Presence of microplastics and nanoplastics in food, with particular focus on seafood

Acknowledgements: The Panel wishes to thank the members of the Working Group on the presence of microplastics and nanoplastics in food, with particular focus on seafood: Francesco Cubadda, Christer Hogstrand, Peter Hollman, Hendrik Van Loveren, Anne-Katrine Lundebye and Annette Petersen for the preparatory work on this statement, the hearing expert: Stephanie Wright and EFSA staff member: Karen Mackay for the support provided to this statement.

Joint interpretation of magnetotelluric, seismic, and well-log data in Hontomín (Spain)

Acknowledgements. This work is dedicated to the memory of Andrés Pérez-Estaún, brilliant scientist, colleague, and friend. The authors sincerely thank Ian Ferguson and an anonymous reviewer for their useful comments on the manuscript. Xènia Ogaya is currently supported in the Dublin Institute for Advanced Studies by a Science Foundation Ireland grant IRECCSEM (SFI grant 12/IP/1313). Juan Alcalde is funded by NERC grant NE/M007251/1, on interpretational uncertainty. Juanjo Ledo, Pilar Queralt and Alex Marcuello thank Ministerio de Economía y Competitividad and EU Feder Funds through grant CGL2014- 54118-C2-1-R. Funding for this Project has been partially provided by the Spanish Ministry of Industry, Tourism and Trade, through the CIUDEN-CSIC-Inst. Jaume Almera agreement (ALM-09-027: Characterization, Development and Validation of Seismic Techniques applied to CO2 Geological Storage Sites), the CIUDEN-Fundació Bosch i Gimpera agreement (ALM-09-009 Development and Adaptation of Electromagnetic techniques: Characterisation of Storage Sites) and the project PIERCO2 (Progress In Electromagnetic Research for CO2 geological reservoirs CGL2009-07604). The CIUDEN project is co-financed by the European Union through the Technological Development Plant of Compostilla OXYCFB300 Project (European Energy Programme for Recovery).

Pressure radiation from a perforated duct exit region

Acknowledgements The authors are grateful to the following bodies that provided financial support for the project: (i) China Scholarship Council, (ii) National Natural Science Foundation of China (Grant no. U1334201) and (iii) UK Engineering and Physical Sciences Research Council (Grant no. EP/G069441/1).

Electrically Tunable Selective Reflection of Light from Ultraviolet to Visible and Infrared by Heliconical Cholesterics

Acknowledgements This work was supported by NSF DMR-1410378 and DMR-1121288. We thank V. Borshch for helping with preparation of illustrations, to Y. K. Kim for the help in experiments, V. A. Belyakov and S. V. Shiyanovskii for useful discussions.

Account book of the executors of the Henry Flynt estate, 1760-1767

The volume contains acknowledgements of the disbursements of Harvard Tutor Henry Flynt's estate written in the hands of the respective beneficiaries. The entries begin on February 27, 1760 following Flynt's death on February 13, 1760, and continue through May 9, 1767. Each receipt includes the date, name of the executors, description of the property, beneficiary's name, and signature. The beneficiaries include the wife of Sol. Davy, Dorothy Jackson, Edmund Quincy, J. Henry Quincy, Esther and Stephen Richard (received by attorney Nicholas Boylston), Dorothy Skinner (also received for her by her husband Richard Skinner), John Wendell, Edmund Wendell, Katherine Wendell, and Oliver Wendell, as well as Harvard College (received by Harvard Treasurer Thomas Hubbard), and the Deacons of the First Church of Cambridge. The volume also includes a loose document titled "Account from Messrs Edmund & Josiah Quincy Settled & Ballanced March 31, 1749."

Genetically engineered silk and genipin-enhanced fibrin hydrogel for annulus fibrosus repair

INTRODUCTION: Around 80% of people are affected by low back pain at least once in their life, often caused by trauma provoking intervertebral disc (IVD) herniation and/or IVD degeneration. Apart from some promising approaches for nucleus pulposus repair, so far no treatment or repair is available for the outer fibrous tissue, annulus fibrosus (AF). We aimed for sealing and repairing an AF injury in a bovine IVD organ culture model in vitro over 14 days under different loading conditions. For this purpose, a silk fleece composite from Bombyx mori silk was combined with genipin-enhanced fibrin hydrogel [1]. METHODS: Bovine IVDs of 12-17 months old animals were isolated by first removing all surrounding tissue, followed by cutting out the IVDs [2]. Culturing of discs occurred in high glucose Dulbecco's Modified Eagle Medium (HG-DMEM) supplemented with 5% serum as previously described. On the next day, injury was induced using a 2mm biopsy punch (Polymed, Switzerland). The formed cavity was filled with (0.4%) genipin-enhanced human based fibrin hydrogel (35- 55mg/mL human fibrinogen, Baxter, Austria) and sealed with a silk fleece-membrane composite (Spintec Engineering, Germany). Different culture conditions were applied: free swelling, static diurnal load of 0.2MPa for 8h/d and complex loading at 0.2MPa compression combined with ± 2° torsion at 0.2Hz for 8h/d. Complex loading was applied by a custom built 2 degree of freedom bioreactor [3]. After 14 days of culture cell activity was determined with resazurin assay. Additionally, glycosaminoglycan (dimethyl-methylene blue), DNA (Hoechst) and collagen content (hydroxy-proline) were determined. Finally, real-time qPCR of major IVD marker genes was performed. RESULTS: The silk seal closing the injury site could successfully withstand the forces of all three loading conditions with no misplacement over the two weeks’ culture. Nevertheless, disc height of the repaired discs did not significantly differ from the injured group. The disc phenotype could be maintained as demonstrated by biochemical analysis of gene expression, cell activity, DNA-, collagen- and GAG content. The silk itself was evaluated to be highly biocompatible for hMSC, as revealed by cytotoxicity assays. DISCUSSION & CONCLUSIONS: The silk can be considered a highly-elastic and biocompatible material for AF closure and the genipin-enhanced fibrin hydrogel has also good biomechanical properties. However, the cyto-compatibility of genipin seems rather poor and other hydrogels and/or cross-linkers should be looked into. REFERENCES: 1 C.C. Guterl et al. (2014) Characterization of Mechanics and Cytocompatibility of Fibrin Genipin Annulus Fibrosus Sealant with the Addition of Cell Adhesion Molecules, Tissue Eng Part A 2 S.C. Chan, B. Gantenbein-Ritter (2012) Preparation of intact bovine tail intervertebral discs for organ culture, J Vis Exp 3 B Gantenbein et al. (2015) Organ Culture Bioreactors - Platforms to Study Human Intervertebral Disc Degeneration and Regenerative Therapy, Curr Stem Cell Res Ther [epub ahead of print] ACKNOWLEDGEMENTS: This project is supported by the Gebert Rüf Stiftung project # GRS-028/13.

