Re-assessment of monoclonal antibodies against diclofenac for their application in the analysis of environmental waters.

The non-steroidal anti-inflammatory drug (NSAID) diclofenac (DCF) is an important environmental contaminant occurring in surface waters all over the world, because, after excretion, it is not adequately removed from wastewater in sewage treatment plants. To be able to monitor this pollutant, highly efficient analytical methods are needed, including immunoassays. In a medical research project, monoclonal antibodies against diclofenac and its metabolites had been produced. Based on this monoclonal anti-DCF antibody, a new indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed and applied for environmental samples. The introduction of a spacer between diclofenac and the carrier protein in the coating conjugate led to higher sensitivity. With a test midpoint of 3 µg L-1 and a measurement range of 1-30 µg L-1, the system is not sensitive enough for direct analysis of surface water. However, this assay is quite robust against matrix influences and can be used for wastewater. Without adjustment of the calibration, organic solvents up to 5%, natural organic matter (NOM) up to 10 mg L-1, humic acids up to 2.5 mg L-1, and salt concentrations up to 6 g L-1 NaCl and 75 mg L-1 CaCl2 are tolerated. The antibody is also stable in a pH range from 3 to 12. Cross-reactivity (CR) of 1% or less was determined for the metabolites 4'-hydroxydiclofenac (4'-OH-DCF), 5-hydroxydiclofenac (5-OH-DCF), DCF lactam, and other NSAIDs. Relevant cross-reactivity occurred only with an amide derivative of DCF, 6-aminohexanoic acid (DCF-Ahx), aceclofenac (ACF) and DCF methyl ester (DCF-Me) with 150%, 61% and 44%, respectively. These substances, however, have not been found in samples. Only DCF-acyl glucuronide with a cross-reactivity of 57% is of some relevance. For the first time, photodegradation products were tested for cross-reactivity. With the ELISA based on this antibody, water samples were analysed. In sewage treatment plant effluents, concentrations in the range of 1.9-5.2 µg L-1 were determined directly, with recoveries compared to HPLC-MS/MS averaging 136%. Concentrations in lakes ranged from 3 to 4.4 ng L-1 and were, after pre-concentration, determined with an average recovery of 100%.

Metabolism and Recovery of Epithionitriles from Glucosinolates-A Human Intervention Study.

SCOPE: Epithionitriles can be main glucosinolate hydrolysis products in Brassica vegetables such as cabbage or pak choi. Here, for the first time, the bioavailability and metabolism of longer-chain epithionitriles (C4-C5) is studied in a human intervention study. METHODS AND RESULTS: After consumption of a white cabbage or pak choi sprouts beverage, rich in either 1-cyano-2,3-epithiopropane (CETP) or 1-cyano-3,4-epithiobutane (CETB) and 1-cyano-4,5-epithiopentane (CETPent), blood and urine samples of nine participants are taken and the metabolites are analyzed. The corresponding N-acetyl-S-(cyano-(methylthio)alkyl)-l-cysteine metabolites are identified and quantified by isotope dilution method using UHPLC-TOF-MS. The standards for N-acetyl-S-(cyano-(methylthio)alkyl)-l-cysteine metabolites from CETB and CETPent are synthesized for the first time and their structure confirmed by NMR spectroscopy. In contrast to the metabolites of CETP and CETPent, the expected metabolite of CETB is not detectable. The recoveries of the CETP and CETPent metabolites are 28 ± 9% for CETP and 12 ± 3% for CETPent in urine within 24 h. CONCLUSION: CETP and CETPent are quickly uptaken, metabolized via the mercapturic acid pathway, and excreted via urine, while for CETB the corresponding metabolite is not detectable. Therefore, an additional metabolization pathway seems to exist.

Assessing Bioavailability and Bioactivity of 4-Hydroxythiazolidine-2-Thiones, Newly Discovered Glucosinolate Degradation Products Formed During Domestic Boiling of Cabbage.

