Elliptical supra-cellular topographies regulate stem cells migratory pattern and osteogenic differentiation

In living systems, the extracellular environment is structured in a hierarchal order assembling into tissues in a myriad of shapes and complex geometries. Residing within the extracellular matrix, cells are presented and influenced by geometrical cues at several scales. While there is an emerging body of evidence that substrate with symmetric supra-cellular scale geometries (e.g. cylinders and spheres) influence the cell behavior, the effect of physiologically relevant, non-symmetric geometries with varying mean curvatures remain unexplored. In this study, we systematically explore the migratory and differentiation behavior of adipose derived stem cells (ADSCs) on arrays of elliptical cylinders (up to 80 × cell size) with varying mean curvature made from hydroxyapatite. Here, we report a new substrate-driven cell response, which we term “ridge-effect” that leads to osteogenic differentiation and nuclear deformation of cells adhered on regions of highest mean curvature at the ridge of the elliptical cylinders. This phenomenon is observed in both expansion and osteogenic medium. Live imaging combined with functional analysis shows that cells travel along-side the zero mean curvature direction on elliptical cylinders and significantly promote expression of collagen I and osteocalcin compared to a flat surface, in the absence of osteogenic supplements. Altogether, this work identifies supra-cellular scale topographies, and suggest the “ridge-effect” as a physical cue for guiding cellular mechanoresponse and promoting osteogenic differentiation. This knowledge could be utilized as an important biomaterial design parameter for the development of biomedical interfaces and bone scaffolds in tissue engineering and regenerative medicine

Development and application of an electrochemical plate coupled with immunomagnetic beads (ELIME) array for salmonella enterica detection in meat samples

Salmonella is one of the main organisms causing outbreaks of foodborne illness, and meat is one of the major vehicles of salmonellosis throughout the world. A novel analytical immunosensor array, based on a 96-well electrochemical plate coupled with immunomagnetic beads (ELIME array), is proposed for the detection of Salmonella in meat samples. After an optimization study, using Salmonella enterica serotype Enteritidis as reference antigen, the ability of the method to interact with a large number of Salmonella serovars commonly present in food was evaluated. The assay was then used to analyze samples of pork, chicken, beef, and turkey experimentally inoculated with Salmonella as well as real samples. The results were compared with those from the International Standard of Organization (ISO) culture method. The comparison showed that the ELIME array is able to detect a low number of Salmonella cells (1-10 CFU per 25 g) after only 6 h of incubation in a pre-enrichment broth. The investigation revealed a very good agreement between culture and ELIME array methods for meat samples, reducing the time for performing the analysis and obtaining the results quickly

Hippo pathway effectors control cardiac progenitor cell fate by acting as dynamic sensors of substrate mechanics and nanostructure

Stem cell responsiveness to extracellular matrix (ECM) composition and mechanical cues has been the subject of a number of investigations so far, yet the molecular mechanisms underlying stem cell mechano-biology still need full clarification. Here we demonstrate that the paralog proteins YAP and TAZ exert a crucial role in adult cardiac progenitor cell mechano-sensing and fate decision. Cardiac progenitors respond to dynamic modifications in substrate rigidity and nanopattern by promptly changing YAP/TAZ intracellular localization. We identify a novel activity of YAP and TAZ in the regulation of tubulogenesis in 3D environments and highlight a role for YAP/TAZ in cardiac progenitor proliferation and differentiation. Furthermore, we show that YAP/TAZ expression is triggered in the heart cells located at the infarct border zone. Our results suggest a fundamental role for the YAP/TAZ axis in the response of resident progenitor cells to the modifications in microenvironment nanostructure and mechanics, thereby contributing to the maintenance of myocardial homeostasis in the adult heart. These proteins are indicated as potential targets to control cardiac progenitor cell fate by materials design

Defined Microenvironments Trigger In Vitro Gastrulation in Human Pluripotent Stem Cells

