1B15.1 is a 316-amino acid protein with 91% amino acid identity to AKR1B10; AKR1B15.2 has a prolonged N terminus and consists of 344 amino acid residues. The two gene products differ in their expression level, subcellular localization, and activity. In contrast with other AKR enzymes, which are mostly cytosolic, AKR1B15.1 co-localizes with the mitochondria. Kinetic studies show that AKR1B15.1 is predominantly a reductive enzyme that catalyzes the reduction of androgens and estrogens with high positional selectivity (17β-hydroxysteroid dehydrogenase activity) as well as 3-keto-acyl-CoA conjugates and exhibits strong cofactor selectivity toward NADP(H). In accordance with its substrate spectrum, the enzyme is expressed at the highest levels in steroid-sensitive tissues, namely placenta, testis, and adipose tissue. Placental and adipose expression could be reproduced in the BeWo and SGBS cell lines, respectively. In contrast, AKR1B15.2 localizes to the cytosol and displays no enzymatic activity with the substrates tested. Collectively, these results demonstrate the existence of a novel catalytically active AKR, which is associated with mitochondria and expressed mainly in steroid-sensitive tissues., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)

Spirocyclic benzopyrans 2 interact with high affinity and selectivity with σ₁ receptors. Bioisosteric replacement of the benzene ring of the benzopyran substructure with the electron rich thiophene ring (3) led to increased σ₁ affinity. Herein the synthesis and pharmacological evaluation of electron deficient pyridine bioisosteres 4 are reported. Homologation of the aldehyde 6 to afford the pyridylacetaldehyde derivative 8 was performed by a Wittig reaction. Bromine lithium exchange of the bromopyridine 8, addition to 1-benzylpiperidin-4-one and cyclization led to the spirocyclic pyrranopyridine 10. Hydrogenolytic removal of the N-benzyl moiety of 10 provided the secondary amine 11, which allowed the introduction of various N-substituents (12a-d). Cyclization of the hydroxy acetal 9 with HCl led to various modifications of the substituent in 3'-position. Generally the σ₁ affinity of the pyridine derivatives is reduced compared with those of the benzene and thiophene derivatives 2 and 3. However, the relationships between the structure and the σ₁ affinity follow the same rules as for the benzene and thiophene derivatives. The most promising σ₁ ligand within this class of compounds is the pyranopyridine 15 with a double bond in the pyran ring revealing a Ki-value of 4.6 nM and a very high selectivity (>217-fold) over the σ₂ subtype., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The quest for realization of non-Abelian phases of matter, driven by their possible use in fault-tolerant topological quantum computing, has been spearheaded by recent developments in p-wave superconductors. The chiral p(x)+ip(y)-wave superconductor in two-dimensions exhibiting Majorana modes provides the simplest phase supporting non-Abelian quasiparticles and can be seen as the blueprint of fractional topological order. Alternatively, Kitaev's Majorana wire has emerged as an ideal toy model to understand Majorana modes. Here we present a way to make the transition from Kitaev's Majorana wires to two-dimensional p-wave superconductors in a system with cold atomic gases in an optical lattice. The main idea is based on an approach to generate p-wave interactions by coupling orbital degrees of freedom with strong s-wave interactions. We demonstrate how this design can induce Majorana modes at edge dislocations in the optical lattice, and we provide an experimentally feasible protocol for the observation of the non-Abelian statistics.

