Identification of Proteins Targeted by the Thioredoxin Superfamily in Plasmodium falciparum

The malarial parasite Plasmodium falciparum possesses a functional thioredoxin and glutathione system comprising the dithiol-containing redox proteins thioredoxin (Trx) and glutaredoxin (Grx), as well as plasmoredoxin (Plrx), which is exclusively found in Plasmodium species. All three proteins belong to the thioredoxin superfamily and share a conserved Cys-X-X-Cys motif at the active site. Only a few of their target proteins, which are likely to be involved in redox reactions, are currently known. The aim of the present study was to extend our knowledge of the Trx-, Grx-, and Plrx-interactome in Plasmodium. Based on the reaction mechanism, we generated active site mutants of Trx and Grx lacking the resolving cysteine residue. These mutants were bound to affinity columns to trap target proteins from P. falciparum cell extracts after formation of intermolecular disulfide bonds. Covalently linked proteins were eluted with dithiothreitol and analyzed by mass spectrometry. For Trx and Grx, we were able to isolate 17 putatively redox-regulated proteins each. Furthermore, the approach was successfully established for Plrx, leading to the identification of 21 potential target proteins. In addition to confirming known interaction partners, we captured potential target proteins involved in various processes including protein biosynthesis, energy metabolism, and signal transduction. The identification of three enzymes involved in S-adenosylmethionine (SAM) metabolism furthermore suggests that redox control is required to balance the metabolic fluxes of SAM between methyl-group transfer reactions and polyamine synthesis. To substantiate our data, the binding of the redoxins to S-adenosyl-L-homocysteine hydrolase and ornithine aminotransferase (OAT) were verified using BIAcore surface plasmon resonance. In enzymatic assays, Trx was furthermore shown to enhance the activity of OAT. Our approach led to the discovery of several putatively redox-regulated proteins, thereby contributing to our understanding of the redox interactome in malarial parasites

LebensRäume - Wohn- und Lebensbedingungen aus Sicht der Bewohnerinnen und Bewohner

In der jährlichen Bevölkerungsumfrage erhebt das Bundesamt für Bauwesen und Raumordnung (BBR) seit 1985 (in Ostdeutschland seit 1990) Einstellungen, Meinungen und Wissensbestände der Deutschen mit Blick auf die Wohn- und Lebensbedingungen in ihrer Region. Ein im Kern gleich bleibender Fragenkatalog gewährleistet die langfristige vergleichende Beobachtung gesellschaftlicher Entwicklungen aus Sicht der befragten Bürgerinnen und Bürger. Neben der Beschreibung regionaler Unterschiede und deren Beurteilung dient die Befragung vor allem der Untersuchung systematischer Zusammenhänge von Lebensbedingungen, Bewertungen und berichteten Verhaltensweisen. Sie ist Teil des Raumbezogenen Informationssystems des BBR. In diesem Informationssystem ist die Bevölkerungsumfrage die einzige Datenquelle, die es erlaubt, Zusammenhänge auf der Ebene von Personen herauszufinden (subjektive Indikatoren). Alle anderen Komponenten der Raumbeobachtung können Analysen nur auf der Ebene von Aggregatstatistiken vornehmen (objektive Indikatoren). Die subjektiven Einsichten der Befragten helfen dabei, regionalstatistische Zusammenhänge zu erklären. Die im vorliegenden Band präsentierten Analysen ergänzen frühere Veröffentlichungen. Ein besonderer Schwerpunkt liegt zudem auf den Lebensbedingungen im Wohngebiet (Sonderfragen 2003). Der Anhang versammelt das Fragenprogramm der verschiedenen Erhebungsjahre und Zeitreihen-Tabellen. Die Datensätze der Umfragen 2000 bis 2012 sind im ZACAT des GESIS Leibniz Instituts für Sozialwissenschaften unter der Studiennummer ZA5611 online verfügbar (s. https://doi.org/10.4232/1.12069)

Evaluation of a multidisciplinary lipid clinic to improve the care of individuals with severe lipid conditions: A RE-AIM framework analysis

