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Ausgewählte Lyrik
(2022)
Microglia are the resident immune cells of the central nervous system (CNS) and play a major role in the regulation of brain homeostasis. To maintain their cellular protein homeostasis, microglia express standard proteasomes and immunoproteasomes (IP), a proteasome isoform that preserves protein homeostasis also in non-immune cells under challenging conditions. The impact of IP on microglia function in innate immunity of the CNS is however not well described. Here, we establish that IP impairment leads to proteotoxic stress and triggers the unfolded and integrated stress responses in mouse and human microglia models. Using proteomic analysis, we demonstrate that IP deficiency in microglia results in profound alterations of the ubiquitin-modified proteome among which proteins involved in the regulation of stress and immune responses. In line with this, molecular analysis revealed chronic activation of NF-κB signaling in IP-deficient microglia without further stimulus. In addition, we show that IP impairment alters microglial function based on markers for phagocytosis and motility. At the molecular level IP impairment activates interferon signaling promoted by the activation of the cytosolic stress response protein kinase R. The presented data highlight the importance of IP function for the proteostatic potential as well as for precision proteolysis to control stress and immune signaling in microglia function.
Aim
Periprosthetic joint infections are a devastating complication after arthroplasty, leading to rejection of the prosthesis. The prevention of septic loosening may be possible by an antimicrobial coating of the implant surface. Poly (hexamethylene) biguanide hydrochloride [PHMB] seems to be a suitable antiseptic agent for this purpose since previous studies revealed a low cytotoxicity and a long-lasting microbicidal effect of Ti6Al4V alloy coated with PHMB. To preclude an excessive activation of the immune system, possible inflammatory effects on macrophages upon contact with PHMB-coated surfaces alone and after killing of S. epidermidis and P. aeruginosa are analyzed.
Methods
THP-1 monocytes were differentiated to M0 macrophages by phorbol 12-myristate 13-acetate and seeded onto Ti6Al4V surfaces coated with various amounts of PHMB. Next to microscopic immunofluorescence analysis of labeled macrophages after adhesion on the coated surface, measurement of intracellular reactive oxygen species and analysis of cytokine secretion at different time points without and with previous bacterial contamination were conducted.
Results
No influence on morphology of macrophages and only slight increases in iROS generation were detected. The cytokine secretion pattern depends on the surface treatment procedure and the amount of adsorbed PHMB. The PHMB coating resulted in a high reduction of viable bacteria, resulting in no significant differences in cytokine secretion as reaction to coated surfaces with and without bacterial burden.
Conclusion
Ti6Al4V specimens after alkaline treatment followed by coating with 5–7 μg PHMB and specimens treated with H2O2 before PHMB-coating (4 μg) had the smallest influence on the macrophage phienotype and thus are considered as the surface with the best cytocompatibility to macrophages tested in the present study.
Background
We investigated the association between low cardiorespiratory fitness and liver fat content (LFC) in the general population.
Materials and Methods
We evaluated data from 2151 adults (51.1% women) from two population-based cohorts of the Study of Health in Pomerania (SHIP-2 and SHIP-TREND-0). We analysed the cross-sectional associations of peak oxygen uptake (VO2peak) with LFC, assessed by magnetic resonance imaging proton density fat fraction, as well as serum gamma-glutamyltransferase (GGT) and aminotransferase concentrations by multivariable regression models.
Results
We observed significant inverse associations of VO2peak with LFC and serum GGT, but not with serum aminotransferase levels. Specifically, a 1 L/min lower VO2peak was associated with a 1.09% (95% confidence interval [CI]: 0.45-1.73; P = .002) higher LFC and a 0.18 μkatal/L (95% CI: 0.09-0.26; P < .001) higher GGT levels. The adjusted odds ratio (OR) for the risk of prevalent hepatic steatosis (HS) by a 1 L/min decrease in VO2peak was 1.61 (95% CI: 1.22-2.13; P = .001). Compared to subjects with high VO2peak, obese and overweight individuals with low VO2peak had 1.78% (95% CI: 0.32-3.25; P = .017) and 0.94% (95% CI: 0.15-1.74; P = .021) higher mean LFC, respectively. Compared to those with high VO2peak, low VO2peak was independently associated with a higher risk of prevalent HS in the obese (adjusted-OR 2.29, 95% CI=1.48-3.56; P < .001) and overweight (adjusted OR 1.57, 95% CI=1.16-2.14; P = .04) groups.
Conclusions
Lower VO2peak was significantly associated with greater LFC and higher serum GGT levels in a population-based cohort of adult individuals. Our results suggest that low VO2peak might be a risk factor for HS.
Target Mechanisms of the Cyanotoxin Cylindrospermopsin in Immortalized Human Airway Epithelial Cells
(2022)
Cylindrospermopsin (CYN) is a cyanobacterial toxin that occurs in aquatic environments worldwide. It is known for its delayed effects in animals and humans such as inhibition of protein synthesis or genotoxicity. The molecular targets and the cell physiological mechanisms of CYN, however, are not well studied. As inhalation of CYN-containing aerosols has been identified as a relevant route of CYN uptake, we analyzed the effects of CYN on protein expression in cultures of immortalized human bronchial epithelial cells (16HBE14o−) using a proteomic approach. Proteins whose expression levels were affected by CYN belonged to several functional clusters, mainly regulation of protein stability, cellular adhesion and integration in the extracellular matrix, cell proliferation, cell cycle regulation, and completion of cytokinesis. With a few exceptions of upregulated proteins (e.g., ITI inhibitor of serine endopeptidases and mRNA stabilizer PABPC1), CYN mediated the downregulation of many proteins. Among these, centrosomal protein 55 (CEP55) and osteonectin (SPARC) were significantly reduced in their abundance. Results of the detailed semi-quantitative Western blot analyses of SPARC, claudin-6, and CEP55 supported the findings from the proteomic study that epithelial cell adhesion, attenuation of cell proliferation, delayed completion of mitosis, as well as induction of genomic instability are major effects of CYN in eukaryotic cells.
The pore forming alpha-toxin (hemolysin A, Hla) of Staphylococcus aureus (S. aureus) is a major virulence factor with relevance for the pathogenicity of this bacterium, which is involved in many cases of pneumonia and sepsis in humans. Until now, the presence of Hla in the body fluids of potentially infected humans could only be shown indirectly, e.g., by the presence of antibodies against Hla in serum samples or by hemolysis testing on blood agar plates of bacterial culture supernatants of the clinical isolates. In addition, nothing was known about the concentrations of Hla actually reached in the body fluids of the infected hosts. Western blot analyses on 36 samples of deep tracheal aspirates (DTA) isolated from 22 hospitalized sepsis patients using primary antibodies against different epitopes of the Hla molecule resulted in the identification of six samples from five patients containing monomeric Hla (approx. 33 kDa). Two of these samples showed also signals at the molecular mass of heptameric Hla (232 kDa). Semiquantitative analyses of the samples revealed that the concentrations of monomeric Hla ranged from 16 to 3200 ng/mL. This is, to our knowledge, the first study directly showing the presence of S. aureus Hla in samples of airway surface liquid in human patients.
Metabolic syndrome is a significant worldwide public health challenge and is inextricably linked to adverse renal and cardiovascular outcomes. The inhibition of the transient receptor potential cation channel subfamily C member 6 (TRPC6) has been found to ameliorate renal outcomes in the unilateral ureteral obstruction (UUO) of accelerated renal fibrosis. Therefore, the pharmacological inhibition of TPRC6 could be a promising therapeutic intervention in the progressive tubulo-interstitial fibrosis in hypertension and metabolic syndrome. In the present study, we hypothesized that the novel selective TRPC6 inhibitor SH045 (larixyl N-methylcarbamate) ameliorates UUO-accelerated renal fibrosis in a New Zealand obese (NZO) mouse model, which is a polygenic model of metabolic syndrome. The in vivo inhibition of TRPC6 by SH045 markedly decreased the mRNA expression of pro-fibrotic markers (Col1α1, Col3α1, Col4α1, Acta2, Ccn2, Fn1) and chemokines (Cxcl1, Ccl5, Ccr2) in UUO kidneys of NZO mice compared to kidneys of vehicle-treated animals. Renal expressions of intercellular adhesion molecule 1 (ICAM-1) and α-smooth muscle actin (α-SMA) were diminished in SH045- versus vehicle-treated UUO mice. Furthermore, renal inflammatory cell infiltration (F4/80+ and CD4+) and tubulointerstitial fibrosis (Sirius red and fibronectin staining) were ameliorated in SH045-treated NZO mice. We conclude that the pharmacological inhibition of TRPC6 might be a promising antifibrotic therapeutic method to treat progressive tubulo-interstitial fibrosis in hypertension and metabolic syndrome.
This study deals with attitudes and opinions of the population on the Ukrainian Black Sea coast in 2020/2021, a good year before the Russian invasion of Ukraine. The basis is a sociological survey conducted in the regions of Odesa, Mykolajiv and Kherson with 1,200 respondents. These regions are a clear target of the Russian offensive against Ukraine, on the grounds of an alleged ideological and linguistic affinity of these areas with Russia. The attitudes and opinions sought in the study cover three domains: firstly, attitudes and opinions on language policy, secondly, on Ukrainian autonomy and identity and the role played by languages in this context, and thirdly, Ukraine’s geopolitical orientation. Respondents were grouped according to their “ethnic/national” self-identification and language preferences, and a comparison was made. Overall, the attitudes of the respondents showed a clear identification with Ukraine and the Ukrainian language and a skepticism towards the Russian state, but by no means a hostile attitude towards the Russian language. A picture emerges that clearly contradicts Russian propaganda and also puts into perspective the difference between southern Ukraine and the center and the west, which has often been emphasized in the past.
Genetic variants in α-actinin-2 (ACTN2) are associated with several forms of (cardio)myopathy. We previously reported a heterozygous missense (c.740C>T) ACTN2 gene variant, associated with hypertrophic cardiomyopathy, and characterized by an electro-mechanical phenotype in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Here, we created with CRISPR/Cas9 genetic tools two heterozygous functional knock-out hiPSC lines with a second wild-type (ACTN2wt) and missense ACTN2 (ACTN2mut) allele, respectively. We evaluated their impact on cardiomyocyte structure and function, using a combination of different technologies, including immunofluorescence and live cell imaging, RNA-seq, and mass spectrometry. This study showed that ACTN2mut presents a higher percentage of multinucleation, protein aggregation, hypertrophy, myofibrillar disarray, and activation of both the ubiquitin-proteasome system and the autophagy-lysosomal pathway as compared to ACTN2wt in 2D-cultured hiPSC-CMs. Furthermore, the expression of ACTN2mut was associated with a marked reduction of sarcomere-associated protein levels in 2D-cultured hiPSC-CMs and force impairment in engineered heart tissues. In conclusion, our study highlights the activation of proteolytic systems in ACTN2mut hiPSC-CMs likely to cope with ACTN2 aggregation and therefore directs towards proteopathy as an additional cellular pathology caused by this ACTN2 variant, which may contribute to human ACTN2-associated cardiomyopathies.
Abstract
Climate change is increasing the frequency and intensity of drought events in many boreal forests. Trees are sessile organisms with a long generation time, which makes them vulnerable to fast climate change and hinders fast adaptations. Therefore, it is important to know how forests cope with drought stress and to explore the genetic basis of these reactions. We investigated three natural populations of white spruce (Picea glauca) in Alaska, located at one drought‐limited and two cold‐limited treelines with a paired plot design of one forest and one treeline plot. We obtained individual increment cores from 458 trees and climate data to assess dendrophenotypes, in particular the growth reaction to drought stress. To explore the genetic basis of these dendrophenotypes, we genotyped the individual trees at 3000 single nucleotide polymorphisms in candidate genes and performed genotype–phenotype association analysis using linear mixed models and Bayesian sparse linear mixed models. Growth reaction to drought stress differed in contrasting treeline populations. Therefore, the populations are likely to be unevenly affected by climate change. We identified 40 genes associated with dendrophenotypic traits that differed among the treeline populations. Most genes were identified in the drought‐limited site, indicating comparatively strong selection pressure of drought‐tolerant phenotypes. Contrasting patterns of drought‐associated genes among sampled sites and in comparison to Canadian populations in a previous study suggest that drought adaptation acts on a local scale. Our results highlight genes that are associated with wood traits which in turn are critical for the establishment and persistence of future forests under climate change.
Forests influence the climate of our Earth and provide habitat and food for many species and resources for human use. These valuable ecosystems are threatened by fast environmental changes caused by human-induced climte change. Negative growth responses and higher tree mortality rates were associated with increasing physiological stress induced by global warming. Especially boreal forests at high latitudes in the arctic region are threatened, a region predicted to undergo the highest increase in temperature during the next decades. Therefore, it is important to assess the adaptation potential in trees. For this purpose, I studied natural populations of white spruce (Picea glauca (Moench) Voss) in Alaska. In this thesis, I present three scientific papers in which my co-authors and I studied the phenotypic plasticity and genetic basis of tree growth, wood anatomy and drought tolerance as well as the genetic structure of white spruce populations in contrasting environments. We established three sites representing two cold-limited treelines and one drought-limited treeline with a paired plot design including one plot located at the treeline and one plot located in a closed-canopy forest, respectively. Additionally, the study design included one forest plot as reference. Within the entire project, in total 3,000 trees were measured, genotyped and dendrochronological data was obtained. I used several approaches to estimate the neutral and adaptive genetic diversity and phenotypic plasticity of white spruce as a model organism to explore the adaptation potential of trees to climate change.
In the first chapter, I combined neutral genetic markers with dendrochronological and climatic data to investigate population structure and individual growth of white spruce. Several individual-based dendrochronological approaches were applied to test the influence of genetic similarity and microenvironment on growth performance. The white spruce populations of the different sites showed high gene flow and high genetic diversity within and low genetic differentiation among populations, rather explained by geographic distance. The individual growth performances showed a high plasticity rather influenced by microenvironment than genetic similarity.
In the second chapter, I investigated the populations of the drought and cold-limited treeline sites to decipher the underlying genetic structure of drought tolerance using different genotype-phenotype association analyses. Based on tree-ring series and climatic data, growth declines caused by drought stress were identified and the individual reaction to the drought stress event was determined. A subset of 458 trees was genotyped, using SNPs in candidate genes and associated with the individual drought response. Most of the associations were revealed by an approach which took into account small-effect size SNPs and their interactions. Populations of the contrasting treelines responded differently to drought stress events. Populations further showed divergent genetic structures associated with drought responsive traits, most of them in the drought-limited site, indicating divergent selection pressure.
In the third chapter, my co-authors and I studied xylem anatomical traits at one of the cold-limited treeline sites to investigate whether genetic or spatial grouping affected the anatomy and growth of white spruce. Annual growth and xylem anatomy were compared between spatial groups and between genetic groups and individuals. Overall, wood traits were rather influenced by spatial than genetic grouping. Genetic effects were only found in earlywood hydraulic diameter and latewood density. Environmental conditions indirectly influenced traits related to water transport.
In conclusion, white spruce showed a high genetic diversity within and a low genetic differentiation among populations influenced by high gene flow rates. Genetic differences among populations are rather caused by geographical distance and therefore genetic drift. Differing selection pressure at the treeline ecotones presumably lead to divergent genetic structures underlying drought-tolerant phenotypes among the populations. Thus, adaptation to drought most likely acts on a local scale and involves small frequency shifts in several interacting genes. The identified genes with adaptive growth traits can be used to further exlore local adaptation in white spruce. Tree growth and wood anatomical traits are rather influenced by the environment than genetics and showed a high phentoypic plasticity. The high genetic diverstiy and phenotypic plasticity of white spruce may help the species to cope with rapid environmental changes. Still, additional work is needed to further explore adaptation processes to estimate how tree species reacted to rapid climate change. The presented thesis shed some light on the adaptation potential of trees by the example of white spruce using several approaches.
Durch das CEREC®-Verfahren mit dem Intraoralscanner Omnicam (Fa. Dentsply Sirona, York, USA) ist es möglich, digitale Abformungen zu erstellen und anschließend digital Restaurationen zu konstruieren. Für die korrekte Herstellung von prothetischen Restaurationen ist die Okklusion essenziell. Diese wird während der digitalen Aufzeichnung durch den Bukkalscan registriert. Im Anschluss wird das digitale okklusale Kontaktpunktmuster sichtbar.
In dieser Studie wurde die Genauigkeit der digitalen Okklusionsbestimmung im Vergleich zu Bissregistraten und intraoralen Kontaktpunktmuster untersucht.
Getestet wurden dazu 30 Probanden einer Altersgruppe zwischen 18 - 30 Jahren. Es handelte sich um 15 Frauen und 15 Männer mit nahezu naturgesundem Gebiss. Es wurden bei allen Probanden zwei zeitlich versetzte Bukkalscans durchgeführt und die durch Artikulationspapier markierten Kontaktpunkte auf den Kauflächen fotografiert. Anschließend erfolge die Herstellung eines Bissregistrates aus A-Silikon. Es erfolgten zeitgleich Elektromyogrammmessungen, um Muskelaktivität während der unterschiedlichen Registrierungsmethoden festzustellen.
Es konnte festgestellt werden, dass beim Vergleich der digitalen Okklusion zum intraoralen Kontaktpunktbild über Artikulationspapier zu 6,69 % weniger Kontaktpunkte durch CEREC® errechnet wurden. Die Kontaktpunktanzahl von der digitalen Okklusion zu den Bissregistraten unterschied sich zu 39,45 % deutlich. Mit dem Korrelationskoeffizienten nach Pearson wurde belegt, dass eine große Korrelation von r = 0,79 zwischen dem ersten Bukkalscan und dem klinischen Kontaktpunktmuster besteht. Beim Vergleich zum Bissregistrat lag der Korrelationskoeffizient bei r = 0,59. Das Ergebnis der Studie war, dass die Übereinstimmung bei der Anzahl der digitalen zu den intraoralen Kontaktpunkten größer war als zur Kontaktpunktanzahl bei Bissregistraten.
Auch bei den Ergebnissen des Elektromyogramms ist ein Unterschied zu erkennen. Es konnte festgestellt werden, dass während der Herstellung der Bissregistrate eine wesentlich höhere Muskelaktivität aufgezeichnet wurde. So wurden 22,99 % der maximalen Muskelaktivität während der Herstellung des Bissregistrates aufgebracht, während die Aktivität beim Bukkalscan im Durchschnitt (vor und nach dem Scannen der Zahnreihen) bei 14,38 % lag. Generell konnte kein Zusammenhang zwischen Muskelaktivität und Kontaktpunktanzahl erkannt werden.
Diese Studie ergab, dass die digitale Okklusion (generiert durch die CEREC® Omnicam) im Vergleich zum klinischen Kontaktpunktmuster verlässlich und für die Herstellung von Restaurationen im digitalen Workflow geeignet ist. Dem Verwender der CEREC® Omnicam wird empfohlen, die Kontaktpunkte vor dem Scan mittels Artikulationspapier zu markieren. So kann die digitale Okklusion am Computer überprüft und auf die Restauration übertragen werden.
Proteasomes comprise a family of proteasomal complexes essential for maintaining protein homeostasis. Accordingly, proteasomes represent promising therapeutic targets in multiple human diseases. Several proteasome inhibitors are approved for treating hematological cancers. However, their side effects impede their efficacy and broader therapeutic applications. Therefore, understanding the biology of the different proteasome complexes present in the cell is crucial for developing tailor-made inhibitors against specific proteasome complexes. Here, we will discuss the structure, biology, and function of the alternative Proteasome Activator 200 (PA200), also known as PSME4, and summarize the current evidence for its dysregulation in different human diseases. We hereby aim to stimulate research on this enigmatic proteasome regulator that has the potential to serve as a therapeutic target in cancer.
