Interaction between a fast rotating sunspot and ephemeral regions as the origin of the major solar event on 2006 December 13

The major solar event on 2006 December 13 is characterized by the approximately simultaneous occurrence of a heap of hot ejecta, a great two-ribbon flare and an extended Earth-directed coronal mass ejection. We examine the magnetic field and sunspot evolution in active region NOAA AR 10930, the source region of the event, while it transited the solar disk centre from Dec. 10 to Dec. 13. We find that the obvious changes in the active region associated with the event are the development of magnetic shear, the appearance of ephemeral regions and fast rotation of a smaller sunspot. Around the area of the magnetic neutral line of the active region, interaction between the fast rotating sunspot and the ephemeral regions triggers continual brightening and finally the major flare. It is indicative that only after the sunspot rotates up to 200$^{\circ}$ does the major event take place. The sunspot rotates at least 240$^{\circ}$ about its centre, the largest sunspot rotation angle which has been reported.Comment: 4 pages, 6 figures, ApJ Letters inpres

Core-shell silk hydrogels with spatially tuned conformations as drug delivery system

Hydrogels of spatially controlled physicochemical properties are appealing platforms for tissue engineering and drug delivery. In this study, core-shell silk fibroin (SF) hydrogels of spatially controlled conformation were developed. The core-shell structure in the hydrogels was formed by means of soaking the preformed (enzymatically crosslinked) random coil SF hydrogels in methanol. When increasing the methanol treatment time from 1 to 10 minutes, the thickness of the shell layer can be tuned from about 200 to around 850 µm as measured in wet status. After lyophilization of the rehydrated core-shell hydrogels, the shell layer displayed compact morphology and the core layer presented porous structure, when observed by scanning electron microscopy. The conformation of the hydrogels was evaluated by Fourier transform infrared spectroscopy in wet status. The results revealed that the shell layer possessed dominant β-sheet conformation and the core layer maintained mainly random coil conformation. Enzymatic degradation data showed that the shell layers presented superior stability to the core layer. The mechanical analysis displayed that the compressive modulus of the core-shell hydrogels ranged from around 25 kPa to about 1.1 MPa by increasing the immersion time in methanol. When incorporated with albumin, the core-shell SF hydrogels demonstrated slower and more controllable release profiles compared with the non-treated hydrogel. These core-shell SF hydrogels of highly tuned properties are useful systems as drug delivery system and may be applied as cartilage substitute.This study was funded by the Portuguese Foundation for Science and Technology (FCT) projects Tissue2Tissue (PTDC/CTM/105703/2008) and OsteoCart (PTDC/CTM-BPC/115977/2009), as well as the European Union’s FP7 Programme under grant agreement no. REGPOT-CT2012-316331-POLARIS. Le-Ping Yan was awarded a FCT PhD scholarship (SFRH/BD/ 64717/2009). The FCT distinctions attributed to J.M. Oliveira and A.L. Oliveira under the Investigador FCT program (IF/ 00423/2012) and (IF/00411/2013) are also greatly acknowledged, respectively

De novo bone formation on macro/microporous silk and silk/nano-sized calcium phosphate scaffolds

Macro/micro porous silk/nano-sized calcium phosphate scaffolds (SC16) with bioactive and superior physicochemical properties have been recently developed. In this study, we aim at evaluating the new bone formation ability of the SC 16 scaffolds in vivo, using silk fibroin scaffolds (S16) as control. The CaP distribution profile in the scaffolds was characterized by Micro-Computed Tomography. The in vitro mineralization behavior was examined by immersion in Simulated Body Fluid solution from 1 to 14 days. The long-term hydration degree and weight loss ratio of the scaffolds were evaluated by immersion in an Isotonic Saline Solution from 1 month to 1 year. In vivo osteogenesis properties of the scaffolds were screened by implantation into the rat femur defects for 3 weeks. The results showed that the CaP phase distributed homogeneously in the SC16 scaffolds. Mineralization was only observed in SC16 scaffolds, and both scaffolds gradually degraded with time. The staining of the explants showed that new bone formation with higher density was observed in the SC16 scaffolds as compared to S16 scaffolds, guiding the growth of new bone directly onto its surface. These results demonstrated that the SC16 hybrid scaffolds are osteoconductive and can be good candidates for bone tissue engineering as promoted superior de novo bone formation.This study was supported by the Portuguese Foundation for Science and Technology (FCT) projects OsteoCart (PTDC/CTM-BPC/115977/2009) and Tissue2Tissue (PTDC/CTM/105703/2008). Research leading to these results has received funding from the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement no REGPOT-CT2012-316331-POLARIS. Le-Ping Yan is an FCT PhD scholarship holder (SFRH/BD/64717/2009)

