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Staphylococcus aureussuperantigens (SAgs) are among the most potent T cell mitogensknown.They stimulate large fractions of T cells by cross-linking their T cell receptor withmajor histocompatibility complex class-II molecules on antigen presenting cells, resulting in Tcell proliferation and massive cytokine release. To date, 26 different SAgs have been described in thespeciesS. aureus; they comprise the toxic shock syndrome toxin (TSST-1), as well as 25 staphylococcalenterotoxins (SEs) or enterotoxin-like proteins (SEls). SAgs can cause staphylococcal food poisoningand toxic shock syndrome and contribute to the clinical symptoms of staphylococcal infection. Inaddition, there is growing evidence that SAgs are involved in allergic diseases. This review providesan overview on recent epidemiological data on the involvement ofS. aureusSAgs and anti-SAg-IgEin allergy, demonstrating that being sensitized to SEs—in contrast to inhalant allergens—is associatedwith a severe disease course in patients with chronic airway inflammation. The mechanisms by whichSAgs trigger or amplify allergic immune responses, however, are not yet fully understood. Here, wediscuss known and hypothetical pathways by which SAgs can drive an atopic disease
Oral administration of drugs is the most common, convenient, safest and economical route of drug administration. There is lack of established tools to study the function of transporters in the intestinal absorption of drugs. Because of its favorable physico-chemical, pharmacokinetic and pharmacodynamic characteristics, trospium could be potentially used as a probe substrate to study the function of drug transporters. Therefore, this study was conducted to examine the suitability of trospium chloride as a probe drug to study the function of multidrug transporters in the human body. To this end, two randomized, controlled, four-period, cross-over pharmacokinetic drug interaction studies of oral and intravenous trospium with co-medication of oral clarithromycin or ranitidine were performed in 24 healthy subjects to mechanistically characterize the role of P-gp, OATP1A2, OCT1, OCT2, MATE1 and MATE2-K in the absorption and disposition of trospium. The contribution of the drug transporters in the absorption and disposition of trospium were examined in isolated systems using in vitro uptake and inhibition assays in transporter transfected human cell lines.
OCT1 (Vmax = 0.8 ± 0.1 nmol/min × mg) is a high capacity transporter of trospium compared to OCT2 (Vmax = 0.04 ± 0.01 nmol/min × mg). But the OCT2 (Km = 0.5 ± 0.1 µM) transporter demonstrated a high affinity in the transport of trospium compared to OCT1 (Km = 17.4 ± 2.1 µM). OCT1 genetic alleles *2, *3, *4 and *7 resulted in significant loss of activity and the alleles *5 and *6 caused complete loss of uptake of trospium. The common OCT2 genetic allele Ser270 caused slight but significant increase in activity of OCT2.
Ranitidine inhibits OCT1 (IC50 = 186 ± 25 µM), MATE1 (IC50 = 134 ± 37 µM) and MATE2-K (IC50 = 35 ± 11 µM)-mediated uptake of trospium in vitro. But it is a weak inhibitor of OCT2 transporter (IC50 = 482 ± 105 µM). Using FDA and EMA in vitro to in vivo extrapolation models, ranitidine was predicted to have a potential inhibition effect on intestinal OCT1 ([I]2/IC50 ~40), renal MATE1 ([I]1/IC50 ~0.02) and MATE2-K ([I]1/IC50 ~0.1) transporters in vivo. Clarithromycin was predicted to cause DDI by inhibiting P-gp-mediated efflux of trospium at the intestine ([I]2/IC50 of ~310) and hepatocytes ([I]3/IC50 ~1). Therefore, co-medication of oral clarithromycin was expected to result in an increase in oral absorption and hepatic clearance of trospium but not changes in distribution volume.
In healthy subjects, oral trospium is slowly (MAT ~10 h) and poorly (F ~10 %) absorbed from the jejunum and cecum/ascending colon, widely distributed into the body (Vss = 5 - 6 l/kg) and slowly eliminated (t1/2 = 9 - 10 h) majorly via renal glomerular filtration and tubular secretion (CLR ~500 ml/min). After co-medication of clarithromycin (inhibitor of P-gp), on the contrary to our IVIVE prediction, we found a non-expected but significant expansion of the shallow and deep distribution spaces for trospium by ~27 %. A single dose administration of trospium with co-medication of ranitidine (inhibitor of OCT1) resulted in no effect on the intestinal absorption of trospium. But the renal clearance of trospium decreased slightly (15 %) but significantly.
Intravenously administered trospium (2 mg TC) might be a suitable probe drug to evaluate the effects of a P-gp inhibitor on distribution of a drug. Oral trospium chloride can be selected for DDI studies with new chemical entities (NCE) with predicted inhibitory potential on OCT1 and P-gp and which are available after oral absorption along the small intestine and in the cecum/ascending colon. Another kind of application of trospium chloride might be pharmacogenomics studies in subjects with functionally relevant polymorphisms of P-gp and OCT1 or in patients with suspected transport failure due to intestinal diseases. The function of the efflux transporters MATE1 and MATE2-K in the PTC of the kidneys can be well assessed with the probe drug trospium by measuring its renal clearance.
Background: Stroke patients are at risk of acquiring secondary infections due to stroke-induced immune suppression (SIIS). Immunosuppressive cells comprise myeloid-derived suppressor cells (MDSCs) and immunosuppressive interleukin 10 (IL-10)-producing monocytes. MDSCs represent a small but heterogeneous population of monocytic, polymorphonuclear (or granulocytic), and early progenitor cells (“early” MDSC), which can expand extensively in pathophysiological conditions. MDSCs have been shown to exert strong immune-suppressive effects. The role of IL-10-producing immunosuppressive monocytes after stroke has not been investigated, but monocytes are impaired in oxidative burst and downregulate human leukocyte antigen—DR isotype (HLA-DR) on the cell surface.
Objectives: The objective of this work was to investigate the regulation and function of MDSCs as well as the immunosuppressive IL-10-producing monocytes in experimental and human stroke.
Methods: This longitudinal, monocentric, non-interventional prospective explorative study used multicolor flow cytometry to identify MDSC subpopulations and IL-10 expression in monocytes in the peripheral blood of 19 healthy controls and 27 patients on days 1, 3, and 5 post-stroke. Quantification of intracellular STAT3p and Arginase-1 by geometric mean fluorescence intensity was used to assess the functionality of MDSCs. In experimental stroke induced by electrocoagulation in middle-aged mice, monocytic (CD11b+Ly6G−Ly6Chigh) and polymorphonuclear (CD11b+Ly6G+Ly6Clow) MDSCs in the spleen were analyzed by flow cytometry.
Results: Compared to the controls, stroke patients showed a relative increase in monocytic MDSCs (percentage of CD11b+ cells) in whole blood without evidence for an altered function. The other MDSC subgroups did not differ from the control. Also, in experimental stroke, monocytic, and in addition, polymorphonuclear MDSCs were increased. The numbers of IL-10-positive monocytes did not differ between the patients and controls. However, we provide a new insight into monocytic function post-stroke since we can report that a differential regulation of HLA-DR and PD-L1 was found depending on the IL-10 production of monocytes. IL-10-positive monocytes are more activated post-stroke, as indicated by their increased HLA-DR expression.
Conclusions: MDSC and IL-10+ monocytes can induce immunosuppression within days after stroke.
Der aktuelle demografische Wandel in Deutschland zeigt eine erhöhte Lebenserwartung und damit einen Anstieg an altersassoziierten Erkrankungen wie dem Schlaganfall. Eine mögliche Folge ist die Armparese, welche eine gravierende Behinderung bei der Ausführung alltäglicher Handlungen darstellt. Dadurch kommt der motorischen Rehabilitation mit dem Ziel der Wiederherstellung der Alltagstauglichkeit eine besonders wichtige Rolle zu.
Unter zahlreichen Therapiekonzepten ist für das Arm-Fähigkeits-Training (AFT), welches einzeln verschiedene sensomotorische Armfähigkeiten anspricht und das motorische Lernen induziert, eine gute Wirksamkeit belegt.
In Studien konnte gezeigt werden, dass eine nicht-invasive Hirnstimulation in Form einer repetitiven transkraniellen Magnetstimulation (rTMS), genauer der intermittierenden Theta-Burst-Stimulation (iTBS), vorübergehend die lokal kortikale Erregbarkeit des stimulierten Areals erhöhen (Huang et al., 2005) und dadurch gegebenenfalls auch das nachfolgende trainingsinduzierte Lernen beeinflussen kann. Das Wissen über mögliche „Priming“-Effekte von iTBS auf das motorische Lernen bei Gesunden kann helfen, zielgerichtete therapeutische Anwendungen für Patienten nach einem Schlaganfall zu entwickeln.
Ziel dieser Untersuchung war es festzustellen, ob das exzitatorische „Priming“ mit iTBS über dem primären motorischen Kortex (M1) oder dem primären somatosensorischen Kortex (S1) unmittelbar vor einer täglichen Trainingseinheit mit AFT (über vier Tage) für den linken Arm bei gesunden rechtshändigen Probanden die sensomotorische Lerndynamik verbessern kann.