The utilization of fluorescein-guided biopsies has recently been discussed to improve and expedite operative techniques in the detection of tumor-positive tissue, as well as to avoid making sampling errors. In this study, we aimed to report our experience with fluorescein-guided biopsies and elucidate distribution patterns in different histopathological diagnoses in order to develop strategies to increase the efficiency and accuracy of this technique. We report on 45 fluorescence-guided stereotactic biopsies in 44 patients (15 female, 29 male) at our institution from March 2016 to March 2021, including 25 frame-based stereotactic biopsies and 20 frameless image-guided biopsies using VarioGuide®. A total number of 347 biopsy samples with a median of 8 samples (range: 4–18) per patient were evaluated for intraoperative fluorescein uptake and correlated to definitive histopathology. The median age at surgery was 63 years (range: 18–87). Of the acquired specimens, 63% were fluorescein positive. Final histopathology included glioblastoma (n = 16), B-cell non-Hodgkin lymphoma (n = 10), astrocytoma, IDH-mutant WHO grade III (n = 6), astrocytoma, IDH-mutant WHO grade II (n = 1), oligodendroglioma, IDH-mutant and 1p/19q-codeleted WHO grade II (n = 2), reactive CNS tissue/inflammation (n = 4), post-transplantation lymphoproliferative disorder (PTLD; n = 2), ependymoma (n = 1), infection (toxoplasmosis; n = 1), multiple sclerosis (n = 1), and metastasis (n = 1). The sensitivity for high-grade gliomas was 85%, and the specificity was 70%. For contrast-enhancing lesions, the specificity of fluorescein was 84%. The number needed to sample for contrast-enhancing lesions was three, and the overall number needed to sample for final histopathological diagnosis was five. Interestingly, in the astrocytoma, IDH-mutant WHO grade III group, 22/46 (48%) demonstrated fluorescein uptake despite no evidence for gadolinium uptake, and 73% of these were tumor-positive. In our patient series, fluorescein-guided stereotactic biopsy increases the likelihood of definitive neuropathological diagnosis, and the number needed to sample can be reduced by 50% in contrast-enhancing lesions.
Amine transaminases (ATAs) are powerful biocatalysts for the stereoselective synthesis of chiral amines. However, wild-type ATAs usually show pH optima at slightly alkaline values and exhibit low catalytic activity under physiological conditions. For efficient asymmetric synthesis ATAs are commonly used in combination with lactate dehydrogenase (LDH, optimal pH: 7.5) and glucose dehydrogenase (GDH, optimal pH: 7.75) to shift the equilibrium towards the synthesis of the target chiral amine and hence their pH optima should fit to each other. Based on a protein structure alignment, variants of (R)-selective transaminases were rationally designed, produced in E. coli, purified and subjected to biochemical characterization. This resulted in the discovery of the variant E49Q of the ATA from Aspergillus fumigatus, for which the pH optimum was successfully shifted from pH 8.5 to 7.5 and this variant furthermore had a two times higher specific activity than the wild-type protein at pH 7.5. A possible mechanism for this shift of the optimal pH is proposed. Asymmetric synthesis of (R)-1-phenylethylamine from acetophenone in combination with LDH and GDH confirmed that the variant E49Q shows superior performance at pH 7.5 compared to the wild-type enzyme.
With the aim to discover and create suitable biocatalysts for the synthesis of chiral amines in a faster and more efficient way, this thesis includes protein engineering studies (Article I), explores transaminase substrate specificities (Articles II and IV), and an ultrahigh-throughput growth system-based for the directed evolution of amine-forming enzymes (Article III).
The protein engineering studies described in Article I deal with the creation of a (R)-amine transaminase activity in the α-amino acid transaminase scaffold to expand our knowledge of the evolutionary relationship between amine transaminase and α-amino acid transaminase. Article II describes the broadening of the limited substrate scope of transaminases to enable the conversion of bulky substrates. In Article III, a growth selection system is described for an ultra-high throughput screening strategy to accelerate the identification of desired mutants, which can be widely applied to the directed evolution of amine-forming enzymes.
Abstract
The KV7 potassium channel openers flupirtine and retigabine have been valuable options in the therapy of pain and epilepsy. However, as a result of adverse reactions, both drugs are currently no longer in therapeutic use. The flupirtine‐induced liver injury and the retigabine linked tissue discolouration do not appear related at first glance; nevertheless, both events can be attributed to the triaminoaryl scaffold, which is affected by oxidation leading to elusive reactive quinone diimine or azaquinone diimine metabolites. Since the mechanism of action, i. e. KV7 channel opening, seems not to be involved in toxicity, this study aimed to further develop safer replacements for flupirtine and retigabine. In a ligand‐based design strategy, replacing amino substituents of the triaminoaryl core with alkyl substituents led to carba analogues with improved oxidation resistance and negligible risk of quinoid metabolite formation. In addition to these improved safety features, some of the novel analogues exhibited significantly improved KV7.2/3 channel opening activity, indicated by an up to 13‐fold increase in potency and an efficacy of up to 176 % compared to flupirtine, thus being attractive candidates for further development.
KV7 channel openers have proven their therapeutic value in the treatment of pain as well as epilepsy and, moreover, they hold the potential to expand into additional indications with unmet medical needs. However, the clinically validated but meanwhile discontinued KV7 channel openers flupirtine and retigabine bear an oxidation‐sensitive triaminoraryl scaffold, which is suspected of causing adverse drug reactions via the formation of quinoid oxidation products. Here, we report the design and synthesis of nicotinamide analogs and related compounds that remediate the liability in the chemical structure of flupirtine and retigabine. Optimization of a nicotinamide lead structure yielded analogs with excellent KV7.2/3 opening activity, as evidenced by EC50 values approaching the single‐digit nanomolar range. On the other hand, weighted KV7.2/3 opening activity data including inactive compounds allowed for the establishment of structure–activity relationships and a plausible binding mode hypothesis verified by docking and molecular dynamics simulations.
The potassium channel opening drugs flupirtine and retigabine have been withdrawn from the market due to occasional drug-induced liver injury (DILI) and tissue discoloration, respectively. While the mechanism underlying DILI after prolonged flupirtine use is not entirely understood, evidence indicates that both drugs are metabolized in an initial step to reactive ortho- and/or para-azaquinone diimines or ortho- and/or para-quinone diimines, respectively. Aiming to develop safer alternatives for the treatment of pain and epilepsy, we have attempted to separate activity from toxicity by employing a drug design strategy of avoiding the detrimental oxidation of the central aromatic ring by shifting oxidation toward the formation of benign metabolites. In the present investigation, an alternative retrometabolic design strategy was followed. The nitrogen atom, which could be involved in the formation of both ortho- or para-quinone diimines of the lead structures, was shifted away from the central ring, yielding a substitution pattern with nitrogen substituents in the meta position only. Evaluation of KV7.2/3 opening activity of the 11 new specially designed derivatives revealed surprisingly steep structure–activity relationship data with inactive compounds and an activity cliff that led to the identification of an apparent “magic methyl” effect in the case of N-(4-fluorobenzyl)-6-[(4-fluorobenzyl)amino]-2-methoxy-4-methylnicotinamide. This flupirtine analogue showed potent KV7.2/3 opening activity, being six times as active as flupirtine itself, and by design is devoid of the potential for azaquinone diimine formation.
The transcription factor EB (TFEB) promotes protein degradation by the autophagy and lysosomal pathway (ALP) and overexpression of TFEB was suggested for the treatment of ALP-related diseases that often affect the heart. However, TFEB-mediated ALP induction may perturb cardiac stress response. We used adeno-associated viral vectors type 9 (AAV9) to overexpress TFEB (AAV9-Tfeb) or Luciferase-control (AAV9-Luc) in cardiomyocytes of 12-week-old male mice. Mice were subjected to transverse aortic constriction (TAC, 27G; AAV9-Luc: n = 9; AAV9-Tfeb: n = 14) or sham (AAV9-Luc: n = 9; AAV9-Tfeb: n = 9) surgery for 28 days. Heart morphology, echocardiography, gene expression, and protein levels were monitored. AAV9-Tfeb had no effect on cardiac structure and function in sham animals. TAC resulted in compensated left ventricular hypertrophy in AAV9-Luc mice. AAV9-Tfeb TAC mice showed a reduced LV ejection fraction and increased left ventricular diameters. Morphological, histological, and real-time PCR analyses showed increased heart weights, exaggerated fibrosis, and higher expression of stress markers and remodeling genes in AAV9-Tfeb TAC compared to AAV9-Luc TAC. RNA-sequencing, real-time PCR and Western Blot revealed a stronger ALP activation in the hearts of AAV9-Tfeb TAC mice. Cardiomyocyte-specific TFEB-overexpression promoted ALP gene expression during TAC, which was associated with heart failure. Treatment of ALP-related diseases by overexpression of TFEB warrants careful consideration.
Fast screening of enzyme variants is crucial for tailoring biocatalysts for the asymmetric synthesis of non-natural chiral chemicals, such as amines. However, most existing screening methods either are limited by the throughput or require specialized equipment. Herein, we report a simple, high-throughput, low-equipment dependent, and generally applicable growth selection system for engineering amine-forming or converting enzymes and apply it to improve biocatalysts belonging to three different enzyme classes. This results in (i) an amine transaminase variant with 110-fold increased specific activity for the asymmetric synthesis of the chiral amine intermediate of Linagliptin; (ii) a 270-fold improved monoamine oxidase to prepare the chiral amine intermediate of Cinacalcet by deracemization; and (iii) an ammonia lyase variant with a 26-fold increased activity in the asymmetric synthesis of a non-natural amino acid. Our growth selection system is adaptable to different enzyme classes, varying levels of enzyme activities, and thus a flexible tool for various stages of an engineering campaign.
Body-size variability results from a variety of extrinsic and intrinsic factors (environmental and biological influences) underpinned by phylogeny. In ostracodes it is assumed that body size is predominantly controlled by ecological conditions, but investigations have mostly focused on local or regional study areas. In this study, we investigate the geographical size variability (length, height, and width) of Holocene and Recent valves of the salinity-tolerant ostracode species Cyprideis torosa within a large geographical area (31°–51° latitude, and 12°–96° longitude). It is shown that distant local size clusters of Cyprideis torosa are framed within two large-scale geographical patterns. One pattern describes the separation of two different size classes (i.e., morphotypes) at around ∼42° N. The co-occurrence of both size morphotypes in the same habitats excludes an environmental control on the distribution of the morphotypes but rather could point to the existence of two differentiated lineages. Generally, correlations between valve size and environmental parameters (salinity, geographical positions) strongly depend on the taxonomic resolution. While latitude explains the overall size variability of C. torosa sensu lato (i.e., undifferentiated for morphotypes), salinity-size correlations are restricted to the morphotype scale. Another large-scale pattern represents a continuous increase in valve size of C. torosa with latitude according to the macroecological pattern referred as Bergmann trend. Existing explanations for Bergmann trends insufficiently clarify the size cline of C. torosa which might be because these models are restricted to intraspecific levels. The observed size-latitude relationship of C. torosa may, therefore, result from interspecific divergence (i.e., size ordered spatially may result from interspecific divergence sorting) while environmental influence is of minor importance. Our results imply that geographical body-size patterns of ostracodes are not straightforward and are probably not caused by universal mechanisms. Consideration of phylogenetic relationships of ostracodes is therefore necessary before attempting to identify the role of environmental controls on body size variability.
Calcitic valves of non-marine ostracodes are important geochemical archives. Investigations of the relationship between the ranges of oxygen and carbon isotope values of modern ostracode populations and their host water provide important information on local or regional conditions and influences. Here we present the first δ18Oostracode and δ13C of the freshwater ostracode species Cytheridella ilosvayi along with the isotopic composition of the waters in which the ostracodes calcified, δDwater, δ18Owater, δ13CDIC values—covering a large geographical range (Florida to Brazil). With this data we extended a newly developed approach based on the estimation of δ18O values of monthly equilibrium calcites as references for the interpretation of δ18Oostracode values. The expected apparent oxygen isotope fractionation between CaCO3 and H2O is correlated with temperature with smaller values occurring at higher temperatures as valid at isotope equilibrium (δ18Ocalcite_eq). Uncertainties about the expected equilibrium calcites derive from incomplete knowledge of high-frequency variations of the water bodies caused by interplay of mixing, evaporation, and temperature. Coincidence between δ18Oostracode and δ18Ocalcite_eq is restricted to few months indicating a seasonal calcification of Cytheridella. There is a characteristic pattern in its difference between mean δ18Oostracode and δ18Ocalcite_eq which implies that Cytheridella provides a synchronous life cycle in its geographical range with two calcification periods in spring (May, June) and autumn (October). This ubiquitous life cycle of Cytheridella in the entire study area is considered to be phylogenetically inherited. It might have originally been adapted to environmental conditions but has been conserved during the migration and radiation of the group over the Neotropical realm.
Myxomycetes or Myxogastria (supergroup Amoebozoa) are one of several Protistean groups dispersing via airborne spores. The model organism for the group, so far exclusively studied in a laboratory environment, is Physarum polycephalum. Here, molecular evolution, distribution and the ecology of spores dispersal was investigated for the non-model species Physarum albescens. This nivicolous myxomycete fruits with snow melt in most mountain ranges of the northern hemisphere and disperses via spherical, dark-colored and melanin-rich spores. Fruit body development and subsequent spore dispersal occurs within a short time window of a few days. At this time, the fruiting plasmodium is fully exposed to the harsh environment if the protecting snow melts away. The spores, with a diameter of 10–13 µm of the typical size for myxomycetes, can potentially reach all suitable habitats worldwide, which led to the assumption that not only Ph. albescens but most myxomycete species should be ubiquitously distributed over the world.
In the first part of this study (article 1), the question was, if spore dispersal can realize a gene flow sufficient to meet the above-mentioned assumption. A total of 324 accessions of Ph. albescens, collected all over the northern hemisphere, was sequenced for 1-3 genetic markers (SSU, EF1A, COI), and 98 specimens were further analyzed using the genotyping by sequencing technique. As a result, at least 18 reproductively isolated units, which can be seen as cryptic biological species, emerged as phylogroups in a three-gene phylogeny, but as well in a SNP-based phylogeny and were confirmed by a recombination analysis between the three markers. However, this evolutive radiation is not simply caused by geographic fragmentation due to low dispersal capability: within a certain region, multiple phylogroups coexisted next to each other, although some appeared to be regional endemics. Most likely, mutations in mating-type genes, as shown to exist for the cultivable Ph. polycephalum, are the main drivers of speciation. This challenges the hypothesis of ubiquitous distribution of Ph. albescens and corroborates the results of the few available studies for other myxomycete species. In addition, groups of clonal specimens, mostly but not always restricted to a certain slope or valley indicated that sexual and asexual reproduction coexists in the natural populations of Ph. albescens.
In the second part (articles 2), the fundamental niche for Ph. albescens was described using all available records for the species. The resulting set of 537 unique occurrence points was subjected to a correlative spatial approach using the software MaxEnt. In dependence on the predictor variables three species distribution models emerged which differed only in details. The first consisted of only 19 bioclimatic variables and an elevation map from the WorldClim dataset. The second was corrected for pseudo-absences resulting from missing survey activities, and the third was expanded with an additional categorical environment variable on snow cover. High mean AUC (area under the curve) values above 0.97 could be reached with all three models. Variables for snow cover, precipitation of the coldest quarter (of the year), and elevation correlated highly to predict the distribution of Ph. albescens. Only in mid-northern latitudes, elevation alone was a good predictor, but it would cause false-positive predictions in arid mountain ranges and failed to explain occurrence in lowland sites at higher latitudes. Mountains in humid climates showed the highest incidences, confirming recent studies that long-lasting snow covers combined with mild summers are crucial for the ecological guild of nivicolous myxomycetes, with Ph. albescens as a typical species.
Spore size is crucial for dispersal ability and should thus be a character under strong selection. In addition, spores carrying two nuclei with opposite mating types should have a colonization advantage. This was the hypothesis for the last part of this study (articles 3 and 4), which investigated this trait in a quantitative manner. This required a method to analyze thousands of spores automatically (article 3) and with high precision for size and the number of nuclei enclosed. Human errors should be excluded, to reveal even subtle differences in the resulting spore size distributions. Two challenges had to be met for this approach. First, a preparation technique was developed to reduce false segmentations due to overlaying spores by aligning spores on one common plane with a high-frequency vibration device. Second, the segmentation process was automated to allow separating spores that are densely packed in the respective images. A machine learning algorithm was set up and trained to reliable identify and measure dark-colored spores. The technique produced consistent results with high accuracy, and the large number of spores allowed to compile spore size distributions, to check for the constancy of this character, which is impossible with manual measurements limited to low numbers.
The resulting spore size distributions, obtained from over 80 specimens (article 4), were mostly narrow, which is in accordance with our hypothesis. Spore size was as well fairly constant within fructifications from one colony. However, mean spore size within different accessions of Ph. albescens showed large variation (ca. 10%, a range often indicated to key out different morphospecies of myxomycetes), and this was explained only by a minor part with differences between biospecies. Not much smaller (8%) was the variation within a group of clonal specimens collected within 25 m distance. This points to a strong influence of environmental factors even at a micro spatial scale, perhaps caused by microclimatic differences and high phenotypic plasticity for spore size. The influence of large-scale covariates like altitude or latitude was negligible. However, spore size correlated with the variance in this trait, indicating that oversized spores may be caused by detrimental environmental conditions. Two aberrations in spore development were found: First, a few specimens showed a multimodal distribution for spore size with two or even three discernible spore populations. The estimated volumes of those populations correspond to a multiple of the first and most abundant conspicuous spore size population. Second, not all spores were uninucleate as to be expected for meiotic products. This was revealed by fluorescence signals from staining the same spores with DAPI, with a second machine learning algorithm trained to identify the nuclei in a spore. A few specimens showed a significant proportion of binucleated spores in the size range of normal-sized ones, and these specimens were not the ones with multimodal spore size distributions. This indicates that the negative impacts (inbreeding) of multinucleate spores should outweigh a possible colonization advantage and is in accordance with the high genetic diversity found in the worldwide population of Ph. albescens, indicating predominantly sexual reproduction in wild populations of myxomycetes.
Introduction
Supplementation with spermidine may support healthy aging, but elevated spermidine tissue levels were shown to be an indicator of Alzheimer's disease (AD).
Methods
Data from 659 participants (age range: 21–81 years) of the population-based Study of Health in Pomerania TREND were included. We investigated the association between spermidine plasma levels and markers of brain aging (hippocampal volume, AD score, global cortical thickness [CT], and white matter hyperintensities [WMH]).
Results
Higher spermidine levels were significantly associated with lower hippocampal volume (ß = −0.076; 95% confidence interval [CI]: −0.13 to −0.02; q = 0.026), higher AD score (ß = 0.118; 95% CI: 0.05 to 0.19; q = 0.006), lower global CT (ß = −0.104; 95% CI: −0.17 to −0.04; q = 0.014), but not WMH volume. Sensitivity analysis revealed no substantial changes after excluding participants with cancer, depression, or hemolysis.
Discussion
Elevated spermidine plasma levels are associated with advanced brain aging and might serve as potential early biomarker for AD and vascular brain pathology.
Thrombozyten haben neben ihrer Funktion in der Hämostase eine wichtige Rolle in der Immunabwehr. Sie interagieren hierbei mit Komponenten des angeborenen und des adaptiven Immunsystems und sind in der Lage, direkte anti-mikrobielle Einflüsse zu vermitteln. Die Interaktion von Thrombozyten mit Gram-positiven Bakterien unterscheidet sich von jener mit Gram-negativen Erregern. Bei beiden Gruppen von Bakterien scheint die Aktivierung von Thrombozyten und Freisetzung anti-mikrobieller Peptiden aus den Granula ein wichtiger Bestandteil der direkten Pathogenabwehr durch Thrombozyten zu sein. Hierbei führt die Interaktion mit S. aureus direkt zu einer starken pathogen-induzierten Thrombozytenaktivierung, während bei Gram-negativen Organismen wie E. coli eine Verstärkung durch die Opsonierung mit PF4 und anti-PF4/H IgG notwendig scheint. Vermutlich ist die Bindung von PF4 und anti-PF4/H IgG an Gram-positive Bakterien von größerer Bedeutung für die Opsonierung für andere Immunzellen als für den direkten bakteriziden Effekt der Thrombozyten.
Der Gram-positive S. pneumoniae führt durch Funktionsstörung und Exposition von Phosphatidylserin zu einer Schädigung der Thrombozyten. Dieser schädigende Effekt auf Thrombozyten durch S. pneumoniae wird unter anderem durch Pneumolysin, ein porenbildendes Toxin der Pneumokokken, vermittelt. Dieses induziert bereits in geringen Konzentrationen die Porenbildung in der Thrombozytenmembran und führt zur Induktion von Apoptose.
In der Arbeit konnten die initialen Fragestellungen folgendermaßen beantwortet werden:
1.Thrombozyten können einen direkten schädigenden Effekt auf Gram-positive Bakterien vermitteln.
2.PF4 und anti-PF4/Polyanion IgG spielen in der direkten Thrombozyten-vermittelten Pathogenabwehr bei Gram-positiven Erregern, trotz der Bindung an Gram-positive Bakterien, eine untergeordnete Rolle. Sie verstärken weder die Thrombozyten-aktivierung noch den anti-bakteriellen Effekt.
3.Die Auswirkung der Co-Inkubation mit Bakterien auf die Thrombozyten ist heterogen und abhängig vom untersuchten Bakterienstamm. Es kommt zur Aktivierung der Thrombozyten durch S. aureus und zur Schädigung der Thrombozyten durch S. pneumoniae.