BACKGROUND: Individuals with complex dyslipidemia, or those with medication intolerance, are often difficult to manage in primary care. They require the additional attention, expertise, and adherence counseling that occurs in multidisciplinary lipid clinics (MDLCs). We conducted a program evaluation of the first year of a newly implemented MDLC utilizing the RE-AIM (reach, effectiveness, adoption, implementation, and maintenance) framework to provide empirical data not only on program effectiveness, but also on components important to local sustainability and future generalizability. METHODS: The purpose of the MDLC is to increase the uptake of guideline-based care for lipid conditions. Established in 2019, the MDLC provides care via a centralized clinic location within the healthcare system. Primary care providers and cardiologists were invited to refer individuals with lipid conditions. Using a pre/post-study design, we evaluated the implementation outcomes from the MDLC using the RE-AIM framework. RESULTS: In 2019, 420 referrals were made to the MDLC (reach). Referrals were made by 19% (148) of the 796 active cardiology and primary care providers, with an average of 35 patient referrals per month in 2019 (SD 12) (adoption). The MDLC saw 83 patients in 2019 (reach). Additionally, 50% (41/82) had at least one follow-up MDLC visit, and 12% (10/82) had two or more follow-up visits in 2019 (implementation). In patients seen by the MDLC, we found an improved diagnosis of specific lipid conditions (FH (familial hypercholesterolemia), hypertriglyceridemia, and dyslipidemia), increased prescribing of evidence-based therapies, high rates of medication prior authorization approvals, and significant reductions in lipid levels by lipid condition subgroup (effectiveness). Over time, the operations team decided to transition from in-person follow-up to telehealth appointments to increase capacity and sustain the clinic (maintenance). CONCLUSIONS: Despite limited reach and adoption of the MDLC, we found a large intervention effect that included improved diagnosis, increased prescribing of guideline-recommended treatments, and clinically significant reduction of lipid levels. Attention to factors including solutions to decrease the large burden of unseen referrals, discussion of the appropriate number and duration of visits, and sustainability of the clinic model could aid in enhancing the success of the MDLC and improving outcomes for more patients throughout the system

Melanocortin-1 receptor signaling markedly induces the expression of the NR4A nuclear receptor subgroup in melanocytic cells

The melanocortin-1 receptor (MCIR) is a G-protein-coupled receptor expressed primarily in melanocytes and is known to play a pivotal role in the regulation of pigmentation in mammals. In humans MC1R has been found to be highly polymorphic with several functional variants associated with the phenotype of red hair color and fair skin, cutaneous UV sensitivity, and increased risk of developing melanoma and non-melanoma skin cancer. Recent evidence suggests that MC1R plays a photo-protective role in melanocytes in response to UV irradiation. Relatively few genetic targets of MC1R signaling have been identified independent of the pigmentation pathway. Here we show that MC1R signaling in B16 mouse melanoma cells and primary human melanocytes rapidly, and transiently, induces the transcription of the NR4A subfamily of orphan nuclear receptors. Furthermore, primary human melanocytes harboring homozygous RHC variant MC1R alleles exhibited an impaired induction of NR4A genes in response to the potent MC1R agonist (Nle4,D-Phe7)-α-melanocyte-stimulating hormone. Using small interference RNA-mediated attenuation of NR4A1 and NR4A2 expression in melanocytes, the ability to remove cyclobutane pyrimidine dimers following UV irradiation appeared to be impaired in the context of MC1R signaling. These data identify the NR4A receptor family as potential mediators of an MC1R-coordinated DNA damage response to UV exposure in melanocytic cells

