Enhanced Immunomodulation in Inflammatory Environments Favors Human Cardiac Mesenchymal Stromal-Like Cells for Allogeneic Cell Therapies

Rising numbers of patients with cardiovascular diseases and limited availability of donor hearts require new and improved therapy strategies. Human atrial appendage-derived cells (hAACs) are promising candidates for an allogeneic cell-based treatment. In this study, we evaluated their inductive and modulatory capacity regarding immune responses and underlying key mechanisms in vitro. For this, cryopreserved hAACs were either cultured in the presence of interferon-gamma (IFNγ) or left unstimulated. The expression of characteristic mesenchymal stromal cell markers (CD29, CD44, CD73, CD105, CD166) was revealed by flow cytometry that also highlighted a predominant negativity for CD90. A low immunogeneic phenotype in an inflammatory milieu was shown by lacking expression of co-stimulatory molecules and upregulation of the inhibitory ligands PD-L1 and PD-L2, despite de novo expression of HLA-DR. Co-cultures of hAACs with allogeneic peripheral blood mononuclear cells, proved their low immunogeneic state by absence of induced T cell proliferation and activation. Additionally, elevated levels of IL-1β, IL-33, and IL-10 were detectable in those cell culture supernatants. Furthermore, the immunomodulatory potential of hAACs was assessed in co-cultures with αCD3/αCD28-activated peripheral blood mononuclear cells. Here, a strong inhibition of T cell proliferation and reduction of pro-inflammatory cytokines (IFNγ, TNFα, TNFβ, IL-17A, IL-2) were observable after pre-stimulation of hAACs with IFNγ. Transwell experiments confirmed that mostly soluble factors are responsible for these suppressive effects. We were able to identify indolamin-2,3-dioxygenase (IDO) as a potential key player through a genome-wide gene expression analysis and could demonstrate its involvement in the observed immunological responses. While the application of blocking antibodies against both PD-1 ligands did not affect the immunomodulation by hAACs, 1-methyl-L-tryptophan as specific inhibitor of IDO was able to restore proliferation and to lower apoptosis of T cells. In conclusion, hAACs represent a cardiac-derived mesenchymal stromal-like cell type with a high potential for the application in an allogeneic setting, since they do not trigger T cell responses and even increase their immunomodulatory potential in inflammatory environments

Cardiac Extracellular Vesicles (EVs) Released in the Presence or Absence of Inflammatory Cues Support Angiogenesis in Different Manners

Cells release extracellular vesicles (EVs) to communicate in a paracrine manner with other cells, and thereby influence processes, such as angiogenesis. The conditioned medium of human cardiac-derived adherent proliferating (CardAP) cells was recently shown to enhance angiogenesis. To elucidate whether their released EVs are involved, we isolated them by differential centrifugation from the conditioned medium derived either in the presence or absence of a pro-inflammatory cytokine cocktail. Murine recipient cells internalized CardAP-EVs as determined by an intracellular detection of human proteins, such as CD63, by a novel flow cytometry method for studying EV-cell interaction. Moreover, endothelial cells treated for 24 h with either unstimulated or cytokine stimulated CardAP-EVs exhibited a higher tube formation capability on Matrigel. Interestingly, unstimulated CardAP-EVs caused endothelial cells to release significantly more vascular endothelial growth factor and interleukin (IL)-6, while cytokine stimulated CardAP-EVs significantly enhanced the release of IL-6 and IL-8. By nCounter® miRNA expression assay (NanoString Technologies) we identified microRNA 302d-3p to be enhanced in unstimulated CardAP-EVs compared to their cytokine stimulated counterparts, which was verified by quantitative polymerase chain reaction. This study demonstrates that both CardAP-EVs are pro-angiogenic by inducing different factors from endothelial cells. This would allow to select potent targets for a safe and efficient therapeutic application

Importance of Oral Hygiene and Maintaining Oral Health in Persons With Disabilities

This paper aims to approximate and facilitate the routine of daily oral hygiene for persons with disabilities, as well as to emphasize the importance of educating persons with disabilities and their caregivers about oral health as an essential part of overall health. Desk research of electronic databases was conducted with the aim of writing this paper, using the following keywords: ‘oral hygiene’, ‘dental plaque’, ‘oral health’ and ‘persons with disabilities’. Literature research has shown that persons with disabilities have poor oral hygiene, as well as that there is a lack of education among them and their caregivers about the importance of oral health and proper oral hygiene. Poor oral hygiene can affect a person’s quality of life due to discomfort during eating, bad breath, poor self-esteem, pain, and disturbed sleep, which is a result of caries or other diseases of the oral cavity. Maintaining oral health is an essential part of overall health