Zu diesem Zweck wurde ein Training des linken, nicht-dominanten Arms von 18 jungen und gesunden Probanden mithilfe von acht unterschiedlichen motorischen Aufgaben (AFT) einmal pro Tag für insgesamt fünf Tage durchgeführt. Mit Ausnahme des ersten Tages (Baseline) erfolgte das Training nach der Applikation einer exzitatorischen Form der repetitiven transkraniellen Magnetstimulation (iTBS). Die Stimulation wurde je nach randomisierter Gruppenzuordnung entweder über M1 oder S1 rechts oder als Sham-Stimulation, um einen möglichen Placebo-Effekt auszuschließen, über M1 rechts durchgeführt.
Die Hauptkomponentenanalyse der Daten zum motorischen Verhalten ergab acht
unabhängige motorische Komponenten, die den acht trainierten Aufgaben entsprachen. AFT induzierte motorisches Lernen über alle Fähigkeiten hinweg mit einem Generalisationseffekt auf eine nicht-trainierte Aufgabe der Fingergeschicklichkeitm(Nine-Hole-Peg-Test,ccNHPT).
Probanden, die iTBS (entweder über M1 oder S1) erhielten, zeigten im Vergleich zur Sham-Stimulation sowohl eine bessere Leistung bei den AFT-Aufgaben während der Trainingsdauer als auch eine größere Verbesserung der nicht-trainierten Fingergeschicklichkeitsaufgabe (NHPT) für den trainierten linken Arm nach Trainingsende.
Daraus resultiert, dass die exzitatorische repetitive transkranielle Magnetstimulation in Form von iTBS über M1 oder S1 das motorische Lernen über verschiedene sensomotorische Fähigkeiten hinweg verbessern kann.
Auch wenn die verstärkenden Effekte eines exzitatorischen „Priming“ absolut gesehen klein waren, so geben sie dennoch Grund zur Annahme, dass darin auch ein therapeutisches Potenzial für die Armrehabilitation nach Schlaganfall liegt. Ob das so ist, wäre jedoch mit geeigneten klinischen Studien zu untersuchen.
Die Ergebnisse des Promotionsvorhabens wurden in einer Peer-Review-Zeitschrift publiziert (Platz et al., 2018a).
Abstract
Introduction
Transabdominal ultrasound (US) and magnetic resonance imaging (MRI) are commonly used for the examination of the pancreas in clinical routine. We therefore were interested in the concordance of these two imaging methods for the size measurement of the pancreas and how age, gender, and body mass index (BMI) affect the organ size.
Methods
A total of 342 participants from the Study of Health in Pomerania underwent whole‐body MRI and transabdominal US on the same day, and the diameter of the pancreatic head, body, and tail were measured. The agreement between US and MRI measurements was assessed by Bland and Altman plots. Intraclass correlation coefficients were used to compare observers. A multivariable regression model was applied using the independent variables age, gender, and body mass index.
Results
Compared to MRI, abdominal US returned smaller values for each segment of the pancreas, with a high level of inconsistency between these two methods. The mean difference was 0.39, 0.18, and 0.54 cm for the head, body, and tail, respectively. A high interobserver variability was detected for US. Multivariable analysis showed that pancreatic size in all three segments increased with BMI in both genders whereas pancreatic head and tail size decreased with age, an effect more marked in women.
Conclusions
Agreement of pancreatic size measurements is poor between US and MRI. These limitations should be considered when evaluating morphologic features for pathologic conditions or setting limits of normal size. Adjustments for BMI, gender, and age may also be warranted.
Das trockene Auge ist eine häufige multikausale Augenerkrankung, welche nicht selten mit einer erheblichen Einschränkung der Lebensqualität einhergeht. Therapie der Wahl ist die Anwendung von Tränensubstitutionspräparaten. Ziel dieser pharmakologischen und zytomorphologischen Untersuchung war die Evaluation der Wirkung und des protektiven Effekts von fünf kommerziellen, auf Hyaluronsäure basierenden Medikamenten und einem Testpräparat an humanen konjunktivalen (Chang) und kornealen (pRSV-T) Epithelzellen. Dafür wurden die Zellen jeder Zelllinie mit einer Zellzahl von 1,5x104 jeweils in einer 96-well-Platte bis zum konfluenten Wachstum inkubiert. Für den Zellversuch wurden die Zellen jeder Vertiefung für 20 Minuten jeweils mit 50μl der zu testenden Proben inkubiert. Als Positivkontrolle wurde das Medium in den Vertiefungen belassen und als Negativkontrolle diente die gleiche Menge an PBS. Die Zellen wurden einem standardisierten Stressfaktor (Luft) für eine der Austrocknungszeiten 0, 15, 30 oder 45 Minuten ausgesetzt. Anschließend erfolgte eine vierstündige Inkubation mit dem Farbstoff alamarBlueR. Aus den mit dem ELISA-Reader gemessenen Absorptionswerten wurde daraufhin die Überlebensrate fur jede Austrocknungszeit ermittelt. Bei dem LIVE/DEADR Viability/ Cytotoxicity Kit wirkte auf die Zellen nach Austrocknung bei Raumtemperatur für 20 Minuten eine Propidium-Jodit-Calcein-Losung ein. Für die Auswertung wurde die Relation von vitalen und avitalen Zellen unter dem Fluoreszenzmikroskop evaluiert. Probe 2 imponierte im zeitlichen Verlauf für beide Zelllinien mit einer sehr geringen Verlustrate vitaler Zellen. Verglichen dazu wies das Testpräparat einen starken Zellzahlabfall auf. Alle weiteren Proben zeigten insgesamt eine gute protektive Wirkung auf die Zellen. Es fiel auf, dass diese umso großer war, je hoher Hyaluronsäure im Präparat konzentriert war. Probe 2 mit 0,3% Hyaluronsäure zeigte die besten Überlebensraten. Über das Testpräparat lassen sich aufgrund fehlender Informationen über dessen pharmakologische Zusammensetzung nur beschrankt Aussagen machen. Sowohl die Chang- als auch die pRSV-T-Zellen zeigten allerdings unter dessen Wirkung nach 20 Minuten Einwirkzeit und nach 0 und 15 Minuten Austrocknung eine überdurchschnittlich hohe Verlustrate, sodass hier ein toxischer Effekt nicht unwahrscheinlich ist. Bei allen untersuchten Proben wurde mit zunehmender Expositionszeit ein Abfall vitaler Zellen verzeichnet. Schlussfolgernd lasst sich sagen, dass Zellkulturverfahren geeignete Methoden sind, um modellhaft den Effekt von ophthalmo-pharmakologischen Zubereitungen für die Augenoberflache zu evaluieren. Grundsätzlich ist eine höhere Konzentration von Hyaluronsäure in Benetzungsmitteln mit einem höheren Austrocknungsschutz und einer antiinflammatorischen Wirkung assoziiert. Auch die Wahl des Puffers spielt eine wichtige Rolle in Bezug auf die Protektion. Für ein optimales Benetzungsmittel kann additiv Natriumchlorid eingesetzt werden. Zellen in vitro reagieren auf verschiedene Expositionen meist wegen der nicht zu simulierenden Barrierefunktion der Augenoberflache deutlich empfindlicher als intaktes Oberflächenepithel, dennoch lassen sich für die klinische Praxis einige Tendenzen ableiten.
National oral health survey on refugees in Germany 2016/2017: caries and subsequent complications
(2020)
Objectives To assess oral health, caries prevalence, and subsequent complications among recently arrived refugees in Germany and to ompare these findings with the German resident population. Methods This multicenter cross-sectional study recruited 544 refugees aged 3–75+ years; they were examined at ten registration institutions in four federal states in Germany by two calibrated dentists. The refugees were screened for caries (dmft/DMFT) and its complications pufa/PUFA); this data was compared to the resident population via the presentative national oral health surveys). Results The deciduous dentition of the 3-year-old refugees had a mean dmft value of 2.62 ± 3.6 compared with 0.48 dmft in the German resident population, and caries increased to 5.22 ± 3.4 for 6–7-year-olds (Germany: 1.73 dmft). Few refugee children had naturally healthy teeth (7% in 6–7-year-olds, Germany: 56%). In the permanent dentition, the gap in caries prevalence between refugees and the German population decreased with age (35–44-year-olds: 10.55 ± 7.1 DMFT; Germany: 11.2), but refugees exhibited more caries defects (35–44-year-olds DT = 3.13 ± 3.0; Germany: 0.5). German residents had more restorations (35–44-year-olds FT = 4.21 ± 4.6). Regarding complications, the 6–7-year-olds exhibited the highest pufa index (0.86 ± 1.4) which decreased in adolescence (13–17-year-olds, 0.18 ± 0.6) and increased in adults (45–64-year-olds, 0.45 ± 0.8). Conclusion The refugees had high caries experience, often untreated caries teeth and more complications compared with the German resident population, especially in children. Closing this gap by extending preventive systems to the refugees would decrease future treatment needs. Clinical relevance European countries should be prepared for the higher dental treatment needs in recent refugees, especially in
children.
Ausgewählte Lyrik
(2020)
In fixed orthodontic treatments debonding of brackets during treatment is an unpleasant occurrence for the clinician and the patients and resultes in an increase in treatment costs and duration. For Damon Q brackets recycling would considered as an economic saving option which could be done with using of in office methods such as the sandblasting.
A sample of sixty sound bovine first upper central incisers, were collected, cleaned, and mounted in acrylic blocks for shear bond strength testing.
The total sample was equally divided into two main groups. Each group had 30 teeth and 30 brackets.