Glucosinolates are plant secondary metabolites found in cruciferous vegetables (Brassicaceae) that are valued for their potential health benefits. Frequently consumed representatives of these vegetables, for example, are white or red cabbage, which are typically boiled before consumption. Recently, 3-alk(en)yl-4-hydroxythiazolidine-2-thiones were identified as a class of thermal glucosinolate degradation products that are formed during the boiling of cabbage. Since these newly discovered compounds are frequently consumed, this raises questions about their potential uptake and their possible bioactive functions. Therefore, 3-allyl-4-hydroxythiazolidine-2-thione (allyl HTT) and 4-hydroxy-3-(4-(methylsulfinyl) butyl)thiazolidine-2-thione (4-MSOB HTT) as degradation products of the respective glucosinolates sinigrin and glucoraphanin were investigated. After consumption of boiled red cabbage broth, recoveries of consumed amounts of the degradation products in urine collected for 24 h were 18 ± 5% for allyl HTT and 21 ± 4% for 4-MSOB HTT (mean ± SD, n = 3). To investigate the stability of the degradation products during uptake and to elucidate the uptake mechanism, both an in vitro stomach and an in vitro intestinal model were applied. The results indicate that the uptake of allyl HTT and 4-MSOB HTT occurs by passive diffusion. Both compounds show no acute cell toxicity, no antioxidant potential, and no change in NAD(P)H dehydrogenase quinone 1 (NQO1) activity up to 100 µM. However, inhibition of glycogen synthase kinases-3 (GSK-3) in the range of 20% for allyl HTT for the isoform GSK-3ß and 29% for 4-MSOB HTT for the isoform GSK-3α at a concentration of 100 µM was found. Neither health-promoting nor toxic effects of 3-alk(en)yl-4-hydroxythiazolidine-2-thiones were found in the four tested assays carried out in this study, which contrasts with the properties of other glucosinolate degradation products, such as isothiocyanates.

Novel transformation products from glucosinolate-derived thioglucose and isothiocyanates formed during cooking.

Glucosinolates are plant secondary metabolites occurring in Brassicaceae plants. Upon tissue disruption, these compounds can be enzymatically hydrolyzed into isothiocyanates, which are very reactive and can react with nucleophiles during thermal processes such as boiling. Here, a novel type of glucosinolate degradation product was identified resulting from the reaction of thioglucose with the isothiocyanates sulforaphane or allyl isothiocyanate during aqueous heating. The two heterocyclic compounds 4-hydroxy-3-(4-(methylsulfinyl)butyl)thiazolidine-2-thione and 3-allyl-4-hydroxythiazolidine-2-thione were isolated and their structure elucidated by NMR spectroscopy and high-resolution mass spectrometry. Based on a set of chemical experiments, a reaction mechanism was proposed. Finally, the formation of the two 3-alk(en)yl-4-hydroxythiazolidine-2-thiones was quantified in boiled cabbage samples using a standard addition method in which 92 pmol/g and 19 pmol/g fresh weight of the sulforaphane and allyl isothiocyanate derivatives were found, respectively.

[Association, expectations and barriers of the use of exoskeletons in small and medium-sized enterprises]. / Assoziation, Erwartungen und Barrieren eines Exoskeletteinsatzes in kleinen mittelständischen Unternehmen.

Background: In the manufacturing industry, work-related musculoskeletal disorders (MSD) result in sick days and have substantial economic consequences for the enterprise and the national economy. Exoskeletons can support the body when handling heavy loads and enduring enforced postures. Exoskeletons are being piloted particularly in large companies in the automotive industry; however, exoskeletons have so far attracted little interest in small and medium-sized enterprises (SME) and their use has so far barely been scientifically examined. The aim of this work was to determine barriers to exoskeleton implementation and expectations for their use in the manufacturing sector. Method: Semi-structured guided interviews in six manufacturing companies were carried out and analyzed. Results: In the enterprises a variety of activities up to the loading limits are carried out. Exoskeletons are generally expected to facilitate work and provide economic advantages. There are concerns with respect to their use due to cost factors, uncertain benefits and wearing discomfort. Particularly uncertainties about the effects of exoskeletons become evident. Conclusion: The presented interview results are one step in an interdisciplinary process of further developing and implementing exoskeletons in the manufacturing industry. Concerns and unawareness of potential enterprises and users must be addressed, also to achieve a high user acceptance. Further studies that survey the identification of needs with better discriminatory power could provide additional insights.