Pentathiepins are cyclic polysulfides that exert antiproliferative and cytotoxic activity in cancer cells, induce oxidative stress and apoptosis, and potently inhibit GPx1. These properties render this class of compounds promising candidates for the development of anticancer drugs. However, the biological effects and how they intertwine to promote high cytotoxicity have not been systematically assessed throughout a panel of cancer cell lines from distinct tissues of origin. In this thesis, six novel pentathiepins were analyzed and constitute the second generation of compounds with additional properties such as fluorescence or improved water solubility to facilitate cellular testing. All compounds underwent extensive biological evaluation in 14 human cancer cell lines. These studies included investigations of the inhibitory potential with regards to GPx1 and cell proliferation, examined the cytotoxicity in human cancer cell lines, as well as the induction of oxidative stress and DNA strand breaks. Furthermore, selected hallmarks of apoptosis, ferroptosis, and autophagy were studied. Experimental approaches regarding these cellular mechanisms included observing morphological changes, detecting phosphatidyl serine exposure and caspase activity, and quantifying cleaved PARP1 and levels of LC3B II. In addition, the analysis of the cell cycle aimed to identify aberrations or arrests in cell division.
Five of the six tested pentathiepins proved to be potent inhibitors of the GPx1, while all six exerted high cytotoxic and antiproliferative activity, although to different extents. There was a clear connection observed between the potential to provoke oxidative stress and damage to DNA in the form of single- and double-strand breaks both extra- and intracellularly. Furthermore, various experiments supported apoptosis but not ferroptosis as the mechanism of cell death in four different cell lines. In particular, the externalization of PS, the detection of activated caspases, and the cleavage of PARP1 corroborated this conclusion. Additionally, indications for autophagy were found, but more investigations are required to verify the current data. The findings of this dissertation are mainly in line with the postulated mechanism of action proposed for pentathiepins and a previous publication from our group that described their biological activity. However, the influence of modulators such as oxygen and GSH on the biological effects was ambiguous and dependent on the compound. The expression profile of the cell lines concerning GPx1 and CAT did not influence the cellular response toward the treatment, whereas the cell doubling time correlated with the cytotoxicity.
As the various pentathiepins give rise to different biological responses, modulation of the biological effects depends on the distinct chemical structures fused to the sulfur ring. This may allow for future optimization of the anticancer activity of pentathiepins. An analysis of the structure-activity relationships revealed that the piperazine scaffold was associated with superior biological activity compared to the pyrrolo-pyrazine backbone. Furthermore, substituents with electron-withdrawing properties or those providing a free electron pair, such as fluorine or morpholine, were advantageous. These findings should help design and synthesize the next generation of pentathiepins, thereby expanding the library of compounds, allowing for the further deduction of structure-activity relationships and an improved understanding of their mechanism of action.
Simple Summary
This multicenter study investigated the extent of patient’s decision regret (PatR) in patients with prostate cancer comparing different surgical modalities. Robot-assisted radical prostatectomy has replaced open radical prostatectomy as the surgical standard of care in many countries worldwide. However, a broad scientific basis evaluating the difference in patient-relevant outcomes between both approaches is still lacking. In this context, PatR is increasingly moving into the scientific focus. Our study shows a critical PatR in slightly more than one third of patients about 15 months after surgery. Patients who underwent robot-assisted surgery, and also patients without postoperative urinary stress incontinence, report significantly lower PatR. Likewise, this difference was also demonstrated for patients who decided together with their treating physician on the specific surgical procedure (consensual decision making). Our study helps to further establish PatR as an important endpoint in the setting of radical prostatectomy and identifies criteria which may be addressed to reduce PatR.
Abstract
Patient’s regret (PatR) concerning the choice of therapy represents a crucial endpoint for treatment evaluation after radical prostatectomy (RP) for prostate cancer (PCA). This study aims to compare PatR following robot-assisted (RARP) and open surgical approach (ORP). A survey comprising perioperative-functional criteria was sent to 1000 patients in 20 German centers at a median of 15 months after RP. Surgery-related items were collected from participating centers. To calculate PatR differences between approaches, a multivariate regressive base model (MVBM) was established incorporating surgical approach and demographic, center-specific, and tumor-specific criteria not primarily affected by surgical approach. An extended model (MVEM) was further adjusted by variables potentially affected by surgical approach. PatR was based on five validated questions ranging 0–100 (cutoff >15 defined as critical PatR). The response rate was 75.0%. After exclusion of patients with laparoscopic RP or stage M1b/c, the study cohort comprised 277/365 ORP/RARP patients. ORP/RARP patients had a median PatR of 15/10 (p < 0.001) and 46.2%/28.1% had a PatR >15, respectively (p < 0.001). Based on the MVBM, RARP patients showed PatR >15 relative 46.8% less frequently (p < 0.001). Consensual decision making regarding surgical approach independently reduced PatR. With the MVEM, the independent impact of both surgical approach and of consensual decision making was confirmed. This study involving centers of different care levels showed significantly lower PatR following RARP.
The multidrug resistance protein 4 (MRP4) is highly expressed in platelets and several lines of evidence point to an impact on platelet function. MRP4 represents a transporter for cyclic nucleotides as well as for certain lipid mediators. The aim of the present study was to comprehensively characterize the effect of a short-time specific pharmacological inhibition of MRP4 on signaling pathways in platelets. Transport assays in isolated membrane vesicles showed a concentrationdependent inhibition of MRP4-mediated transport of cyclic nucleotides, thromboxane (Tx)B2 and fluorescein (FITC)- labeled sphingosine-1-phosphate (S1P) by the selective MRP4 inhibitor Ceefourin-1. In ex vivo aggregometry studies in human platelets, Ceefourin-1 significantly inhibited platelet aggregation by about 30-50% when ADP or collagen was used as activating agents, respectively. Ceefourin-1 significantly lowered the ADP-induced activation of integrin aIIbb3, indicated by binding of FITC-fibrinogen (about 50% reduction at 50 mM Ceefourin-1), and reduced calcium influx. Furthermore, pre-incubation with Ceefourin-1 significantly increased PGE1- and cinaciguat-induced vasodilatorstimulated phosphoprotein (VASP) phosphorylation, indicating increased cytosolic cAMP as well as cGMP concentrations, respectively. The release of TxB2 from activated human platelets was also attenuated. Finally, selective MRP4 inhibition significantly reduced both the total area covered by thrombi and the average thrombus size by about 40% in a flow chamber model. In conclusion, selective MRP4 inhibition causes reduced platelet adhesion and thrombus formation under flow conditions. This finding is mechanistically supported by inhibition of integrin aIIbb3 activation, elevated VASP phosphorylation and reduced calcium influx, based on inhibited cyclic nucleotide and thromboxane transport as well as possible further mechanisms.
Urbanization is a major contributor to the loss of biodiversity. Its rapid progress is mostly at the expense of natural ecosystems and the species inhabiting them. While some species can adjust quickly and thrive in cities, many others cannot. To support biodiversity conservation and guide management decisions in urban areas, it is important to find robust methods to estimate the urban affinity of species (i.e. their tendency to live in urban areas) and understand how it is associated with their traits. Since previous studies mainly relied on discrete classifications of species' urban affinity, often involving inconsistent assessments or variable parameters, their results were difficult to compare. To address this issue, we developed and evaluated a set of continuous indices that quantify species' urban affinity based on publicly available occurrence data. We investigated the extent to which a species' position along the urban affinity gradient depends on the chosen index and how this choice affects inferences about the relationship between urban affinity and a set of morphological, sensory and functional traits. While these indices are applicable to a wide range of taxonomic groups, we examined their performance using a global set of 356 bat species. As bats vary in sensitivity to anthropogenic disturbances, they provide an interesting case study. We found that different types of indices resulted in different rankings of species on the urban affinity spectrum, but this had little effect on the association of traits with urban affinity. Our results suggest that bat species predisposed to urban life are characterized by low echolocation call frequencies, relatively long call durations, small body size and flexibility in the selection of the roost type. We conclude that simple indices are appropriate and practical, and propose to apply them to more taxa to improve our understanding of how urbanization favours or filters species with particular traits.
(1) Background: The aim of this study was to systematically compare TEM sections of mineralized human enamel and dentine prepared by focused ion beam (in situ lift-out) technique and ultramicrotomy through a combination of microscopic examination methods (scanning electron microscopy and transmission electron microscopy). In contrast with published studies, we compared the TEM preparation methods using the same specimen blocks as those for the ultramicrotomy and FIB technique. (2) Methods: A further evaluation of TEM sample preparation was obtained by confocal laser scanning microscopy and atomic force microscopy. In addition, ultramicrotome- and focused ion beam-induced artefacts are illustrated. (3) Results: The FIB technique exposed a major difference between non-decalcified enamel and dentine concerning the ultrastructural morphology compared to ultramicrotome-prepared sections. We found that ultramicrotomy was useful for cutting mineralized dentine, with the possibility of mechanical artefacts, but offers limited options for the preparation of mineralized enamel. FIB preparation produced high-quality TEM sections, showing the anisotropic ultrastructural morphology in detail, with minor structural artefacts. Our results show that the solution of artificial saliva and glutardialdehyde (2.5% by volume) is a very suitable fixative for human mineralized tissue. (4) Conclusions: The protocol that we developed has strong potential for the preparation of mineralized biomaterials for TEM imaging and analysis.
The study of host-pathogen interactions is central to a better understanding of the human microbiome, infections and the inner workings of immune cells. One focal point of this research is how the human immune system recognises both harmful and harmless antigens, integrates the resulting signals and forms a response, and how, conversely, microbes can manipulate this reaction.
In this thesis, Pseudomonas aeruginosa (P. aeruginosa), a critical pathogen in chronic and nosocomial infections, was in the focus. The aim was to search for bacterial proteins that favour a type 2 immune response, as it is orchestrated by CD4+ type 2 T helper cells (Th2 cells). The humoral arm of a type 2 response is dominated by IgG4 and IgE. Such immune responses are typically directed against multicellular pathogens like helminths and other parasites. However, type 2 immune responses are suboptimal for the defence against extracellular bacteria like P. aeruginosa. Previous research suggests that some bacterial proteins may promote a switch to such an insufficient immune response as a mechanism of immune evasion.
To optimise the sensitivity of the search for type 2 response inducing proteins of P. aeruginosa, cystic fibrosis (CF) patients were studied, as many are exposed to the pathogen in their airways over prolonged time periods. As such, the humoral immune response of 9 CF patients to their own P. aeruginosa strain was examined. For this, the secretomes of 9 clinical P. aeruginosa isolates from CF patients and the P. aeruginosa reference strain PAO-1 were studied by 2D-immunoblotting for their ability to be bound by IgG4 and IgG1 from respective patient sera. IgG4 served as a proxy for IgE, as assays analysing IgE binding suffer from low sensitivity because of low serum concentrations of IgE. Antibody reactive P. aeruginosa proteins were then identified by liquid chromatography tandem mass spectrometry and the results were compared with proteomics data from literature.
In total, 308 distinct protein spots were analysed. These belonged to 17 bacterial proteins, which comprise the entire known P. aeruginosa secretome. Of these spots, 232 were bound by IgG4, and 24 by IgG1 only. Notably proteases like serralysin and P. aeruginosa elastase presented with an IgG4 bias. This is concordant with previous research linking proteases to a type 2 immune response. Moreover, structural proteins like
agellins were also immunodominant. Flagellins are known as common targets of immune detection in bacteria. These proteins also demonstrated a clear IgG4 bias.
Thus, the search for secreted P. aeruginosa proteins that elicit an IgG4-dominated antibody response was successful. It remains to be shown whether these bacterial proteins are also recognized by IgE and Th2 cells, meaning whether they are truly driving a type 2 immune response in CF patients. It is also an open question whether the observed IgG4 bias in the antibody response to the exoproteome of P. aeruginosa is specific to CF or a general feature of the human immune response to the pathogen.
Given the increasing prevalence of chronic kidney disease (CKD) and its impact on health care, it is important to better understand the multiple factors influencing health-related quality of life (HRQOL), particularly since they have been shown to affect CKD outcomes. Determinants of HRQOL as measured by the validated Kidney Disease Quality of Life questionnaire (KDQOL) and the Patient Health Questionnaire depression screener (PHQ-9) were assessed in a routine CKD patient sample, the Greifswald Approach to Individualized Medicine (GANI_MED) renal cohort (N = 160), including a wide range of self-reported data, sociodemographic and laboratory measures. Compared to the general population, CKD patients had lower HRQOL indices. Dialysis was associated with (1) low levels of physical functioning, (2) increased impairments by symptoms and problems, and (3) more effects and burden of kidney disease. HRQOL is seriously affected in CKD patients. However, impairments were found irrespective of eGFR decline and albuminuria. Rather, the comorbid conditions of depression and diabetes predicted a lower HRQOL (physical component score). Further studies should address whether recognizing and treating depression may not only improve HRQOL but also promote survival and lower hospitalization rates of CKD patients.
The COVID-19 pandemic is one of the most serious health and economic crises of the 21st century. From a psychological point of view, the COVID-19 pandemic and its consequences can be conceptualized as a multidimensional and potentially toxic stressor for mental health in the general population. This selective literature review provides an overview of longitudinal studies published until June 2021 that have investigated the impact of the COVID-19 pandemic on mental health in the European population. Risk and protective factors identified in the studies are summarized. Forty-two studies that met inclusion and search criteria (COVID-19, mental health, longitudinal, and Europe) in PubMed, PsycInfo, and Web of Science databases indicate differential effects of the pandemic on mental distress, depression, and anxiety, depending on samples and methods used. Age-specific (e.g., young age), social (e.g., female, ethnical minority, loneliness), as well as physical and mental health-related factors (e.g., pre-pandemic illness) were identified as risk factors for poor mental health. The studies point to several protective factors such as social support, higher cognitive ability, resilience, and self-efficacy. Increasing evidence supports the assumption of the pandemic being a multidimensional stressor on mental health, with some populations appearing more vulnerable than others, although inconsistencies arise. Whether the pandemic will lead to an increase in the prevalence of mental disorders is an open question. Further high-quality longitudinal and multi-national studies and meta-analyses are needed to draw the complete picture of the consequences of the pandemic on mental health.
The release of DNA by cells during extracellular trap (ET) formation is a defense function of neutrophils and monocytes. Neutrophil ET (NET) formation in term infants is reduced compared to adults. Objective: The aim was to quantify NET and monocyte ET (MET) release and the respective key enzymes myeloperoxidase (MPO) and neutrophil elastase (NE) in preterm infants. In this prospective explorative study, ET induction was stimulated by N-formylmethionine-leucyl-phenylalanine (fMLP), phorbol 12-myristate 13-acetate (PMA), lipopolysaccharide (LPS), and lipoteichoic acid (LTA) in the cord blood of preterm infants (n = 55, 23–36 weeks) compared to term infants and adults. METs were quantified by microscopy, and NETs by microscopy and flow cytometry. We also determined the MPO levels within NETs and the intracellular concentrations of NE and MPO in neutrophils. The percentage of neutrophils releasing ET was significantly reduced for preterm infants compared to adults for all stimulants, and with a 68% further reduction for PMA compared to term infants (p = 0.0141). The NET area was not reduced except for when fMLP was administered. The amount of MPO in NET-producing cells was reduced in preterm infants compared to term infants. For preterm infants, but not term infants, the percentage of monocytes releasing ETs was significantly reduced compared to healthy adults for LTA and LPS stimulation. Conclusion: In preterm infants, ETs are measurable parts of the innate immune system, but are released in a reduced percentage of cells compared to adults.
Abstract
Objective
This study was undertaken to calculate epilepsy‐related direct, indirect, and total costs in adult patients with active epilepsy (ongoing unprovoked seizures) in Germany and to analyze cost components and dynamics compared to previous studies from 2003, 2008, and 2013. This analysis was part of the Epi2020 study.
Methods
Direct and indirect costs related to epilepsy were calculated with a multicenter survey using an established and validated questionnaire with a bottom‐up design and human capital approach over a 3‐month period in late 2020. Epilepsy‐specific costs in the German health care sector from 2003, 2008, and 2013 were corrected for inflation to allow for a valid comparison.
Results
Data on the disease‐specific costs for 253 patients in 2020 were analyzed. The mean total costs were calculated at €5551 (±€5805, median = €2611, range = €274–€21 667) per 3 months, comprising mean direct costs of €1861 (±€1905, median = €1276, range = €327–€13 158) and mean indirect costs of €3690 (±€5298, median = €0, range = €0–€11 925). The main direct cost components were hospitalization (42.4%), antiseizure medication (42.2%), and outpatient care (6.2%). Productivity losses due to early retirement (53.6%), part‐time work or unemployment (30.8%), and seizure‐related off‐days (15.6%) were the main reasons for indirect costs. However, compared to 2013, there was no significant increase of direct costs (−10.0%), and indirect costs significantly increased (p < .028, +35.1%), resulting in a significant increase in total epilepsy‐related costs (p < .047, +20.2%). Compared to the 2013 study population, a significant increase of cost of illness could be observed (p = .047).
Significance
The present study shows that disease‐related costs in adult patients with active epilepsy increased from 2013 to 2020. As direct costs have remained constant, this increase is attributable to an increase in indirect costs. These findings highlight the impact of productivity loss caused by early retirement, unemployment, working time reduction, and seizure‐related days off.
The switch from working in-office to working from home in the context of the COVID-19 pandemic had a significant impact on people’s mobility behavior. In view of the need for action arising from the ongoing challenge of climate change, these changes should be seen as an opportunity to reduce emissions in the traffic sector. The aim of this study was to analyze changes in work-related mobility that occurred during the COVID-19 pandemic using the case of a multinational medium-sized retail chain situated in semi-rural Germany. The case study allowed us to examine those changes in connection with individual attitudes and perspectives of the company and its employees. Thus, we quantitatively recorded the mobility behavior of the company’s employees, followed by an expert interview to ascertain the company’s perspective. We found a reduction in the frequency of commuting and business trips made by employees, which seemed to continue beyond the COVID-19 crisis. However, according to our findings these changes were not based on individual motivation to act in a climate-aware manner but are subject to the framework conditions created by employers for the adoption of climate-friendly behavior. The results of this work could be used by companies and policymakers to create such favorable framework conditions.
Objective
Acute pancreatitis (AP) is an inflammatory disorder, the severe form of which is burdened with multi-organ dysfunction and high mortality. The pathogenesis of life –threatening organ complications, such as respiratory and renal failure, is unknown.
Design
Organ dysfunction was investigated in a mouse model of AP. The influence of monocytes and neutrophils on multi organ dysfunction syndrome (MODS) was investigated in vivo by antibody depletion. Using real-time-fluorescence and deformability-cytometry (RT-DC) analysis we determined the mechanical properties of neutrophils and monocytes during AP. Furthermore, blood samples of pancreatitis patients were used to characterize severity-dependent chemokine profiles according to the revised Atlanta classification.
Results
Similar to AP in humans, severe disease in the mouse model associates with organ dysfunction mainly of lung and kidney, which is triggered by a mobilisation of Ly6g-/CD11b+/Ly6c hi monocytes, but not of Ly6g+/CD11b+ neutrophils. Monocyte depletion by anti-CCR2 antibody treatment ameliorated lung function (oxygen consumption) without interfering with the systemic immune response. RT-DC analysis of circulation monocytes showed a significant increase in cell size during SAP, but without a compensatory increase in elasticity. Patient chemokine profiles show a correlation of AP severity with monocyte attracting chemokines like MCP-1 or MIG and with leukocyte mobilisation.
Conclusion
In AP, the physical properties of mobilized monocytes, especially their large size, result in an obstruction of the fine capillary systems of the lung and of the kidney glomeruli. A selective depletion of monocytes may represent a treatment strategy for pancreatitis as well as for other inflammation-related disorders.
Abstract
The neritid snail Theodoxus fluviatilis has formed regional subgroups in northern Europe, where it appears in both freshwater (FW) and brackish water (BW) in coastal areas of the Baltic Sea. These ecotypes show clear differences in osmotolerance and in the modes of accumulating organic osmolytes under hyperosmotic stress. We reasoned that the expression patterns of soluble proteins in the two ecotypes may differ as well. BW snails have to deal with a higher salinity (up to 20‰) than FW snails (0.5‰) and also cope with frequent fluctuations in environmental salinity that occur after heavy rains or evaporation caused by extended periods of intense sunshine. Therefore, the protein expression patterns of specimens collected at five different FW and BW sites were analyzed using 2D SDS‐PAGE, mass spectrometry, and sequence comparisons based on a transcriptome database for Theodoxus fluviatilis. We identified 89 differentially expressed proteins. The differences in the expression between FW and BW snails may be due to phenotypic plasticity, but may also be determined by local genetic adaptations. Among the differentially expressed proteins, 19 proteins seem to be of special interest as they may be involved in mediating the higher tolerance of BW animals towards environmental change compared with FW animals.