Gastrulation is a stage in embryo development where three germ layers arise to dictate the human body plan. In vitro models of gastrulation have been demonstrated by treating pluripotent stem cells with soluble morphogens to trigger differentiation. However, in vivo gastrulation is a multistage process coordinated through feedback between soluble gradients and biophysical forces, with the multipotent epiblast transforming to the primitive streak followed by germ layer segregation. Here, the authors show how constraining pluripotent stem cells to hydrogel islands triggers morphogenesis that mirrors the stages preceding in vivo gastrulation, without the need for exogenous supplements. Within hours of initial seeding, cells display a contractile phenotype at the boundary, which leads to enhanced proliferation, yes-associated protein (YAP) translocation, epithelial to mesenchymal transition, and emergence of SRY-box transcription factor 17 (SOX17)+ T/BRACHYURY+ cells. Molecular profiling and pathway analysis reveals a role for mechanotransduction-coupled wingless-type (WNT) signaling in orchestrating differentiation, which bears similarities to processes observed in whole organism models of development. After two days, the colonies form multilayered aggregates, which can be removed for further growth and differentiation. This approach demonstrates how materials alone can initiate gastrulation, thereby providing in vitro models of development and a tool to support organoid bioengineering efforts.Pallavi Srivastava, Sara Romanazzo, Chantal Kopecky, Stephanie Nemec, Jake Ireland, Thomas G. Molley, Kang Lin, Pavithra B. Jayathilaka, Elvis Pandzic, Avani Yeola, Vashe Chandrakanthan, John Pimanda, and Kristopher Kilia

Substrate Stiffness Modulates Gene Expression and Phenotype in Neonatal Cardiomyocytes In Vitro

Biomaterials to be used as cell delivery systems for cardiac tissue engineering should be able to comply with cardiac muscle contractile activity, while favoring cell survival and neo-angiogenesis in a hostile environment. Biocompatible synthetic materials can be tailored to mimic cardiac tissue three-dimensional organization in the micro- and nanoscales. Nonetheless, they usually display mechanical properties that are far from those of the native myocardium and thus could affect host cell survival and activity. In the present investigation, inert poly-ɛ-caprolactone planar layers were manufactured to change the surface stiffness (with Young's modulus ranging from 1 to 133 MPa) without changing matrix chemistry. These substrates were challenged with neonatal murine cardiomyocytes to study the possible effect of substrate stiffness on such cell behavior without changing biological cues. Interestingly, softer substrates (0.91±0.08 and 1.53±0.16 MPa) were found to harbor mostly mature cardiomyocytes having assembled sarcomeres, as shown by the expression of alpha actinin and myosin heavy chain in typical striations and the upregulation of sarcomeric actin mRNA. On the other hand, a preferential expression of immature cardiac cell genes (Nkx-2.5) and proteins (GATA-4) in cardiac cells grown onto stiffer materials (49.67±2.56 and 133.23±8.67 MPa) was detected. This result could not be ascribed to significant differences in cell adhesion or proliferation induced by the substrates, but to the stabilization of cardiomyocyte differentiated phenotype induced by softer layers. In fact, cardiac cell electromechanical coupling was shown to be more organized on softer surfaces, as highlighted by connexin 43 distribution. Moreover, a differential regulation of genes involved in extracellular matrix remodeling was detected on soft films (0.91±0.08 MPa) as compared with the stiffest (133.23±8.67 MPa). Finally, the upregulation of a number of genes involved in inflammatory processes was detected when the stiffest polymer is used. These events highlight the differences in cell mechanosensitivity in a heterogeneous cell preparation and are likely to contribute to the differences encountered in cardiac cell phenotype induced by substrate stiffness.Peer reviewe

Surgical treatment of gingival overgrowth with 10 years of follow-up

<p>Abstract</p> <p>Background</p> <p>In some pathological conditions, gingivitis caused by plaque accumulation can be more severe, with the result of an overgrowth. Nevertheless, the overgrowth involves the gingival margin with extension to the inter-dental papilla. The lesion may involve the inter-proximal spaces, and become so extensive that the teeth are displaced and their crowns covered. Severe overgrowth may lead to impairment in aesthetic and masticatory functions, requiring surgical excision of the excessive tissue. Aim of this study is to describe an operative protocol for the surgical treatment of localized gingival overgrowth analyzing the surgical technique, times and follow-up.</p> <p>Methods</p> <p>A total of 20 patients were enrolled and underwent initial, non surgical, periodontal treatment and training sessions on home oral hygiene training. The treatment plan involved radical exeresis of the mass followed by positioning of an autograft of connective tissue and keratinized gingiva.</p> <p>Results</p> <p>During 10 years of follow-up, all the grafts appeared well vascularized, aesthetically satisfactory, and without relapse.</p> <p>Conclusions</p> <p>Periodontal examinations, surgical procedures, and dental hygiene with follow-up are an essential part of the treatment protocol. However, additional effort is needed from the patient. Hopefully, the final treatment result makes it all worthwhile.</p

