Pneumolysin induced mitochondrial dysfunction leads to release of mitochondrial DNA

Abstract Streptococcus pneumoniae (S.pn.) is the most common bacterial pathogen causing community acquired pneumonia. The pore-forming toxin pneumolysin (PLY) is the major virulence factor of S.pn. and supposed to affect alveolar epithelial cells thereby activating the immune system by liberation of danger-associated molecular patterns (DAMP). To test this hypothesis, we established a novel live-cell imaging based assay to analyse mitochondrial function and associated release of mitochondrial DNA (mtDNA) as DAMP in real-time. We first revealed that bacterially released PLY caused significant changes of the cellular ATP homeostasis and led to morphologic alterations of mitochondria in human alveolar epithelial cells in vitro and, by use of spectral live-tissue imaging, in human alveoli. This was accompanied by strong mitochondrial calcium influx and loss of mitochondrial membrane potential resulting in opening of the mitochondrial permeability transition pore and mtDNA release without activation of intrinsic apoptosis. Moreover, our data indicate cellular mtDNA liberation via microvesicles, which may contribute to S.pn. related pro-inflammatory immune activation in the human alveolar compartment

About the impact of erectile disorders on the partnership-related satisfaction

Die Prävalenz von Erektionsstörungen schwankt in den großen Studien zwischen 17,8% und 52%, damit ist die Erektionsstörung die zweithäufigste sexuelle Funktionsstörung des Mannes. Die betroffenen Männer sind mit ihrem Sexualleben unzufriedener als nicht Erkrankte und geben seltener eine hohe partner- schaftsbezogene Lebensqualität an. Wie es um die partnerschaftliche Beziehungszufriedenheit derjenigen steht, die aufgrund ihrer Erektionsstörung professionelle Hilfe in einer andrologischen Sprechstunde suchen und ob es einen Zusammenhang zwischen dem Schweregrad der Erkrankung und der partnerschaftlichen Zufriedenheit gibt, soll vorliegende Arbeit klären. Auch wurde untersucht, ob der Leidensdruck bezüglich der Erkrankung mit der Beziehungszufriedenheit in emotionaler Hinsicht korreliert ist. Hierfür wurden in einem Zeitraum von sieben Monaten n = 37 Männer befragt, die die andrologische Spezialsprechstunde der Charité und des Klinikums am Urban aufgrund einer Erektionsstörung besuchten. Empirische Instrumente dieser sexualmedizinischen Querschnittfragebogenuntersuchung waren eine verkürzte Form des FSEV und der Fragebogen „Ressourcen in Sexualität und Partnerschaft“. Mittels Methoden der deskriptiven Statistik wurde die Studienpopulation detailliert beschrieben. Für die Überprüfung der aufgestellten Arbeitshypothese (H 1 = es besteht ein Zusammenhang zwischen dem Schweregrad der Erkrankung und der partnerschaftlichen Zufriedenheit) wurde der Rangkorrelationskoeffizient nach Kendall (Kendalls Tau) verwendet. Die befragten Männer waren zwischen 33 und 76 Jahren alt, 34 der 37 Befragten lebten zum Zeitpunkt der Befragung in einer festen Partnerschaft. Nur n = 4 Männer waren mit ihrer Partnerschaft in sexueller Hinsicht ziemlich bis sehr zufrieden, demgegenüber gaben jedoch n = 27 Männer an, mit ihrer Partnerschaft in emotionaler Hinsicht sehr zufrieden zu sein. Der überwiegende Anteil der Hilfe Suchenden lebte also in einer emotional stabilen Partnerschaft. Weiter wurde herausgefunden, dass mit zunehmendem Schweregrad der Erektionsstörung die partnerschaftliche Zufriedenheit in sexueller Hinsicht abnahm. Ein Zusammenhang zwischen dem Leidensdruck der Erkrankung und der partnerschaftlichen Zufriedenheit in emotionaler Hinsicht konnte mathematisch nicht nachgewiesen werden. Es ist bekannt, dass eine Kombination aus medikamentöser Therapie und Sexualtherapie bei der Behandlung der Erektionsstörung erfolgversprechender ist als die alleinige Gabe von Medikamenten. Wenn nun, wie die vorliegende Studie belegen konnte, die meisten in einer andrologischen Sprechstunde Hilfe suchenden Männer eine hohe emotionale Beziehungszufriedenheit aufweisen, so sollte diese im Rahmen paarinterventioneller therapeutischer Bemühungen als Ressource stärker nutzbar gemacht werden.The prevalence of erectile dysfunction reported in the larger studies varies between 17.8% and 52%. Accordingly, the erectile dysfunction is the second most sexual dysfunction related to men. The affected men are less satisfied with their sexual life than non-affected men and mention less often a high partnership-related life quality. This work elaborates on the partnership- related satisfaction of men who seek professional help in andrological consultation hours due to their erectile dysfunction as well as on the correlation between the severity of the disorder and the degree of partnership-related satisfaction. Another topic of the work was the question whether the psychological strain related to the disorder is correlated to the partnership-related satisfaction on an emotional level. For this purpose we interviewed n=37 men over a period of seven months who visited the specialized andrological consultation hours at “Charité” (Berlin) and the “Klinikum am Urban” due to an erectile dysfunction. The survey was conducted via a cross sectional questionnaire which was based on an abbreviated version of the FSEV and the questionnaire “Ressources in Sexuality and Partnership”. We described the population of the survey by means of descriptive statistics in detail. In order to challenge the working hypothesis (H 1 = the severity of the disorder is correlated with the in-partnership satisfaction) we used the rank correlation coefficient by Kendall (Kendall Tau). The age of the participants varied between 33 and 76 years. 34 out of 37 participants lived in a permanent partnership during the survey. Only n=4 men stated that they are “quite” to “very much” satisfied with respect to their sexual life while n=27 men mentioned an emotional satisfaction related to their partnership. Thus, the majority of the help-seeking men lived in an emotionally stable partnership. Another result was a decreasing in-partnership satisfaction related to sexuality with increasing severity of the disorder. A connection between psychological strain and the partnership satisfaction related to the emotional level could not be found by mathematical means. It is well known that a medicamentous therapy in combination with a sexual therapy is a more promising treatment of erectile dysfunctions than a medicamentous therapy alone. In conclusion and in accordance with the findings of the present study that most help-seeking men mentioned a high emotional in-partnership satisfaction, the partnership itself should be increasingly used as resource during the therapeutically treatment