In der vorliegenden Arbeit wurde der Einfluss von kaltem Atmosphärendruckplasma (CAP) auf die Oberfläche des dentalen Restaurationsmaterials Amalgam untersucht.
Bisher liegen in der Literatur noch keine vergleichbaren Untersuchungen vor.
Die Amalgamprüfkörper wurden aus dem kupferhaltigen Non-Gamma-2-Amalgam Amalcap plus (Ivoclar Vivadent, Schaan, Liechtenstein) hergestellt, welches in zylinderförmige Polyoxymethylenformen kondensiert wurde. Die Kaltplasma-Behandlung erfolgte mit dem kINPen® (neoplas GmbH, Greifswald, Deutschland) in 3 Behandlungsgruppen. Gruppe A wurde 60 Sekunden rasternd, Gruppe B 30 Sekunden stationär und Gruppe C 5 Minuten stationär mit CAP behandelt.
Die Amalgamprüfkörperoberflächen wurden jeweils vor und nach der Plasmabehandlung untersucht. Die mikroskopische Dokumentation erfolgte hierbei mittels Lichtmikroskopie und Rasterelektronenmikroskopie (REM), die chemische Analyse durch die Röntgenphotoelektronenspektroskopie (XPS). Für die Beurteilung der Oberflächenrauheit wurden Weißlichtinterferometer und Profilometer eingesetzt. Zur Ermittlung der Hydrophilität kam die Kontaktwinkelmessung zum Einsatz.
Optisch traten in Behandlungsgruppe A und B keine Veränderungen auf. Lediglich bei den 5 Minuten plasmabehandelten Amalgamproben waren leichte Veränderungen erkennbar, die vor allem im Bereich der Poren und der optisch dunkleren Cu6Sn5-Phase lokalisiert waren und sich mikroskopisch braun-orange beziehungsweise dunkelgrau bis schwarz darstellten. Mittels Focused Ion Beam (FIB) präparierte Querschnitte zeigten hierbei keine Veränderungen in der Tiefe, sodass anzunehmen ist, dass die Effekte sich auf die direkt mit dem Plasma in Kontakt stehende Oberfläche beschränken.
Eine Veränderung der Oberflächenrauheit konnte in keiner Behandlungsgruppe festgestellt werden.
Direkt nach der Plasmabehandlung zeigte sich in allen Behandlungsgruppen eine Hydrophilisierung der Oberfläche. Diese war in Gruppe C am stärksten ausgeprägt. Allerdings haben sich bereits nach 2 Tagen die Ausgangswerte wieder eingestellt, der Effekt war also reversibel. Elementar hat durch die Plasmabehandlung im Rahmen eines Reinigungseffekts der oberflächliche Kohlenstoffanteil deutlich abgenommen, der Sauerstoffanteil hingegen zugenommen. Gleichzeitig stieg das Zinn/Quecksilberverhältnis zugunsten des Zinns. Zusätzlich ist es in Gruppe A zu einer Verschiebung des Cu 2p-Peaks in Richtung einer geringeren Bindungsenergie gekommen, das Kupfer lag also nicht mehr einwertig, sondern zunehmend zweiwertig vor.
Mittels energiedispersiver Röntgenspektroskopie (EDX) war es zudem möglich, die an der Oberfläche optisch differenzierbaren Phasen des Amalgams zu charakterisieren.
Insgesamt kann festgestellt werden, dass es in durchgeführten Untersuchungen keine Hinweise gab, die darauf schließen lassen, dass es durch die Einwirkung von CAP auf Amalgamrestaurationen zu unerwünschten Effekten bezüglich der Oberflächeneigenschaften kommt, die sich negativ auf Patienten oder Behandler auswirken könnten. Um die Übertragbarkeit der Ergebnisse auf die klinische Praxis sicherzustellen, sind allerdings noch weitere Untersuchungen notwendig.
Infolge des Verlustes von exokriner und endokriner Pankreasfunktion besteht bei Patienten mit chronischer Pankreatitis ein erhöhtes Mangelernährungsrisiko. Zu Früherkennung, Diagnostik und Therapie von Mangelernährung bei dieser Krankheitsentität liegen indes nur unzureichende Erkenntnisse vor. Ziel dieser Arbeit ist es daher, die bestehenden Wissenslücken hinsichtlich Risikofaktoren, Früherkennung und Behandlung von Mangelernährung bei chronischer Pankreatitis zu schließen.
Zur Identifizierung von Behandlungsansätzen erfolgte mittels einer systematischen Übersichtsarbeit mit Metaanalyse eine Auswertung der Evidenz aus randomisierten kontrollierten Studien. Risikofaktoren, Phänotyp und gesundheitliche Folgen der Mangelernährung wurden in einer prospektiven multizentrischen Querschnittstudie untersucht. Schließlich erfolgte die Testung der Umsetzbarkeit und des Effekts einer intensivierten ernährungsmedizinischen Therapie im Rahmen einer einarmigen Machbarkeitsstudie.
Die systematische Übersichtsarbeit mit Metaanalyse ergab, dass hinsichtlich der Behandlung von Mangelernährung bei chronischer Pankreatitis nur sehr begrenzt Daten aus hochwertigen randomisierten kontrollierten Studien vorliegen. Aktuelle Empfehlungen basieren daher in der Regel auf Erkenntnissen aus Beobachtungsstudien oder Expertenmeinungen. In der Querschnittstudie zeigte sich, dass eine Mangelernährung bei etwa zwei Dritteln der Patienten vorlag. Die mangelernährten Individuen wiesen häufiger eine kürzere Krankheitsdauer sowie eine höhere Krankheitsschwere auf. Der Phänotyp der Mangelernährung war insbesondere durch den Verlust von Muskelmasse nicht aber -funktion charakterisiert und trat bei Vorliegen einer schweren Mangelernährung ausgeprägter in Erscheinung als bei moderatem Schweregrad. Infolge der intensivierten Ernährungstherapie konnten diese pathologischen Veränderungen teilweise umkehrt werden. Neben einer Gewichtssteigerung resultierte die Intervention in einer Zunahme an Muskelmasse sowie einer Verbesserung des Krankheitsschweregrades. Hinsichtlich der Umsetzbarkeit konnte für die meisten Interventionskomponenten eine hohe Compliance beobachtet werden. Zudem ergab sich nur eine geringe Dropout-Rate im Interventionsverlauf.
Mangelernährung stellt in der Tat eine häufige Komplikation der chronischen Pankreatitis dar und ist insbesondere durch eine verringerte Muskelmasse charakterisiert. Hierdurch ist die Muskelfunktion zwar kaum kompromittiert, allerdings geht dieser Zustand mit einer höheren Krankheitsschwere einher. Eine intensivierte ernährungsmedizinische Therapie bei mangelernährten Patienten mit chronischer Pankreatitis ist umsetzbar und verbessert effektiv den Ernährungszustand, aber auch funktionale und prognostische Parameter. Anschließende Untersuchungen sind indes erforderlich, um Früherkennung und Behandlung weiter zu optimieren und die notwendige Evidenz für starke Empfehlungen zu generieren.
Background
In acute pancreatitis, secondary infection of pancreatic necrosis is a complication that mostly necessitates interventional therapy. A reliable prediction of infected necrotizing pancreatitis would enable an early identification of patients at risk, which however, is not possible yet.
Methods
This study aims to identify parameters that are useful for the prediction of infected necrosis and to develop a prediction model for early detection. We conducted a retrospective analysis from the hospital information and reimbursement data system and screened 705 patients hospitalized with diagnosis of acute pancreatitis who underwent contrast-enhanced computed tomography and additional diagnostic puncture or drainage of necrotic collections. Both clinical and laboratory parameters were analyzed for an association with a microbiologically confirmed infected pancreatic necrosis. A prediction model was developed using a logistic regression analysis with stepwise inclusion of significant variables. The model quality was tested by receiver operating characteristics analysis and compared to single parameters and APACHE II score.
Results
We identified a total of 89 patients with necrotizing pancreatitis, diagnosed by computed tomography, who additionally received biopsy or drainage. Out of these, 59 individuals had an infected necrosis. Eleven parameters showed a significant association with an infection including C-reactive protein, albumin, creatinine, and alcoholic etiology, which were independent variables in a predictive model. This model showed an area under the curve of 0.819, a sensitivity of 0.692 (95%-CI [0.547–0.809]), and a specificity of 0.840 (95%-CI [0.631–0.947]), outperforming single laboratory markers and APACHE II score. Even in cases of missing values predictability was reliable.
Conclusion
A model consisting of a few single blood parameters and etiology of pancreatitis might help for differentiation between infected and non-infected pancreatic necrosis and assist medical therapy in acute necrotizing pancreatitis.
Background/Aims
Patients with chronic pancreatitis (CP) have an increased risk of malnutrition, a condition linked to reduced muscle mass and physical performance. We have investigated the risk factors, phenotypic presentation, and health implications associated with malnutrition in CP.
Materials and Methods
In a multicenter cross-sectional study we recruited patients with confirmed CP and healthy volunteers as a control group. Malnutrition was diagnosed according to the criteria proposed by the Global Leadership Initiative on Malnutrition. We performed detailed examinations of body composition and physical function as well as testing of routine blood parameters and markers of inflammation.
Results
We included 66 patients [mean (±SD) age: 56.0 (±14.5) years; 51 males] and an equal number of age- and sex-matched controls. Moderate malnutrition was diagnosed in 21% (n = 14) and severe malnutrition in 42% (n = 28) of patients. Besides weight loss malnourished patients showed lower fat and skeletal muscle mass compared to both non-malnourished subjects and healthy controls. Only in severe malnutrition, blood parameters reflected elevated inflammation and reduced muscle reserves. Handgrip strength in patients did not differ by nutritional status but there was a significant correlation (rho = 0.705, p < 0.001) with skeletal muscle mass. Although 20 patients (30%) had pathologically reduced skeletal muscle mass, only two individuals (3%) had sarcopenia with concomitantly reduced handgrip strength.
Conclusion
Malnutrition is a frequent complication of CP characterized by loss of skeletal muscle mass. As this condition becomes evident only at an advanced stage, regular testing for altered body composition is recommended. Suitable biomarkers and the link between loss of muscle mass and physical function require further investigation.
Clinical Trial Registration
[https://clinicaltrials.gov/ct2/show/NCT04474743], identifier [NCT04474743].
Background
The focal form of CHI is caused by an autosomal recessive pathogenic variant affecting the paternal homologue of genes ABCC8 or KCNJ11 and a second somatic event specifically occurring in the affected islet of Langerhans. The approach of this study was to integrate the genetic changes occurring in pancreatic focal lesions of CHI at the genomic and transcriptional level.
Research Design and Methods
Patients receiving therapeutic surgery and with proven ABCC8 or KCNJ11 pathogenic variants were selected and analyzed for loss of heterozygosity (LOH), changes in copy number and uniparental disomy (UPD) on the short am of chromosome 11 by molecular microarray analysis and methylation-specific MLPA. Gene expression was analyzed by RT-PCR and Massive Analysis of cDNA Ends (MACE).
Results
Both genes, ABCC8 and KCNJ11, are located in proximity to the Beckwith-Wiedemann (BWS) imprinting control region on chromosome 11p15. Somatic paternal uniparental isodisomy (UPD) at chromosome 11p was identified as second genetic event in focal lesions resulting in LOH and monoallelic expression of the mutated ABCC8/KCNJ11 alleles. Of five patients with samples available for microarray analysis, the breakpoints of UPD on chromosome 11p were different. Samples of two patients were analyzed further for changes in gene expression. Profound downregulation of growth suppressing genes CDKN1 and H19 was detected in focal lesions whereas growth promoting gene ASCL2 and pancreatic transcription factors of the endocrine cell lineage were upregulated.
Conclusions
Paternal UPD on the short arm of chromosome 11 appears to be the major second genetic event specifically within focal lesions of CHI but no common breakpoint for UDP can be delineated. We show for the first time upregulation of growth promoting ASCL2 (achaete-scute homolog 2) suggestive of a driving factor in postnatal focal expansion in addition to downregulation of growth suppressing genes CDKN1C and H19.
Simple Summary
Neurotoxicity is an on-target side effect of GD2-directed immunotherapy due to the expression of GD2 on healthy cells. Patients with high-risk neuroblastoma who receive treatment with anti-GD2 immunotherapy, therefore, require close observation and supportive management to improve treatment tolerance and avoid the persistence of neurological symptoms. This study reports on the incidence, clinical course and management of patients who experienced neurotoxicity due to treatment with the anti-GD2 antibody dinutuximab beta, given with or without interleukin-2, in two clinical trials. The majority of severe neurotoxic events were observed in patients treated with dinutuximab beta combined with interleukin-2, with most patients recovering following supportive management. Given the increased risk of neurotoxic events and the lack of clinical benefit reported for the combination treatment in clinical trials, adding interleukin-2 to dinutuximab beta therapy is not recommended. The clinical experiences described here may aid clinicians in managing neurotoxicity associated with dinutuximab beta more effectively.
Abstract
Neurotoxicity is an off-tumour, on-target side effect of GD2-directed immunotherapy with monoclonal antibodies. Here, we report the frequency, management and outcome of patients enrolled in two prospective clinical trials who experienced severe neurotoxicity during immunotherapy with the anti-GD2 antibody dinutuximab beta (DB) administered as short-term infusion (HR-NBL1/SIOPEN study, randomisation R2, EudraCT 2006-001489-17) or as long-term infusion (HR-NBL1/SIOPEN study, randomisation R4, EudraCT 2006-001489-17 and LTI/SIOPEN study, EudraCT 2009-018077-31), either alone or with subcutaneous interleukin-2 (scIL-2). The total number of patients included in this analysis was 1102. Overall, 44/1102 patients (4.0%) experienced Grade 3/4 neurotoxicities (HR-NBL1 R2, 21/406; HR-NBL1 R4, 8/408; LTI study, 15/288), including 27 patients with severe neurotoxicities (2.5%). Events occurred predominantly in patients receiving combined treatment with DB and scIL-2. Neurotoxicity was treated using dexamethasone, prednisolone, intravenous immunoglobulins and, in two patients, plasmapheresis, which was highly effective. While neurological recovery was observed in 16 of 21 patients with severe neurotoxicities, 5/1102 (0.45%) patients experienced persistent and severe neurological deficits. In conclusion, severe neurotoxicity is most commonly observed in patients receiving DB with scIL-2. Considering the lack of clinical benefit for IL-2 in clinical trials so far, the administration of IL-2 alongside DB is not recommended.
Simple Summary
Paratuberculosis is a disease which affects ruminants worldwide. Many countries have implemented certification and monitoring systems to control the disease, particularly in dairy herds. Monitoring herds certified as paratuberculosis non-suspect is an important component of paratuberculosis herd certification programs. The challenge is to detect the introduction or reintroduction of the infectious agent as early as possible with reasonable efforts but high certainty. In our study, we evaluated different low-cost testing schemes in herds where the share of infected animals was low, resulting in a low within-herd prevalence of animals shedding the bacteria that causes paratuberculosis in their feces. The test methods used were repeated pooled milk samples and fecal samples from the barn environment. Our study showed that numerous repetitions of different samples are necessary to monitor such herds with sufficiently high certainty. In the case of herds with a very low prevalence, our study showed that a combination of different sampling approaches is required.
Abstract
An easy-to-use and affordable surveillance system is crucial for paratuberculosis control. The use of environmental samples and milk pools has been proven to be effective for the detection of Mycobacterium avium subsp. paratuberculosis (MAP)-infected herds, but not for monitoring dairy herds certified as MAP non-suspect. We aimed to evaluate methods for the repeated testing of large dairy herds with a very low prevalence of MAP shedders, using different sets of environmental samples or pooled milk samples, collected monthly over a period of one year in 36 herds with known MAP shedder prevalence. Environmental samples were analyzed by bacterial culture and fecal PCR, and pools of 25 and 50 individual milk samples were analyzed by ELISA for MAP-specific antibodies. We estimated the cumulative sensitivity and specificity for up to twelve sampling events by adapting a Bayesian latent class model and taking into account the between- and within-test correlation. Our study revealed that at least seven repeated samplings of feces from the barn environment are necessary to achieve a sensitivity of 95% in herds with a within-herd shedder prevalence of at least 2%. The detection of herds with a prevalence of less than 2% is more challenging and, in addition to numerous repetitions, requires a combination of different samples.
We present the first systematic literature review on stress and burnout in K−12 teachers during the COVID-19 pandemic. Based on a systematic literature search, we identified 17 studies that included 9,874 K−12 teachers from around the world. These studies showed some indication that burnout did increase during the COVID-19 pandemic. There were, however, almost no differences in the levels of stress and burnout experienced by K−12 teachers compared to individuals employed in other occupational fields. School principals' leadership styles emerged as an organizational characteristic that is highly relevant for K−12 teachers' levels of stress and burnout. Individual teacher characteristics associated with burnout were K−12 teachers' personality, self-efficacy in online teaching, and perceived vulnerability to COVID-19. In order to reduce stress, there was an indication that stress-management training in combination with training in technology use for teaching may be superior to stress-management training alone. Future research needs to adopt more longitudinal designs and examine the interplay between individual and organizational characteristics in the development of teacher stress and burnout during the COVID-19 pandemic and beyond.
Untersuchungen zur Immunantwort gegen potenzielle Vakzinkandidaten von Streptococcus pneumoniae
(2022)
Das Gram-positive Bakterium Streptococcus pneumoniae (Pneumokokken) stellt vor allem bei der sehr jungen und älteren sowie immungeschwächten Bevölkerung einen immer mehr an Bedeutung gewinnenden Krankheitserreger dar. Krankheitsbilder wie die Pneumokokken-Meningitis, -Pneumonie und -Sepsis, um nur einige zu nennen, können z.T. schwere Verläufe nehmen. Neue Serotyp-unabhängige Impfstoffe sollen flächendeckenden Schutz vor Infektionen mit S. pneumoniae bieten. Das Hauptaugenmerk der Impfstoffforschung liegt unter anderem auf der Untersuchung von Pneumokokken Protein-basierten Impfstoffen als Bestandteil eines Konjugat- bzw. Multikomponenten Impfstoffes. In diesem Zusammenhang wurde in dieser Arbeit die Immunantwort humaner Immunzellen gegenüber den vier Pneumokokken Oberflächen-Lipoproteinen MetQ, DacB, PnrA und PsaA untersucht. Diese Lipoproteine wurden sowohl in lipidierter als auch in nicht-lipidierter Form als heterologe Proteine generiert, um den Einfluss der Lipidierung auf die Immunreaktion beurteilen zu können. Zur Untersuchung der Reaktion des menschlichen Immunsystems wurden humane Blut-Monozyten (hPBMCs) isoliert und zu pro-inflammatorischen Makrophagen (M1) einerseits und anti-inflammatorischen Makrophagen (M2a) andererseits differenziert. Die Zellen wurden mit den lipidierten bzw. nicht-lipidierten Proteinen stimuliert. In den Überständen wurden zur Beurteilung der abgelaufenen Immunreaktion die Konzentrationen von IL-1β, IL-2, IL-6, IL-8 und TNF bestimmt. Eine Produktion von IL-1β und IL-2 durch die Makrophagen konnte hierbei nicht nachgewiesen werden. M1 Makrophagen zeichneten sich durch eine kaum messbare Cytokin-Produktion auf beide Proteinstimuli aus. Anti-inflammatorische Makrophagen zeigten eine signifikant verstärkte IL-6, IL-8 und TNF Produktion (p< 0,05) nach Stimulation mit allen vier lipidierten Proteinen im Gegensatz zur Stimulation mit nicht-lipidierten Proteinen. Das stärkere Stimulationspotenzial der lipidierten Proteine konnte auf deren Agonismus am TLR2 zurückgeführt werden. Weiterhin konnte die starke Reaktion der M2a Makrophagen mit deren Plastizität sowie mit dem Potenzial von Lipoproteinen zur Umpolarisation von Makrophagen aufgrund des TLR2 Agonismus erklärt werden. Die Ergebnisse zeigen, dass die eingesetzten lipidierten Proteine einen suffizienten und signifikant potenteren Immunstimulus im menschlichen Organismus darstellen, als die gleichen Proteine in nicht-lipidierter Form. Somit könnten diese lipidierten Proteine geeignete Vakzinkandidaten gegen S. pneumoniae darstellen.