Jet-Powered Molecular Hydrogen Emission from Radio Galaxies

H2 pure-rotational emission lines are detected from warm (100-1500 K) molecular gas in 17/55 (31% of) radio galaxies at redshift z<0.22 observed with the Spitzer IR Spectrograph. The summed H2 0-0 S(0)-S(3) line luminosities are L(H2)=7E38-2E42 erg/s, yielding warm H2 masses up to 2E10 Msun. These radio galaxies, of both FR radio morphological types, help to firmly establish the new class of radio-selected molecular hydrogen emission galaxies (radio MOHEGs). MOHEGs have extremely large H2 to 7.7 micron PAH emission ratios: L(H2)/L(PAH7.7) = 0.04-4, up to a factor 300 greater than the median value for normal star-forming galaxies. In spite of large H2 masses, MOHEGs appear to be inefficient at forming stars, perhaps because the molecular gas is kinematically unsettled and turbulent. Low-luminosity mid-IR continuum emission together with low-ionization emission line spectra indicate low-luminosity AGNs in all but 3 radio MOHEGs. The AGN X-ray emission measured with Chandra is not luminous enough to power the H2 emission from MOHEGs. Nearly all radio MOHEGs belong to clusters or close pairs, including 4 cool core clusters (Perseus, Hydra, A 2052, and A 2199). We suggest that the H2 in radio MOHEGs is delivered in galaxy collisions or cooling flows, then heated by radio jet feedback in the form of kinetic energy dissipation by shocks or cosmic rays.Comment: ApJ in press, 40 pages, 18 figures, 14 table

Probiotic Sonicates Selectively Induce Mucosal Immune Cells Apoptosis through Ceramide Generation via Neutral Sphingomyelinase

This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al.[Background]: Probiotics appear to be beneficial in inflammatory bowel disease, but their mechanism of action is incompletely understood. We investigated whether probiotic-derived sphingomyelinase mediates this beneficial effect. [Methodology/Principal Findings]: Neutral sphingomyelinase (NSMase) activity was measured in sonicates of the probiotic L. brevis (LB) and S. thermophilus (ST) and the non-probiotic E. coli (EC) and E. faecalis (EF). Lamina propria mononuclear cells (LPMC) were obtained from patients with Crohn's disease (CD) and Ulcerative Colitis (UC), and peripheral blood mononuclear cells (PBMC) from healthy volunteers, analysing LPMC and PBMC apoptosis susceptibility, reactive oxygen species (ROS) generation and JNK activation. In some experiments, sonicates were preincubated with GSH or GW4869, a specific NSMase inhibitor. NSMase activity of LB and ST was 10-fold that of EC and EF sonicates. LB and ST sonicates induced significantly more apoptosis of CD and UC than control LPMC, whereas EC and EF sonicates failed to induce apoptosis. Pre-stimulation with anti-CD3/CD28 induced a significant and time-dependent increase in LB-induced apoptosis of LPMC and PBMC. Exposure to LB sonicates resulted in JNK activation and ROS production by LPMC. NSMase activity of LB sonicates was completely abrogated by GW4869, causing a dose-dependent reduction of LB-induced apoptosis. LB and ST selectively induced immune cell apoptosis, an effect dependent on the degree of cell activation and mediated by bacterial NSMase. [Conclusions]: These results suggest that induction of immune cell apoptosis is a mechanism of action of some probiotics, and that NSMase-mediated ceramide generation contributes to the therapeutic effects of probiotics.The funding sources included grants from Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Ministerio de Ciencia e Innovación (SAF2005-00280 and SAF2008-03676 to MS, FIS2009-00056 to AM, SAF2009-11417 to JCF), Fundación Ramón Areces (to MS), the National Institutes of Health (DK30399 and DK50984 to CF) and the Research Center for Liver and Pancreatic Diseases funded by the United States National Institute for Alcohol Abuse and Alcoholism (P50 AA 11999 to JCF).Peer reviewe

Physician and Patient Predictors of Evidence-Based Prescribing in Heart Failure: A Multilevel Study

