Metabolic Profiling of S-praziquantel: Structure Elucidation Using the Crystalline Sponge Method in Combination with Mass Spectrometry and Nuclear Magnetic Resonance

Praziquantel (PZQ) is the drug of choice for treatment of the neglected tropical disease schistosomiasis. Although the drug has been extensively used over several decades and its metabolism well studied (several oxidative metabolites are known from literature), the knowledge of the complete structure of some of its metabolites remains elusive. Conventional techniques, such as nuclear magnetic resonance or liquid chromatography mass spectrometry were used in the past to investigate phase I and phase II metabolites of PZQ. These techniques are either limited to provide the complete molecular structure (liquid chromatography mass spectrometry) or require large amount of sample material (NMR), which are not always available when in vitro systems are used for investigation of the metabolites. In this study, we describe new structures of S-PZQ metabolites generated in vitro from human liver microsomes using the crystalline sponge method. After chromatographic separation and purification of the oxidative metabolites, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis was conducted to narrow down the position of oxidation to a certain part of the molecule. To determine the exact position of hydroxylation, singe-crystal X-ray diffraction analysis of the crystalline sponges and absorbed analyte was used to identify the structure of S-PZQ and its metabolites. The crystalline sponge method allowed for complete structure elucidation of the known metabolites S-trans-4'-hydroxy-PZQ (M1), S-cis-4'-hydroxy-PZQ (M2) and S-/R-11b-hydroxy-PZQ (M6) as well as the unknown metabolites S-9-hydroxy-PZQ (M3) and S-7-hydroxy-S-PZQ (M4). For comparison of structural elucidation techniques, one metabolite (M3) was additionally analyzed using NMR. SIGNIFICANCE STATEMENT: The information content of the metabolic pathway of praziquantel is still limited. The crystalline sponge method allowed the complete structural elucidation of three known and two unknown metabolites of S-praziquantel, using only trace amounts of analyte material, as demonstrated in this study

Controversial Aspects of Diagnostics and Therapy of Arthritis of the Temporomandibular Joint in Rheumatoid and Juvenile Idiopathic Arthritis: An Analysis of Evidence- and Consensus-Based Recommendations Based on an Interdisciplinary Guideline Project

Introduction: Due to potentially severe sequelae (impaired growth, condylar resorption, and ankylosis) early diagnosis of chronic rheumatic arthritis of the temporomandibular joint (TMJ) and timely onset of therapy are essential. Aim: Owing to very limited evidence the aim of the study was to identify and discuss controversial topics in the guideline development to promote further focused research. Methods: Through a systematic literature search, 394 out of 3771 publications were included in a German interdisciplinary guideline draft. Two workgroups (1: oral and maxillofacial surgery, 2: interdisciplinary) voted on 77 recommendations/statements, in 2 independent anonymized and blinded consensus phases (Delphi process). Results: The voting results were relatively homogenous, except for a greater proportion of abstentions amongst the interdisciplinary group (p < 0.001). Eighty four percent of recommendations/statements were approved in the first round, 89% with strong consensus. Fourteen recommendations/statements (18.2%) required a prolonged consensus phase and further discussion. Discussion: Contrast-enhanced MRI was confirmed as the method of choice for the diagnosis of TMJ arthritis. Intraarticular corticosteroid injection is to be limited to therapy refractory cases and single injection only. In adults, alloplastic joint replacement is preferable to autologous replacement. In children/adolescents, autologous reconstruction may be performed lacking viable alternatives. Alloplastic options are currently still considered experimental

Feeding, survival, and reproduction of two populations of Eurytemora (Copepoda) exposed to local toxic cyanobacteria

Studying the responses of crustacean zooplankton to harmful algal blooms is important for understanding changes in lower food webs following eutrophication in the Laurentian Great Lakes and other regions around the world. Here we examine responses to toxic cyanobacteria by crustacean copepods of the genus Eurytemora from eutrophic coastal regions of Lake Michigan (Green Bay) and the Baltic Sea (Gulf of Finland). We measured grazing, survivorship, reproduction, and juvenile (nauplius) size in short-term laboratory experiments. Females were incubated with representative non-toxic food and mixtures of non-toxic food with either cyanobacteria or cyanobacteria filtrate. Eurytemora from both locations were affected negatively by cyanobacteria filtrates, even with non-toxic food available. Eurytemora carolleeae from Green Bay exhibited reduced grazing rates when exposed to filtrates, but this effect was not observed when animals were fed the cyanobacteria and filtrate along with non-toxic food. Eurytemora sp. from the Baltic Sea given filtrates and non-toxic food also exhibited decreased grazing rates, as well as decreased adult survival and nauplius size. Similarly, when cyanobacterial cells were included along with filtrate and non-toxic food these effects were not observed. Our results also demonstrated a significant trade-off between offspring quantity and quality for both groups of animals, being more pronounced when food quality was manipulated by the presence of cyanobacterial cells. These findings further our knowledge of how a widely distributed group like Eurytemora may respond in the face of changing local selection pressures from natural and anthropogenic stressors. (C) 2017 International Association for Great Lakes Research. Published by Elsevier B.V. All rights reserved.Peer reviewe