Bradyrhizobium diazoefficiens, a bacterial symbiont of soybean and other leguminous plants, enters a nodulation‐promoting genetic programme in the presence of host‐produced flavonoids and related signalling compounds. Here, we describe the crystal structure of an isoflavonoid‐responsive regulator (FrrA) from Bradyrhizobium, as well as cocrystal structures with inducing and noninducing ligands (genistein and naringenin, respectively). The structures reveal a TetR‐like fold whose DNA‐binding domain is capable of adopting a range of orientations. A single molecule of either genistein or naringenin is asymmetrically bound in a central cavity of the FrrA homodimer, mainly via C–H contacts to the π‐system of the ligands. Strikingly, however, the interaction does not provoke any conformational changes in the repressor. Both the flexible positioning of the DNA‐binding domain and the absence of structural change upon ligand binding are corroborated by small‐angle X‐ray scattering (SAXS) experiments in solution. Together with a model of the promoter‐bound state of FrrA our results suggest that inducers act as a wedge, preventing the DNA‐binding domains from moving close enough together to interact with successive positions of the major groove of the palindromic operator.
Die Sicherheit und Wirksamkeit der Arzneimitteltherapie wird maßgeblich von Transportproteinen beeinflusst. Die zelluläre Lokalisation von Transportern hat hierbei wesentlichen Einfluss darauf, ob diese als funktionelle Aufnahme- oder Effluxtransporter fungieren. Für den menschlichen Darm ist die Lokalisation einiger Transporter noch unklar. Ein Beispiel hierfür ist der organic cation transporter (OCT1), welcher für die intestinale Aufnahme zahlreicher kationischer Arzneistoffe, wie beispielsweise Morphin verantwortlich gemacht wird. Bisher gibt es allerdings widersprüchliche Aussagen über die exakte Lokalisation dieses Transporters in der Zellmembran von Enterozyten. Folglich ist die tatsächliche Bedeutung dieses Proteins für die Absorption von Arzneistoffen bis heute ungeklärt.
Daher war das Ziel dieser Arbeit die Expression, Lokalisation und Funktion von OCT1 in Enterozyten anhand verschiedener labortechnischer Methoden näher zu charakterisieren.
Mittels Immunfluoreszenzfärbung wurde versucht die Lokalisation von OCT1 im Zellmodell zu bestimmen. Ebenfalls im Zellmodell erfolgte die Untersuchung des vektoriellen Transportes von Morphin mittels Transwellassay. Diese, sowie entsprechende Analysen vitalen intestinalen Gewebes in der Ussing-Kammer, wurden genutzt, um indirekt Rückschlüsse auf die Transporterlokalisation zu ziehen.
Trotz eindeutiger und der Hypothese entsprechender Expression und Funktion in MDCKII-OCT1/P-gp-Zellen, konnten im Rahmen dieser Arbeit keine eindeutigen Ergebnisse bezüglich der Lokalisation von OCT1 in Caco-2-Zellen generiert werden.
Caco-2-Zellen sollten als Zellmodell für Enterozyten, insbesondere hinsichtlich der Charakterisierung von OCT1, neu bewertet werden, da aktuellen Erkenntnissen entsprechend möglicherweise keine signifikante Expression von OCT1 in diesen Zellen vorliegt. Auch das genutzte OCT1-Modellsubstrat Morphin ist möglicherweise problematisch. Es ist darauf hinzuweisen, dass es sich bei den vorliegenden Daten aufgrund der geringen Versuchszahl nur um vorläufige Ergebnisse handeln kann, welche in zukünftigen Arbeiten verifiziert werden sollten.
Zusammenfassend kann festgehalten werden, dass die vorliegende Arbeit zwar keine neuen Erkenntnisse bezüglich der Lokalisation von OCT1 in Enterozyten erbringen konnte, jedoch die Bedeutung eines kritischen Umgangs mit etablierten Methoden und deren Ergebnissen unterstreicht.
Background: Unwanted drug-drug interactions (DDIs), as caused by the upregulation of clinically relevant drug metabolizing enzymes and transporter proteins in intestine and liver, have the potential to threaten the therapeutic efficacy and safety of drugs. The molecular mechanism of this undesired but frequently occurring scenario of polypharmacy is based on the activation of nuclear receptors such as the pregnane X receptor (PXR) or the constitutive androstane receptor (CAR) by perpetrator agents such as rifampin, phenytoin or St. John’s wort. However, the expression pattern of nuclear receptors in human intestine and liver remains uncertain, which makes it difficult to predict the extent of potential DDIs. Thus, it was the aim of this study to characterize the gene expression and protein abundance of clinically relevant nuclear receptors, i.e., the aryl hydrocarbon receptor (AhR), CAR, farnesoid X receptor (FXR), glucocorticoid receptor (GR), hepatocyte nuclear factor 4 alpha (HNF4α), PXR and small heterodimer partner (SHP), in the aforementioned organs. Methods: Gene expression analysis was performed by quantitative real-time PCR of jejunal, ileal, colonic and liver samples from eight human subjects. In parallel, a targeted proteomic method was developed and validated in order to determine the respective protein amounts of nuclear receptors in human intestinal and liver samples. The LC-MS/MS method was validated according to the current bioanalytical guidelines and met the criteria regarding linearity (0.1–50 nmol/L), within-day and between-day accuracy and precision, as well as the stability criteria. Results: The developed method was successfully validated and applied to determine the abundance of nuclear receptors in human intestinal and liver samples. Gene expression and protein abundance data demonstrated marked differences in human intestine and liver. On the protein level, only AhR and HNF4α could be detected in gut and liver, which corresponds to their highest gene expression. In transfected cell lines, PXR and CAR could be quantified. Conclusions: The substantially different expression pattern of nuclear receptors in human intestinal and liver tissue may explain the different extent of unwanted DDIs in the dependence on the administration route of drugs.
The relevance of cold atmospheric plasmas (CAPs) in biomedicine has recently grown. The potential of CAPs has been discussed in multiple scientific works, highlighting its effectiveness in promoting wound healing, limiting cancer progression, and for sterilization of surfaces. Main bioactive molecules, such as reactive oxygen and nitrogen species (RONS), are proposed as key candidates in these processes. Indeed, the generation of cold plasma induces noble gas ionization which, reacting with atmospheric air molecules, generates species such as singlet oxygen, atomic oxygen radicals, nitric oxide radicals. Although molecular simulations have been conducted, the mechanism of action on biological molecules, as well as the possibility to tune plasmas to produce specific species cocktails (e.g., with different degree of oxidation power) has been not fully unleashed. In this dissertation, presented in form of 5 published scientific articles, focus has been placed on the interaction of plasmas with peptides and proteins, which are main biological effectors in cellular compartments. Precisely, through the development of liquid chromatography coupled mass spectrometry (LC-MS) methods, the effects of plasmas on peptides and proteins in form of oxidative post-translational modifications (oxPTMs) has been investigated. The characterization of these oxPTMs has been performed by treating peptide or protein aqueous solutions and on porcine skin tissues. It has been found that, introducing small amounts of different gases (oxygen, nitrogen, or both) or even water molecules, can made CAPs tunable tools to produce oxygen-species dominating effects versus nitrogen-species dominating effects. In addition to this, it was found that the amino acid position in a peptide or protein influences the quality and quantity of the resulting oxPTMs. Besides this, other important parameters like driven gases, admixture gases or treatment duration were identified as relevant factors for the modification of amino acids in the peptide structure. By comparing the effects between peptide solutions and complex matrices such as porcine skin, water has been identified as a valid vehicle to transport and amplify the plasma chemistry. In an experimental study, the inactivation of a protein (PLA2) was observed after CAP treatment and together with simulation studies, the specific dioxidation of tryptophane W128 was detected as a potential explanation for this inactivation, indicating the strong impact of plasma on biological targets. In summary, oxidative modifications found in peptide solutions were observed also in complex protein structures and sample matrices. In conclusion, this work provides a starting point for future studies of oxidative modifications in complex models and may thus be helpful for further investigations in the fields of plasma medicine and redox chemistry.
This study aims to describe social network and social participation and to assess associations with depressive symptoms in older persons with increased risk for dementia in Germany. We conducted a cross-sectional observational study in primary care patients (aged 60–77) as part of a multicenter cluster-randomized controlled trial (AgeWell.de). We present descriptive and multivariate analyses for social networks (Lubben Social Network Scale and subscales) and social participation (item list of social activities) and analyze associations of these variables with depressive symptoms (Geriatric Depression Scale). Of 1030 included patients, 17.2% were at risk for social isolation (Lubben Social Network Scale < 12). Looking at the subscales, a reduced non-family network was found almost twice as often as a reduced family network. Patients with depressive symptoms had significantly smaller social networks than patients without depression (p < 0.001). They rather engaged in social activities of low involvement level or no weekly social activity at all (p < 0.001). The study shows associations of depressive symptoms with a decreased social network and less social participation in elderly participants. Sufficient non-family contacts and weekly social activities seem to play an important role in mental health and should be encouraged in elderly primary care patients.
Abstract
Modeling of metabolite kinetics after oral administration of ketamine is of special interest because of the higher concentrations of active metabolites because of the hepatic first‐pass effect. This holds especially in view of the potential analgesic and antidepressant effects of 2R,6R‐ and 2S,6S‐hydroxynorketamine at low doses of ketamine. Therefore, a 9‐compartment model was developed to analyze the pharmacokinetics of ketamine enantiomers and their metabolites after racemic ketamine administered intravenously (5 mg) and as 4 doses (10, 20, 40, and 80 mg) of a prolonged‐release formulation (PR‐ketamine). Using a population approach, the serum concentration‐time data of the enantiomers of ketamine, norketamine, dehydronorketamine, and 2,6‐hydroxynorketamine obtained in 15 healthy volunteers could be adequately fitted. The estimated model parameters were used to simulate serum concentration‐time profiles; after multiple dosing of PR‐ketamine (2 daily doses of 20 mg), the steady‐state concentrations of R‐ and S‐ketamine were 1.4 and 1.3 ng/mL, respectively. The steady‐state concentration of 2R,6R‐hydroxynorketamine exceeded those of R‐norketamine (4‐fold), R‐dehydonorketamine (8‐fold), and R‐ketamine (46‐fold), whereas that of 2S,6S‐hydroxynorketamine exceeded that of S‐ketamine by 14‐fold. The model may be useful for identifying dosing regimens aiming at optimal plasma concentrations of 2,6‐hydroxynorketamines.
The relationship between Alzheimer's-related brain atrophy patterns and sleep macro-architecture
(2022)
Introduction
Sleep is increasingly recognized as a major risk factor for neurodegenerative disorders such as Alzheimer's disease (AD).
Methods
Using an magnetic resonance imaging (MRI)–based AD score based on clinical data from the Alzheimer's Disease Neuroimaging Initiative 1 (ADNI1) case-control cohort, we investigated the associations between polysomnography-based sleep macro-architecture and AD-related brain atrophy patterns in 712 pre-symptomatic, healthy subjects from the population-based Study of Health in Pomerania.
Results
We identified a robust inverse association between slow-wave sleep and the AD marker (estimate: −0.019; 95% confidence interval: −0.03 to −0.0076; false discovery rate [FDR] = 0.0041), as well as with gray matter (GM) thicknesses in typical individual cortical AD-signature regions. No effects were identified regarding rapid eye movement or non–rapid eye movement (NREM) stage 2 sleep, and NREM stage 1 was positively associated with GM thickness, mainly in the prefrontal cortical regions.
Discussion
There is a cross-sectional relationship between AD-related neurodegenerative patterns and the proportion of sleep spent in slow-wave sleep.
Zusammenfassung
Die vorliegende Studie befasst sich mit der Morphologie des menschlichen Tuber maxillae unter Berücksichtigung alters- und geschlechtsspezifischer Unterschiede. Grundlage dieser Arbeit waren DVT-Datensätze von 118 erwachsenen Probanden beiderlei Geschlechts. Basierend auf den Rohdatensätzen wurden, mit Hilfe der NewTom 3G Software (QR NNT Version 2.11 Professional ©), Axialschnitte parallel zur Ebene des Nasenhöhlenbodens angefertigt. Die Schichtstärke der Axialschnitte betrug 3,0 mm. Die daraus resultierenden axialen DVT-Schichtaufnahmen waren die Basis für alle weiteren Untersuchungen. Im Folgenden konnte auf der Grundlage definierter Messpunkte und Hilfslinien, Streckenmessungen am Tuber maxillae vorgenommen werden. Diese Streckenmessungen waren Ausgangspunkt für die Definition unterschiedlicher Formentypen des Tuber maxillae.
Um geschlechts- und altersspezifische Unterschiede zwischen den gewonnenen Messdaten und Formtypen des Tuber maxillae beurteilen zu können, wurde zunächst eine deskriptive Statistik erstellt. Darauf aufbauend erfolgte ein t-Test, durch den Geschlechtsunterschiede dargestellt werden konnten. Mit Hilfe einer einfaktoriellen Varianzanalyse (ANOVA) untersuchten wir die gewonnenen Daten in Bezug zu den drei Altersgruppen der erwachsenen Probanden. Des Weiteren nahmen wir eine Korrelations- und Regressionsanalyse unter Berücksichtigung verschiedener Gesichtsschädelmaße vor. Mit Hilfe der Bestimmung eines Symmetriequotienten erfolgte die Beurteilung von Seitenunterschieden.
Anhand der Streckenmessungen des Tuber maxillae zeigte sich, dass der Tuber maxillae bei den Männern höher ist als bei den Frauen. Breite und Länge des Tuber maxillae sind jedoch geschlechtsunabhängig. Obgleich keine signifikanten Altersunterschiede in Bezug auf die Höhe des Tuber maxillae nachgewiesen werden konnten, wurden Unterschiede zwischen den Altersgruppen 2 und 3 beobachtet. So nahm die Höhe des Tuber maxillae bis zur Altersgruppe 2 zunächst zu, um bei den älteren Probanden in der Altersgruppe 3 wieder leicht abzunehmen. Das Symmetrieverhalten des Tuber maxillae zeigte im Bezug zur Höhe symmetrische Höhenmaße. Anders verhielt es sich bei der Breite und Länge des Tuber maxillae. Sowohl für die Breite als auch für die Länge zeigten sich bei über der
Hälfte der Probanden Symmetrieunterschiede zwischen dem rechten und linken Tuber maxillae. Weiterhin konnte diese Studie verschiedene Korrelationen zwischen den Maßen des Tuber maxillae und verschiedenen Schädelmaßen aufzeigen. Eine weiterführende Regressionsanalyse bestätigte einen signifikanten Zusammenhang, insbesondere zwischen der Höhe des linken Tuber maxillae und dem Volumen des linken Sinus maxillaris.
Basierend auf den in dieser Studie ermittelten Maßen des Tuber maxillae konnten dem Tuber maxillae sowohl in der Norma lateralis als auch in der Norma horizontalis drei verschiedene Formtypen zugeordnet werden. Für die Formbestimmung in der Norma lateralis wurden jeweils die größten Längen des Tuber maxillae von kranial nach kaudal auf Millimeterpapier übertragen und an der anterioren Begrenzung des Tuber maxillae ausgerichtet. Mit Hilfe der so gewonnen Skizzen definierten wir die Formtypen I, II und III. Beim Typ I beschreibt die Dorsalfläche des Tuber maxillae einen annährend geraden Verlauf von kranial nach kaudal. Der Typ II hingegen ist gekennzeichnet durch eine tendenzielle Zunahme der Dorsalfläche von kranial nach kaudal. Beim Typ III zeigt sich ein konkaver Verlauf der Dorsalfläche des Tuber maxillae. Unsere Untersuchungen zeigten, dass in der Norma lateralis der Typ III bei den Männern am häufigsten vorkam. Bei den weiblichen Probanden zeigte sich dagegen häufiger der Typ II.
Die Formbestimmung des Tuber maxillae in der Norma horizontalis erfolgte an drei definierten Horizontalschnitten. Der erste Horizontalschnitt befindet sich auf Höhe des mittleren Nasenganges, der zweite Horizontalschnitt auf Höhe des Foramen palatinum majus und der dritte Horizontalschnitt in Höhe der Wurzelspitzen der Oberkiefermolaren. Die Einteilung in die Formen erfolgte in ähnlicher Weise wie in der Norma lateralis. Die größten Längen des Tuber maxillae wurden von lateral nach medial auf Millimeterpapier übertragen und wieder an der anterioren Begrenzung des Tuber maxillae ausgerichtet. Anhand der gewonnenen Skizzen definierten wir die Formtypen a, b und c für die Norma horizontalis. Beim Typ a befindet sich die stärkste Ausprägung der Dorsalfläche im medialen Drittel des Tuber maxillae. Der Typ b zeigt seine stärkste Ausprägung im mittleren Drittel des Tuber maxillae. Beim Typ c befindet sich die stärkste Ausprägung im lateralen Drittel des Tuber maxillae. Es zeigte sich, dass in Höhe des mittleren Nasenganges bei den Männern der Typ a am häufigsten auftrat. In Höhe des Foramen palatinum majus und auf Höhe der Wurzelspitzen der oberen Molaren ermittelten wir den Typ b am häufigsten. Bei den Frauen zeigte sich auf Höhe des mittleren Nasenganges der
Typ a am häufigsten. Gleiches war auf Höhe des Foramen palatinum majus zu beobachten. In der Ebene der Wurzelspitzen der oberen Molaren zeigte sich häufiger der Typ b. Anhand der hier gewonnenen Daten ist es möglich, die Form des Tuber maxillae zu quantifizieren. Die Untersuchungsergebnisse können für verschiedene klinische Fachgebiete, wie die Mund-Kiefer-Gesichtschirurgie (MKG), die Prothetik und die Kieferorthopädie, von Interesse sein. In der MKG beispielweise können sie von Bedeutung bei der Wahl des passenden dentalen Implantatsystems in der Tuberregion sein. Aber auch bei der Gewinnung von Knochen aus der Tuberregion kann die Kenntnis über Form und Größe des Tuber maxillae von Nutzen sein. Die hier vorliegende Studie gibt Aufschluss über die Morphologie des Tuber maxillae beim Menschen. Weiterhin liefert sie Grundlagen für weiterführende Studien an anderen menschlichen Populationen.
Background
Although chronic kidney disease (CKD) is highly prevalent in the general population, little research has been conducted on CKD management in ambulatory care.
Objective was to assess management and quality of care by evaluating CKD coding in ambulatory care, patient diagnosis awareness, frequency of monitoring and whether appropriate patients are referred to nephrology.
Methods
Clinical data from the population-based cohort Study of Health in Pomerania (SHIP-START) were matched with claims data of the Association of Statutory Health Insurance Physicians. Quality of care was evaluated according international and German recommendations.
Results
Data from 1778 participants (56% female, mean age 59 years) were analysed. 10% had eGFR < 60 ml/min/1.73m2 (mean age 74 years), 15% had albuminuria. 21% had CKD as defined by KDIGO. 20% of these were coded and 7% self-reported having CKD. Coding increased with GFR stage (G3a 20%, G3b 61%, G4 75%, G5 100%). Serum creatinine and urinary dip stick testing were billed in the majority of all participants regardless of renal function. Testing frequency partially surpassed recommendations. Nephrology consultation was billed in few cases with stage G3b-G4.
Conclusion
CKD coding increased with stage and was performed reliably in stages ≥ G4, while CKD awareness was low. Adherence to monitoring and referral criteria varied, depending on the applicability of monitoring criteria. For assessing quality of care, consent on monitoring, patient education, referral criteria and coordination of care needs to be established, accounting for patient related factors, including age and comorbidity.
Trial registration
This study was prospectively registered as DRKS00009812 in the German Clinical Trials Register (DRKS).
Operationsziel
Durchführung einer periazetabulären Osteotomie (PAO) über einen minimal-invasiven Zugang zur dreidimensionalen Korrektur der Orientierung der Hüftgelenkpfanne.
Indikationen
Symptomatische Hüftdysplasie in der Adoleszenz nach dem Schluss der Y‑Wachstumsfuge und beim Erwachsenen.
Kontraindikationen
Fortgeschrittene Arthrose (Arthrosegrad ≥ 2 nach Tönnis), präformierte Sekundärpfanne, offene Y‑Wachstumsfuge.
Operationstechnik
Über einen minimal-invasiven Zugang wird eine periazetabuläre Osteotomie durchgeführt.
Ergebnisse
Insgesamt wurden 39 Patienten über 3,5 (3 bis 4,5) Jahre nachbeobachtet. Der laterale Centrum-Erker-Winkel nach Wiberg konnte signifikant von 16,1° (7–24°) auf 30,5° (25–37°) (p < 0,0001), der Tragflächenwinkel von 13,2° (2–25,3°) auf 2,8° (−3–13°) (p < 0,0001) korrigiert werden. Die mittlere Operationszeit betrug 88 (57 bis 142) Minuten. Es traten keine schweren Komplikationen auf.