The first group had 30 teeth bonded with metal Damon Q brackets, the second group had 30 teeth bonded with metal Mini-Mono® brackets.The study included bonding and rebonding experiments. Therefore the same brackets with their same teeth were used in bonding and in the rebonding experiments. The bonding and the rebonding procedures were done with using 3M Unitek etching, Grengloo adhasive, and Ortho solo bonding. In addition the rebonding procedure was done after cleaning the teeth and recycling their brackets with sandblasting. All specimens were recycled 5000 times for the bonding and rebonding experiments.
The first and second debonding forces were done in Newton using a Zwick Roell machine.
After that SBS and SRS were computed in MPa. Furthermore all the teeth, after each debonding, were examined under a digital scanning microscope VHX-5000, 50X magnifying, to performe the ARA and ARI.
The collected data was statistically analyzed for descriptive statistics as well as significance of differences among the different bracket types, and their ARI scores, in the bonding and rebonding experiments.
The results showed that SRS was significantly higher than SBS of both types of the brackets, and Damon Q brackets had higher SBS, and SRS than Mini-Mono® brackets, and there was no correlation between SBS, SRS and their ARI, ARA.
Infections with Helicobacter pylori are a global challenge that affects both developed and developing countries. This infection is currently treated using multiple antimicrobials that are mostly absorbed after oral administration and subsequently secreted into the gastric lumen. The eradication rates from the different therapeutic regimens, however, are declining nowadays, primarily due to high antibiotic resistance and possibly the mode of drug delivery. H. pylori is commonly found adhering to epithelial cells, and therefore, intragastric drug delivery may be a more direct treatment option. In this work, we developed a new strategy for the local eradication of H. pylori within the stomach.
Initial in vitro experiments revealed that penicillin G shows promising antibiotic activity against resistant strains of H. pylori with MIC values of 0.125 µg/mL. To provide luminal concentrations above the MIC for an extended time, we decided to follow two different formulation strategies: effervescent granules and HPMC-based hydrogel matrix tablets. Among the granule formulations, only one batch was stable and demonstrated excellent performance with respect to drug content, effervescent action, and drug release. It was therefore selected for further in vitro studies. All matrix tablets showed the desired tablet quality requirements and drug release was scalable in vitro by the HPMC concentration.
In order to quantify PGS in various formulations and media, an HPLC method was developed and validated. Due to the stability concerns, the degradation behavior of PGS was studied at different pH. PGS was found to be unstable at acidic pH values, but its stability was higher at more neutral pH values. Sufficient stability was exhibited at pH values above pH 4.5. Due to the instability of PGS in acidic media, alkalizers were added to the matrix tablets to prevent the degradation of the drug within the tablet. Among the alkalizers tested, NaHCO3 showed the most promising results as it significantly enhanced the stability within the matrix and also the concentration of PGS in the dissolution media. The stabilizing effect was caused mainly by the modulation of the microenvironmental pH rather than a pH change in the dissolution media. As a result, these matrix tablets were selected for further in vitro characterization.
In order to guide formulation development, a flow-through model (FTM), which was able to simulate various physiological conditions of the gastric environment, was developed and applied. In contrast to compendial dissolution methods, the FTM allowed studying the effect of gastric secretion, mixing and emptying on the gastric concentration of the drug in vitro. It could be shown that the granules generated a high initial concentration, which decreased over time. On the contrary, the matrix tablets did not provide such a profile due to the absence of pressure events in the model. Further investigations of the matrix tablets in a dissolution stress test device revealed faster drug release if pressure events of physiological relevance are simulated.
In the last part of this thesis, the two formulation concepts were compared in vivo by using the salivary tracer technique. For this purpose, caffeine was used as a model drug. The in vivo investigations suggested that granules administered in a fed state demonstrated longer gastric retention than in a fasted state. In a fed state, effervescent granules provided longer gastric retention of caffeine in comparison to the matrix tablets. Interestingly, the administration of the granules together with 240 mL of tap water provided an even better gastric retention of caffeine than the smaller volume (20 mL). Additional MRI investigations after 4 h of tablets’ intake revealed that the matrix tablets were already disintegrated in vivo.
In conclusion, effervescent granules dosed after food are expected to better maintain intragastric drug concentration over an extended period compared to matrix tablets. Moreover, the carbon dioxide generated after disintegration supports the mixing of the drug with the chyme and thus, provides a uniform distribution of the drug. By this, bacterial sanctuary sites within the stomach can be avoided. The major challenge could be the stability of PGS in acidic media. This problem could be addressed via concomitant administration of PPIs. H2 blockers could also be recommended to address nocturnal acid-breakthrough during the mid-night. In combination with an acid-reducing agent, PGS granule formulations alone or part of the treatment regimens could enable the local eradication of H. pylori directly within the stomach.
Abstract
Aims
Pinus uncinata is the major treeline‐forming species in the Pyrenees. Yet, the role of its reproduction and dispersal as drivers of treeline dynamics remains unknown. Here we quantify seed production, dispersal and germination changes along the elevation gradient to assess whether they may constrain the foreseen treeline advance in the Pyrenees.
Location
Central Pyrenees, Catalonia, NE Spain.
Methods
We established four plots along an elevation gradient from the closed subalpine forest to the krummholz zone at five study sites. In each plot, we collected cones from five to six trees, measured their length, and triggered their opening in the laboratory to count the number of empty seeds and the number and weight of full seeds. We used the collected seeds in a germination experiment under controlled conditions in growth chambers. Additionally, we installed seed traps along the forest–alpine grassland transition to measure seed rain for three consecutive years in three of the study sites.
Results
The number of full seeds per cone decreased along the elevation gradient and was correlated with cone length. However, the proportion of full seeds per cone and their weight did not differ between elevation positions. Seed rain decreased drastically with elevation and no seeds arrived into the alpine grassland traps consistently across study years. Although germination success did not significantly differ between elevation provenances (i.e., elevation position of origin), we found significant differences in germination dynamics between study sites and between elevation provenances within sites.
Conclusions
Our results indicate that whereas the viability of Pinus uncinata seeds is not limited by elevation, seed production and dispersal are constraining the ongoing rates of treeline advance in the Pyrenees.
Herein, we report the synthesis of a series of push–pull imines by considering cyclic diamino substituent at the C‐centre and fluoroaryl substituent at the N‐centre. This has been achieved by a selective aromatic nucleophilic substitution of different fluoroarenes by N‐H‐substituted N‐heterocyclic imines (NHIs) at ambient conditions without any additional reagent. Solid‐state molecular structure analysis reveals the elongation of the central C–N bond of the imine functionality, which is consistent with the push–pull nature of these imines. The push–pull nature of these imines is further validated by computational studies.
Abstract
Purpose
This study aims to assess the implementation of published research, contraindications, and warnings on the prescription of dual renin‐angiotensin‐hormone system (RAS) blockade in ambulatory care in Germany.
Methods
Cohort study based on health claims data of 6.7 million subjects from 2008 to 2015. Yearly prevalence and incidence for dual RAS blockade with (a) angiotensin‐converting enzyme inhibitors and angiotensin‐receptor blockers (ACEI + ARB) and (b) aliskiren and ACEI or ARB (aliskiren + ACEI/ARB) were calculated. We assessed prescriber specialty and associations between discontinuing dual RAS blockade with specialist (internal medicine, cardiology, nephrology) visits and hospital discharge in the previous year.
Results
A total of 2 984 517 patients were included (age 51.4 ± SD 18.4 y, 48.5% male). Prescription rates for ACEI + ARB decreased from 0.6% (n = 17 907) to 0.4% (n = 12 237) and for aliskiren + ACEI/ARB from 0.23% (n = 6634) to 0.03% (n = 818). Incident prescriptions decreased from 0.23% (n = 6705) to 0.19% (n = 5055) (ACE + ARB) and from 0.1% (n = 2796) to 0.005% (n = 142) (aliskiren + ACE/ARB); 59% of ACEI + ARB and 48% of aliskiren + ACE/ARB combinations were prescribed only by one physician. Of those, 73% (ACEI + ARB) and 58% (aliskiren + ACE/ARB) were primary care providers (PCPs). Discontinuing dual RAS blockade was associated with specialist care and hospital discharge in the previous year (specialist care: RR 1.4, 95% CI, 1.3‐1.6; hospital visit: RR 1.5, 95% CI, 1.3‐1.6).
Conclusions
Our results suggest a delayed uptake of treatment recommendation for ACEI + ARB and a higher impact of Dear Doctor letters addressing PCPs directly compared with published research, contraindications, and warnings. Targeted continuous medical education, practice software alerts, and stronger involvement of pharmacists might improve the implementation of medication safety recommendations in ambulatory care.