The uulmMAC Database-A Multimodal Affective Corpus for Affective Computing in Human-Computer Interaction.

In this paper, we present a multimodal dataset for affective computing research acquired in a human-computer interaction (HCI) setting. An experimental mobile and interactive scenario was designed and implemented based on a gamified generic paradigm for the induction of dialog-based HCI relevant emotional and cognitive load states. It consists of six experimental sequences, inducing Interest, Overload, Normal, Easy, Underload, and Frustration. Each sequence is followed by subjective feedbacks to validate the induction, a respiration baseline to level off the physiological reactions, and a summary of results. Further, prior to the experiment, three questionnaires related to emotion regulation (ERQ), emotional control (TEIQue-SF), and personality traits (TIPI) were collected from each subject to evaluate the stability of the induction paradigm. Based on this HCI scenario, the University of Ulm Multimodal Affective Corpus (uulmMAC), consisting of two homogenous samples of 60 participants and 100 recording sessions was generated. We recorded 16 sensor modalities including 4 × video, 3 × audio, and 7 × biophysiological, depth, and pose streams. Further, additional labels and annotations were also collected. After recording, all data were post-processed and checked for technical and signal quality, resulting in the final uulmMAC dataset of 57 subjects and 95 recording sessions. The evaluation of the reported subjective feedbacks shows significant differences between the sequences, well consistent with the induced states, and the analysis of the questionnaires shows stable results. In summary, our uulmMAC database is a valuable contribution for the field of affective computing and multimodal data analysis: Acquired in a mobile interactive scenario close to real HCI, it consists of a large number of subjects and allows transtemporal investigations. Validated via subjective feedbacks and checked for quality issues, it can be used for affective computing and machine learning applications.

Supported employment in Switzerland-Are we on track?

OBJECTIVES: Supported employment (SE) was introduced to Switzerland in 2002. Since then, an increasing number of SE programs have been launched across Switzerland. This article reviews the state of SE in Switzerland. METHOD: Several Individual Placement and Support (IPS-SE) studies have been conducted in Switzerland. We reviewed the implementation, results, and possible future developments related to all Swiss IPS-SE studies. FINDINGS: Studies have demonstrated that IPS-SE programs can be successfully implemented with excellent results in Switzerland, but most vocational rehabilitation programs do not provide SE. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: While SE is becoming the first choice for vocational rehabilitation in Switzerland, development and potential improvements are underway. (PsycINFO Database Record (c) 2020 APA, all rights reserved).

[Supported Employment in Routine Operation: Evaluation of the Bern Job Coach Placement Program 2005 - 2016]. / Supported Employment im Routinebetrieb: Evaluation des Berner Job Coach Placement-Programms 2005 ­â€Š2016.

OBJECTIVE: This study reports on the evaluation of the Bern Job Coach Placement program, a supported employment intervention, in the routine operation. METHODS: Administrative data of participants from the years 2005 to 2016 were analysed in terms of integration into the general labour market and in terms of achieving a regular job. RESULTS: 46 percent of all participants (N = 420) were able to achieve the primary outcome of being integrated into the general labour market and 33 percent achieved a regular job. DISCUSSION: The Bern Job Coach Placement program has achieved a high rate of job integration even in routine implementation.

Instrument for Assessing the Ability to Identify Emotional Facial Expressions in Healthy Children and in Children With ADHD: The FEEL Test.

OBJECTIVE: This study presents the validation of a computerized assessment tool that studies the ability to recognize emotional facial expressions in children between 8 and 11 years of age: the Facially Expressed Emotion Labeling Test (FEEL Test). METHOD: The two tests composing the protocol were applied using a laptop in the following order: the FEEL Test followed by the Deusto-e-Motion 1.0 Test.The sample consisted of a total of 1,189 schoolchildren aged between 8 and 11 years, 594 boys and 594 girls. A clinical sample of 47 children with ADHD also took part in this study. RESULTS: The Cronbach's α coefficient for the total scale was .82, showing high levels of reliability. The difficulty index of the items ranged between .4 and .7. The statistical analyses showed a high rate of discrimination between those who obtained low scores compared with those who obtained high scores. The test results reflected differences according to age and gender of participants in many of the variables associated with both response accuracy and response speed. Regarding its predictive validity, the test is able to find statistically significant differences in the total test score among a group of children diagnosed with ADHD and a matched control group. CONCLUSION: This article presents the validation of an instrument that assesses the ability to recognize facial expressions in children between 8 and 11 years old and can discriminate and detect differences in gender, age, and possible deficits in social skills within the ADHD.