Thermally treated kaolinite is used to develop a range of alumino‐silicate‐based precursor materials but its behavior during plasma spraying has not been well‐researched. In this study, two types of kaolinite samples were investigated in the form of low defect (KGa‐1b) and high defect (KGa‐2) varieties. The extreme temperatures of the plasma stream (up to 20 000 K) induced flash melting to produce a highly porous alumino‐silicate glass without any crystallization of new Al−Si oxide minerals. The glass is comprised largely of intact or deformed spheres (average diameters 1.14–1.44 μm), which indicates rapid quenching and solidification before impact. The subspherical structures contain up to 40 % closed pore space caused by the rapid escape of water during melting. The low‐density, porous alumino‐silicate glass coatings with predicted specific surface areas (>0.95 m2/g) and hardnesses >1.8 GPa represent a potentially reactive but physically stable substrate ideal for further chemical functionalization.
Advancing Radiation-Detected Resonance Ionization towards Heavier Elements and More Exotic Nuclides
(2022)
RAdiation-Detected Resonance Ionization Spectroscopy (RADRIS) is a versatile method for highly sensitive laser spectroscopy studies of the heaviest actinides. Most of these nuclides need to be produced at accelerator facilities in fusion-evaporation reactions and are studied immediately after their production and separation from the primary beam due to their short half-lives and low production rates of only a few atoms per second or less. Only recently, the first laser spectroscopic investigation of nobelium (Z=102) was performed by applying the RADRIS technique in a buffer-gas-filled stopping cell at the GSI in Darmstadt, Germany. To expand this technique to other nobelium isotopes and for the search for atomic levels in the heaviest actinide element, lawrencium (Z=103), the sensitivity of the RADRIS setup needed to be further improved. Therefore, a new movable double-detector setup was developed, which enhances the overall efficiency by approximately 65% compared to the previously used single-detector setup. Further development work was performed to enable the study of longer-lived (t1/2>1 h) and shorter-lived nuclides (t1/2<1 s) with the RADRIS method. With a new rotatable multi-detector design, the long-lived isotope 254Fm (t1/2=3.2 h) becomes within reach for laser spectroscopy. Upcoming experiments will also tackle the short-lived isotope 251No (t1/2=0.8 s) by applying a newly implemented short RADRIS measurement cycle.
Background and objective
The COVID-19 pandemic started in Wuhan, China, in December 2019. Although there are some doubts about the reporting of cases and deaths in China, it seems that this country was able to control the epidemic more effectively than many other countries. In this paper, we would like to analyze the measures taken in China and compare them with other countries in order to find out what they can learn from China.
Methods
We develop a system dynamics model of the COVID-19 pandemic in Wuhan. Based on a number of simulations we analyze the impact of changing parameters, such as contact rates, on the development of a second wave.
Results
Although China’s health care system seems to be poorly financed and inefficient, the epidemic was brought under control in a comparably short period of time and no second wave was experienced in Wuhan until today. The measures to contain the epidemic do not differ from what was implemented in other countries, but China applied them very early and rigorously. For instance, the consequent implementation of health codes and contact-tracking technology contributed to contain the disease and effectively prevented the second and third waves.
Conclusions
China’s success in fighting COVID-19 is based on a very strict implementation of a set of measures, including digital management. While other countries discuss relaxing the lock-down at a rate of 50 per 100,000 inhabitants, China started local lock-downs at a rate of 1.59 per 100,000. We call for a public debate whether this policy would be feasible for more liberal countries as well.
N6-methyladenosine (m6A) RNA methylation is an emerging epigenetic modification in recent years and epigenetic regulation of the immune response has been demonstrated, but the potential role of m6A modification in GBM tumor microenvironment (TME) cell infiltration and stemness remain unknown. The m6A modification patterns of 310 GBM samples were comprehensively evaluated based on 21 m6A regulators, and we systematically correlated these modification patterns with TME cell infiltration characteristics and stemness characteristics. Construction of m6Ascore to quantify the m6A modification patterns of individual GBM samples using a principal component analysis algorithm. We identified two distinct patterns of m6A modification. The infiltration characteristics of TME cells in these two patterns were highly consistent with the immunophenotype of the GBM, including the immune activation differentiation pattern and the immune desert dedifferentiation pattern. We also identified two modes of regulation of immunity and stemness by m6A methylation. Stromal activation and lack of effective immune infiltration were observed in the high m6Ascore subtype. Pan-cancer analysis results illustrate a significant correlation between m6AScore and tumor clinical outcome, immune infiltration, and stemness. Our work reveals that m6A modifications play an important role in the development of TME and stemness diversity and complexity. Patients with a low m6AScore showed significant therapeutic advantages and clinical benefits. Assessing the m6A modification pattern of individual tumors will help enhance our knowledge of TME infiltration and stemness characteristics, contribute to the development of immunotherapeutic strategies.
The impact of summer drought on peat soil microbiome structure and function-A multi-proxy-comparison
(2022)
Different proxies for changes in structure and/or function of microbiomes have been developed, allowing assessing microbiome dynamics at multiple levels. However, the lack and differences in understanding the microbiome dynamics are due to the differences in the choice of proxies in different studies and the limitations of proxies themselves. Here, using both amplicon and metatranscriptomic sequencings, we compared four different proxies (16/18S rRNA genes, 16/18S rRNA transcripts, mRNA taxonomy and mRNA function) to reveal the impact of a severe summer drought in 2018 on prokaryotic and eukaryotic microbiome structures and functions in two rewetted fen peatlands in northern Germany. We found that both prokaryotic and eukaryotic microbiome compositions were significantly different between dry and wet months. Interestingly, mRNA proxies showed stronger and more significant impacts of drought for prokaryotes, while 18S rRNA transcript and mRNA taxonomy showed stronger drought impacts for eukaryotes. Accordingly, by comparing the accuracy of microbiome changes in predicting dry and wet months under different proxies, we found that mRNA proxies performed better for prokaryotes, while 18S rRNA transcript and mRNA taxonomy performed better for eukaryotes. In both cases, rRNA gene proxies showed much lower to the lowest accuracy, suggesting the drawback of DNA based approaches. To our knowledge, this is the first study comparing all these proxies to reveal the dynamics of both prokaryotic and eukaryotic microbiomes in soils. This study shows that microbiomes are sensitive to (extreme) weather changes in rewetted fens, and the associated microbial changes might contribute to ecological consequences.
Periodontitis is a multifactorial disease. The aim of this explorative study was to investigate the role of Interleukin-(IL)-1, IL-4, GATA-3 and Cyclooxygenase-(COX)-2 polymorphisms after non-surgical periodontal therapy with adjunctive systemic antibiotics (amoxicillin/metronidazole) and subsequent maintenance in a Caucasian population. Analyses were performed using blood samples from periodontitis patients of a multi-center trial (ClinicalTrials.gov NCT00707369=ABPARO-study). Polymorphisms were analyzed using quantitative real-time PCR. Clinical attachment levels (CAL), percentage of sites showing further attachment loss (PSAL) ≥1.3 mm, bleeding on probing (BOP) and plaque score were assessed. Exploratory statistical analysis was performed. A total of 209 samples were genotyped. Patients carrying heterozygous genotypes and single-nucleotide-polymorphisms (SNP) on the GATA-3-IVS4 +1468 gene locus showed less CAL loss than patients carrying wild type. Heterozygous genotypes and SNPs on the IL-1A-889, IL-1B +3954, IL-4-34, IL-4-590, GATA-3-IVS4 +1468 and COX-2-1195 gene loci did not influence CAL. In multivariate analysis, CAL was lower in patients carrying GATA-3 heterozygous genotypes and SNPs than those carrying wild-types. For the first time, effects of different genotypes were analyzed in periodontitis progression after periodontal therapy and during supportive treatment using systemic antibiotics demonstrating a slight association of GATA-3 gene locus with CAL. This result suggests that GATA-3 genotypes are a contributory but non-essential risk factor for periodontal disease progression.
The Mt. Pinatubo eruption in 1991 had a severe impact on the Earth system, with a well-documented warming of the tropical lower stratosphere and a general cooling of the surface. This study focuses on the impact of this event on the mesosphere by analyzing solar occultation temperature data from the Halogen Occultation Experiment (HALOE) instrument on the Upper Atmosphere Research Satellite (UARS). Previous analyses of lidar temperature data found positive temperature anomalies of up to 12.9 K in the upper mesosphere that peaked in 1993 and were attributed to the Pinatubo eruption. Fitting the HALOE data according to a previously published method indicates a maximum warming of the mesosphere region of 4.1 ± 1.4 K and does not confirm significantly higher values reported for that lidar time series. An alternative fit is proposed that assumes a more rapid response of the mesosphere to the volcanic event and approximates the signature of the Pinatubo with an exponential decay function having an e-folding time of 6 months. It suggests a maximum warming of 5.4 ± 3.0 K, if the mesospheric perturbation is assumed to reach its peak 4 months after the eruption. We conclude that the HALOE time series probably captures the decay of a Pinatubo-induced mesospheric warming at the beginning of its measurement period.
Childhood abuse was inconsistently related to whole-brain cortical thickness in former studies. However, both childhood abuse and cortical thickness have been associated with depressive symptoms. We hypothesised that childhood abuse moderates the association between depressive symptoms and cortical thickness. In 1551 individuals of the general population, associations between whole-brain cortical thickness and the interaction of childhood abuse (emotional, physical, and sexual) and depressive symptoms were analysed using an ANCOVA. Linear regression analyses were used to estimate the same effect on the cortical thickness of 34 separate regions (Desikan-Killiany-atlas). A significant interaction effect of childhood abuse and depressive symptoms was observed for whole-brain cortical thickness (F(2, 1534) = 5.28, p = 0.007). A thinner cortex was associated with depressive symptoms in abused (t value = 2.78, p = 0.025) but not in non-abused participants (t value = − 1.50, p = 0.224). Focussing on non-depressed participants, a thicker whole-brain cortex was found in abused compared to non-abused participants (t value = − 2.79, p = 0.025). Similar interaction effects were observed in 12 out of 34 cortical regions. Our results suggest that childhood abuse is associated with reduced cortical thickness in subjects with depressive symptoms. In abused subjects without depressive symptoms, larger cortical thickness might act compensatory and thus reflect resilience against depressive symptoms.
AbstractPlasma medicine refers to the application of nonequilibrium plasmas at approximately body temperature, for therapeutic purposes. Nonequilibrium plasmas are weakly ionized gases which contain charged and neutral species and electric fields, and emit radiation, particularly in the visible and ultraviolet range. Medically-relevant cold atmospheric pressure plasma (CAP) sources and devices are usually dielectric barrier discharges and nonequilibrium atmospheric pressure plasma jets. Plasma diagnostic methods and modelling approaches are used to characterize the densities and fluxes of active plasma species and their interaction with surrounding matter. In addition to the direct application of plasma onto living tissue, the treatment of liquids like water or physiological saline by a CAP source is performed in order to study specific biological activities. A basic understanding of the interaction between plasma and liquids and bio-interfaces is essential to follow biological plasma effects. Charged species, metastable species, and other atomic and molecular reactive species first produced in the main plasma ignition are transported to the discharge afterglow to finally be exposed to the biological targets. Contact with these liquid-dominated bio-interfaces generates other secondary reactive oxygen and nitrogen species (ROS, RNS). Both ROS and RNS possess strong oxidative properties and can trigger redox-related signalling pathways in cells and tissue, leading to various impacts of therapeutic relevance. Dependent on the intensity of plasma exposure, redox balance in cells can be influenced in a way that oxidative eustress leads to stimulation of cellular processes or oxidative distress leads to cell death. Currently, clinical CAP application is realized mainly in wound healing. The use of plasma in cancer treatment (i.e. plasma oncology) is a currently emerging field of research. Future perspectives and challenges in plasma medicine are mainly directed towards the control and optimization of CAP devices, to broaden and establish its medical applications, and to open up new plasma-based therapies in medicine.
The idea of estimating stratospheric aerosol optical thickness from the twilight colours in historic paintings – particularly under conditions of volcanically enhanced stratospheric aerosol loading – is very tantalizing because it would provide information on the stratospheric aerosol loading over a period of several centuries. This idea has in fact been applied in a few studies in order to provide quantitative estimates of the aerosol optical depth after some of the major volcanic eruptions that occurred during the past 500 years. In this study we critically review this approach and come to the conclusion that the uncertainties in the estimated aerosol optical depths are so large that the values have to be considered questionable. We show that several auxiliary parameters – which are typically poorly known for historic eruptions – can have a similar effect on the red–green colour ratio as a change in optical depth typically associated with eruptions such as, for example, Tambora in 1815 or Krakatoa in 1883. Among the effects considered here, uncertainties in the aerosol particle size distribution have the largest impact on the colour ratios and hence the aerosol optical depth estimate. For solar zenith angles exceeding 80∘, uncertainties in the stratospheric ozone amount can also have a significant impact on the colour ratios. In addition, for solar zenith angles exceeding 90∘ the colour ratios exhibit a dramatic dependence on solar zenith angle, rendering the estimation of aerosol optical depth highly challenging. A quantitative determination of the aerosol optical depth may be possible for individual paintings for which all relevant parameters are sufficiently well constrained in order to reduce the related errors.
This work investigated the enzymatic degradation of polyethylene terephthalate (PET) (ArticlesI and II) and polyvinyl alcohol (PVA) (Article III). Physical or chemical degradation of plastic polymers is often performed under extreme conditions like high temperatures or pressure. In comparison to that, recycling of plastics with enzymes can be carried out at ambient temperatures and neutral pH. Enzymes themselves are non- toxic, environmentally friendly, and have been used successfully in a variety of industrial processes.
Enzymatic degradation of polyesters is well studied. Their heteroatomic backbone, which is connecting monomers via ester bonds offers a target for an enzymatic attack. Especially PET, one of the most common polyesters, has been in the focus of research. The first enzyme capable of degrading the polymer was found in 2005. Since then, researchers discovered several enzymes with similar functions and subjected them to enzyme engineering. Improving the enzyme's substrate affinity, activity, and stability aims at making PET recycling more efficient. Article I provides an overview of limitations that enzymatic PET recycling is still facing and the research carried out to overcome them. More precisely, enzyme−substrate interactions, thermostability, catalytic efficiency, and inhibition caused by oligomeric degradation intermediates are summarized and discussed in detail.
Article II further addresses one of the above-mentioned limitations, namely product inhibition of PET hydrolyzing enzymes. We elucidated the crystal structure of TfCa, a carboxylesterase from Thermobifida fusca (T. fusca), and applied semi-rational enzyme engineering. The article discusses the structure-function relationship of TfCa based on the apo-structure as well as ligand-soaked structures. Furthermore, it compares the structures of TfCa and MHETase, another PET hydrolase helper enzyme. Lastly, we determined the substrate profile of the carboxylesterase based on terephthalate-based oligo-esters of various lengths and one ortho-phthalate ester. In a dual enzyme system, TfCa degraded intermediate products derived from the PET hydrolysis of a variant of PETase hydrolase from Ideonella sakaiensis (I. sakaiensis). The dual enzyme system utilized PET more efficiently in comparison to solely PETase due to relieved product inhibition. Since TfCa successfully degraded oligomeric intermediates, the reaction not only released terephthalic acid as the sole product but also increased the overall product yield.
While PET contains an ester bond that can be attacked and hydrolyzed by esterases or lipases, PVA consists of a homoatomic C-C-backbone with repeating 1,3-diol units. The polymer is water soluble with remarkable physical properties such as thermostability and viscosity. PVA is often described as biodegradable, but microbial degradation is slow and frequently involves cost-intensive cofactors. In this study, we present an improved PVA polymer with derivatized side chains and an enzyme cascade that can degrade not only modified but also unmodified PVA in a one-pot reaction. The enzyme cascade consists of a lipase, an alcohol dehydrogenase (ADH), and a Baeyer-Villiger monooxygenase (BVMO). In comparison to the scarcely published research on PVA degradation with free enzyme, this cascade is not only independent from the frequently required cofactor pyrroloquinoline quinone (PQQ) but, in principle, contains an in vitro cofactor recycling mechanism.
Simple Summary
Neuronal plasticity refers to the brain’s ability to adapt in response to activity-dependent changes. This process, among others, allows the brain to acquire memory or to compensate for a neurocognitive deficit. We analyzed adult FTSJ1-deficient mice in order to gain insight into the role of FTSJ1 in neuronal plasticity. These mice displayed alterations in the hippocampus (a brain structure that is involved in memory and learning, among other functions) e.g., in the form of changes in dendritic spines. Changes in dendritic spines are considered to represent a morphological hallmark of altered neuronal plasticity, and thus FTSJ1 deficiency might have a direct effect upon the capacity of the brain to adapt to plastic changes. Long-term potentiation (LTP) is an electrophysiological correlate of neuronal plasticity, and is related to learning and to processes attributed to memory. Here we show that LTP in FTSJ1-deficient mice is reduced, hinting at disturbed neuronal plasticity. These findings suggest that FTSJ1 deficiency has an impact on neuronal plasticity not only morphologically but also on the physiological level.
Abstract
The role of the tRNA methyltransferase FTSJ1 in the brain is largely unknown. We analyzed whether FTSJ1-deficient mice (KO) displayed altered neuronal plasticity. We explored open field behavior (10 KO mice (aged 22–25 weeks)) and 11 age-matched control littermates (WT) and examined mean layer thickness (7 KO; 6 WT) and dendritic spines (5 KO; 5 WT) in the hippocampal area CA1 and the dentate gyrus. Furthermore, long-term potentiation (LTP) within area CA1 was investigated (5 KO; 5 WT), and mass spectrometry (MS) using CA1 tissue (2 each) was performed. Compared to controls, KO mice showed a significant reduction in the mean thickness of apical CA1 layers. Dendritic spine densities were also altered in KO mice. Stable LTP could be induced in the CA1 area of KO mice and remained stable at for at least 1 h, although at a lower level as compared to WTs, while MS data indicated differential abundance of several proteins, which play a role in neuronal plasticity. FTSJ1 has an impact on neuronal plasticity in the murine hippocampal area CA1 at the morphological and physiological levels, which, in conjunction with comparable changes in other cortical areas, might accumulate in disturbed learning and memory functions.
Obstructive sleep apnea is known to be an overall public health problem that, among other things, increases morbidity and mortality. Risk factors as well as symptoms of this multidimensional sleep-related breathing disorder negatively affect quality of life. With our study we aimed to expose the association between obstructive sleep apnea and quality of life in the population of Pomerania, Germany. We utilized data from the population-based Study of Health in Pomerania (SHIP). Information on health status and risk factors about 4420 participants (2275 women) were gathered within the cohort SHIP-TREND, of which 1209 (559 women) underwent an overnight polysomnography and completed sleep questionnaires. The quality of life of the participants was measured using the Short-Form 12 questionnaire. For our study, an ordinal regression analysis with age, sex, body mass index and the Short-Form 12 health survey as predictors for apnea–hypopnea index was computed. The potential factors affecting quality of life are different between physical and mental dimensions of quality of life. Significant effects were found regarding age, sex, body mass index and the Short-Form 12 Mental Component Score, but not the Physical Component Score.
Response of Osteoblasts to Electric Field Line Patterns Emerging from Molecule Stripe Landscapes
(2022)
Molecular surface gradients can constitute electric field landscapes and serve to control local cell adhesion and migration. Cellular responses to electric field landscapes may allow the discovery of routes to improve osseointegration of implants. Flat molecule aggregate landscapes of amine- or carboxyl-teminated dendrimers, amine-containing protein and polyelectrolytes were prepared on glass to provide lateral electric field gradients through their differing zeta potentials compared to the glass substrate. The local as well as the mesoscopic morphological responses of adhered osteoblasts (MG-63) with respect to the stripes were studied by means of Scanning Ion Conductance Microscopy (SICM) and Fluorescence Microscopy, in situ. A distinct spindle shape oriented parallel to the surface pattern as well as a preferential adhesion of the cells on the glass site have been observed at a stripe and spacing width of 20 μm. Excessive ruffling is observed at the spindle poles, where the cells extend. To explain this effect of material preference and electro-deformation, we put forward a retraction mechanism, a localized form of double-sided cathodic taxis.