Macro/microporous silk fibroin scaffolds with potential for articular cartilage and meniscus tissue engineering applications

This study describes the developmental physicochemical properties of silk fibroin scaffolds derived from high concentration aqueous silk fibroin solutions. The silk fibroin scaffolds were prepared with different initial concentrations (8%, 10%, 12% and 16% (wt%)) and obtained by combining the salt-leaching and freeze-drying methodologies. The results indicated that the antiparallel β-pleated sheet (silk-II) conformation was present in the silk fibroin scaffolds. All the scaffolds possessed macro/micro porous structure. Homogeneous porosity distribution was achieved in all the groups of samples. As the silk fibroin concentration increased from 8% to 16%, the mean porosity decreased from 90.8±0.9% to 79.8±0.3%, and the mean interconnectivity decreased from 97.4±0.5% to 92.3±1.3%. The mechanical properties of the scaffolds exhibited a concentration dependence. The dry state compressive modulus increased from 0.81±0.29 MPa to 15.14±1.70 MPa, and the wet state dynamic storage modulus increased around 20-30 folds at each testing frequencies when the silk fibroin concentration increased from 8% to 16%. The water-uptake ratio decreased by means of increasing silk fibroin concentration. The scaffolds present favorable stability as their structure integrity, morphology and mechanical properties were maintained after in vitro degradation for 30 days. Based on these results, the scaffolds developed in this study are herein proposed to be used in meniscus and cartilage tissue engineering scaffolding.Tissue2Tissue project (PTDC/CTM/105703/2008

Combinatory approach for developing silk fibroin-based scaffolds with hierarchical porosity and enhanced performance for cartilage tissue engineering applications

Introduction: The combination of several processing technologies can open the possibility for producing scaffolds with superior performance for tissue engineering (TE) applications. Hydrogels are structurally similar to the natural extracellular matrix microenvironment presenting high elasticity and resistance to compression forces. They have been extensively used in biomedical devices fabrication and for TE applications, including for cartilage defects repair[1]. Recently, it was found that proteins like silk fibroin (SF), presenting tyrosine groups can be used to prepare fast formed hydrogels with controlled gelation properties, via an enzyme-mediated cross-linking reaction using horseradish peroxidase (HRP) and hydrogen peroxide (H2O2)[2],[3]. Moreover, the high versatility, processability and tailored mechanical properties of SF, make this natural polymer attractive for the development of innovative scaffolding strategies for cartilage TE applications[4],[5]. Materials and Methods: The present work proposes a novel route for developing SF-based scaffolds derived from high- concentrated SF (16wt%) enzymatically cross-linked by a HRP/H2O2 complex. The combination of salt-leaching and freeze-drying methodologies was used to prepare macro/microporous SF scaffolds with an interconnected structure and specific features regarding biodegradation and mechanical properties (Fig. 1a). The scaffolds morphology and porosity were analyzed by SEM and micro-CT. The mechanical properties (Instron) and protein conformation (FTIR, XRD) were also assessed. In order to evaluate the scaffolds structural integrity, swelling ratio and degradation profile studies were performed for a period of 30 day. This work also aims to evaluate the in vitro chondrogenic differentiation response by culturing human adipose derived stem cells (hASCs) over 21 days in basal and chondrogenic conditions. Cell behaviour in the presence of the macro/microporous structures will be evaluated through different quantitative (Live/Dead, DNA, GAGs, RT PCR) and qualitative (SEM, histology, immunocytochemistry) assays. Results and Discussion: The macro/microporous SF scaffolds showed high porosity and interconnectivity with the trabecular structures evenly distributed (Fig. 1b,c). A dramatic decrease of compressive modulus was observed for samples in hydrated state. Chemical analysis revealed that SF scaffolds displayed the characteristic peaks for β-sheet conformation. Swelling ratio data demonstrated a large swelling capacity, maintaining their structural integrity for 30 days. As expected, when immersed in protease XIV the degradation rate of SF scaffolds increased. Based on the promising morphology and physicochemical properties of the developed SF scaffolds, in vitro chondrogenic differentiation studies with hASCs are envisioned in order to validate their performance for cartilage regeneration applications. Conclusion: This study proposes an innovative approach to produce fast-formed porous SF scaffolds using enzymatically cross- linked SF hydrogels structured by the combination of salt-leaching and freeze-drying methodologies. The obtained results can provide a valuable reference of SF as a tunable and versatile biomaterial with great potential for applications in cartilage TE scaffolding. Portuguese Foundation for Science and Technology (FCT) project PEst-C/SAU/LA0026/201; ERDP funding through POCTEP Project 0687_NOVOMAR_1_P; Investigator FCT program IF/00423/2012 and IF/00411/2013 References: [1] Xia, L.-W., R. Xie, X.-J. Ju, W. Wang, Q. Chen, and L.-Y. Chu, Nano-structured smart hydrogels with rapid response and high elasticity. Nature communications, 2013. 4. [2] Sofia, S.J., A. Singh, and D.L. Kaplan, Peroxidase-catalyzed crosslinking of functionalized polyaspartic acid polymers. Journal of Macromolecular Science, Part A, 2002. 39(10): p. 1151-1181. [3] Reis, R.L., L.-P. Yan, A.L. Oliveira, J.M. Oliveira, D.R. Pereira, C. Correia, and R.A. Sousa, Hydrogels derived from silk fibroin: Methods and uses thereof. 2014. 107426. [4] Chen, C.-H., J.M.-J. Liu, C.-K. Chua, S.-M. Chou, V.B.-H. Shyu, and J.-P. Chen, Cartilage tissue engineering with silk fibroin scaffolds fabricated by indirect additive manufacturing technology. Materials, 2014. 7(3): p. 2104-2119. [5] Yan, L.-P., J.M. Oliveira, A.L. Oliveira, S.G. Caridade, J.F. Mano, and R.L. Reis, Macro/microporous silk fibroin scaffolds with potential for articular cartilage and meniscus tissue engineering applications. Acta biomaterialia, 2012. 8(1): p. 289-301.Â