Effectiveness of supported employment in non-trial routine implementation: systematic review and meta-analysis.

PURPOSE: While supported employment (SE) programs for people with mental illness have demonstrated their superiority in randomized controlled trials (RCTs) and meta-analyses, little is known about the effectiveness of non-trial routine programs. The primary objective of this study was to estimate a pooled competitive employment rate of non-trial SE programs by means of a meta-analysis. A secondary objective was to compare this result to competitive employment rates of SE programs in RCTs, prevocational training programs in RCTs and in routine implementation. METHODS: A systematic review and a random-effects meta-analysis of proportions were conducted. Quality assessment was provided. Moderator analyses by subgroup comparisons were conducted. RESULTS: Results from 28 samples were included in the meta-analysis. The pooled competitive employment rate for SE routine programs was 0.43 (95% CI 0.37-0.50). The pooled competitive employment rates for comparison conditions were: SE programs in RCTs: 0.50 (95% CI 0.43-0.56); prevocational programs in RCTs: 0.22 (95% CI 0.16-0.28); prevocational programs in routine programs: 0.17 (95% CI 0.11-0.23). SE routine studies conducted prior to 2008 showed a significantly higher competitive employment rate. CONCLUSION: SE routine programs lose only little effectiveness compared to SE programs from RCTs but are much more successful in reintegrating participants into the competitive labor market than prevocational programs. Labor market conditions have to be taken into account when evaluating SE programs.

In situ collection of dust grains falling from Saturn's rings into its atmosphere.

Saturn's main rings are composed of >95% water ice, and the nature of the remaining few percent has remained unclear. The Cassini spacecraft's traversals between Saturn and its innermost D ring allowed its cosmic dust analyzer (CDA) to collect material released from the main rings and to characterize the ring material infall into Saturn. We report the direct in situ detection of material from Saturn's dense rings by the CDA impact mass spectrometer. Most detected grains are a few tens of nanometers in size and dynamically associated with the previously inferred "ring rain." Silicate and water-ice grains were identified, in proportions that vary with latitude. Silicate grains constitute up to 30% of infalling grains, a higher percentage than the bulk silicate content of the rings.

Investigations of the Copper Peptide Hepcidin-25 by LC-MS/MS and NMR.

Hepcidin-25 was identified as the main iron regulator in the human body, and it by binds to the sole iron-exporter ferroportin. Studies showed that the N-terminus of hepcidin is responsible for this interaction, the same N-terminus that encompasses a small copper(II)-binding site known as the ATCUN (amino-terminal Cu(II)- and Ni(II)-binding) motif. Interestingly, this copper-binding property is largely ignored in most papers dealing with hepcidin-25. In this context, detailed investigations of the complex formed between hepcidin-25 and copper could reveal insight into its biological role. The present work focuses on metal-bound hepcidin-25 that can be considered the biologically active form. The first part is devoted to the reversed-phase chromatographic separation of copper-bound and copper-free hepcidin-25 achieved by applying basic mobile phases containing 0.1% ammonia. Further, mass spectrometry (tandem mass spectrometry (MS/MS), high-resolution mass spectrometry (HRMS)) and nuclear magnetic resonance (NMR) spectroscopy were employed to characterize the copper-peptide. Lastly, a three-dimensional (3D) model of hepcidin-25 with bound copper(II) is presented. The identification of metal complexes and potential isoforms and isomers, from which the latter usually are left undetected by mass spectrometry, led to the conclusion that complementary analytical methods are needed to characterize a peptide calibrant or reference material comprehensively. Quantitative nuclear magnetic resonance (qNMR), inductively-coupled plasma mass spectrometry (ICP-MS), ion-mobility spectrometry (IMS) and chiral amino acid analysis (AAA) should be considered among others.

Improved LC-MS/MS method for the quantification of hepcidin-25 in clinical samples.