Quadruplex-duplex (Q–D) junctions constitute unique structural motifs in genomic sequences. Through comprehensive calorimetric as well as high-resolution NMR structural studies, Q–D junctions with a hairpin-type snapback loop coaxially stacked onto an outer G-tetrad were identified to be most effective binding sites for various polycyclic quadruplex ligands. The Q–D interface is readily recognized by intercalation of the ligand aromatic core structure between G-tetrad and the neighboring base pair. Based on the thermodynamic and structural data, guidelines for the design of ligands with enhanced selectivity towards a Q–D interface emerge. Whereas intercalation at Q–D junctions mostly outcompete stacking at the quadruplex free outer tetrad or intercalation between duplex base pairs to varying degrees, ligand side chains considerably contribute to the selectivity for a Q–D target over other binding sites. In contrast to common perceptions, an appended side chain that additionally interacts within the duplex minor groove may confer only poor selectivity. Rather, the Q–D selectivity is suggested to benefit from an extension of the side chain towards the exposed part of the G-tetrad at the junction. The presented results will support the design of selective high-affinity binding ligands for targeting Q–D interfaces in medicinal but also technological applications.
MicroRNAs (miRNA) are ubiquitous non-coding RNAs that have a prominent role in cellular regulation. The expression of many miRNAs is often found deregulated in prostate cancer (PCa) and castration-resistant prostate cancer (CRPC). Although their expression can be associated with PCa and CRPC, their functions and regulatory activity in cancer development are poorly understood. In this study, we used different proteomics tools to analyze the activity of hsa-miR-3687-3p (miR-3687) and hsa-miR-4417-3p (miR-4417), two miRNAs upregulated in CRPC. PCa and CRPC cell lines were transfected with miR-3687 or miR-4417 to overexpress the miRNAs. Cell lysates were analyzed using 2D gel electrophoresis and proteins were subsequently identified using mass spectrometry (Maldi-MS/MS). A whole cell lysate, without 2D-gel separation, was analyzed by ESI-MS/MS. The expression of deregulated proteins found across both methods was further investigated using Western blotting. Gene ontology and cellular process network analysis determined that miR-3687 and miR-4417 are involved in diverse regulatory mechanisms that support the CRPC phenotype, including metabolism and inflammation. Moreover, both miRNAs are associated with extracellular vesicles, which point toward a secretory mechanism. The tumor protein D52 isoform 1 (TD52-IF1), which regulates neuroendocrine trans-differentiation, was found to be substantially deregulated in androgen-insensitive cells by both miR-3687 and miR-4417. These findings show that these miRNAs potentially support the CRPC by truncating the TD52-IF1 expression after the onset of androgen resistance.
Platelets within one individual display heterogeneity in reactivity, size, age, and expression of surface receptors. To investigate the combined intraindividual contribution of platelet size, platelet age, and receptor expression levels on the reactivity of platelets, we studied fractions of large and small platelets from healthy donors separated by using differential centrifugation. Size-separated platelet fractions were perfused over a collagen-coated surface to assess thrombus formation. Multicolor flow cytometry was used to characterize resting and stimulated platelet subpopulations, and platelet age was determined based on RNA and HLA-I labeling. Signal transduction was analyzed by measuring consecutive phosphorylation of serine/threonine-protein kinase Akt. Compared with small platelets, large platelets adhered faster to collagen under flow and formed larger thrombi. Among the large platelets, a highly reactive juvenile platelet subpopulation was identified with high glycoprotein VI (GPVI) expression. Elevated GPVI expression correlated with high HLA-I expression, RNA content, and increased platelet reactivity. There was a stronger difference in Akt phosphorylation and activation upon collagen stimulation between juvenile and older platelets than between large and small platelets. GPVI expression and platelet reactivity decreased throughout platelet storage at 22°C and was better maintained throughout cold storage at 4°C. We further detected higher GPVI expression in platelets of patients with immune thrombocytopenia. Our findings show that high GPVI expression is a feature of highly reactive juvenile platelets, which are predominantly found among the large platelet population, explaining the better performance of large platelets during thrombus formation. These data are important for studies of thrombus formation, platelet storage, and immune thrombocytopenia.
Background
Disseminated Intravascular Coagulation (DIC) is a life-threatening complication of sepsis. In surgical ICUs, DIC is frequently caused by abdominal sepsis, and the disarranged coagulation and complications often lead to death. The severity of sepsis is associated with a higher DIC score according to the parameters proposed by the International Society of Hemostasis and Thrombosis (ISTH) in 2001: platelet count, bleeding time (Quick), D-dimer, and fibrinogen. One problem in studying DIC is finding an adequate animal model that reflects the clinical situation of polymicrobial overwhelming infection.
Aims and methods
We investigated whether a well-established polymicrobial sepsis model of colon ascendens stent peritonitis (CASP) is suited to investigate the complexity of DIC. For this purpose, CASP-operated mice were examined 20 h after the operation with regard to coagulation parameters using cell counts, bleeding times, rotational thromboelastometry (ROTEM), ELISAs for D-dimer and fibrinogen, and platelet accumulation in affected organs via immunohistochemistry to see if the mice develop a coagulation disorder that meets the definition of DIC proposed by the ISTH 2001 consensus conference.
Results
Herein, we showed that the CASP model is an all-encompassing animal model to analyze the complexity of systemic DIC in murine abdominal sepsis. There is highly reproducible thrombocytopenia, a significant prolongation of the bleeding time, and a loss of fibrinogen in plasma. We also observed microvascular thrombosis due to platelet accumulation in the microcirculation of the liver.
Conclusion
The CASP model seems superior to other artificial models, e.g., injecting substances, for inducing DIC. CASP is one of the best true-to-life models for analyzing the complexity of disseminated intravascular coagulation in polymicrobial sepsis.
The hypothalamus–pituitary–adrenal axis is the main physiological stress response system and regulating the release of cortisol. The two corticoid receptors encoded by the genes NR3C1 and NR3C2 are the main players in regulating the physiological response to cortisol. This biological system has been linked to neurocognitive processes and memory, yet the mechanisms remain largely unclear. In two independent general population studies (SHIP, total sample size > 5500), we aim to diseantangle the effects of genetic variation, gene expression and cortisol on verbal memory and memory associated brain volume. Especially for NR3C1 results exhibited a consistent pattern of direct an interactive effects. All three biological layers, genetic variation (rs56149945), gene expression for NR3C1 and cortisol levels, were directly associated with verbal memory. Interactions between these components showed significant effects on verbal memory as well as hippocampal volume. For NR3C2 such a complex association pattern could not be observed. Our analyses revealed that different components of the stress response system are acting together on different aspects of cognition. Complex phenotypes, such as cognition and memory function are regulated by a complex interplay between different genetic and epigenetic features. We promote the glucocorticoid receptor NR3C1 as a main target to focus in the context of verbal memory and provided a mechanistic concept of the interaction between various biological layers spanning NR3C1 function and its effects on memory. Especially the NR3C1 transcript seemed to be a key element in this complex system.
Effectiveness of Acupuncture for Pain Control After Cesarean Delivery: A Randomized Clinical Trial
(2022)
Importance: A pharmacological approach to pain control after cesarean delivery is often insufficient on its own. Acupuncture is a promising method for mitigating postoperative pain and reducing postoperative opioid requirements.
Objective: To evaluate the efficacy and effectiveness of acupuncture as an adjunctive therapy for pain control after cesarean delivery, compared with a placebo intervention and standard care alone.
Design, Setting, and Participants: This single-center, placebo-controlled, patient- and assessor-blinded randomized clinical trial was conducted from January 13, 2015, to June 27, 2018, at a tertiary university hospital in Greifswald, Germany. Participants were women who were scheduled for elective cesarean delivery under spinal anesthesia and were randomized to either the acupuncture group (n = 60) or placebo group (n = 60). Another 60 consecutive patients who met the eligibility criteria and received the standard postoperative analgesia were selected to form a nonrandomized standard care group. The intention-to-treat analysis was performed from August 19, 2019, to September 13, 2019.
Interventions: In addition to standard pain treatment, each patient in the acupuncture group received auricular and body acupuncture with indwelling intradermal needles, whereas patients in the placebo group were treated with nonpenetrating placebo needles.
Main Outcomes and Measures: The primary outcome was pain intensity on movement, which was measured using an 11-item verbal rating scale. Secondary outcomes were analgesia-related adverse effects, analgesics consumption, time to mobilization and Foley catheter removal, quality of patient blinding to randomization, and patient satisfaction with treatment of pain.
Results: A total of 180 female patients (mean [SD] age, 31 [5] years) were included in the intention-to-treat analysis. The mean pain intensity on movement in the acupuncture group on the first postoperative day was lower than in the placebo group (4.7 [1.8] vs 6.0 [2.0] points; Cohen d, 0.73; 95% CI, 0.31-1.01; P = .001) and the standard care group (6.3 [1.3] points; Cohen d, 1.01; 95% CI, 0.63-1.40; P < .001). On the first postoperative day, 59 patients (98%) in the acupuncture group were fully mobilized vs 49 patients (83%) in the placebo group (relative risk [RR], 1.18; 95% CI, 1.06-1.33; P = .01) and 35 patients (58%) in the standard care group (RR, 1.69; 95% CI, 1.36-2.09; P < .001). The Foley catheter was removed in a total of 57 patients (93%) from the acupuncture group vs 43 patients (72%) from the placebo group (RR, 1.33; 95% CI, 1.12-1.57; P = .003) and 42 patients (70%) from the standard care group (RR, 1.37; 95% CI, 1.14-1.62; P = .002). Other parameters were comparable across the 3 study groups.
Conclusions and Relevance: Results of this trial showed that acupuncture was safe and effective in reducing pain and accelerating mobilization of patients after cesarean delivery. With consideration for personnel and time expenditures, acupuncture can be recommended as routine, supplemental therapy for pain control in patients after elective cesarean delivery.
This is the first study to analyze the association of accelerometer-measured patterns of habitual physical activity (PA) and sedentary behavior (SB) with serum BDNF in individuals with coronary heart disease. A total of 30 individuals (M = 69.5 years; 80% men) participated in this pre-post study that aimed to test a multi-behavioral intervention. All participants underwent standardized measurement of anthropometric variables, blood collection, self-administered survey, and accelerometer-based measurement of PA and SB over seven days. Serum BDNF concentrations were measured using enzyme-linked immunosorbent assay kit. We applied separate multiple linear regression analysis to estimate the associations of baseline SB pattern measures, light and moderate-to-vigorous PA with serum BDNF (n = 29). Participants spent 508.7 ± 76.5 min/d in SB, 258.5 ± 71.2 min/d in light PA, and 21.2 ± 15.2 min/d in moderate-to-vigorous PA. Per day, individuals had 15.5 ± 3.2 numbers of 10-to-30 min bouts of SB (average length: 22.2 ± 2.1 min) and 3.4 ± 1.2 numbers of > 30 min bouts of SB (average length: 43.8 ± 2.4 min). Regression analysis revealed no significant associations between any of the accelerometer-based measures and serum BDNF. The findings of this study did not reveal an association of accelerometer-measured PA and SB pattern variables with serum BDNF in individuals with coronary heart disease. In addition, our data revealed a considerable variation of PA and SB which should be considered in future studies.
Linking transcriptional dynamics of CH4-cycling grassland soil microbiomes to seasonal gas fluxes
(2022)
Soil CH4 fluxes are driven by CH4-producing and -consuming microorganisms that determine whether soils are sources or sinks of this potent greenhouse gas. To date, a comprehensive understanding of underlying microbiome dynamics has rarely been obtained in situ. Using quantitative metatranscriptomics, we aimed to link CH4-cycling microbiomes to net surface CH4 fluxes throughout a year in two grassland soils. CH4 fluxes were highly dynamic: both soils were net CH4 sources in autumn and winter and sinks in spring and summer, respectively. Correspondingly, methanogen mRNA abundances per gram soil correlated well with CH4 fluxes. Methanotroph to methanogen mRNA ratios were higher in spring and summer, when the soils acted as net CH4 sinks. CH4 uptake was associated with an increased proportion of USCα and γ pmoA and pmoA2 transcripts. We assume that methanogen transcript abundance may be useful to approximate changes in net surface CH4 emissions from grassland soils. High methanotroph to methanogen ratios would indicate CH4 sink properties. Our study links for the first time the seasonal transcriptional dynamics of CH4-cycling soil microbiomes to gas fluxes in situ. It suggests mRNA transcript abundances as promising indicators of dynamic ecosystem-level processes.
Understanding the effects of temperature and moisture on radial growth is vital for assessing the impacts of climate change on carbon and water cycles. However, studies observing growth at sub-daily temporal scales remain scarce.
We analysed sub-daily growth dynamics and its climatic drivers recorded by point dendrometers for 35 trees of three temperate broadleaved species during the years 2015–2020. We isolated irreversible growth driven by cambial activity from the dendrometer records. Next, we compared the intra-annual growth patterns among species and delimited their climatic optima.
The growth of all species peaked at air temperatures between 12 and 16°C and vapour pressure deficit (VPD) below 0.1 kPa. Acer pseudoplatanus and Fagus sylvatica, both diffuse-porous, sustained growth under suboptimal VPD. Ring-porous Quercus robur experienced a steep decline of growth rates with reduced air humidity. This resulted in multiple irregular growth peaks of Q. robur during the year. By contrast, the growth patterns of the diffuse-porous species were always right-skewed unimodal with a peak in June between day of the year 150–170.
Intra-annual growth patterns are shaped more by VPD than temperature. The different sensitivity of radial growth to VPD is responsible for unimodal growth patterns in both diffuse-porous species and multimodal growth pattern in Q. robur.
A massive shift in agricultural practices over the past decades, to support exceptionally high yields and productivities involving intensive agriculture, have led to unsustainable agriculture practices across the globe. Sustenance of such high yields and productivities demand high use of organic and industrial fertilizers. This acts as a negative pressure on the environment. Excessive use of fertilizers leads to nutrient surplus in the fields, which, as a part of catchment runoff, flows into the water bodies as diffuse pollution. These nutrients through rivers are eventually passed into seas. High nutrients ending up into water bodies cause eutrophication. The situation is worsened when such unsustainable agricultural activities are carried out on drained peatlands. As a result, the nutrients that were not part of the nutrient cycle in the landscape for years begin to leach out due to mineralization of peatlands, thereby putting an additional load of nutrients on the environment, that was already under the negative impact of nutrient surplus. In view of the above, a small lowland catchment of the Ryck river in northeast Germany was assessed for its nitrogen losses from agricultural lands through empirical modelling. Initial empirical modelling resulted in an average annual total nitrogen loss of 14.7 kg ha−1 year−1. After a comparative analysis of these results with procured data, the empirical equation was modified to suit the catchment, yielding more accurate results. The study showed that 75.6% of peatlands in the catchment are under agricultural use. Subsequently, a proposal was made for potential wetland buffer zones in the Ryck catchment. Altogether, 13 peatland sites across 8 sub-catchments were recommended for mitigation of high nutrient runoff. In the end, nutrient efficiency of proposed WBZs in one of the sub-catchments of Ryck has been discussed. The results show that (i) the modified empirical equation can act as a key tool in application-based future strategies for nitrogen reduction in the Ryck catchment, (ii) restoration of peatlands and introduction of WBZs can help in mitigating the nutrient runoff for improved water quality of Ryck, and subsequently (ii) contribute to efficient reduction of riverine loads of nutrients into the Baltic Sea.
Background and study aims: Gastric cancer (GC) is one of the leading causes of malignancy-related death in Vietnam, with increasing incidence of non-cardia early gastric cancer (N-EGC). Data on accurate diagnosis of EGC and treatment by endoscopic submucosal dissection (ESD) in Vietnam are very sparse. The aim of this study was to describe the characteristics of N-EGC and evaluate the effectiveness and the safety of ESD in Central Vietnam.
Patients and methods: We prospectively enrolled patients with N-EGC detected by magnified chromoendoscopy from December 2013 to August, 2018 in Central Vietnam. Selected cases of N-EGC received standardized ESD technique and have been following up carefully as in protocol.
Results: Among 606 GC patients, 46 had N-GEC and underwent ESD. The depth of invasion was pT1a in 33 (71.7 %), pT1b1 in 10 (21.7 %), and pT1b2 in three cases (6.6 %). Mild chronic atrophic gastritis, most being C2 (63 %), and gastritis-like EGC that did not appear malignant was the predominant type. ESD achieved a 97.8 % en bloc resection rate; the mean procedure time was 76 ± 22 minutes (range 24–155), and mean endoscopic tumor size was 23 ± 5 mm (range 13–52) and ESD sample size was 28 ± 7 mm (range 16.5–60). Complications consisted of two patients with bleeding and one with a minor perforation, all of which were successfully managed by endoscopy. The longest and the mean follow-up times were 84 and 64 months, respectively, with no recurrence.
Conclusions: A significant proportion patients with N-EGC have a background mucosa of mild chronic atrophic gastritis. Our results 7 years after starting ESD demonstrate early promising outcomes with the procedure.
Acute pancreatitis (AP) is a major, globally increasing gastrointestinal disease and a biliary origin is the most common cause. However, the effects of bile acids (BAs), given systemically, on the pancreas and on disease severity remains elusive. In this study, we have investigated the roles of different circulating BAs in animal models for AP to elucidate their impact on disease severity and the underlying pathomechanisms. BAs were incubated on isolated acini and AP was induced through repetitive injections of caerulein or L-arginine; pancreatic duct ligation (PDL); or combined biliopancreatic duct ligation (BPDL). Disease severity was assessed using biochemical and histological parameters. Serum cholecystokinin (CCK) concentrations were determined via enzyme immunoassay. The binding of the CCK1 receptor was measured using fluorescence-labeled CCK. In isolated acini, hydrophobic BAs mitigated the damaging effects of CCK. The same BAs further enhanced pancreatitis in L-arginine- and PDL-based pancreatitis, whereas they ameliorated pancreatic damage in the caerulein and BPDL models. Mechanistically, the binding affinity of the CCK1 receptor was significantly reduced by hydrophobic BAs. The hydrophobicity of BAs and the involvement of CCK seem to be relevant in the course of AP. Systemic BAs may affect the severity of AP by interfering with the CCK1 receptor.
Purpose
The continuum of mental health/illness has been subject to scientific debate for decades. While current research indicates that continuum belief interventions can reduce mental health stigma and improve treatment seeking in affected populations, no study has yet systematically examined measures of continuum beliefs.
Methods
This preregistered systematic review summarizes measures of continuum beliefs. Following the PRISMA statement, three scientific databases (PubMed, PsycInfo and PsycArticles via EBSCOhost, Web of Science) are searched, instruments are described and discussed regarding their scope, and methodological quality.
Results
Overall, 7351 records were identified, with 35 studies reporting relevant findings on 11 measures. Most studies examined general population samples and used vignette-based measures. Schizophrenia and depression were most commonly examined, few studies focused on dementia, ADHD, OCD, eating disorders, and problematic alcohol use, or compared continuum beliefs across disorders. Validity was very good for most measures, but reliability was rarely tested. Measures mostly assessed beliefs in the normality of mental health symptoms or the normality of persons with such symptoms but rarely nosological aspects (i.e., categorical v continuous conceptualization of mental disorders).
Conclusions
Current research provides psychometrically sound instruments to examine continuum beliefs for a variety of mental disorders. While studies suggest utility for general population samples and mental health professionals, more research is necessary to corroborate findings, for instance, regarding age (e.g., in adolescents), gender, or type of mental disorder. Future research should also compare self-report ratings, and vignette-based measures, include measures of nosological concepts to fully grasp the continuum concept of mental illness.
Preregistration
PROSPERO: CRD42019123606.
Although disaster research has acknowledged the role of social media in crisis communication, the interplay of new (e.g., mobile apps) and traditional media (e.g., TV, radio) in public warnings has received less attention, particularly from the recipients’ perspective. Therefore, we examined sociodemographic and psychosocial correlates of different types of media use (i.e., traditional, new, mixed) for receiving public warning messages in a population survey (N = 613, 63% female; Mage = 31.56 years). More than two-thirds (68%) reported mixed media use, with 20% relying on new media and 12% on traditional media. Traditional media users were older and reported lower levels of education, while new media users were significantly younger and reported lower trust toward traditional media (i.e., TV). Migrants were more likely to use new but not mixed media. In sum, most participants utilized a mixture of traditional and new media for warning purposes, which has implications for crisis communication. Though, vulnerable populations (e.g., older and less educated participants) mainly rely on traditional media, stressing the need for continued support. Thus, it is paramount to increasingly use mixed methods designs and concurrently examine multiple channels to reflect real-world warning practices and generate ecologically valid results.