Bacillus subtilis has been extensively used as a microbial cell factory for industrial enzymes due to its excellent capacities for protein secretion and large-scale fermentation. This bacterium is also an attractive host for biopharmaceutical production. However, the secretion potential of this organism is not fully utilized yet, mostly due to a limited understanding of critical rearrangements in the membrane proteome upon high-level protein secretion. Recently, it was shown that bottlenecks in heterologous protein secretion can be resolved by genome minimization. Here, we present for the first time absolute membrane protein concentrations of a genome-reduced B. subtilis strain (“midiBacillus”) expressing the immunodominant Staphylococcus aureus antigen A (IsaA). We quantitatively characterize the membrane proteome adaptation of midiBacillus during production stress on the level of molecules per cell for more than 400 membrane proteins, including determination of protein concentrations for ∼61% of the predicted transporters. We demonstrate that ∼30% of proteins with unknown functions display a significant increase in abundance, confirming the crucial role of membrane proteins in vital biological processes. In addition, our results show an increase of proteins dedicated to translational processes in response to IsaA induction. For the first time reported, we provide accumulation rates of a heterologous protein, demonstrating that midiBacillus secretes 2.41 molecules of IsaA per minute. Despite the successful secretion of this protein, it was found that there is still some IsaA accumulation occurring in the cytosol and membrane fraction, leading to a severe secretion stress response, and a clear adjustment of the cell’s array of transporters. This quantitative dataset offers unprecedented insights into bioproduction stress responses in a synthetic microbial cell.
Der Beitrag gibt einen systematischen Überblick zur Netzstabilität im Stromsystem. Ein Schwerpunkt liegt auf der systemischen Darstellung und Beschreibung der Netzstabilität als zentrale Herausforderung für eine sichere und stabile Stromversorgung. Basis für die Betrachtung ist eine sogenannte „Landkarte“. Diese visualisiert die technischen, organisatorischen sowie rechtlichen Grundlagen für die Netzstabilität unterteilt nach Systembilanz und Netzkapazität, wobei hier wiederum zwischen Regelungen, die primär notwendige Investitionen sicherstellen sollen und Ansätzen zur Organisation des Betriebs unterschieden wird. Aufbauend auf der „Landkarte“ werden vier rechtliche Schwerpunktthemen untersucht: (1) Netzbetrieb und Engpassmanagement – zu- und abschaltbare Lasten nach §§ 13 Abs. 6, Abs. 6a EnWG, (2) Netzbetrieb- und Netzkapazität: Flexibilitäten von Letztverbrauchern – Fokus auf § 14a EnWG und (3) Frequenzhaltung – zukünftige Herausforderungen für die Bereitstellung von Momentanreserve sowie (4) neue Lösungsoptionen durch virtuelle Kraftwerke. Die Darstellung wird mit einem Exkurs zur Digitalisierung und der datenrechtlichen Lage in Hinblick auf die Energiewende und die sichere Stromversorgung abgerundet.
In agricultural grasslands, management practice highly determines reproductive success for ground-nesting bird species. The most effective conservation measure is the delay of first mowing dates until broods fledge or bird friendly mowing. Late mowing often implies economical losses for farmers and may increase land use abandonment, which will, in turn, cause habitat deterioration. Thus, grassland bird conservation involves the challenge to protect broods against land use and to promote an appropriate management to sustain habitat quality at the same time. Because of their late and extended breeding season Corncrakes Crex crex are in particular vulnerable to frequent mowing which increases nest destruction, chick mortality and habitat loss.
This thesis aims to gain knowledge on favourable habitat characteristics and brood protection in relation with grassland management to derive implications for the conservation of Corncrake breeding sites in floodplain meadows. Study area is the Lower Oder Valley National Park in northeastern Germany that holds a Corncrake population of 50 to 250 calling males. The study covered two study periods, before (1998-2000) and after (2012-2015) the implementation of new Corncrake conservation measures allowing inferences on the effects of different timing and intensity of mowing for brood protection and habitat conservation.
Breeding was only confirmed on meadows with high forb cover, low sedge cover, low litter heights and a close location to ditches. Radio-tracked females preferred areas with high cover of forbs (> 30%) and a distinct relief heterogeneity, which was associated with increased vegetation diversity. Vegetation characteristics on sites with day calling activity of males showed more similarity with breeding sites than with sites only used for nocturnal calling, supporting the assumption that diurnal calling indicates the occurrence of females. Favoured vegetation structure was best provided by mowing in the preceding year. Low-intensity grazing was less effective in reducing litter and sedge cover, especially when conducted late in the season. In the absence of management, meadows rapidly overgrow and dense litter accumulates from dead plant material in eutrophic floodplains, which increases walking resistance for Corncrakes and may impede prey accessibility. Plant species richness and forb cover declined after land use cessation. Male Corncrakes abandoned calling sites on meadows unmanaged for longer periods.
Besides the availability of suitable nesting sites, food supply and nest predation risk are also related to vegetation structure and may indirectly influence the habitat quality. Faecal samples of Corncrakes consisted mainly of beetles and their larvae, followed by snails, spiders and earthworms. Invertebrate biomass, sampled with pitfall-traps, was twice as high, the numbers of large ground beetles even five times higher on previously unmanaged than on managed meadows. Invertebrate abundance was highest in the first and second years after land use abandonment, but strongly decreased afterwards to a similar level like under annual management. Therefore, unmown refuge strips for Corncrake protection and alternating mowing also enhance invertebrate prey resources in floodplain meadows.
Mammals caused the majority of all observed artificial ground nest predations. Nest predation risk was higher on previously unmanaged than on managed sites. Unmanaged meadows probably attract mammalian predators, because they provide a more favourable vegetation structure for foraging and harbour high numbers of small rodents, increasing also the risk of incidental nest predations. These findings suggest that an annual removal of vegetation, if conducted late in the season to protect grassland birds may reduce predation risk of ground nests in the subsequent year.
Whereas during 1998-2000 half of the study area was managed by the end of July, land use was delayed on meadows occupied by Corncrakes until at least 15 July or 15 August during 2012-2015. On meadows mown between 15 July and 15 August refuge strips were applied. The majority of Corncrake broods were started in the second half of May and mowing postponement until 15 August allows 80% of chicks to fledge without disturbance in the study area. In 65% of broods chicks reach independence (> 14 days old) until 15 July and can be protected by Corncrake friendly mowing because then they are large enough to successfully escape during mowing. Both adults and chicks survived in 10 m wide refuge strips. Because most birds tried to leave the unmown block for the first time when it was up to 30 m wide and only 15 to 30 m wide strips served as temporary habitat for unfledged chicks from mowing to departure, 10 m should be considered as the absolute minimum width for refuge strips.
The strong reduction of land use especially during July should have allowed more chicks to survive until fledging in 2012-2015 than 1998-2000. Besides the protection of nests and higher chick survival, the decline of mowing intensity increased the extent of habitat available for second breeding attempts. In 2012-2015, broods were initiated until late July in the study area. Male Corncrakes showed continuous arrival and departure during the breeding season. Similar departure rates were estimated by a multi-state occupancy model and for radio-tracked males in the same study area and periods, which both left their home ranges spontaneous and due to the impact of mowing. Compared to 1998-2000, total departure of males during June and July was reduced by 50% in 2012-2015, when more calling sites were protected from mowing. Although male Corncrakes show high intra-seasonal dispersal due to their sequential polygamous breeding system, postponed land use should have increased mating opportunities and re-nesting at first breeding sites.
Therefore, future directions of Corncrake conservation in eutrophic floodplains should address the increase of annual late mowing to protect broods and maintain favourable habitat conditions by creating a more flexible mowing regime adjusted to actual occurrence of Corncrakes. This requires expert advice to farmers based on an intense monitoring of calling Corncrakes. Repeated nocturnal surveys during May and June are highly recommended because low detection probability in combination with constant departure substantially underestimated the number of males present. Additionally, diurnal calling activity could improve the identification of breeding sites and timing could be used to estimate chick age in July to select sites for Corncrake friendly mowing. Because currently late mowing dates are unattractive for farmers conservation actions should along with financial compensations for mowing after 15 August promote the utilization of late-cut grass with poor nutrient quality for combustion. Energy production could provide an alternative income for farmers operating in conservation areas with delayed land use dates and will increase their acceptance of Corncrake protection measures.
AbstractFluctuations of electron cyclotron emission (ECE) signals are analyzed for differently heated Wendelstein 7-X plasmas. The fluctuations appear to travel predominantly on flux surfaces and are used as ‘tracers’ in multivariate time series. Different statistical techniques are assessed to reveal the coupling and information entropy-based coupling analysis are conducted. All these techniques provide evidence that the fluctuation analysis allows one to check the consistency of magneto-hydrodynamic (MHD) equilibrium calculations. Expanding the suite of techniques applied in fusion data analysis, partial mutual information (PMI) analysis is introduced. PMI generalizes traditional partial correlation (Frenzel and Pompe Phys. Rev. Lett.
99 204101) and also Schreiber’s transfer entropy (Schreiber 2000 Phys. Rev. Lett.
85 461). The main additional capability of PMI is to allow one to discount for specific spurious data. Since PMI analysis allows one to study the effect of common drivers, the influence of the electron cyclotron resonance heating on the mutual dependencies of simultaneous ECE measurements was assessed. Additionally, MHD mode activity was found to be coupled in a limited volume in the plasma core for different plasmas. The study reveals an experimental test for equilibrium calculations and ECE radiation transport.