Mass spectrometry-based methods play a crucial role in the quantification of the main iron metabolism regulator hepcidin by singling out the bioactive 25-residue peptide from the other naturally occurring N-truncated isoforms (hepcidin-20, -22, -24), which seem to be inactive in iron homeostasis. However, several difficulties arise in the MS analysis of hepcidin due to the "sticky" character of the peptide and the lack of suitable standards. Here, we propose the use of amino- and fluoro-silanized autosampler vials to reduce hepcidin interaction to laboratory glassware surfaces after testing several types of vials for the preparation of stock solutions and serum samples for isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS). Furthermore, we have investigated two sample preparation strategies and two chromatographic separation conditions with the aim of developing a LC-MS/MS method for the sensitive and reliable quantification of hepcidin-25 in serum samples. A chromatographic separation based on usual acidic mobile phases was compared with a novel approach involving the separation of hepcidin-25 with solvents at high pH containing 0.1% of ammonia. Both methods were applied to clinical samples in an intra-laboratory comparison of two LC-MS/MS methods using the same hepcidin-25 calibrators with good correlation of the results. Finally, we recommend a LC-MS/MS-based quantification method with a dynamic range of 0.5-40 µg/L for the assessment of hepcidin-25 in human serum that uses TFA-based mobile phases and silanized glass vials. Graphical abstract Structure of hepcidin-25 (Protein Data Bank, PDB ID 2KEF).

Liquid chromatography-tandem mass spectrometry detection of diclofenac and related compounds in water samples.

A frequently studied environmental contaminant is the active substance diclofenac, which is removed insufficiently in sewage treatment plants. Since its inclusion in the watch list of the EU Water Framework Directive, the concentrations in surface waters will be determined throughout Europe. For this, still, more precise analytical methods are needed. As a reference, HPLC-MS is frequently employed. One of the major metabolites is 4'-hydroxydiclofenac (4'-OH-DCF). Also, diclofenac lactam is important for assessing degradation and transformation. Aceclofenac (ACF), the glycolic acid ester of diclofenac is used as a drug, too, and could potentially be cleaved to yield diclofenac again. In various sewage treatment plant influent samples, diclofenac, 4'-OH-DCF, DCF lactam and ACF could be determined with detection limits of 3 µg/L, 0.2 µg/L, 0.17 µg/L and 10 ng/L, respectively.

The Impact of Sex Differences on Odor Identification and Facial Affect Recognition in Patients with Schizophrenia Spectrum Disorders.

BACKGROUND: Social interactive functions such as facial emotion recognition and smell identification have been shown to differ between women and men. However, little is known about how these differences are mirrored in patients with schizophrenia and how these abilities interact with each other and with other clinical variables in patients vs. healthy controls. METHODS: Standardized instruments were used to assess facial emotion recognition [Facially Expressed Emotion Labelling (FEEL)] and smell identification [University of Pennsylvania Smell Identification Test (UPSIT)] in 51 patients with schizophrenia spectrum disorders and 79 healthy controls; furthermore, working memory functions and clinical variables were assessed. RESULTS: In both the univariate and the multivariate results, illness showed a significant influence on UPSIT and FEEL. The inclusion of age and working memory in the MANOVA resulted in a differential effect with sex and working memory as remaining significant factors. Duration of illness was correlated with both emotion recognition and smell identification in men only, whereas immediate general psychopathology and negative symptoms were associated with emotion recognition only in women. CONCLUSION: Being affected by schizophrenia spectrum disorder impacts one's ability to correctly recognize facial affects and identify odors. Converging evidence suggests a link between the investigated basic and social cognitive abilities in patients with schizophrenia spectrum disorders with a strong contribution of working memory and differential effects of modulators in women vs. men.

Contribution of crop model structure, parameters and climate projections to uncertainty in climate change impact assessments.