Finding Early Farming Communities in southern Mozambique: Using Geophysical Surveys to examine potential new open-air sites

The arrival of Early Farming Communities (EFC) in Mozambique is traditionally defined by the appearance of the “Bantu package”, especially of the so-called Matola pottery at the beginning of the 1st millennium CE. Although many EFC sites are known in Mozambique and South Africa, little is known about their settlement structures. In the case of Mozambique, the well-known Matola, Zitundo and University Campus sites were discovered by chance. The Department of Archaeology and Anthropology (DAA) at the University Eduardo Mondlane has successfully conducted surveys in the Changalane Administrative Post (Namaacha District, Maputo province) for years, documenting new potential EFC and Stone Age sites. Together with the German Archaeological Institute and the University of Hamburg, geophysical surveys were carried out on four sites. The aim was to get an overview of the sites and to locate potential excavation areas such as waste or storage pits, furnaces or huts. Although the method is already of great importance in Europe and is used regularly, only few comparative studies from sub-Saharan Africa are known. Within this region in the southeast and the described context of the continent the method is applied for the first time. A verification of the results in the form of test excavations is planned in the near future.The arrival of Early Farming Communities (EFC) in Mozambique is traditionally defined by the appearance of the “Bantu package”, especially of the so-called Matola pottery at the beginning of the 1st millennium CE. Although many EFC sites are known in Mozambique and South Africa, little is known about their settlement structures. In the case of Mozambique, the well-known Matola, Zitundo and University Campus sites were discovered by chance. The Department of Archaeology and Anthropology (DAA) at the University Eduardo Mondlane has successfully conducted surveys in the Changalane Administrative Post (Namaacha District, Maputo province) for years, documenting new potential EFC and Stone Age sites. Together with the German Archaeological Institute and the University of Hamburg, geophysical surveys were carried out on four sites. The aim was to get an overview of the sites and to locate potential excavation areas such as waste or storage pits, furnaces or huts. Although the method is already of great importance in Europe and is used regularly, only few comparative studies from sub-Saharan Africa are known. Within this region in the southeast and the described context of the continent the method is applied for the first time. A verification of the results in the form of test excavations is planned in the near future.&nbsp

Extracellular vesicles from regenerative human cardiac cells act as potent immune modulators by priming monocytes

Background: Nano-sized vesicles, so called extracellular vesicles (EVs), from regenerative cardiac cells represent a promising new therapeutic approach to treat cardiovascular diseases. However, it is not yet sufficiently understood how cardiac-derived EVs facilitate their protective effects. Therefore, we investigated the immune modulating capabilities of EVs from human cardiac-derived adherent proliferating (CardAP) cells, which are a unique cell type with proven cardioprotective features. Results: Differential centrifugation was used to isolate EVs from conditioned medium of unstimulated or cytokinestimulated (IFNγ, TNFα, IL-1β) CardAP cells. The derived EVs exhibited typical EV-enriched proteins, such as tetraspanins, and diameters mostly of exosomes (< 100 nm). The cytokine stimulation caused CardAP cells to release smaller EVs with a lower integrin ß1 surface expression, while the concentration between both CardAP-EV variants was unaffected. An exposure of either CardAP-EV variant to unstimulated human peripheral blood mononuclear cells (PBMCs) did not induce any T cell proliferation, which indicates a general low immunogenicity. In order to evaluate immune modulating properties, PBMC cultures were stimulated with either Phytohemagglutin or anti-CD3. The treatment of those PBMC cultures with either CardAP-EV variant led to a significant reduction of T cell proliferation, pro-inflammatory cytokine release (IFNγ, TNFα) and increased levels of active TGFβ. Further investigations identified CD14+ cells as major recipient cell subset of CardAP–EVs. This interaction caused a significant lower surface expression of HLA-DR, CD86, and increased expression levels of CD206 and PD-L1. Additionally, EV-primed CD14+ cells released significantly more IL-1RA. Notably, CardAP-EVs failed to modulate anti-CD3 triggered T cell proliferation and pro-inflammatory cytokine release in monocultures of purified CD3+ T cells. Subsequently, the immunosuppressive feature of CardAPEVs was restored when anti-CD3 stimulated purified CD3+ T cells were co-cultured with EV-primed CD14+ cells. Beside attenuated T cell proliferation, those cultures also exhibited a significant increased proportion of regulatory T cells. Conclusions: CardAP-EVs have useful characteristics that could contribute to enhanced regeneration in damaged cardiac tissue by limiting unwanted inflammatory processes. It was shown that the priming of CD14+ immune cells by CardAP-EVs towards a regulatory type is an essential step to attenuate significantly T cell proliferation and proinflammatory cytokine release in vitro