Für eine intakte Filtration des Blutes sind hochspezialisierte Epithelzellen in den Glomeruli der Nieren, die Podozyten, essentiell. Der Verlust oder die Schädigung dieser postmitotischen Epithelzellen bzw. morphologische Veränderungen der komplex geformten Fortsätze dieser Zellen sind die häufigsten Ursachen für den Verlust der Filtrationsfähigkeit der Nieren. Diese besondere 3D-Morphologie der Podozyten hängt entscheidend vom Aktinzytoskelett und von Aktin-bindenden Proteinen ab. Aus der Literatur weiß man, dass das Aktin-bindende Protein Palladin einen entscheidenden Einfluss auf die Nukleation bzw. Polymerisation von Aktinfilamenten ausübt und dass Palladin sowohl die Morphologie als auch die Dynamik von Zellen bestimmt. In der vorliegenden Arbeit wurde die Rolle von Palladin hinsichtlich der Podozytenmorphologie und -funktion in vitro und in vivo erstmals untersucht.
Mittels in vitro Experimenten an kultivierten Podozyten der Maus konnte gezeigt werden, dass ein Knockdown von Palladin zu einer deutlichen Abnahme der Aktinfilamente und kleineren Fokalkontakte führt. Interessanterweise hatte dies aber keinen Einfluss auf die Adhäsionfähigkeit der Podozyten, sogar unter mechanischer Bean¬spruchung. Ferner konnte gezeigt werden, dass Palladin einen entscheidenden Einfluss auf die Expression anderer essentieller Aktin-assoziierter Proteine, wie Synaptopodin und α-Aktinin-4, aufweist.
Dass Palladin eine wichtige Rolle bei der Bildung und Stabilität von Aktinfilamenten spielt, konnte durch die Inkubation von kultivierten Palladin Knockdown-Podozyten mit verschiedenen Inhibitoren der Aktin-Polymerisation gezeigt werden. Die quantitative Auswertung mit Hilfe der Software F_Seg zeigte, dass Palladin Knockdown-Podozyten nach der Inkubation deutlich weniger Aktinfilamente und mehr Aktin-Cluster im Vergleich zu den Kontrollen aufweisen. Der Einsatz eines Migrations-Assays zeigte zudem, dass kultivierte Palladin Knockdown-Podozyten schneller migrieren und vermehrt dynamische Strukturen wie Lamellipodien und sogenannte Ring-Like-Structures (RiLiS) ausbilden.
Um den Einfluss von Palladin auf Podozyten in vivo zu untersuchen, wurden Mäuse generiert, bei denen Palladin spezifisch in den Podozyten ausgeknockt ist. Analysen der Glomeruli-Morphologie dieser Tiere mit Hilfe der Immunfluoreszenz-, Superresolution- und Elektronenmikroskopie (Raster- und Transmissionsmikros-kopie) zeigten eindeutig, dass die glomerulären Kapillaren stark erweitert waren und sich ein stark vergrößerter sub-podozytärer Raum ausgebildet hatte. Ferner waren die für die Filtration des Blutes maßgeblichen Fortsätze der Podozyten stark verbreitert und die Expression des essentiellen Schlitzmembranproteins Nephrin nach dem Knockout von Palladin signifikant reduziert.
Durch den Einsatz eines nephrotoxischen Serums wurde eine Glomerulonephritis induziert, die bei Podozyten-spezifischen Palladin-Knockout Mäusen zu einer stärkeren Schädigung der Glomeruli im Vergleich zu den Kontrolltieren führte. Dies deutet auf eine essentielle Rolle von Palladin für die Morphologie und Funktion der Filtrationsbarriere hin.
Des Weiteren konnte anhand von Nierenbiopsien nachgewiesen werden, dass die Palladin-Expression bei Patienten, die an einer fokal segmentalen Glomerulosklerose bzw. an der diabetischen Nephropathie erkrankt waren, im Vergleich zu den Kontrollnieren deutlich verringert ist.
Zusammenfassend zeigen die Ergebnisse der vorliegenden Arbeit, dass Palladin sowohl in vitro als auch in vivo einen entscheidenden Einfluss auf das Aktin-zytoskelett der Podozyten und somit auf die Funktion dieser hochspezialisierten Epithelzelle hat.
An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation
(2020)
Abstract
Halide assays are important for the study of enzymatic dehalogenation, a topic of great industrial and scientific importance. Here we describe the development of a very sensitive halide assay that can detect less than a picomole of bromide ions, making it very useful for quantifying enzymatic dehalogenation products. Halides are oxidised under mild conditions using the vanadium‐dependent chloroperoxidase from Curvularia inaequalis, forming hypohalous acids that are detected using aminophenyl fluorescein. The assay is up to three orders of magnitude more sensitive than currently available alternatives, with detection limits of 20 nM for bromide and 1 μM for chloride and iodide. We demonstrate that the assay can be used to determine specific activities of dehalogenases and validate this by comparison to a well‐established GC‐MS method. This new assay will facilitate the identification and characterisation of novel dehalogenases and may also be of interest to those studying other halide‐producing enzymes.
In summer 2017, the World Health Organization published 10 facts on asthma, which is known as a major non-communicable disease of high clinical and scientific importance with currently several hundred million people—with many children among them—suffering from air passages inflammation and narrowing. Importantly, the World Health Organization sees asthma as being underdiagnosed and undertreated. Consequently, much more efforts in clinical disease management and research need to be spent on reducing the asthma-related health burden. Particularly, for young approximately 6 months aged patients presenting recurrent bronchitic respiratory symptoms, many parents anxiously ask the doctors for risk prognosis for their children's future life. Therefore, we urgently need to reevaluate if the current diagnostic and treatment measures are in concordance with our yet incomplete knowledge of pathomechanisms on exacerbation. To contribute to this increasing concern worldwide, we established a multicentric pediatric exacerbation study network, still recruiting acute exacerbated asthmatics (children >6 years) and preschool asthmatics/wheezers (children <6 years) since winter 2018 in Germany. The current study that has a currently population comprising 176 study participants aims to discover novel holistic entry points for achieving a better understanding of the poorly understood plasticity of involved molecular pathways and to define biomarkers enabling improved diagnostics and therapeutics. With this study description, we want to present the study design, population, and few ongoing experiments for novel biomarker research.
Clinical Trial Registration: German Clinical Trials Register (Deutsches Register für Klinische Studien, DRKS): DRKS00015738.
Zusammenfassung:
Zielstellung dieser Arbeit war es, den Einfluss von „lone atrial fibrillation“ auf die extrazelluläre und intrazelluläre Signaltransduktion des TGF-beta1-Signalweges zu untersuchen. Dazu wurde das Modell des „acute rapid pacing“ unter Verwendung muriner HL-1-Zellen genutzt. Weiterhin wurde die Einflussnahme von Irbesartan auf die festgestellten Veränderungen geprüft.
„Acute rapid pacing“ führte zu einer erhöhten mRNA-Expression profibrotischer Faktoren wie CTGF, SGK1 und TGF-beta1. Marker für kardiale Schädigung wie MSTN und FSTL3 zeigten ebenfalls eine Erhöhung des mRNA-Gehaltes nach „acute rapid pacing“. Kardial protektive Faktoren wie FSTL1 fanden sich im mRNA-Gehalt dagegen erniedrigt. Auf Proteinebene zeigte sich eine Mehrexpression des Stressmarkers GSK. Die Verwendung von Irbesartan beim „acute rapid pacing“ führte zu einer Reduktion der elevierten mRNA-Gehalte der profibrotischen Faktoren CTGF, SGK1 und TGF-beta1. Gleichfalls sank durch Irbesartan der erhöhte mRNA-Gehalt der kardialen Schädigungsmarker MSTN und FSTL3. Auf den mRNA-Gehalt des kardioprotektiven FSTL1 hatte Irbesartan keinen bedeutenden Einfluss. Auf Proteinebene konnte eine Minderexpression des Stressmarkers GSK festgestellt werden, wenn „rapidly paced“ Zellen mit Irbesartan inkubiert worden waren. Für andere untersuchte, mutmaßliche Modifikatoren wie Endoglin und FSTL5 lassen sich keine relevanten Aussagen ableiten.
In Bezug auf den weiteren Signalweg sprechen die Ergebnisse der mRNA-Werte von ALK1 (Acvrl1), ALK2 (Acvr1) und ALK5 (Tgfbr1) nach „acute rapid pacing“ für eine Aktivierung des Phospho-Smad-1/-5/-8-Schenkels. Die Ergebnisse der Proteinexpression von Phospho-Smad-1/-5/-8 und phospho-Smad-2 nach „acute rapid pacing“ bzw. Inkubation mit TGF-beta1 stützen diese These.
Tgfbr2-mRNA konnte in HL-1-Zellen wiederholt nicht nachgewiesen werden, wurde jedoch in Kardiozyten von Mus musculus detektiert. Dies spricht für relevante Unterschiede im kanonischen TGF-beta-Signalweg zwischen HL-1-Zellen und nativem Mausgewebe.
Die untersuchten Zielgene des TGF-beta-Signalwegs – ID1, ID2 und ID3 – zeigten in Bezug auf ihre mRNA nach „acute rapid pacing“ ein differenziertes Verhalten mit Anstieg von ID1 und Absenkung von ID2 und ID3, was zum Prozess eines Remodelings passt. Irbesartan führte in Bezug auf die mRNA der genannten Zielgene nach „acute rapid pacing“ zu keiner signifikanten Änderung.
Aims
Averaged measurements, but not the progression based on multiple assessments of carotid intima-media thickness, (cIMT) are predictive of cardiovascular disease (CVD) events in individuals. Whether this is true for conventional risk factors is unclear.
