Regulation der Zelladhäsion in Meibomdrüsen und deren Rolle in der Pathogenese des Trockenen Auges

Meibomian glands are modified, holocrine sebaceous glands within the eyelid important for the maintenance of the integrity and health of the ocular surface. These glands produce an oily secret which stabilizes the tear film. Therefore, a dysfunction of the glands (Meibomian Gland Dysfunction, MGD) is nowadays seen as the leading cause of the so- called Dry eye disease (DED). Former in situ research done by our group showed that the number of desmosomes in Meibomian glands increases over time during cell differentiation. What is more, Pemphigus vulgaris, a chronic blistering autoimmune disease caused by autoantibodies against desmosomal cadherins, is very often associated with DED. With this work we firstly wanted to analyse the physiological conditions for the formation of cell-cell-contacts in Meibomian glands and secondly investigate the role of cell cohesion in the pathogenesis of the Meibomian gland dysfunction. We used a stable human meibocyte cell-line (human meibomian gland cells, HMGECs) and started with identifying the perfect conditions for differentiation. We determined two points in time that were associated with an early and a more mature state of differentiation. We then wanted to analyze the exact composition of cell cohesion molecules in Meibomian glands and quantify the adhesion strength during maturation. Desmosomal cadherins were of major interest in the beginning, because they are known to be essential for strong cell cohesion in various tissues. However, this data shows that cell cohesion is regulated differently in Meibomian glands. What is more, there is evidence that the adherens junction component E-cadherin, and not desmosomes, is crucial for the maintenance of physiological processes in Meibocytes

A systematic comparison of pan-Trk immunohistochemistry assays among multiple cancer types

AIMS NTRK rearranged tumours are rare but can be successfully treated using anti-TRK-targeted therapies, making NTRK testing important for treatment choices in patients with advanced cancers. Pan-Trk immunohistochemistry (IHC) has become a valuable and affordable screening tool in many laboratories. Unfortunately, the choice of antibodies and IHC protocols to investigate biomarkers is not standardised. In this study, we compared the performance of four pan-Trk IHC methods, using three different clones, primarily in NTRK fusion-positive tumours. METHODS AND RESULTS We studied the performance of four pan-Trk IHC methods using three different clones: EPR17341 (Abcam and Ventana), EP1058Y (Abcam) and A7H6R (Cell Signaling) in 22 molecularly confirmed NTRK rearranged tumours. Additionally, selected NTRK fusion-negative tumours were further included: NTRK mutated (n = 8) and amplified (n = 15) tumours as well as NTRK fusion-negative tumours driven by other gene fusions, such as ALK, ROS1 and BCOR (n = 20), as well as salivary gland tumours (n = 16). Inter-rater agreement of three pathologists was additionally calculated, including H-score. With clone EPR17341 (Abcam in-house and ready-to-use Ventana protocol), all molecularly confirmed NTRK1-3 rearranged tumours were positively detected by immunohistochemistry, while the other clones missed NTRK2-3 rearranged tumours. For the fusion-negative cohort we found the best performance (least false-positive cases) using the clone A7H6R (Cell Signalling). CONCLUSION Given the therapeutic importance, testing for NTRK rearrangements in daily practice has become necessary and, despite IHC being a fast and affordable tool, using it in routine diagnostics is complicated and requires a high level of expertise

Methacrylated Gelatin as a Scaffold for Mechanically Isolated Stromal Vascular Fraction for Cutaneous Wound Repair

Mechanically processed stromal vascular fraction (mSVF) is a highly interesting cell source for regenerative purposes, including wound healing, and a practical alternative to enzymatically isolated SVF. In the clinical context, SVF benefits from scaffolds that facilitate viability and other cellular properties. In the present work, the feasibility of methacrylated gelatin (GelMA), a stiffness-tunable, light-inducible hydrogel with high biocompatibility is investigated as a scaffold for SVF in an in vitro setting. Lipoaspirates from elective surgical procedures were collected and processed to mSVF and mixed with GelMA precursor solutions. Non-encapsulated mSVF served as a control. Viability was measured over 21 days. Secreted basic fibroblast growth factor (bFGF) levels were measured on days 1, 7 and 21 by ELISA. IHC was performed to detect VEGF-A, perilipin-2, and CD73 expression on days 7 and 21. The impact of GelMA-mSVF on human dermal fibroblasts was measured in a co-culture assay by the same viability assay. The viability of cultured GelMA-mSVF was significantly higher after 21 days (p < 0.01) when compared to mSVF alone. Also, GelMA-mSVF secreted stable levels of bFGF over 21 days. While VEGF-A was primarily expressed on day 21, perilipin-2 and CD73-positive cells were observed on days 7 and 21. Finally, GelMA-mSVF significantly improved fibroblast viability as compared with GelMA alone (p < 0.01). GelMA may be a promising scaffold for mSVF as it maintains cell viability and proliferation with the release of growth factors while facilitating adipogenic differentiation, stromal cell marker expression and fibroblast proliferation