Ion temperature clamping in Wendelstein 7-X electron cyclotron heated plasmas

The neoclassical transport optimization of the Wendelstein 7-X stellarator has not resulted in the predicted high energy confinement of gas fueled electron-cyclotron-resonance-heated (ECRH) plasmas as modelled in (Turkin et al 2011 Phys. Plasmas 18 022505) due to high levels of turbulent heat transport observed in the experiments. The electron-turbulent-heat transport appears non-stiff and is of the electron temperature gradient (ETG)/ion temperature gradient (ITG) type (Weir et al 2021 Nucl. Fusion 61 056001). As a result, the electron temperature Te can be varied freely from 1 keV–10 keV within the range of PECRH = 1–7 MW, with electron density ne values from 0.1–1.5 × 1020 m−3. By contrast, in combination with the broad electron-to-ion energy-exchange heating profile in ECRH plasmas, ion-turbulent-heat transport leads to clamping of the central ion temperature at Ti ∼ 1.5 keV ± 0.2 keV. In a dedicated ECRH power scan at a constant density of 〈ne〉 = 7 × 1019 m−3, an apparent \u27negative ion temperature profile stiffness\u27 was found in the central plasma for (r/a < 0.5), in which the normalized gradient ∇Ti/Ti decreases with increasing ion heat flux. The experiment was conducted in helium, which has a higher radiative density limit compared to hydrogen, allowing a broader power scan. This \u27negative stiffness\u27 is due to a strong exacerbation of turbulent transport with an increasing ratio of Te/Ti in this electron-heated plasma. This finding is consistent with electrostatic microinstabilities, such as ITG-driven turbulence. Theoretical calculations made by both linear and nonlinear gyro-kinetic simulations performed by the GENE code in the W7-X three-dimensional geometry show a strong enhancement of turbulence with an increasing ratio of Te/Ti. The exacerbation of turbulence with increasing Te/Ti is also found in tokamaks and inherently enhances ion heat transport in electron-heated plasmas. This finding strongly affects the prospects of future high-performance gas-fueled ECRH scenarios in W7-X and imposes a requirement for turbulence-suppression techniques

Confinement in electron heated plasmas in Wendelstein 7-X and ASDEX Upgrade; the necessity to control turbulent transport

In electron (cyclotron) heated plasmas, in both ASDEX Upgrade (L-mode) and Wendelstein 7-X, clamping of the ion temperature occurs at Ti ∼ 1.5 keV independent of magnetic configuration. The ions in such plasmas are heated through the energy exchange power as ${n}_{\mathrm{e}}^{2}({T}_{\mathrm{e}}-{T}_{\mathrm{i}})/{T}_{\mathrm{e}}^{3/2}$, which offers a broad ion heating profile, similar to that offered by alpha heating in future thermonuclear fusion reactors. However, the predominant electron heating may put an additional constraint on the ion heat transport, as the ratio Te/Ti > 1 can exacerbates ITG/TEM core turbulence. Therefore, in practical terms the strongly 'stiff' core transport translates into Ti-clamping in electron heated plasmas. Due to this clamping, electron heated L-mode scenarios, with standard gas fueling, in either tokamaks or stellarators may struggle to reach high normalized ion temperature gradients required in a compact fusion reactor. The comparison shows that core heat transport in neoclassically optimized stellarators is driven by the same mechanisms as in tokamaks. The absence of a strong H-mode temperature edge pedestal in stellarators, sofar (which, like in tokamaks, could lift the clamped temperature-gradients in the core), puts a strong requirement on reliable and sustainable core turbulence suppression techniques in stellarators.EC/H2020/633053/EU/Implementation of activities described in the Roadmap to Fusion during Horizon 2020 through a Joint programme of the members of the EUROfusion consortium/Eurato

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

GWAS meta-analysis of over 29,000 people with epilepsy identifies 26 risk loci and subtype-specific genetic architecture

Epilepsy is a highly heritable disorder affecting over 50 million people worldwide, of which about one-third are resistant to current treatments. Here we report a multi-ancestry genome-wide association study including 29,944 cases, stratified into three broad categories and seven subtypes of epilepsy, and 52,538 controls. We identify 26 genome-wide significant loci, 19 of which are specific to genetic generalized epilepsy (GGE). We implicate 29 likely causal genes underlying these 26 loci. SNP-based heritability analyses show that common variants explain between 39.6% and 90% of genetic risk for GGE and its subtypes. Subtype analysis revealed markedly different genetic architectures between focal and generalized epilepsies. Gene-set analyses of GGE signals implicate synaptic processes in both excitatory and inhibitory neurons in the brain. Prioritized candidate genes overlap with monogenic epilepsy genes and with targets of current antiseizure medications. Finally, we leverage our results to identify alternate drugs with predicted efficacy if repurposed for epilepsy treatment

Genome-wide identification and phenotypic characterization of seizure-associated copy number variations in 741,075 individuals

Copy number variants (CNV) are established risk factors for neurodevelopmental disorders with seizures or epilepsy. With the hypothesis that seizure disorders share genetic risk factors, we pooled CNV data from 10,590 individuals with seizure disorders, 16,109 individuals with clinically validated epilepsy, and 492,324 population controls and identified 25 genome-wide significant loci, 22 of which are novel for seizure disorders, such as deletions at 1p36.33, 1q44, 2p21-p16.3, 3q29, 8p23.3-p23.2, 9p24.3, 10q26.3, 15q11.2, 15q12-q13.1, 16p12.2, 17q21.31, duplications at 2q13, 9q34.3, 16p13.3, 17q12, 19p13.3, 20q13.33, and reciprocal CNVs at 16p11.2, and 22q11.21. Using genetic data from additional 248,751 individuals with 23 neuropsychiatric phenotypes, we explored the pleiotropy of these 25 loci. Finally, in a subset of individuals with epilepsy and detailed clinical data available, we performed phenome-wide association analyses between individual CNVs and clinical annotations categorized through the Human Phenotype Ontology (HPO). For six CNVs, we identified 19 significant associations with specific HPO terms and generated, for all CNVs, phenotype signatures across 17 clinical categories relevant for epileptologists. This is the most comprehensive investigation of CNVs in epilepsy and related seizure disorders, with potential implications for clinical practice