Methods and results
An individual participant meta-analysis was used to associate the annualised progression of systolic blood pressure, total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol with future cardiovascular disease risk in 13 prospective cohort studies of the PROG-IMT collaboration (n = 34,072). Follow-up data included information on a combined cardiovascular disease endpoint of myocardial infarction, stroke, or vascular death. In secondary analyses, annualised progression was replaced with average. Log hazard ratios per standard deviation difference were pooled across studies by a random effects meta-analysis. In primary analysis, the annualised progression of total cholesterol was marginally related to a higher cardiovascular disease risk (hazard ratio (HR) 1.04, 95% confidence interval (CI) 1.00 to 1.07). The annualised progression of systolic blood pressure, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol was not associated with future cardiovascular disease risk. In secondary analysis, average systolic blood pressure (HR 1.20 95% CI 1.11 to 1.29) and low-density lipoprotein cholesterol (HR 1.09, 95% CI 1.02 to 1.16) were related to a greater, while high-density lipoprotein cholesterol (HR 0.92, 95% CI 0.88 to 0.97) was related to a lower risk of future cardiovascular disease events.
Conclusion
Averaged measurements of systolic blood pressure, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol displayed significant linear relationships with the risk of future cardiovascular disease events. However, there was no clear association between the annualised progression of these conventional risk factors in individuals with the risk of future clinical endpoints.
In modern-day organic synthesis, transitional metal catalysis has become an essential tool-kit to access the biologically significant complex organic scaffolds. The activation profile of these sophisticated catalytic systems in cross-coupling chemistry and ring-closing processes has been well appreciated and frequently employed by the scientific community.
The present thesis is describing the results of interdisciplinary research involving medicinal chemistry and transitional metal homogeneous catalysis. A molybdenum mediated process was employed to access 32 unprecedented heterocyclic fused poly sulfur ring containing pentathiepins in moderate to good yields as a part of medicinal chemistry. Biologically significant, such as quinoxaline, pyrazine, pyridine, nicotinamide, quinoline, imdazo-pyrazine, pyrrolo-pyrazine, purine, and pyridine sulfonamide scaffolds were functionalized with pentathiepin unit via multi-step organic synthesis. Essentially, the Sonogashira cross-coupling and(Et4N)2[MoO(S4)2] mediated ring-closing steps were commonly employed in all pentathiepin syntheses. The analytically pure samples were characterized by 1H, 13C, 19F-NMR, FTIR, ESI-MS, CHNS, and X-ray single-crystal diffraction analysis. Notably, all pentathiepins exhibited an ABX3 multiplet pattern between δ: 4.2-4.5 ppm with the integration of 2H for the ethoxy functional group's methylene protons substituted on the five-membered ring of pentathiepin, which was later considered as a fingerprint for pentathiepin formation. The mechanistic investigations via control experiments suggest that the tetra sulfur ring Mo(IV) precursor (Et4N)2[MoO(S4)2] is vital along with elemental sulfur for the pentathiepin formation, and the Mo(IV) complex regenerates in the reaction. Furthermore, For the first time, the GPx1 enzyme inhibitor properties of novel fused heterocyclic pentathiepins were established, where these probes exhibited 9-12 folds higher potency than mercaptosuccinic acid. Notably, <1 µM concentration of quinoxaline, pyrazine, and quinoline fused pentathiepins were potent enough to inhibit 50% of GPx1 enzyme activity. Additionally, cytotoxicity, antimicrobial and antifungal studies were conducted for all pentathiepins. In anticancer investigations, the IC50 concentrations for all pentathiepins were ranging between 0.22 to 4.7 µM.
The second half of the thesis introduces a novel water-soluble Pd/PTABS as a potent catalyst for C-X (X = N, O, and S) cross-coupling chloroheteroarenes and halonucleosides. The novel, mild and efficient Pd/PTABS catalytic system was successfully employed at low catalytic loadings (1 mol%) for the amination (C−N), etherification (C−O), and thioetherification (C−S) of chloroheteroarenes at ambient to moderate temperatures. The Pd/PTABS catalyst is well-tolerating various heterocyclic scaffolds, and under the optimized catalytic conditions, various secondary amines, electron-rich or electron-poor phenols, thiophenols, and alkylthiols, were efficiently employed as nucleophilic coupling partners. Notably, the catalyst offered tremendous regio and chemoselectivity with excellent temperature control. Besides, novel sulfones and sulfoximines were prepared from the thioethers obtained via Pd/PTABS. The catalyst was employed efficiently for synthesizing biologically significant known drugs or drug candidates such as alogliptin (anti-diabetic agent), XRK 469 (antitumor agent), and Imuran-Azathioprine (immunosuppressive) in competitive yields. Preliminary DFT investigations were performed, and based on the DFT analysis, the electropositive character of the phosphorous atom in quaternary ammonium salts of PTABS supports the heteroatom directed C−Cl activation hypothesis.
Entropy Ratio and Entropy Concentration Coefficient, with Application to the COVID-19 Pandemic
(2020)
Der rundblättrige Sonnentau (Drosera rotundifolia L.) ist typisch für nährstoffarme Hochmoore und nimmt eine besondere Rolle im Moor-Ökosystem ein. Die Pflanzenart gilt in vielen europäischen Ländern als gefährdet bzw. stark gefährdet. Ihre Gefährdung lässt sich auf drei Ursachen zurückführen:
1) Seit Jahrzehnten führt die Bewirtschaftung der europäischen Moore und die damit einhergehende Entwässerung und Düngung zu einem deutlichen Rückgang der von Drosera-Arten bevorzugten oligotrophen, nassen und sauren Standorte.
2) Bereits im Mittelalter waren Drosera-Arten als Heilpflanzen bekannt und wurden hauptsächlich zur Behandlung von Atemwegserkrankungen (Asthma, Bronchitis, Keuchhusten etc.) eingesetzt.
3) Obwohl seit den 1920er Jahren bereits immer wieder Kultivierungsversuche mit Drosera-Arten durchgeführt wurden, konnte bisher keine Methode für den großflächigen Anbau von Sonnentau realisiert werden, um die von der Pharmaindustrie benötigten Mengen des Drosera-Rohstoffs zu produzieren. Daher werden bis heute europäische und nicht europäische Drosera-Arten immer noch in großen Mengen in natürlichen Mooren gesammelt.
Die zunehmende Zerstörung der natürlichen Moore und die Sammlung für arzneiliche Zwecke stellen zusammen eine ernsthafte Bedrohung für den Erhalt von D. rotundifolia dar. Die Torfmooskultivierungsflächen in Deutschland sind in vieler Hinsicht vergleichbar mit intakten Hochmooren. Das nährstoffarme Milieu der kultivierten Torfmoose dient als Lebensraum für heimische Drosera-Arten, wie Drosera rotundifolia L. und Drosera intermedia Hayne. Daher bieten diese Kulturflächen eine neue Alternative für den Anbau von Drosera-Arten.
In vier Studien wurde die Eignung von Torfmoosrasen für den Drosera-Anbau untersucht, mit Schwerpunkt auf den Anbau von Drosera rotundifolia auf Torfmoos- kultivierungsflächen. In der ersten Studie wurde das Wissen über die Morphologie, Verbreitung, Ökologie, Reproduktion, Nutzung, den Schutz und den Anbau von D. rotundifolia erstmals zusammenfassend diskutiert, um eine wissenschaftliche Grundlage für einen erfolgreichen Anbau auf Torfmoosrasen zu schaffen. Basierend auf diesen Kenntnissen konzentriert sich die zweite Studie auf die Keimfähigkeit von D. rotundifolia und die Überlebensrate von jungen Drosera-Pflanzen auf Torfmoosrasen unter natürlichen, naturnahen und künstlichen Bedingungen. Die dritte Studie fokussiert auf den Gehalt pharmakologisch wirksamer Inhaltsstoffe angebauter und „wild wachsender“ D. rotundifolia- sowie D. intermedia-Pflanzen auf Torfmooskultivierungsflächen. Die vierte Studie untersucht die Biomasseproduktivität und den Ertrag, d. h. den Biomasseanteil der geerntet wird, von beiden o. g. Drosera-Arten auf Torfmooskultivierungsflächen.
Die generierten Daten und Erkenntnisse der vier Studien wurden in vier wissenschaftlichen Artikeln zusammengefasst, wovon zwei bereits veröffentlicht und zwei eingereicht sind.
Die wichtigsten Ergebnisse dieser Studien sind die Folgenden:
I) Drosera rotundifolia ist sehr stark mit Sphagnum-dominierten Pflanzengemeinschaften verbunden, welche durch Entwässerung europaweit zurückgegangen bzw. verschwunden sind. Dadurch ist D. rotundifolia in den meisten europäischen Ländern eine seltene und geschützte Pflanzenart geworden.
II) Verschiedene Drosera-Arten, u. a. D. rotundifolia, D. intermedia, D. anglica und D. madagascariensis, werden immer noch von Pharmaunternehmen verwendet. Die Pflanzen werden in der freien Natur gesammelt, weil deren Anbau zeitaufwendig und (noch) nicht effizient ist. Daher ist die Entwicklung von Anbaumethoden erforderlich.
III) Die selbstentwickelte „Torf-Gefäß-Methode“ ergab sich als die meist geeignete Drosera-Anbau-Methode durch das spezielle Mikroklima des Sphagnum- Rasens, das konkurrenzarme Milieu und den permanent nassen Sphagnum- Torf in den Pflanzgefäßen.