Problemlösen in der Mechanik: eine Untersuchung mit Studierenden

Sowohl im schulischen als auch universitären Umfeld nehmen Probleme in Form von Übungsaufgaben oder Prüfungen einen wichtigen Platz ein, da so die Leistungen der Lernenden in Physik eingeordnet werden können (vgl. z.B. Fischer & Draxler [1] oder Kühn [2]).Um die Leistung von Studierenden im Bereich „Mechanik“ zu messen, wurde ein Papier-und-Bleistift-Test entwickelt, der auf dem Modell des wissenszentrierten Problemlösens (nach Friege [11]) basiert. Nach diesem Modell lässt sich das Problemlösen in vier Phasen unterteilen, die unterschiedliche Anforderungen an den Bearbeitenden stellen. Die einzelnen Items des Tests wurden so entworfen, dass jeweils nur eine Phase eines Problems bearbeitet wird. Um der Komplexität des Problemlösens mit einer großen Variation von Aufgaben gerecht werden zu können, wurde ein spezielles Testheft-Design verwendet, das die Anzahl der zu bearbeitenden Aufgaben pro Testheft auf ein praktikables Maß reduziert.Ausgewertet wurde der Test mit Hilfe eines Rasch-Modells, das die Leistungen der Studierenden gut beschreibt. Über die Itemschwierigkeit lassen sich mehrere, qualitativ unterschiedliche Stufen finden, welche die Fähigkeiten der Studierenden mit entsprechendem Personenparameter widerspiegeln. Es wurden Gruppenvergleiche durchgeführt, die im Hinblick auf ein Modell zur Erklärung des Erfolgs beim Problemlösen erste Hinweise liefern

Nicht-harmonische Schwingungen am Huygens-Raebiger Pendel

In der Lehramtsausbildung haben im physikalischen Praktikum zunehmend computergestützte Versuche ihren Platz gefunden. Hierbei ist nicht nur die computergestützte Messwertaufnahme von Vorteil. Die Studierenden haben auch bei der Auswertung die Möglichkeit sich mit vergleichsweise geringem Aufwand mit mathematischen Modellierungen von physikalischen Sachverhalten zu beschäftigen. Aus einer Reihe entwickelter Versuche für das physikalische Praktikum an der Pädagogischen Hochschule Freiburg beschäftigt sich ein Versuch mit den Unterschieden zwischen einer harmonischen Schwingung und einer nicht-harmonischen Schwingung. Es soll mit diesem Versuch gezeigt werden, dass für große Winkel ein physikalisches Pendel keine harmonische Schwingung vollzieht und dass dieser Effekt bei längeren Pendeln größer ist als bei kürzeren. In diesem Zusammenhang werden die Grenzen einer mathematischen Modellierung mit Hilfe eines CAS veranschaulicht. Als physikalisches Pendel wird das Huygens-Raebiger Pendel verwendet – ein Pendel das sowohl gekoppelt als ein Pendel schwingen kann, als auch entkoppelt als sieben „kleine“, sich durch die Länge der Stange unterscheidende, Pendel. Im Folgenden werden der Versuch und die Modellierung der Messergebnisse vorgestellt

Nicht-harmonische Schwingungen am Huygens-Raebiger Pendel

In der Lehramtsausbildung haben im physikalischen Praktikum zunehmend computergestützte Versuche ihren Platz gefunden. Hierbei ist nicht nur die computergestützte Messwertaufnahme von Vorteil. Die Studierenden haben auch bei der Auswertung die Möglichkeit sich mit vergleichsweise geringem Aufwand mit mathematischen Modellierungen von physikalischen Sachverhalten zu beschäftigen. Aus einer Reihe entwickelter Versuche für das physikalische Praktikum an der Pädagogischen Hochschule Freiburg beschäftigt sich ein Versuch mit den Unterschieden zwischen einer harmonischen Schwingung und einer nicht-harmonischen Schwingung. Es soll mit diesem Versuch gezeigt werden, dass für große Winkel ein physikalisches Pendel keine harmonische Schwingung vollzieht und dass dieser Effekt bei längeren Pendeln größer ist als bei kürzeren. In diesem Zusammenhang werden die Grenzen einer mathematischen Modellierung mit Hilfe eines CAS veranschaulicht. Als physikalisches Pendel wird das Huygens-Raebiger Pendel verwendet – ein Pendel das sowohl gekoppelt als ein Pendel schwingen kann, als auch entkoppelt als sieben „kleine“, sich durch die Länge der Stange unterscheidende, Pendel. Im Folgenden werden der Versuch und die Modellierung der Messergebnisse vorgestellt