Ruolo dell'infermiere nella prevenzione del linfedema, secondario a carcinoma mammario

I trattamenti efficaci per il cancro al seno hanno portato ad un aumento significativo del numero di sopravvissuti. Molte di queste donne, però, devono adeguarsi agli effetti avversi del cancro e dei trattamenti che hanno salvato le loro vite. Il più comune effetto avverso correlato al trattamento del cancro al seno è il linfedema dell’arto superiore. Il linfedema è una condizione cronica determinata da un anomalo accumulo di liquidi, determinato da un inadeguato drenaggio linfatico che si manifesta clinicamente con la presenza di edema, infiammazione, fibrosi ed indurimento dei tessuti interessati. Queste condizioni possono portare ad una riduzione della mobilità, dolore e disagio, può essere stimolata una crescita batterica con conseguente aumento del rischio di infezioni. Tutte queste condizioni tendono a determinare un progressivo scadere della qualità della vita che in alcuni casi può essere fortemente limitata proprio dalla riduzione della mobilità delle parti interessate. L’obiettivo di questa revisione bibliografica è di fornire indicazioni riguardanti le strategie di prevenzione del linfedema secondario a carcinoma mammario effettuate dall’infermiere, individuando quali sono gli interventi ritenuti efficaci a questo scopo, sulla base di un’analisi sistematica della letteratura corrente. L’analisi del materiale ha quindi posto in evidenza il ruolo dell’infermiere nella prevenzione del linfedema, non solo come operatore sanitario che agisce in collaborazione con il personale medico, ma anche e soprattutto, come professionista responsabile dell’educazione terapeutica del paziente: infatti, l’infermiere è l’operatore sanitario più a stretto contatto con il paziente e che quindi ricopre un ruolo fondamentale nell’attuazione corretta delle misure preventive

Methacrylated Gelatin as a Scaffold for Mechanically Isolated Stromal Vascular Fraction for Cutaneous Wound Repair

Mechanically processed stromal vascular fraction (mSVF) is a highly interesting cell source for regenerative purposes, including wound healing, and a practical alternative to enzymatically isolated SVF. In the clinical context, SVF benefits from scaffolds that facilitate viability and other cellular properties. In the present work, the feasibility of methacrylated gelatin (GelMA), a stiffness-tunable, light-inducible hydrogel with high biocompatibility is investigated as a scaffold for SVF in an in vitro setting. Lipoaspirates from elective surgical procedures were collected and processed to mSVF and mixed with GelMA precursor solutions. Non-encapsulated mSVF served as a control. Viability was measured over 21 days. Secreted basic fibroblast growth factor (bFGF) levels were measured on days 1, 7 and 21 by ELISA. IHC was performed to detect VEGF-A, perilipin-2, and CD73 expression on days 7 and 21. The impact of GelMA-mSVF on human dermal fibroblasts was measured in a co-culture assay by the same viability assay. The viability of cultured GelMA-mSVF was significantly higher after 21 days (p p < 0.01). GelMA may be a promising scaffold for mSVF as it maintains cell viability and proliferation with the release of growth factors while facilitating adipogenic differentiation, stromal cell marker expression and fibroblast proliferation

Methacrylated Gelatin as a Scaffold for Mechanically Isolated Stromal Vascular Fraction for Cutaneous Wound Repair

Mechanically processed stromal vascular fraction (mSVF) is a highly interesting cell source for regenerative purposes, including wound healing, and a practical alternative to enzymatically isolated SVF. In the clinical context, SVF benefits from scaffolds that facilitate viability and other cellular properties. In the present work, the feasibility of methacrylated gelatin (GelMA), a stiffness-tunable, light-inducible hydrogel with high biocompatibility is investigated as a scaffold for SVF in an in vitro setting. Lipoaspirates from elective surgical procedures were collected and processed to mSVF and mixed with GelMA precursor solutions. Non-encapsulated mSVF served as a control. Viability was measured over 21 days. Secreted basic fibroblast growth factor (bFGF) levels were measured on days 1, 7 and 21 by ELISA. IHC was performed to detect VEGF-A, perilipin-2, and CD73 expression on days 7 and 21. The impact of GelMA-mSVF on human dermal fibroblasts was measured in a co-culture assay by the same viability assay. The viability of cultured GelMA-mSVF was significantly higher after 21 days (p < 0.01) when compared to mSVF alone. Also, GelMA-mSVF secreted stable levels of bFGF over 21 days. While VEGF-A was primarily expressed on day 21, perilipin-2 and CD73-positive cells were observed on days 7 and 21. Finally, GelMA-mSVF significantly improved fibroblast viability as compared with GelMA alone (p < 0.01). GelMA may be a promising scaffold for mSVF as it maintains cell viability and proliferation with the release of growth factors while facilitating adipogenic differentiation, stromal cell marker expression and fibroblast proliferation.ISSN:1422-006

Seminari di topografia antica e medievale per Letizia Ermini Pani

Atti della giornata di studi, Roma, Istituto Nazionale di Studi Romani, 4 dicembre 201