IV) In den Feldversuchen wurden bei der Aussaat sehr niedrige Keimungsraten < 1 % registriert. Deshalb sind für den Anbau mit Aussaat große Mengen an Samen erforderlich.
V) Die Entfernung von Gefäßpflanzen zeigte im ersten Jahr eine positive Korrelation mit der Anzahl der Drosera-Keimlinge und führte im zweiten Jahr zu einer höheren Anzahl überlebender Drosera-Pflanzen.
VI) Auf Torfmooskultivierungsflächen wachsende Drosera-rotundifolia-Pflanzen wiesen eine 7- bis 8-mal höhere Konzentration von 7-Methyljuglon auf als D. madagascariensis, die hauptsächlich für ‘Droserae herba’ verwendet wird.
VII) Für Drosera rotundifolia gab es bezüglich der Tageszeit keine signifikanten Unterschiede in den Konzentrationen bioaktiver Inhaltsstoffe. Dies bedeutet, sie kann ganztägig zwischen 7 und 16 Uhr gesammelt werden. Die höchsten Konzentrationen bioaktiver Inhaltsstoffe wurden für D. rotundifolia und D. intermedia bei 13 bis 24 Monate alten blühenden Pflanzen festgestellt
VIII) Im Vergleich zu natürlichen Mooren Mittel- und Nordeuropas, zeigte D. rotundifolia auf den Torfmooskultivierungsflächen eine 3-34 Mal höhere Biomasseproduktivität (275 kg ha-1 a-1) und einen 2-21 Mal höheren Ertrag (214 kg ha-1 a-1).
IX) Der höchste Ertrag von D. rotundifolia und D. intermedia wurde im Juli und August dokumentiert. In diesen Monaten erreichen die Pflanzen ihr höchstes Gewicht. Auf Torfmooskultivierungsflächen erreichte D. rotundifolia einen viermal höheren Ertrag als D. intermedia. Deshalb ist D. rotundifolia für den Anbau zu bevorzugen.
X) Für eine langfristige nachhaltige Produktion von Drosera wird die Ernte von mindestens 12 Monate alten Pflanzen empfohlen.
Purpose: To assess the comparative efficacy and the long-term cost-utility of alternative minimally invasive glaucoma surgeries (MIGSs) when combined with cataract surgery in patients with primary open-angle glaucoma (POAG).
Methods: Treatment effects, as measured by the 1-year reduction in intraocular pressure (IOP), were estimated with an adjusted indirect treatment comparison. Evidence from randomized clinical trials was identified for four different MIGS methods. A disease-transition model was developed by capturing clinically relevant POAG stages and the expected natural disease evolution. Outcomes of the disease-transition model were the comparative utility [quality-adjusted life years (QALYs)], cost and cost-utility of included strategies in a lifetime horizon.
Results: Estimated 1-year IOP reductions were: cataract surgery - 2.05 mmHg (95% CI - 3.38; - 0.72), one trabecular micro-bypass stent - 3.15 mmHg (95% CI - 5.66; - 0.64), two trabecular micro-bypass stents - 4.85 mmHg (95% CI - 7.71; - 1.99) and intracanalicular scaffold - 2.25 mmHg (95% CI - 4.87; 0.37). Discounted outcomes from the disease-transition model appraised the strategy of two trabecular micro-bypass stents with cataract surgery in the moderate POAG stage as the one providing the greatest added value, with 10,955€ per additional QALY. Improved outcomes were seen when assessing MIGS in the moderate POAG stage.
Conclusions: When indirectly comparing alternative MIGS methods combined with cataract surgery, the option of two trabecular micro-bypass stents showed both a superior efficacy and long-term cost-utility from a German perspective. Moreover, outcomes of the disease-transition model suggest POAG patients to beneficiate the most from an earlier intervention in the moderate stage contrary to waiting until an advanced disease is present.
Abstract
Environmentally‐friendly processes for the manufacturing of valuable industrial compounds like ω‐hydroxy fatty acids (ω‐OHFAs) are highly desirable. Herein, we present such an approach by establishing a two‐step enzymatic cascade reaction for the production of 2,15,16‐trihydroxy hexadecanoic acid (THA). Starting with the easily accessible natural compound ustilagic acid (UA) that is secreted by the corn smut fungus Ustilago maydis, the recombinantly expressed esterase BS2 from Bacillus subtilis and the commercial β‐glucosidase from almonds were applied yielding 86 % product. Both hydrolases do not require expensive cofactors, making the process economically attractive. Additionally, no harmful solvents are required, so that the product THA can be labelled natural to be used in food and cosmetic products.
Bruxismus wird in SB und WB unterteilt und sind als verschiedene Entitäten zu betrachten. Die Ätiologie von SB ist noch nicht vollständig bekannt, als gesichert gilt, dass es ein multifaktorielles Geschehen ist. Die Behandlung der Patienten in der zahnärztlichen Praxis ist vielfältig, meist jedoch symptomatisch ausgerichtet. Am häufigsten wird eine Art der Aufbissschienen in der Therapie verwendet. CES und die Vorgängerversionen des in der vorliegenden Studie verwendeten GrindCare 4 zeigten sich erfolgreich in der Therapie von Bruxismus. Ziel der vorliegenden Studie war es, die Effektivität der Biofeedbacktherapie des neu auf dem Markt erschienen GrindCare 4 an einem Patientenkollektiv in der zahnärztlichen Praxis zu beurteilen.
In der vorliegenden Therapieverlaufsstudie wurden 23 Patienten untersucht. Die Probanden stammten sowohl aus Angehörigen des ZZMK als auch aus Patienten die mit dem Wunsch der Behandlung von Bruxismussymptomen in das ZZMK kamen und zur Teilnahme an der klinischen Untersuchung eingeladen wurden. 13 Probanden (männlich/weiblich (m/w) = 4/9; Mittelwert (MW) = 35,0 Jahr) durchliefen die Studie im 21-tägigen Studienprotokoll. Ebenfalls 13 Probanden wurden zur 42-tägigen Studie eingeladen, zehn (m/w= 2/8; MW = 36,7 Jahre) konnten diese erfolgreich abschließen.
Der Untersuchungsteil dieser Studie unterteilt sich in drei Abschnitte:
Im ersten Abschnitt wurde ein anamnestischer Befund sowie ein detaillierter Funktionsbefund nach den Richtlinien RDC/TMD erhoben. Zudem wurde ein Befund nach dem Bruxismus- Status nach Lange und Bernhardt angefertigt. Die Patienten wurden aufgeklärt und das GrindCare- Gerät wurde mit Anweisungen bezüglich der Anwendung an den Patienten ausgehändigt. Das Studienprotokoll sah eine Messung der Häufigkeit der Muskelkontraktionen mittels GrindCare vor. Die Messung dauerte vier Nächte im 21-tägigen Protokoll und sieben Nächte im 42-tägigen Protokoll an. Der zweite Abschnitt, der eine Therapie mit GrindCare mit Biofeedback umfasst, begann nach dieser Messphase. Die Therapie wurde je nach Studienprotokoll zwei bzw. vier Wochen durchgeführt. Im letzten Abschnitt nach der Therapie wurde mit Hilfe von GrindCare weitere drei bzw. sieben Nächte die Häufigkeit der Muskelkontraktionen gemessen und ausgewertet. Darauf folgte eine zweite klinische Untersuchung mittels klinischer Funktionsanalyse.
Die Auswertung der Daten erfolgte mittels nicht parametrischer Tests.
In der Anwendung des Gerätes kam es vermehrt zu Schwierigkeiten in der Anwendung. Vor allem kam es zum vorzeitigen Ablösen der Gelpads. Es zeigte sich keine signifikante Veränderung der EMG-Ereignisse im Studienteil der zweiwöchigen Therapiephase. Eine signifikante Verringerung der EMG-Ereignisse konnte erst nach längerer Biofeedbacktherapie im 42-tägigen Studienteil gezeigt werden. Die Werte konnten in der vierten Woche Biofeedback um 40% und in der abschließenden Messphase um 32% gegenüber der ersten Messphase reduziert werden. Die Anzahl der druckdolenten Palpationsstellen sank in beiden Studienteilen signifikant (21-tägiger Studienabschnitt p=0,010; 42-tägiger Studienabschnitt p=0,035). Der durch die Probanden berichtete myofasziale Schmerz und die Einschränkung ihrer Leistungsfähigkeit, sowie die Jaw Disability konnte in der vorliegenden Studie nicht signifikant vermindert werden.
Die EMG-Ereignisse scheinen nur nach längerer Therapie mittels Biofeedback abzunehmen, eine zweiwöchige Interventionsphase ist nicht ausreichend. Die in der abschließenden Messphase weiterhin bestehende Reduktion der EMG-Werte bei vierwöchiger Therapie deutet auf einen möglichen Lerneffekt hin. Eine Verringerung der Bruxismussymptome ist nicht gesichert. Nur die Anzahl der druckdolenten Palpationsstellen konnte signifikant reduziert werden.
Der Hersteller hat das Studiengerät zur Überarbeitung unter anderem aufgrund der vorliegenden Ergebnisse zum Zeitpunkt der Veröffentlichung vom Markt genommen. Weitere Studien mit einer größeren Kohorte und einer Kontrollgruppe sind notwendig, um die Ergebnisse zu sichern. Eine längere Therapiedauer und ein ausgedehntes Follow-up könnten sinnvoll sein.