Dominant cataracts result from incongruous mixing of wild-type lens connexins

Gap junctions are composed of proteins called connexins (Cx) and facilitate both ionic and biochemical modes of intercellular communication. In the lens, Cx46 and Cx50 provide the gap junctional coupling needed for homeostasis and growth. In mice, deletion of Cx46 produced severe cataracts, whereas knockout of Cx50 resulted in significantly reduced lens growth and milder cataracts. Genetic replacement of Cx50 with Cx46 by knockin rescued clarity but not growth. By mating knockin and knockout mice, we show that heterozygous replacement of Cx50 with Cx46 rescued growth but produced dominant cataracts that resulted from disruption of lens fiber morphology and crystallin precipitation. Impedance measurements revealed normal levels of ionic gap junctional coupling, whereas the passage of fluorescent dyes that mimic biochemical coupling was altered in heterozygous knockin lenses. In addition, double heterozygous knockout lenses retained normal growth and clarity, whereas knockover lenses, where native Cx46 was deleted and homozygously knocked into the Cx50 locus, displayed significantly deficient growth but maintained clarity. Together, these findings suggest that unique biochemical modes of gap junctional communication influence lens clarity and lens growth, and this biochemical coupling is modulated by the connexin composition of the gap junction channels

Mediator complex component MED13 regulates zygotic genome activation and is required for postimplantation development in the mouse

Understanding factors that regulate zygotic genome activation (ZGA) is critical for determining how cells are reprogrammed to become totipotent or pluripotent. There is limited information regarding how this process occurs physiologically in early mammalian embryos. Here, we identify a mediator complex subunit, MED13, as translated during mouse oocyte maturation and transcribed early from the zygotic genome. Knockdown and conditional knockout approaches demonstrate that MED13 is essential for ZGA in the mouse, in part by regulating expression of the embryo-specific chromatin remodeling complex, esBAF. The role of MED13 in ZGA is mediated in part by interactions with E2F transcription factors. In addition to MED13, its paralog, MED13L, is required for successful preimplantation embryo development. MED13L partially compensates for loss of MED13 function in preimplantation knockout embryos, but postimplantation development is not rescued by MED13L. Our data demonstrate an essential role for MED13 in supporting chromatin reprogramming and directed transcription of essential genes during ZGA.Fil: Miao, Yi Liang. National Institutes of Health; Estados UnidosFil: Gambini, Andres. National Institutes of Health; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zhang, Yingpei. National Institutes of Health; Estados UnidosFil: Padilla Banks, Elizabeth. National Institutes of Health; Estados UnidosFil: Jefferson, Wendy N.. National Institutes of Health; Estados UnidosFil: Bernhardt, Miranda L.. National Institutes of Health; Estados UnidosFil: Huang, Weichun. National Institutes of Health; Estados UnidosFil: Li, Leping. National Institutes of Health; Estados UnidosFil: Williams, Carmen J.. National Institutes of Health; Estados Unido

Genipin-cross-linked collagen/chitosan biomimetic scaffolds for articular cartilage tissue engineering applications