Problemlösen in der Mechanik: eine Untersuchung mit Studierenden

Sowohl im schulischen als auch universitären Umfeld nehmen Probleme in Form von Übungsaufgaben oder Prüfungen einen wichtigen Platz ein, da so die Leistungen der Lernenden in Physik eingeordnet werden können (vgl. z.B. Fischer & Draxler [1] oder Kühn [2]).Um die Leistung von Studierenden im Bereich „Mechanik“ zu messen, wurde ein Papier-und-Bleistift-Test entwickelt, der auf dem Modell des wissenszentrierten Problemlösens (nach Friege [11]) basiert. Nach diesem Modell lässt sich das Problemlösen in vier Phasen unterteilen, die unterschiedliche Anforderungen an den Bearbeitenden stellen. Die einzelnen Items des Tests wurden so entworfen, dass jeweils nur eine Phase eines Problems bearbeitet wird. Um der Komplexität des Problemlösens mit einer großen Variation von Aufgaben gerecht werden zu können, wurde ein spezielles Testheft-Design verwendet, das die Anzahl der zu bearbeitenden Aufgaben pro Testheft auf ein praktikables Maß reduziert.Ausgewertet wurde der Test mit Hilfe eines Rasch-Modells, das die Leistungen der Studierenden gut beschreibt. Über die Itemschwierigkeit lassen sich mehrere, qualitativ unterschiedliche Stufen finden, welche die Fähigkeiten der Studierenden mit entsprechendem Personenparameter widerspiegeln. Es wurden Gruppenvergleiche durchgeführt, die im Hinblick auf ein Modell zur Erklärung des Erfolgs beim Problemlösen erste Hinweise liefern

Climatological and radiative properties of midlatitude cirrus clouds derived by automatic evaluation of lidar measurements

Cirrus, i.e., high, thin clouds that are fully glaciated, play an important role in the Earth's radiation budget as they interact with both long- and shortwave radiation and affect the water vapor budget of the upper troposphere and stratosphere. Here, we present a climatology of midlatitude cirrus clouds measured with the same type of ground-based lidar at three midlatitude research stations: at the Swiss high alpine Jungfraujoch station (3580 m a.s.l.), in Zürich (Switzerland, 510 m a.s.l.), and in Jülich (Germany, 100 m a.s.l.). The analysis is based on 13 000 h of measurements from 2010 to 2014. To automatically evaluate this extensive data set, we have developed the Fast LIdar Cirrus Algorithm (FLICA), which combines a pixel-based cloud-detection scheme with the classic lidar evaluation techniques. We find mean cirrus optical depths of 0.12 on Jungfraujoch and of 0.14 and 0.17 in Zürich and Jülich, respectively. Above Jungfraujoch, subvisible cirrus clouds (τ < 0.03) have been observed during 6 % of the observation time, whereas above Zürich and Jülich fewer clouds of that type were observed. Cirrus have been observed up to altitudes of 14.4 km a.s.l. above Jungfraujoch, whereas they have only been observed to about 1 km lower at the other stations. These features highlight the advantage of the high-altitude station Jungfraujoch, which is often in the free troposphere above the polluted boundary layer, thus enabling lidar measurements of thinner and higher clouds. In addition, the measurements suggest a change in cloud morphology at Jungfraujoch above ∼ 13 km, possibly because high particle number densities form in the observed cirrus clouds, when many ice crystals nucleate in the high supersaturations following rapid uplifts in lee waves above mountainous terrain. The retrieved optical properties are used as input for a radiative transfer model to estimate the net cloud radiative forcing, CRFNET, for the analyzed cirrus clouds. All cirrus detected here have a positive CRFNET. This confirms that these thin, high cirrus have a warming effect on the Earth's climate, whereas cooling clouds typically have cloud edges too low in altitude to satisfy the FLICA criterion of temperatures below −38 °C. We find CRFNET = 0.9 W m−2 for Jungfraujoch and 1.0 W m−2 (1.7 W m−2) for Zürich (Jülich). Further, we calculate that subvisible cirrus (τ < 0.03) contribute about 5 %, thin cirrus (0.03 < τ < 0.3) about 45 %, and opaque cirrus (0.3 < τ) about 50 % of the total cirrus radiative forcing