Abstract
This paper focuses on the expectation formation process of professional forecasters by relying on survey data on forecasts regarding gross domestic product growth, consumer price index inflation and 3‐month interest rates for a broad set of countries. We examine the interrelation between macroeconomic forecasts and also the impact of uncertainty on forecasts by allowing for cross‐country interdependencies and time variation in the coefficients. We find that professional forecasts are often in line with the Taylor rule and identify significant expectation spillovers from monetary policy in the USA.
Abstract
This article takes a novel look at the relationship between government activity, partisan preferences and varieties of capitalism. Evidence from panel regressions for 25 EU countries from 1990 to 2014 suggests that there are major divides among European countries in terms of the drivers of government activity, that is, government spending and government regulation. The European divide appears to be even more pronounced between liberal and coordinated economic systems than between the classical geographical divide of east and west, which is typically used in most contributions. While both divides apply to the determinants of government activity in general, a reversal of the classical partisan effect for the east is to be found only in specific cases and, is most likely in government spending in liberal eastern countries.
The role of glutathione peroxidases (GPx) in cancer and their influence on tumor prognosisand the development of anticancer drug resistance has been extensively and controversially discussed.The aim of this study was to evaluate the influence of GPx1 expression on anticancer drug cytotoxicity.For this purpose, a GPx1 knockout of the near-haploid human cancer cell line HAP-1 was generatedand compared to the native cell line with regards to morphology, growth and metabolic rates,and oxidative stress defenses. Furthermore, the IC50values of two peroxides and 16 widely usedanticancer drugs were determined in both cell lines. Here we report that the knockout of GPx1 in HAP-1cells has no significant effect on cell size, viability, growth and metabolic rates. Significant increasesin the cytotoxic potency of hydrogen peroxide andtert-butylhydroperoxide, the anticancer drugscisplatin and carboplatin as well as the alkylating agents lomustine and temozolomide were found.While a concentration dependent increases in intracellular reactive oxygen species (ROS) levelswere observed for both HAP-1 cell lines treated with either cisplatin, lomustine or temozolamide,no significant enhancement in ROS levels was observed in the GPx1 knockout compared to the nativecell line except at the highest concentration of temozolamide. On the other hand, a ca. 50% decreasein glutathione levels was noted in the GPx1 knockout relative to the native line, suggesting thatfactors other than ROS levels alone play a role in the increased cytotoxic activity of these drugs in theGPx1 knockout cells.
Abstract
Over the last years, there has been an enormous increase in the knowledge on koi herpesvirus (KHV), koi herpesvirus disease (KHVD), pathogenesis and virus variants. Different KHV lineages have clearly been identified, possible genomic changes during replication in different cell cultures at different temperatures but also in several hosts have been identified, a persistent stage of infection has been specified and it has been shown that infection with KHV is not host specific at all, but KHVD is. Additionally, it has been shown that it is possible to combat KHVD by immunization with inactivated and attenuated live vaccines using different delivery systems but also to benefit from alternative treatments with e.g. exopolysaccharids obtained from Arthrospira platensis.
Abstract
Head motion during magnetic resonance imaging (MRI) induces image artifacts that affect virtually every brain measure. In parallel, cross‐sectional observations indicate a correlation of head motion with age, psychiatric disease status and obesity, raising the possibility of a systematic artifact‐induced bias in neuroimaging outcomes in these conditions, due to the differences in head motion. Yet, a causal link between obesity and head motion has not been tested in an experimental design. Here, we show that a change in body mass index (BMI) (i.e., weight loss after bariatric surgery) systematically decreases head motion during MRI. In this setting, reduced imaging artifacts due to lower head motion might result in biased estimates of neural differences induced by changes in BMI. Overall, our finding urges the need to rigorously control for head motion during MRI to enable valid results of neuroimaging outcomes in populations that differ in head motion due to obesity or other conditions.
Die traumatische Dissektion des Truncus coeliacus ist eine der seltenen Gefäßverletzungen. In der Literatur finden sich hierzu bislang lediglich 14 Einzelfallberichte. Im Zeitraum von September 1997 bis Dezember 2012 wurden 5.194 Patienten unter dem Verdacht auf ein erlittenes Polytrauma im Unfallkrankenhaus Berlin behandelt. Hiervon wurde bei neun Patienten die Diagnose einer traumatischen Dissektion des Truncus coeliacus gestellt. In diesen neun Fällen wurden die Unfallmechanismen und Verletzungsmuster, die bildgebenden Untersuchungen sowie die erfolgten therapeutischen Maßnahmen retrospektiv ausgewertet. Die Ergebnisse wurden im Hinblick auf mögliche Aussagen zur Ätiopathogenese, zu etwaigen Markerverletzungen, zur Bildgebung im Akut- und Langzeitverlauf sowie zur Behandlung dieser seltenen Verletzung mit den publizierten Daten zu traumatischen und spontanen Viszeralarteriendissektionen in der Literatur verglichen.
Es zeigte sich ein heterogenes Verteilungsmuster an Unfallursachen, wobei vermutlich Scherkräfte am Übergang des fixierten zum mobilen Gefäßabschnitt für die Dissektion verantwortlich sind. Eine vorbestehende Stenose des Truncus coeliacus durch das Ligamentum arcuatum scheint die Dissektion zu begünstigen.
Aufgrund auffälliger Koinzidenzen knöcherner Wirbelsäulenverletzungen im thorako-lumbalen Übergang besitzen diese beim polytraumatisierten Patienten möglicherweise den Charakter von Markerverletzungen für eine potenziell vorliegende traumatische Dissektion des Truncus coeliacus.
In der Akutphase stellt die CT-Angiographie den Goldstandard für die bildgebende Diagnostik dar, während in späteren Verlaufskontrollen auch die MR-Angiographie sowie duplexsonographische Untersuchungen eingesetzt werden können.
Die Behandlung ist primär konservativ. In Abhängigkeit der Hämodynamik des Patienten und der kollateralen Blutversorgung im Bereich der Viszeralarterien stellen Zeichen einer Organischämie das wichtigste Kriterium für die Notwendigkeit einer endovaskulären oder operativen Intervention dar.
Aneurysmen im Bereich der peripankreatischen arteriellen Arkaden sind als mögliche vaskuläre Spätkomplikationen zu berücksichtigen.
Rūdolfs Blaumanis (1863–1908) ist ein bedeutender Autor aus den Anfängen der modernen lettischen Literatur. Er ist hauptsächlich dem literarischen Realismus verpflichtet und gilt als ein Meister der Erzählung. Durch seine Komödie »Schneidertage auf Lüttwalden« (1902) lernen wir ihn allerdings von einer anderen Seite kennen. Rūdolfs Blaumanis ist auch ein bedeutender lettischer Bühnenautor.
Auf dem Lüttwaldenhof steht eine Hochzeit an, und deshalb holt man den Schneider her. Doch die Schneidertage bringen so einiges durcheinander, denn sie sind Tage, an denen man sich etwas traut. Zusammen mit den Schneidersleuten kommen die Juden, und alle Gäste treffen auf die ohnehin facettenreichen Bewohner des Lüttwaldenhofs. Wie gut ist es da, dass Bäuerin und Lüttwalden-Witwe Antonia selbst gerne das Tanzbein schwingt …
Investigation of In-Vitro Adaptation toward Sodium Bituminosulfonate in Staphylococcus aureus
(2020)
Staphylococcus aureus can cause life-threatening diseases, and hospital- as well as community-associated antibiotic-resistant strains are an emerging global public health problem. Therefore, prophylactic vaccines or immune-based therapies are considered as alternative treatment opportunities. To develop such novel treatment approaches, a better understanding of the bacterial virulence and immune evasion mechanisms and their potential effects on immune-based therapies is essential. One important staphylococcal virulence factor is alpha-toxin, which is able to disrupt the epithelial barrier in order to establish infection. In addition, alpha-toxin has been reported to modulate other cell types including immune cells. Since CD4+ T cell-mediated immunity is required for protection against S. aureus infection, we were interested in the ability of alpha-toxin to directly modulate CD4+ T cells. To address this, murine naïve CD4+ T cells were differentiated in vitro into effector T cell subsets in the presence of alpha-toxin. Interestingly, alpha-toxin induced death of Th1-polarized cells, while cells polarized under Th17 conditions showed a high resistance toward increasing concentrations of this toxin. These effects could neither be explained by differential expression of the cellular alpha-toxin receptor ADAM10 nor by differential activation of caspases, but might result from an increased susceptibility of Th1 cells toward Ca2+-mediated activation-induced cell death. In accordance with the in vitro findings, an alpha-toxin-dependent decrease of Th1 and concomitant increase of Th17 cells was observed in vivo during S. aureus bacteremia. Interestingly, corresponding subsets of innate lymphoid cells and γδ T cells were similarly affected, suggesting a more general effect of alpha-toxin on the modulation of type 1 and type 3 immune responses. In conclusion, we have identified a novel alpha-toxin-dependent immunomodulatory strategy of S. aureus, which can directly act on CD4+ T cells and might be exploited for the development of novel immune-based therapeutic approaches to treat infections with antibiotic-resistant S. aureus strains.