Background: Type 2 immune responses directed by Th2 cells and characterized by the signature cytokines IL4, IL5, and IL13 play major pathogenic roles in atopic diseases. Single nucleotide polymorphisms in the human Th2 cytokine locus in particular in a locus control region within the DNA repair gene RAD50, containing several RAD50 DNase1-hypersensitive sites (RHS), have been robustly associated with atopic traits in genome-wide association studies (GWAS). Functional variants in IL13 have been intensely studied, whereas no causative variants for the IL13-independent RAD50 signal have been identified yet. This study aimed to characterize the functional impact of the atopy-associated polymorphism rs2240032 located in the human RHS7 on cis-regulatory activity and differential binding of transcription factors., Methods: Differential transcription factor binding was analyzed by electrophoretic mobility shift assays (EMSAs) with Jurkat T-cell nuclear extracts. Identification of differentially binding factors was performed using mass spectrometry (LC-MS/MS). Reporter vector constructs carrying either the major or minor allele of rs2240032 were tested for regulating transcriptional activity in Jurkat and HeLa cells., Results: The variant rs2240032 impacts transcriptional activity and allele-specific binding of SMAD3, SP1, and additional putative protein complex partners. We further demonstrate that rs2240032 is located in an RHS7 subunit which itself encompasses repressor activity and might be important for the fine-tuning of transcription regulation within this region., Conclusion: The human RHS7 critically contributes to the regulation of gene transcription, and the common atopy-associated polymorphism rs2240032 impacts transcriptional activity and transcription factor binding., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Regulatory B cells have been identified that strongly reduce allergic and auto-immune inflammation in experimental models by producing IL-10. Recently, several human regulatory B-cell subsets with an impaired function in auto-immunity have been described, but there is no information on regulatory B cells in allergic asthma., Objective: In this study, the frequency and function of IL-10 producing B-cell subsets in allergic asthma were investigated., Methods: Isolated peripheral blood B cells from 13 patients with allergic asthma and matched healthy controls were analyzed for the expression of different regulatory B-cell markers. Next, the B cells were activated by lipopolysaccharide (LPS), CpG or through the B-cell receptor, followed by co-culture with endogenous memory CD4(+) T cells and house dust mite allergen DerP1., Results: Lower number of IL-10 producing B cells were found in patients in response to LPS, however, this was not the case when B cells were activated through the B-cell receptor or by CpG. Further dissection showed that only the CD24(hi)CD27(+) B-cell subset was reduced in number and IL-10 production to LPS. In response to DerP1, CD4(+) T cells from patients co-cultured with LPS-primed total B cells produced less IL-10 compared to similar cultures from controls. These results are in line with the finding that sorted CD24(hi)CD27(+) B cells are responsible for the induction of IL-10(+) CD4(+) T cells., Conclusions: Taken together, these data indicate that CD24(hi)CD27(+) B cells from allergic asthma patients produce less IL-10 in response to LPS leading to a weaker IL-10 induction in T cells in response to DerP1, which may play a role in allergic asthma., (© 2013 John Wiley & Sons Ltd.)

Digital cameras would be colorblind if they did not have pixelated color filters integrated into their image sensors. Integration of conventional fixed filters, however, comes at the expense of an inability to modify the camera's spectral properties. Instead, we demonstrate a micropolarizer-based camera that can reconfigure its spectral response. Color is encoded into a linear polarization state by a chiral dispersive element and then read out in a single exposure. The polarization encoded color camera is capable of capturing three-color images at wavelengths spanning the visible to the near infrared.

Near-infrared spectroscopy (NIRS) can monitor changes in cerebral regional oxygen saturation (rSO2) and tissue hemoglobin content (HbT). The relation between cerebral NIRS readings and vital parameters has not been analyzed before at a fine temporal scale. This study analyzed this relation during cardiopulmonary bypass (CPB) surgery in 10 children (0-9 years, 1,770 min of data records) by using a novel random-coefficient model. The analysis indicated that a small number of patients is sufficient for obtaining significant results with this model. Changes of vital parameters explained 84.7 % of rSO2 changes and 90.7 % of HbT changes. Cerebral rSO2 correlated positively with perfusion pressure and inversely with body temperature (P < 0.05). Cerebral HbT correlated positively with perfusion pressure, central venous pressure, and temperature and inversely with arterial oxygen saturation (P < 0.05). During hypothermic circulatory arrest, the half-life of the exponential rSO2 decay correlated to the rSO2 reserve (P = 0.016). In conclusion, NIRS readings of cerebral hemoglobin content and tissue oxygen saturation correlate well to vital parameters during CPB surgery in children. NIRS may therefore become a monitoring device for the neuroprotective optimization of those vital parameters.