This was the first study to use genipin to cross-link collagen and chitosan.In this study, genipin-cross-linked collagen/chitosan biodegradable porous scaffolds were prepared for articular cartilage regeneration. The influence of chitosan amount and genipin concentration on the scaffolds physicochemical properties was evaluated. The morphologies of the scaffolds were characterized by scanning electron microscope (SEM) and cross-linking degree was investigated by ninhydrin assay. Additionally, the mechanical properties of the scaffolds were assessed under dynamic compression. To study the swelling ratio and the biostability of the collagen/chitosan scaffold, in vitro tests were also carried out by immersion of the scaffolds in PBS solution or digestion in collagenase, respectively. The results showed that the morphologies of the scaffolds underwent a fiber-like to a sheet-like structural transition by increasing chitosan amount. Genipin cross-linking remarkably changed the morphologies and pore sizes of the scaffolds when chitosan amount was less than 25%. Either by increasing the chitosan ratio or performing cross-linking treatment, the swelling ratio of the scaffolds can be tailored. The ninhydrin assay demonstrated that the addition of chitosan could obviously increase the cross-linking efficiency. The degradation studies indicated that genipin cross-linking can effectively enhance the biostability of the scaffolds. The biocompatibility of the scaffolds was evaluated by culturing rabbit chondrocytes in vitro. This study demonstrated that a good viability of the chondrocytes seeded on the scaffold was achieved. The SEM analysis has revealed that the chondrocytes adhered well to the surface of the scaffolds and contacted each other. These results suggest that the genipin-cross-linked collagen/chitosan matrix may be a promising formulation for articular cartilage scaffolding.Key Projects in the National Science and Technology Pillar Program in the Eleventh Five-year Plan Period. Grant Number: 2006BA116B04Guangdong Natural Science Foundation. Grant Number: 07300602Natural Science Foundation Team Project of Guangdong. Grant Number: 4205786State Key Program of National Natural Science of China. Grant Number: 50732003National Basic Research Program of China. Grant Number: 2005CB62390

Silk-Fibroin/Methacrylated Gellan Gum Hydrogel as an novel scaffold for application in meniscus cell-based tissue engineering