BACKGROUND: The management of patients with heart failure (HF) needs to account for changeable and complex individual clinical characteristics. The use of renin angiotensin system inhibitors (RAAS-I) to target doses is recommended by guidelines. But physicians seemingly do not sufficiently follow this recommendation, while little is known about the physician and patient predictors of adherence. METHODS: To examine the coherence of primary care (PC) physicians' knowledge and self-perceived competencies regarding RAAS-I with their respective prescribing behavior being related to patient-associated barriers. Cross-sectional follow-up study after a randomized medical educational intervention trial with a seven month observation period. PC physicians (n = 37) and patients with systolic HF (n = 168) from practices in Baden-Wuerttemberg. Measurements were knowledge (blueprint-based multiple choice test), self-perceived competencies (questionnaire on global confidence in the therapy and on frequency of use of RAAS-I), and patient variables (age, gender, NYHA functional status, blood pressure, potassium level, renal function). Prescribing was collected from the trials' documentation. The target variable consisted of ≥50% of recommended RAAS-I dosage being investigated by two-level logistic regression models. RESULTS: Patients (69% male, mean age 68.8 years) showed symptomatic and objectified left ventricular (NYHA II vs. III/IV: 51% vs. 49% and mean LVEF 33.3%) and renal (GFR<50%: 22%) impairment. Mean percentage of RAAS-I target dose was 47%, 59% of patients receiving ≥50%. Determinants of improved prescribing of RAAS-I were patient age (OR 0.95, CI 0.92-0.99, p = 0.01), physician's global self-confidence at follow-up (OR 1.09, CI 1.02-1.05, p = 0.01) and NYHA class (II vs. III/IV) (OR 0.63, CI 0.38-1.05, p = 0.08). CONCLUSIONS: A change in physician's confidence as a predictor of RAAS-I dose increase is a new finding that might reflect an intervention effect of improved physicians' intention and that might foster novel strategies to improve safe evidence-based prescribing. These should include targeting knowledge, attitudes and skills

First results from the JWST Early Release Science Program Q3D: Ionization cone, clumpy star formation and shocks in a z=3z=3 extremely red quasar host

Massive galaxies formed most actively at redshifts $z=1-3$ during the period known as `cosmic noon.' Here we present an emission-line study of an extremely red quasar SDSSJ165202.64+172852.3 host galaxy at $z=2.94$, based on observations with the Near Infrared Spectrograph (NIRSpec) integral field unit (IFU) on board JWST. We use standard emission-line diagnostic ratios to map the sources of gas ionization across the host and a swarm of companion galaxies. The quasar dominates the photoionization, but we also discover shock-excited regions orthogonal to the ionization cone and the quasar-driven outflow. These shocks could be merger-induced or -- more likely, given the presence of a powerful galactic-scale quasar outflow -- these are signatures of wide-angle outflows that can reach parts of the galaxy that are not directly illuminated by the quasar. Finally, the kinematically narrow emission associated with the host galaxy presents as a collection of 1 kpc-scale clumps forming stars at a rate of at least 200 $M_{\odot}$ yr$^{-1}$. The ISM within these clumps shows high electron densities, reaching up to 3,000 cm$^{-3}$ with metallicities ranging from half to a third solar with a positive metallicity gradient and V band extinctions up to 3 magnitudes. The star formation conditions are far more extreme in these regions than in local star-forming galaxies but consistent with that of massive galaxies at cosmic noon. JWST observations reveal an archetypical rapidly forming massive galaxy undergoing a merger, a clumpy starburst, an episode of obscured near-Eddington quasar activity, and an extremely powerful quasar outflow simultaneously.Comment: 19 pages, 8 figures. Accepted for publication in Ap

Recommendations for whole genome sequencing in diagnostics for rare diseases

In 2016, guidelines for diagnostic Next Generation Sequencing (NGS) have been published by EuroGentest in order to assist laboratories in the implementation and accreditation of NGS in a diagnostic setting. These guidelines mainly focused on Whole Exome Sequencing (WES) and targeted (gene panels) sequencing detecting small germline variants (Single Nucleotide Variants (SNVs) and insertions/deletions (indels)). Since then, Whole Genome Sequencing (WGS) has been increasingly introduced in the diagnosis of rare diseases as WGS allows the simultaneous detection of SNVs, Structural Variants (SVs) and other types of variants such as repeat expansions. The use of WGS in diagnostics warrants the re-evaluation and update of previously published guidelines. This work was jointly initiated by EuroGentest and the Horizon2020 project Solve-RD. Statements from the 2016 guidelines have been reviewed in the context of WGS and updated where necessary. The aim of these recommendations is primarily to list the points to consider for clinical (laboratory) geneticists, bioinformaticians, and (non-)geneticists, to provide technical advice, aid clinical decision-making and the reporting of the results