Contributions from the DISC to accomplish the Aeolus mission objectives

The Aeolus Data Innovation and Science Cluster (DISC) supports the Aeolus mission with a wide range of activities from instrument and product quality monitoring over retrieval algorithm improvements to numerical weather prediction (NWP) impact assessments for wind and aerosols. The Aeolus DISC provides support to ESA, Cal/Val teams, numerical weather prediction (NWP) centers, and scientific users for instrument special operations and calibration, for the re-processing of Aeolus products from the past and through the provision of bi-annual updates of the L1A, L1B, L2A and L2B operational processors. The Aeolus DISC is coordinated by DLR with partners from ECMWF, KNMI, Météo-France, TROPOS, DoRIT, ABB, s&t, serco, OLA, Physics Solutions, IB Reissig and Les Myriades involving more than 40 scientists and engineers. The presentation will highlight the Aeolus DISC activities with a focus for the year 2021 and early 2022 since the last Aeolus workshop in November 2020. This covers the evolution of the instrument performance including investigations of the cause of the on-going signal loss and the achieved improvement via dedicated laser tests in 2021. In addition, refinements of algorithms and correction of the wind bias will be discussed - including a known remaining seasonal bias in October and March as encountered during the re-processing campaigns. Finally, the strategy for the on-going and future re-processing campaigns will be addressed to inform the scientific community about the availability and quality of the re-processed data products. The Aeolus mission has fully achieved its mission objectives including the unprecedented demonstration of direct-detection Doppler wind lidar technology and high-power laser operation in space in the ultraviolet spectral region over its planned full mission lifetime of 3 years and 3 months. Aeolus wind products have clearly demonstrated positive impact on forecasts using several NWP models. Since early 2020, and thus only 1.5 years after launch, the Aeolus wind products are used in operation at various NWP centers worldwide. This was achieved even despite the larger than expected wind random errors due to lower initial atmospheric signal levels and the observed signal losses during the operation of the first and second laser. In addition to this incredible success, first scientific studies demonstrated the use of Aeolus for atmospheric dynamics research in the stratosphere and for the analysis of aerosol transport. These achievements of the Aeolus mission and its success were only possible with the essential and critical contributions from the Aeolus DISC. This demonstrates the need and potential for setting up such scientific consortia covering a wide range of expertise from instrument, processors, and scientific use of products for Earth Explorer type missions. The invaluable experience gained by the Aeolus DISC during the more then 3 years of Aeolus mission in orbit (preceded by a period of 20 years before launch by a similar study team) is a pre-requisite for a successful preparation of an operational follow-on Aeolus-2 mission

Risks to human and animal health related to the presence of deoxynivalenol and its acetylated and modified forms in food and feed

Deoxynivalenol (DON) is a mycotoxin primarily produced by Fusarium fungi, occurring predominantly in cereal grains. Following the request of the European Commission, the CONTAM Panel assessed the risk to animal and human health related to DON, 3-acetyl-DON (3-Ac-DON), 15-acetyl-DON (15-Ac-DON) and DON-3-glucoside in food and feed. A total of 27,537, 13,892, 7,270 and 2,266 analytical data for DON, 3-Ac-DON, 15-Ac-DON and DON-3-glucoside, respectively, in food, feed and unprocessed grains collected from 2007 to 2014 were used. For human exposure, grains and grain-based products were main sources, whereas in farm and companion animals, cereal grains, cereal by-products and forage maize contributed most. DON is rapidly absorbed, distributed, and excreted. Since 3-Ac-DON and 15-Ac-DON are largely deacetylated and DON-3-glucoside cleaved in the intestines the same toxic effects as DON can be expected. The TDI of 1 μg/kg bw per day, that was established for DON based on reduced body weight gain in mice, was therefore used as a group-TDI for the sum of DON, 3-Ac-DON, 15-Ac-DON and DON-3-glucoside. In order to assess acute human health risk, epidemiological data from mycotoxicoses were assessed and a group-ARfD of 8 μg/kg bw per eating occasion was calculated. Estimates of acute dietary exposures were below this dose and did not raise a health concern in humans. The estimated mean chronic dietary exposure was above the group-TDI in infants, toddlers and other children, and at high exposure also in adolescents and adults, indicating a potential health concern. Based on estimated mean dietary concentrations in ruminants, poultry, rabbits, dogs and cats, most farmed fish species and horses, adverse effects are not expected. At the high dietary concentrations, there is a potential risk for chronic adverse effects in pigs and fish and for acute adverse effects in cats and farmed mink