Background: Metabolomics is a versatile unbiased method to search for biomarkers of human disease. In particular, one approach in cancer therapy is to promote apoptosis in tumour cells; this could be improved with specific biomarkers of apoptosis for monitoring treatment. We recently observed specific metabolic patterns in apoptotic cell lines; however, in that study, apoptosis was only induced with one pro-apoptotic agent, staurosporine., Objective: The aim of this study was to find novel biomarkers of apoptosis by verifying our previous findings using two further pro-apoptotic agents, 5-fluorouracil and etoposide, that are commonly used in anticancer treatment., Methods: Metabolic parameters were assessed in HepG2 and HEK293 cells using the newborn screening assay adapted for cell culture approaches, quantifying the levels of amino acids and acylcarnitines with mass spectrometry., Results: We were able to identify apoptosis-specific changes in the metabolite profile. Moreover, the amino acids alanine and glutamate were both significantly up-regulated in apoptotic HepG2 and HEK293 cells irrespective of the apoptosis inducer., Conclusion: Our observations clearly indicate the potential of metabolomics in detecting metabolic biomarkers applicable in theranostics and for monitoring drug efficacy., (© 2013 The Association for the Publication of the Journal of Internal Medicine.)

Zebrafish, Danio rerio, has long been used as a model organism in developmental biology. Nowadays, due to their advantages compared to other model animals, the fish gain popularity and are also increasingly used in endocrinology. This review focuses on an important aspect of endocrinology in zebrafish by summarizing the progress in steroid hormone related research. We present the state of the art of research on steroidogenesis, the action of steroid hormones, and steroid catabolism and cover the incremental usage of zebrafish as a test animal in endocrine disruption research. By this approach, we demonstrate that some aspects of steroid hormone research are well characterized (e.g., expression patterns of the genes involved), while other aspects such as functional analyses of enzymes, steroid hormone elimination, or the impact of steroid hormones on embryonic development or sex differentiation have not been extensively studied and are poorly understood. This article is part of a Special Issue entitled 'CSR 2013'., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ultrasound in the 20-1000 kHz range show unique propagation characteristics in fluid media and possess energy that can break down fruit matrices to facilitate the extraction of valuable bioactive compounds. Red raspberries carry significant amounts of specific antioxidants, including ellagitannins and anthocyanins that are important for human health. The objective of this study was to investigate the effects of ultrasound frequencies associated with cavitation (20 kHz) and microstreaming (490 and 986 kHz) on total antioxidant activity (AOA), total phenolics content (TPC), and total monomeric anthocyanin content (ACY) of red raspberry puree prepared from crushed berries. The pureed fruit was subjected to high-intensity (20 kHz) and higher frequency-low intensity (490 and 986 kHz) ultrasound for 30 min. The temperature of treated purees increased to a maximum of 56 °C with 986 kHz. Sonication at 20 and 490 kHz significantly (p<0.05) affected the AOA, ACY, and TPC of red raspberry puree, while 986 kHz had no significant effect on ACY and AOA (p<0.05). In all cases, ultrasound treatment had significant and positive effect on at least one of the measured parameters up to 30 min. Sonication beyond 10 min (and up to 30 min) using 20 kHz either produced no change or caused a drop in AOA and ACY. However, for 986 and 20 kHz, TPC, increased by 10% and 9.5%, respectively after 30 min (p<0.05) compared to the control. At 20 kHz, AOA and ACY increased by 17.3% and 12.6% after 10 min. It was demonstrated that 20 kHz ultrasound treatment, when limited to 10 min, was the most effective for extraction of bioactive compounds in red raspberry compared to 490 and 986 kHz although the effect could be similar at the higher frequencies if different amplitudes are used., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Inhibition of 17β-HSD2 is an attractive mechanism for the treatment of osteoporosis. We report here the optimization of human 17β-HSD2 inhibitors in the 2,5-thiophene amide class by varying the size of the linker (n equals 0 and 2) between the amide moiety and the phenyl group. While none of the phenethylamides (n = 2) were active, most of the anilides (n = 0) turned out to moderately or strongly inhibit 17β-HSD2. The four most active compounds showed an IC₅₀ of around 60 nM and a very good selectivity toward 17β-HSD1, 17β-HSD4, 17β-HSD5, 11β-HSD1, 11β-HSD2 and the estrogen receptors α and β. The investigated compounds inhibited monkey 17β-HSD2 moderately, and one of them showed good inhibitory activity on mouse 17β-HSD2. SAR studies allowed a first characterization of the human 17β-HSD2 active site, which is predicted to be considerably larger than that of 17β-HSD1.