Introduction: Knee meniscus injury is highly prevalent and there is a demand for new cost-effective treatment solutions. Tissue engineering (TE) and regenerative medicine strategies using acellular scaffolds are being used in clinical application for total or partial meniscus replacement [1]. Although this strategy has been considered as a safe and promising approach, progressive volume reduction of the implant and early failure have been described. Advances in the field of meniscus TE are required and greatly depend on increased knowledge of meniscus biology, improvement of biomaterials and cell-based therapies [2]. Advanced scaffolds for meniscus TE should possess adequate mechanics, biodegradability and biocompatibility, and also be able to mimic and preserve the asymmetric vascular network of this complex tissue, i.e. enable controlling the segmental vascularization during the regeneration process. Silk fibroin scaffolds derived from Bombyx mori cocoon have been recognized as a versatile biomaterial for application in meniscus TE [3]. The purpose of this study is to: 1) contribute to the knowledge of meniscus aiming future clinical applications (namely, the aspects dealing with the characterization of cellular phenotypes and density, biomechanics and extracellular matrix composition) and 2) to develop an alternative and viable silk fibroin scaffold possessing adequate properties for either use in acellular or cellular approaches for partial and/or total meniscus replacement, and combine it with the methacrylated gellan gum hydrogel (iGG-MA) hydrogel, which is able to prevent the ingrowth of endothelial cells and blood vessels into the hydrogels [4,5]. Patients & Methods: Morphologically intact menisci were collected from 44 human donors (12 male, 32 female). All menisci (30 lateral and 14 medial) were divided into anterior, middle and posterior segments prior to mechanical, biological or histological characterization. Human meniscus cells (HMC´s) were isolated using an enzymatic standard protocol. HMC´s phenotype was characterized by flow cytometry analysis. Haematoxylin and eosin (H&E), safranin-O and collagen I staining were performed for segmental characterization of the extracellular matrix. For the evaluation of the viscoelastic properties, dynamic mechanical analysis (DMA) was performed using fresh tissue samples. The three segments of menisci were cut in cylindrical shapes with 4 mm diameter and analyzed at 37ºC in PBS (pH 7.4). The microstructure of freeze-dried meniscus was investigated by micro-computed tomography (micro-CT) analysis. Silk-based scaffolds (10 and 12 wt%) were produced by means of combining salt leaching and freeze-drying methods [3], in order to match human tissue biological and biomechanical features. HMC’s were seeded onto the different silk scaffolds at a cell density of 5x104 cells/disc. Then, the cell-laden scaffolds were cultured in static conditions, for times of culturing up to 21 days. After specific times of culturing (from 1 day up to 21 days), HMC´s adhesion, viability and proliferation were investigated by scanning electron microscopy (SEM), calcein-AM assay and DNA quantification tests, respectively. In addition, the mechanical properties of the cell-loaded scaffolds were evaluated by DMA. The HMC’s-laden hydrogel/silk scaffolds were produced by encapsulating the HMC’s into a 2 wt% iGG-MA hydrogel, followed by impregnation onto the 12 wt% silk scaffold. A chorioallantoic membrane (CAM) assay was used to investigate in vivo the control of segmental vascularization of the acellular and cell-laden hydrogel/silk scaffolds by the effect of iGG-MA hydrogel, until day 14 of embryonic development. Results & Discussion: The biological characterization of this meniscus tissue, although not yet completely accomplished, has evolved significantly in the last few years. In this work, DMA analysis has shown that medial meniscus has significantly higher stiffness (E' and Tan d) than lateral meniscus. There is also significant regional variation form anterior to posterior menisci segments regarding biomechanical features. Age, gender and bone mass index (BMI) also influences meniscus stiffness. The FACS analysis revealed that cells isolated from the human meniscus are a mixed population of cells, i.e. fibrochondrocyte-like and MSCs (cells are positive for CD105, CD73 and CD90, and lack CD34 and CD45). HMC’s maintained their phenotype for 21 days when cultured in tissue culture polystyrene plate (2D). The micro-CT analysis revealed that the human freeze-dried meniscus possessed a mean porosity of 58.0±20.3% and interconectivity of 41.9%±53.7. The mean pore size and trabeculae thickness was 220.7±111.5 µm and 159.7±78.6 µm, respectively. The knowledge arising from the present study allowed us to develop a novel polymeric scaffold made of silk fibroin, which was subsequently characterized without cells and after cell-loading. SEM analysis revealed that the HMC´s adhered to the surface of the scaffolds. The viability assay and DNA quantification showed that HMC´s were viable and proliferated well when cultured onto both silk-10 and silk-12 scaffolds, until 21 days. DMA analysis has shown that the moduli of the acellular scaffolds immersed in culture medium for 14 days were 27.6 ± 7.9 kPa and 61.1 ± 0.4 at 10 Hz, for silk-10 e silk-12, respectively. By its turn, the moduli determined at 10 Hz of the cell-laden scaffolds cultured after 14 days of culturing were 48.2± 19.8 and 140.1 ± 15.6 kPa, for silk-10 and silk-12, respectively. The in vivo study allowed investigating the number of macroscopic blood vessels converging to the implants. The evaluation of possible inflammation and endothelial cells ingrowths was performed by histological (H&E staining) and immunohistochemical methods (SNA-lectin staining). Results have shown that iGG-MA hydrogel prevented the endothelial cells adhesion and blood vessels infiltration into the HMC’s hydrogel/silk scaffolds, after 4 days of implantation, even in the presence of VEGF

Bioactive macro/micro porous silk fibroin/Nano-sized calcium phosphate scaffolds with potential for bone tissue engineering applications

Aim: The development of novel silk/nano-sized calcium phosphate (silk/nano-CaP) scaffolds with highly dispersed CaP nanoparticles in the silk fibroin (SF) matrix for bone tissue engineering. Materials & methods: Nano-CaP was incorporated in a concentrated aqueous SF solution (16 wt.%) by using an in situ synthesis method. The silk/nano-CaP scaffolds were then prepared through a combination of salt-leaching/ lyophilization approaches. Results: The CaP particles presented good affinity to SF and their size was inferior to 200 nm when theoretical CaP/silk ratios were between 4 and 16 wt.%, as determined by scanning electron microscopy. The CaP particles displayed a uniform distribution in the scaffolds at both microscopic and macroscopic scales as observed by backscattered scanning electron microscopy and micro-computed tomography, respectively. The prepared scaffolds presented self-mineralization capability and no cytotoxicity confirmed by in vitro bioactivity tests and cell viability assays, respectively. Conclusion: These results indicated that the produced silk/nano-CaP scaffolds could be suitable candidates for bone-tissueengineering applications.This study was funded by the Portuguese Foundation for Science and Technology (FCT) through the projects Tissue2Tissue (PTDC/CTM/105703/2008) and Osteo Cart (PTDC/CTM-BPC/115977/2009). The funding from Foundation Luso-Americana is greatly acknowledged. L-P Yan gives thanks for his PhD scholarship from FCT (SFRHIBD/64717/2009). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or potions, expert testimony, grants or patents received or pending, or royalties