Non-Viral Gene Therapy to Cells of the bovine Intervertebral Disc

Background. Low back pain is an increasing global health problem, which is associated with intervertebral disc (IVD) damage and degeneration. Major changes occur in the nucleus pulposus (NP), with the degradation of the extracellular matrix (ECM).1 Further studies showed that growth factors from transforming growth factor β (TGFβ) and bone morphogenic proteins (BMP) family may induce chondrogenic differentiation of mesenchymal stem cells (MSC).2 Focusing on non-viral gene therapies and their possible translation into the clinics, we investigated if GDF6 (syn. BMP13 or CDMP2) can induce regeneration of degraded NP. We hypothesized that IVD transfected with plasmid over-expressing GDF6 also up-regulates other NP- and chondrogenic cell markers and enhances ECM deposition. Methods. Bovine nucleus pulposus (bNPC) and annulus fibrosus cells (bAFC) were harvested from bovine coccygeal IVD. Primary cells were then electroporized with plasmid GDF6 (Origene, vector RG211366) by optimizing parameters using the Neon Transfection system (Life Technologies, Basel). After transfection, cells were cultured in 2D monolayer or 3D alginate beads for 7, 14 or 21 days. Transfection efficiency of pGDF6 was analyzed by immunohistochemistry and fluorescent microscopy. Cell phenotype was quantified by real-time RT-PCR. To test a non-viral gene therapy applied directly to 3D whole organ culture, coccygeal bovine IVDs were harvested as previously described. Bovine IVDs were transfected by injection of plasmid GDF6 into the center. Electroporation was performed with ECM830 Square Wave Electroporation System (Harvard Apparatus, MA) using 2-needle array electrode or tweezertrodes. 72 h after tranfection discs were fixed and cryosectioned and analyzed by immunofluorescence against GDF6. Results. RT-PCR and immunohistochemistry confirmed up-regulation of GFP and GDF6 in the primary bNPC/bAFC culture. The GFP-tagged GDF6 protein, however, was not visible, possibly due to failure of dimer formation as a result of fusion structure. Organ IVD culture transfection revealed GDF6 positive staining in the center of the disc using 2-needle array electrode. Results from tweezertrodes did not show any GDF6 positive cells. Conclusion. Non-viral transfection is an appealing approach for gene therapy as it fulfills the translational safety aspects of transiency and lacks the toxic effects of viral transduction. We identified novel parameters to successfully transfect primary bovine IVD cells. For transfection of whole IVD explants electroporation parameters need to be further optimized. Acknowledgements. This project was funded by the Lindenhof Foundation (Funds “Research & Teaching”) Project no. 13-02-F. The imaging part of this study was performed with the facility of the Microscopy Imaging Center (MIC), University of Bern. References. Roughly PJ (2004): Spine (Phila), 29:2691-2699 Clarke LE, McConell JC, Sherratt MJ, Derby B, Richardson SM, Hoyland JA (2014), Arthritis Research & Therapy, 16:R67

The friction of an oscillating bearing,

"The work described has been carried out in the Engineering Department of the National Physical Laboratory."--Acknowledgements.

Health education; a program for girls in secondary schools.

"Acknowledgements" signed: Esther Sherman.

Skilled nursing facilities : available data show average nursing staff time changed little after Medicare payment increase : report to congressional committees.

"GAO-03-176."

Growing older in Illinois : needs, challenges, opportunities.

"This report was written by the Heartland Center on Aging, Disability and Long Term Care, School of Public and Environmental Affairs, Indiana University and the National Center for Senior Living, South Bend, Indiana. May, 1991." Acknowledgements.

Think small! / curated by Robert Sill.

"The catalog was made possible in part by financial support from Chase Bank and the Illinois State Museum."

The Pilgrim hymnal with responsive readings.

Preface -- Contents -- Order of Subjects -- The hymns -- Sentences and canticles -- Canticles -- Responsive readings and other aids to worship -- Editorial notes -- Acknowledgements -- Indexes of tunes -- Indexes of hymns -- Index of sentences and canticles.

A history of education in the United States,

"With acknowledgements to J. E. Miller, A.M., and to other members of my educational seminar for valuable assistance in the preparation of material."

Federal employees' health benefits : effects of using pharmacy benefit managers on health plans, enrollees, and pharmacies : report to the Honorable Byron L. Dorgan, U.S. Senate.

"GAO-03-196."