Proof-of-Concept Organ-on-Chip Study: Topical Cinnamaldehyde Exposure of Reconstructed Human Skin with Integrated Neopapillae Cultured under Dynamic Flow

Pharmaceutical and personal care industries require human representative models for testing to ensure the safety of their products. A major route of penetration into our body after substance exposure is via the skin. Our aim was to generate robust culture conditions for a next generation human skin-on-chip model containing neopapillae and to establish proof-of-concept testing with the sensitizer, cinnamaldehyde. Reconstructed human skin consisting of a stratified and differentiated epidermis on a fibroblast populated hydrogel containing neopapillae spheroids (RhS-NP), were cultured air-exposed and under dynamic flow for 10 days. The robustness of three independent experiments, each with up to 21 intra-experiment replicates, was investigated. The epidermis was seen to invaginate into the hydrogel towards the neopapille spheroids. Daily measurements of lactate dehydrogenase (LDH) and glucose levels within the culture medium demonstrated high viability and stable metabolic activity throughout the culture period in all three independent experiments and in the replicates within an experiment. Topical cinnamaldehyde exposure to RhS-NP resulted in dose-dependent cytotoxicity (increased LDH release) and elevated cytokine secretion of contact sensitizer specific IL-18, pro-inflammatory IL-1β, inflammatory IL-23 and IFN-γ, as well as anti-inflammatory IL-10 and IL-12p70. This study demonstrates the robustness and feasibility of complex next generation skin models for investigating skin immunotoxicity