Climate change impact assessments are plagued with uncertainties from many sources, such as climate projections or the inadequacies in structure and parameters of the impact model. Previous studies tried to account for the uncertainty from one or two of these. Here, we developed a triple-ensemble probabilistic assessment using seven crop models, multiple sets of model parameters and eight contrasting climate projections together to comprehensively account for uncertainties from these three important sources. We demonstrated the approach in assessing climate change impact on barley growth and yield at Jokioinen, Finland in the Boreal climatic zone and Lleida, Spain in the Mediterranean climatic zone, for the 2050s. We further quantified and compared the contribution of crop model structure, crop model parameters and climate projections to the total variance of ensemble output using Analysis of Variance (ANOVA). Based on the triple-ensemble probabilistic assessment, the median of simulated yield change was -4% and +16%, and the probability of decreasing yield was 63% and 31% in the 2050s, at Jokioinen and Lleida, respectively, relative to 1981-2010. The contribution of crop model structure to the total variance of ensemble output was larger than that from downscaled climate projections and model parameters. The relative contribution of crop model parameters and downscaled climate projections to the total variance of ensemble output varied greatly among the seven crop models and between the two sites. The contribution of downscaled climate projections was on average larger than that of crop model parameters. This information on the uncertainty from different sources can be quite useful for model users to decide where to put the most effort when preparing or choosing models or parameters for impact analyses. We concluded that the triple-ensemble probabilistic approach that accounts for the uncertainties from multiple important sources provide more comprehensive information for quantifying uncertainties in climate change impact assessments as compared to the conventional approaches that are deterministic or only account for the uncertainties from one or two of the uncertainty sources.

Effects of Sustained Competitive Employment on Psychiatric Hospitalizations and Quality of Life.

OBJECTIVE: There is extensive evidence for the effectiveness of supported employment among people with severe mental illness. However, less research has been conducted to evaluate the effects of sustained competitive employment on nonvocational outcomes. The aim of this study was to evaluate the effects of sustained competitive work on quality of life and psychiatric hospitalizations. METHODS: As part of a randomized controlled trial, a mediation analysis was used to compare the direct and indirect effects of supported employment versus a traditional vocational program on sustained competitive employment, days of psychiatric hospitalization, and quality of life among 85 participants over five years. RESULTS: The five-year follow-up indicated that the effects of supported employment on reduced days of hospitalization and increased quality of life were fully mediated by the program's effects on increasing sustained competitive employment. CONCLUSIONS: The rehabilitative and therapeutic dimensions of functional health conditions are interrelated in the long term. The achievement of sustained competitive employment may be a key factor in improving social and psychiatric outcomes for people with severe and persistent mental disorders.

Preference for Independent Housing of Persons with Mental Disorders: Systematic Review and Meta-analysis.

Choice of housing has become an important political and therapeutic goal for psychiatric rehabilitation. We conducted a systematic review and meta-analysis of proportions of studies on preference for independent housing. A subgroup analysis compared studies with homeless and non-homeless consumers. The meta-analysis included 8 studies with 3134 consumers. The overall proportion of consumers who had expressed a preference for living independently was 0.84. There were only marginal differences between studies with homeless and non-homeless consumers. In a given service planning area, the rate of independent housing settings should exceed the rate of more institutionalized settings by a wide margin.

Isolation, Co-Crystallization and Structure-Based Characterization of Anabaenopeptins as Highly Potent Inhibitors of Activated Thrombin Activatable Fibrinolysis Inhibitor (TAFIa).

Mature thrombin activatable fibrinolysis inhibitor (TAFIa) is a carboxypeptidase that stabilizes fibrin clots by removing C-terminal arginines and lysines from partially degraded fibrin. Inhibition of TAFIa stimulates the degradation of fibrin clots and may help to prevent thrombosis. Applying a lead finding approach based on literature-mining, we discovered that anabaenopeptins, cyclic peptides produced by cyanobacteria, were potent inhibitors of TAFIa with IC50 values as low as 1.5 nM. We describe the isolation and structure elucidation of 20 anabaenopeptins, including 13 novel congeners, as well as their pronounced structure-activity relationships (SAR) with respect to inhibition of TAFIa. Crystal structures of the anabaenopeptins B, C and F bound to the surrogate protease carboxypeptidase B revealed the binding modes of these large (~850 Da) compounds in detail and explained the observed SAR, i.e. the strong dependence of the potency on a basic (Arg, Lys) exocyclic residue that addressed the S1' binding pocket, and a broad tolerance towards substitutions in the pentacyclic ring that acted as a plug of the active site.