In recent years, online radicalization has received increasing attention from researchers and policymakers, for instance, by analyzing online communication of radical groups and linking it to individual and collective pathways of radicalization into violent extremism. But these efforts often focus on radical individuals or groups as senders of radicalizing messages, while empirical research on the recipient is scarce. To study the impact of radicalized online content on vulnerable individuals, this study compared cognitive and affective appraisal and visual processing (via eye tracking) of three political Internet memes (empowering a right-wing group, inciting violence against out-groups, and emphasizing unity among human beings) between a right-wing group and a control group. We examined associations between socio-political attitudes, appraisal ratings, and visual attention metrics (total dwell time, number of fixations). The results show that right-wing participants perceived in-group memes (empowerment, violence) more positively and messages of overarching similarities much more negatively than controls. In addition, right-wing participants and participants in the control group with a high support for violence directed their attention towards graphical cues of violence (e.g., weapons), differentness, and right-wing groups (e.g., runes), regardless of the overall message of the meme. These findings point to selective exposure effects and have implications for the design and distribution of de-radicalizing messages and counter narratives to optimize the efficacy of prevention of online radicalization.
Introduction
Privacy concerns are an important barrier to adoption and continued use of digital technologies, particularly in the health sector. With the introduction of mobile health applications (mHealth apps), the construct of app information privacy concerns has received increased attention. However, few validated measures exist to capture said concerns in population samples, although they can help to improve public health efforts.
Methods
Using a cross-sectional survey of German adults (mean age = 35.62; 63.5% female), this study examined psychometric properties of the app information privacy concerns scale (AIPC). Analyses comprised confirmatory factor analysis, factorial validity (exploratory factor analysis), internal consistency, convergent validity (i.e., correlations with privacy victimhood, and app privacy concerns), and discriminant validity (i.e., daily app use, adoption intentions, and attitudes toward COVID-19 contact tracing app use).
Results
The analysis did not support the proposed three-factor structure of the AIPC (i.e., anxiety, personal attitude, and requirements). Instead, a four-factor model was preferable that differentiated requirements regarding disclosure policies, and personal control. In addition, factors mirroring anxiety and personal attitude were extracted, but shared a significant overlap. However, these factors showed good reliability, convergent and discriminant validity.
Discussion
The findings underline the role of app information privacy concerns as a significant barrier to mHealth app use. In this context, anxiety and personal attitudes seemed particularly relevant, which has implications for health communication. Moreover, the observed differentiation of external (disclosure) and internal (control) requirements aligns with health behavior change models and thus is a promising area for future research.
Sphingosine-1-phosphate (S1P) is a versatile signaling lipid involved in the regulation of numerous cellular processes. S1P regulates cellular proliferation, migration, and apoptosis as well as the function of immune cells. S1P is generated from sphingosine (Sph), which derives from the ceramide metabolism. In particular, high concentrations of S1P are present in the blood. This originates mainly from erythrocytes, endothelial cells (ECs), and platelets. While erythrocytes function as a storage pool for circulating S1P, platelets can rapidly generate S1P de novo, store it in large quantities, and release it when the platelet is activated. Platelets can thus provide S1P in a short time when needed or in the case of an injury with subsequent platelet activation and thereby regulate local cellular responses. In addition, platelet-dependently generated and released S1P may also influence long-term immune cell functions in various disease processes, such as inflammation-driven vascular diseases. In this review, the metabolism and release of platelet S1P are presented, and the autocrine versus paracrine functions of platelet-derived S1P and its relevance in various disease processes are discussed. New pharmacological approaches that target the auto- or paracrine effects of S1P may be therapeutically helpful in the future for pathological processes involving S1P.
Background
Despite the current undersupply of cochlear implants (CIs) with simultaneously increasing indication, CI implantation numbers in Germany still are at a relatively low level.
Methods
As there are hardly any solid forecasts available in the literature, we develop a System Dynamics model that forecasts the number and costs of CI implantations in adults for 40 years from a social health insurance (SHI) perspective.
Results
CI demand will grow marginally by demographic changes causing average annual costs of about 538 million €. Medical-technical progress with following relaxed indication criteria and patients’ increasing willingness for implantation will increase implantation numbers significantly with average annual costs of 765 million €.
Conclusion
CI demand by adults will increase in the future, thus will the costs for CI supply. Continuous research and development in CI technology and supply is crucial to ensure long-term financing of the growing CI demand through cost-reducing innovations.
Neuroinflammatory mechanisms and maladaptive neuroplasticity underlie the progression of complex regional pain syndrome (CRPS), which is prototypical of central neuropathic pain conditions. While cortical maladaptive alterations are well described, little is known about the contribution of the brainstem to the pathophysiology. This study investigates the role of pain-modulatory brainstem pathways in CRPS using the nociceptive blink reflex (nBR), which not only provides a direct read-out of brainstem excitability and habituation to painful stimuli but may also be suitable for use as a diagnostic biomarker for CRPS. Thirteen patients with CRPS and thirteen healthy controls (HCs) participated in this prospective case-control study investigating the polysynaptic trigemino-cervical (R2) nBR response. The R2 area and its habituation were assessed following repeated supraorbital electrical stimulation. Between-group comparisons included evaluations of diagnostic characteristics as a potential biomarker for the disease. Patients with CRPS showed a substantial decrease in habituation on the stimulated (Cohen’s d: 1.3; p = 0.012) and the non-stimulated side (Cohen’s d: 1.1; p = 0.04). This is the first study to reveal altered nBR habituation as a pathophysiological mechanism and potential diagnostic biomarker in CRPS. We confirmed previous findings of altered nBR excitability, but the diagnostic accuracy was inferior. Future studies should investigate the nBR as a marker of progression to central mechanisms in CRPS and as a biomarker to predict treatment response or prognosis.
Hintergrund
Migräne ist eine hochprävalente Erkrankung, die bei betroffenen Patient*innen eine hohe Belastung sowohl durch die Kopfschmerzen an sich als auch durch die Einschränkung ihres Sozial- und Berufslebens hervorruft. Die Therapie besteht aus einer Akuttherapie der Kopfschmerzattacken sowie einer prophylaktischen Therapie zur Reduktion der Kopfschmerzfrequenz und -schwere. In der Prophylaxe stehen mit Antikörpern gegen das Calcitonin-gene-related-peptide (CGRP) und dessen Rezeptor erstmalig für die Migräne entwickelte gezielte prophylaktische Therapien zur Verfügung. Es stellt sich jedoch hierbei die Frage, ob CGRP-Antikörper lediglich symptomatisch in der Peripherie des trigemino-vaskulären-Systems wirken oder auch im zentralen Nervensystem die zugrundeliegenden pathophysiologischen Mechanismen beeinflussen, was einer krankheitsmodifizierenden Wirkung entspräche. Ziel unserer Studie war es, die Nullhypothese einer rein symptomatischen Wirkung gegen die Alternativhypothese einer Krankheitsmodifikation und somit zentralnervösen Wirkung, zu prüfen, indem bei Patient*innen mit episodischer Migräne der nozizeptive Blinkreflex vor und nach der Behandlung mit CGRP-Antikörpern untersucht wurde.
Methoden
22 Patient*innen mit episodischer Migräne (21 Frauen, 46,2 ± 13,8 Jahre alt) und 22 alters- und geschlechts-gematchte Kontrollen wurden im Rahmen dieser prospektiven Beobachtungsstudie eingeschlossen. Sie erhielten einen umfassenden Fragebogen zur Erhebung demografischer Charakteristika sowie der Kopfschmerzanamnese. Es erfolgte eine Messung des Blinkreflexes (10 Durchgänge à 6 Stimuli) vor (V0) und 3 Monate (V3) nach der Behandlung mit CGRP-Antikörpern (Kontrollen wurden einmalig gemessen). Im Rahmen der Messung wurden wiederholt schmerzhafte Stimuli supraorbital appliziert, die direkte Rückschlüsse auf die zentralnervöse Erregbarkeit des Hirnstamms als pathophysiologisch zentralen Mechanismus im Rahmen der Migräneentstehung zulassen. Die Area-under-the-curve (AUC) der R2-Komponente der Muskelsummenaktionspotentiale des Blinkreflexes sowie das Habituationsverhalten (Regressionskoeffizient über mehrere Blöcke) der stimulierten sowie nicht-stimulierten Seite wurden über 10 Blöcke hinweg evaluiert (primärer Endpunkt). Es wurde jeweils zuerst ein Test auf globale Veränderungen durchgeführt, der dann durch post-hoc-Analysen weiter spezifiziert wurde.
Ergebnisse
Alle Patient*innen zeigten eine signifikante Reduktion der Kopfschmerztage/Monat (V0: 12,4±3,3, V3: 6,6 ± 4,9) nach Beginn der Behandlung mit einem CGRP-/Rezeptorantikörper. Auf der stimulierten Seite reduzierte sich die AUC signifikant in den Blöcken eins, zwei sowie acht (Fglobal=5,86, p<0,001; block 1: R2a_s: -28%, p<0,001). Auf der nicht-stimulierten Seite zeigten sich Block eins, zwei, drei, acht sowie zehn als signifikant reduziert (Fglobal=8,22, p<0,001, block 1: R2a_ns: -22%, p=0,003). Die Veränderung der Habituation erwies sich in den Blöcken sechs, sieben, acht und zehn auf der nicht-stimulierten Seite als signifikant (Fglobal=3,07, p<0,001; block 6: R2h_ns: r=-1,36, p=0,007). Weder die AUC noch die Habituation des ersten Messtermins (V0) korrelierte mit dem späteren klinischen Ansprechen, sodass kein Prädiktor für das Therapieansprechen detektiert werden konnte (binär logistische Regression; alle Prädiktoren p>0,05).
Diskussion & Zusammenfassung
Die Ergebnisse dieser Studie zeigen, dass die dreimonatige Therapie mit CGRP-Antikörpern die Erregbarkeit des Hirnstamms als Antwort auf wiederholte schmerzhafte Stimuli bei Patient*innen mit Migräne normalisiert und liefert somit Hinweise für ein krankheitsmodifizierendes Potenzial. Veränderungen der Habituation korrelierten signifikant mit der Verringerung der Kopfschmerz-Frequenz, weitere Studien sind jedoch nötig, um zu eruieren, ob Parameter als Prädiktor geeignet sind um eine Voraussage über das Therapieansprechen und das Risiko einer Verschlechterung nach Beendigung der Therapie zu ermöglichen.
Die Pankreatitis ist gekennzeichnet durch den Selbstverdau des Organs. Dabei werden pankreatische Proteasen aktiviert und es entstehen lokale Entzündungsherde. Diese aktivieren die Immunantwort durch Ausschüttung pro- inflammatorischer Mediatoren und der Rekrutierung von Immunzellen in das geschädigte Gewebe. Als erste Schutzinstanz reagiert das angeborene Immunsystem inklusive der Neutrophilen, Granulozyten und Makrophagen. Es ist bekannt, dass diese Immunzellen Mustererkennungsrezeptoren nutzen, um die Infektion zu erkennen. Zu diesen gehören die Toll-like Rezeptoren, welche u.a. über den MyD88/IRAK Signalweg den Transkriptionsfaktor NF-κB aktivieren können. In dieser Signalkaskade existiert ein negativer Feedback Regulator IRAK-M, auch bekannt als IRAK-3. Dieser ist in der Lage die Signalweiterleitung zu inhibieren. In dieser Arbeit wurde untersucht ob und inwieweit IRAK-M Einfluss auf den Verlauf einer experimentell induzierten Pankreatitis in Mäusen hat. Bisherige Studien zeigten die Expression von IRAK-M in verschiedenen Zelltypen und Geweben, jedoch nicht im Pankreas sowie den Azinuszellen. In dieser Arbeit konnte nachgewiesen werden, dass IRAK-M in Pankreasgewebe sowie isolierten Azini von C57BL/6 Mäusen exprimiert wird. Die Stimulation von isolierten C57BL/6-Azinuszellen mit CCK hatte eine Expressionserhöhung von IRAK-M zur Folge. Es konnte zudem gezeigt werden, dass Toll-like Rezeptoren (TLR), insbesondere 2, 3, 4 und 9, in bzw. auf Azini exprimiert werden. Die TLR1, 2, 3, 7 und 9 zeigten ein höheres Expressionslevel in den Azinuszellen der defizienten Tiere. Die Caerulein induzierte akute Pankreatitis zeigte einen milderen Verlauf in Bezug auf die Schweregradmarker Amylase und Lipase im Serum der IRAK-M -/- Tiere sowie einen geringen lokalen pankreatischen Schaden. Die Entzündungsreaktion erhöhte die MPO-Aktivität in der Lunge der defizienten Tiere. Zudem zeigten die Tiere eine erhöhte T-Zellaktivierung und Sekretion pro-inflammatorischer Zytokine wie TNFα und IL12 sowie des anti-inflammatorischen Zytokins IL10. Tendenziell wanderten mehr Neutrophile, M1- sowie M2- Makrophagen in das Pankreasgewebe während der Entzündung. Der Transkriptionsfaktor NF-κB konnte nach 8h akuter Pankreatitis, transloziert im Zellerkern, vermehrt in den IRAK-M defizienten Tieren nachgewiesen werden. Die Untersuchung von IRAK-M -/- BMDM zeigte, dass das Zytokin Milieu zur Differenzierung des M1 -Phänotyps dominierend war. Zudem lag eine verstärkte Phagozytose vor und die Makrophagen wiesen eine verstärkte Sekretion und Expression von TNFα, IL6, IL10 und IL12 auf. Nach 3d schwerer akuter Pankreatitis (SAP) wurde eine höhere Konzentration an Serumlipase sowie ein stärkerer pankreatischer Schaden beobachtet. Die MPO-Aktivität in der Lunge der defizienten Tiere war vermindert. Dennoch konnten vermehrt pro-inflammatorische Zytokine wie TNFα, IL6, IL12 und MCP1 im Serum gemessen werden. Die T-Zellen der defizienten Tiere zeigten zudem eine erhöhte Aktivierung. Die Visualisierung von infiltrierten Zellen im Pankreas zeigte keine Unterschiede. Der Vergleich der beiden experimentellen Pankreatitis-Modelle zeigte, dass die Caerulein induzierte Pankreatitis bei den IRAK-M -/- Tieren zu einer lokal begrenzten Entzündung im Pankreas führte. Wohingegen die Pankreatitis nach Gangligatur einen deutlich stärkeren Schaden aufwies und in einer signifikant erhöhten Zytokinsekretion resultierte. Der Vergleich von IL6 zeigte, dass die defizienten Tieren das 14-fache ins Serum sekretieren nach SAP, während die Kontrolltiere nur einen Anstieg um das 4,5- fache zeigten. Somit lässt sich zusammenfassen, dass eine kontrollierte und in Maßen ablaufende Immunantwort protektiv bzw. förderlich für den Krankheitsverlauf ist. Andererseits kann eine überschießende Immunantwort zu systemischen Komplikationen sowie Multiorganversagen führen. IRAK-M nimmt dabei eine regulierende Rolle ein und verhindert u.a., dass die Immunreaktion überschießt.
Pollen productivity estimates (PPEs) are a key parameter for quantitative land-cover reconstructions from pollen data. PPEs are commonly estimated using modern pollen-vegetation data sets and the extended R-value (ERV) model. Prominent discrepancies in the existing studies question the reliability of the approach. We here propose an implementation of the ERV model in the R environment for statistical computing, which allows for simplified application and testing. Using simulated pollen-vegetation data sets, we explore sensitivity of ERV application to (1) number of sites, (2) vegetation structure, (3) basin size, (4) noise in the data, and (5) dispersal model selection. The simulations show that noise in the (pollen) data and dispersal model selection are critical factors in ERV application. Pollen count errors imply prominent PPE errors mainly for taxa with low counts, usually low pollen producers. Applied with an unsuited dispersal model, ERV tends to produce wrong PPEs for additional taxa. In a comparison of the still widely applied Prentice model and a Lagrangian stochastic model (LSM), errors are highest for taxa with high and low fall speed of pollen. The errors reflect the too high influence of fall speed in the Prentice model. ERV studies often use local scale pollen data from for example, moss polsters. Describing pollen dispersal on his local scale is particularly complex due to a range of disturbing factors, including differential release height. Considering the importance of the dispersal model in the approach, and the very large uncertainties in dispersal on short distance, we advise to carry out ERV studies with pollen data from open areas or basins that lack local pollen deposition of the taxa of interest.
Lake‐level reconstructions are a key tool in hydro‐climate reconstructions, based on the assumption that lake‐level changes primarily reflect climatic changes. Although it is known that land cover changes can affect evapotranspiration and groundwater formation, this factor commonly receives little attention in the interpretation of past lake‐level changes. To address this issue in more detail, we explore the effects of land cover change on Holocene lake‐level fluctuations in Lake Tiefer See in the lowlands of northeastern Germany. We reconstruct lake‐level changes based on the analysis of 28 sediment records from different water depths and from the shore. We compare the results with land cover changes inferred from pollen data. We also apply hydrological modelling to quantify effects of land cover change on evapotranspiration and the lake level. Our reconstruction shows an overall lake‐level amplitude of about 10 m during the Holocene, with the highest fluctuations during the Early and Late Holocene. Only smaller fluctuations during the Middle Holocene can unambiguously be attributed to climatic fluctuations because the land cover was stable during that period. Fluctuations during the Early and Late Holocene are at least partly related to changes in natural and anthropogenic land cover. For several intervals the reconstructed lake‐level changes agree well with variations in modelled groundwater recharge inferred from land cover changes. In general, the observed amplitudes of lake‐level fluctuations are larger than expected from climatic changes alone and thus underline that land cover changes in lake catchments must be considered in climatic interpretations of past lake‐level fluctuations.
Introduction
With the worldwide increase of life expectancy leading to a higher proportion of older adults experiencing age-associated deterioration of cognitive abilities, the development of effective and widely accessible prevention and therapeutic measures has become a priority and challenge for modern medicine. Combined interventions of cognitive training and transcranial direct current stimulation (tDCS) have shown promising results for counteracting age-associated cognitive decline. However, access to clinical centres for repeated sessions is challenging, particularly in rural areas and for older adults with reduced mobility, and lack of clinical personnel and hospital space prevents extended interventions in larger cohorts. A home-based and remotely supervised application of tDCS would make the treatment more accessible for participants and relieve clinical resources. So far, studies assessing feasibility of combined interventions with a focus on cognition in a home-based setting are rare. With this study, we aim to provide evidence for the feasibility and the effects of a multisession home-based cognitive training in combination with tDCS on cognitive functions of healthy older adults.
Methods and analysis
The TrainStim-Home trial is a monocentric, randomised, double-blind, placebo-controlled study. Thirty healthy participants, aged 60–80 years, will receive 2 weeks of combined cognitive training and anodal tDCS over left dorsolateral prefrontal cortex (target intervention), compared with cognitive training plus sham stimulation. The cognitive training will comprise a letter updating task, and the participants will be stimulated for 20 min with 1.5 mA. The intervention sessions will take place at the participants’ home, and primary outcome will be the feasibility, operationalised by two-thirds successfully completed sessions per participant. Additionally, performance in the training task and an untrained task will be analysed.
Ethics and dissemination
Ethical approval was granted by the ethics committee of the University Medicine Greifswald. Results will be available through publications in peer-reviewed journals and presentations at national and international conferences.Trial registration numberNCT04817124.
Neural mechanisms of behavioral improvement induced by repeated transcranial direct current stimulation (tDCS) combined with cognitive training are yet unclear. Previously, we reported behavioral effects of a 3-day visuospatial memory training with concurrent anodal tDCS over the right temporoparietal cortex in older adults. To investigate intervention-induced neural alterations we here used functional magnetic resonance imaging (fMRI) and diffusion tensor imaging (DTI) datasets available from 35 participants of this previous study, acquired before and after the intervention. To delineate changes in whole-brain functional network architecture, we employed eigenvector centrality mapping. Gray matter alterations were analyzed using DTI-derived mean diffusivity (MD). Network centrality in the bilateral posterior temporooccipital cortex was reduced after anodal compared to sham stimulation. This focal effect is indicative of decreased functional connectivity of the brain region underneath the anodal electrode and its left-hemispheric homolog with other “relevant” (i.e., highly connected) brain regions, thereby providing evidence for reorganizational processes within the brain's network architecture. Examining local MD changes in these clusters, an interaction between stimulation condition and training success indicated a decrease of MD in the right (stimulated) temporooccipital cluster in individuals who showed superior behavioral training benefits. Using a data-driven whole-brain network approach, we provide evidence for targeted neuromodulatory effects of a combined tDCS-and-training intervention. We show for the first time that gray matter alterations of microstructure (assessed by DTI-derived MD) may be involved in tDCS-enhanced cognitive training. Increased knowledge on how combined interventions modulate neural networks in older adults, will help the development of specific therapeutic interventions against age-associated cognitive decline.