A Multi-Organ-on-Chip Approach to Investigate How Oral Exposure to Metals Can Cause Systemic Toxicity Leading to Langerhans Cell Activation in Skin

Investigating systemic toxicity in vitro is still a huge challenge. Here, a multi-organ-on-chip approach is presented as a typical case of topical exposure of oral mucosa to metals, which are known to activate the immune system and in turn may result in skin inflammation. Reconstructed human gingiva (RHG) and reconstructed human skin containing MUTZ-3-derived Langerhans cells (MUTZ-LC) in the epidermis (RHS-LC) were incorporated into a HUMIMIC Chip3plus, connected by dynamic flow and cultured for a total period of 72 h. Three independent experiments were performed each with an intra-experiment replicate in order to assess the donor and technical variations. After an initial culture period of 24 h to achieve stable dynamic culture conditions, nickel sulfate was applied topically to RHG for 24 h, and LC activation (maturation and migration) was determined in RHS-LC after an additional 24 h incubation time. A stable dynamic culture of RHG and RHS-LC was achieved as indicated by the assessment of glucose uptake, lactate production, and lactate dehydrogenase release into the microfluidics compartment. Nickel exposure resulted in no major histological changes within RHG or RHS-LC, or cytokine release into the microfluidics compartment, but did result in an increased activation of LC as observed by the increased mRNA levels of CD1a, CD207, HLA-DR, and CD86 in the dermal compartment (hydrogel of RHS-LC (PCR)). This is the first study to describe systemic toxicity and immune cell activation in a multi-organ setting and can provide a framework for studying other organoids in the future

Using a contextualized sensemaking model for interaction design: A case study of tumor contouring

Sensemaking theories help designers understand the cognitive processes of a user when he/she performs a complicated task. This paper introduces a two-step approach of incorporating sensemaking support within the design of health information systems by: (1) modeling the sensemaking process of physicians while performing a task, and (2) identifying software interaction design requirements that support sensemaking based on this model. The two-step approach is presented based on a case study of the tumor contouring clinical task for radiotherapy planning. In the first step of the approach, a contextualized sensemaking model was developed to describe the sensemaking process based on the goal, the workflow and the context of the task. In the second step, based on a research software prototype, an experiment was conducted where three contouring tasks were performed by eight physicians respectively. Four types of navigation interactions and five types of interaction sequence patterns were identified by analyzing<br/>the gathered interaction log data from those twenty-four cases. Further in-depth study on each of the navigation interactions and interaction sequence patterns in relation to the contextualized sensemaking model revealed five main areas for design improvements to increase sensemaking support. Outcomes of the case study indicate that the proposed two-step approach was beneficial for gaining a deeper understanding of the sensemaking process during the task, as well as for identifying design requirements for<br/>better sensemaking support

Distinct Patterns of University Students Study Crafting and the Relationships to Exhaustion, Well-Being, and Engagement

Job crafting has been established as a bottom-up work design instrument for promoting health and well-being in the workplace. In recent years, the concepts of job crafting have been applied to the university student context, proving to be positively related to student well-being. Building on person-centered analyses from the employment context, we assessed approach study crafting strategy combinations and the relationships to students’ exhaustion, study engagement, and general well-being. Data from 2,882 German university students were examined, collected online during the summer term in 2020. Using latent profile analysis, we found five distinct crafting groups, which showed discriminate validity with regard to emotional exhaustion, engagement, and well-being. The results underscore the positive role of study crafting for students’ health and well-being. They further indicate a less important role of increasing social resources for emotional exhaustion when combined with a moderate increase in structural resources and a moderate increase in challenging demands. Our findings imply that interventions to promote study crafting should be considered to promote student health and well-being