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In vitro mammalian cell culture has served as an invaluable tool in cell biology for several decades. Classically, monolayer cultures of adherent cells were grown on flat and rigid two-dimensional (2D) substrates, such as polystyrene or glass. However, many cells, when isolated from tissues and placed onto stiff planar 2D cell culture surfaces, such as tissue culture plastic, become progressively flatter, divide aberrantly, and lose their differentiated phenotype1,2. While these two-dimensional cell culture studies have played a pivotal role in furthering our understanding of many biological processes, they do not emulate in vivo conditions.
In vitro mammalian cell culture has served as an invaluable tool in cell biology for several decades. Classically, monolayer cultures of adherent cells were grown on flat and rigid two-dimensional (2D) substrates, such as polystyrene or glass. However, many cells, when isolated from tissues and placed onto stiff planar 2D cell culture surfaces, such as tissue culture plastic, become progressively flatter, divide aberrantly, and lose their differentiated phenotype1,2. While these two-dimensional cell culture studies have played a pivotal role in furthering our understanding of many biological processes, they do not emulate in vivo conditions.
In vitro mammalian cell culture has served as an invaluable tool in cell biology for several decades. Classically, monolayer cultures of adherent cells were grown on flat and rigid two-dimensional (2D) substrates, such as polystyrene or glass. However, many cells, when isolated from tissues and placed onto stiff planar 2D cell culture surfaces, such as tissue culture plastic, become progressively flatter, divide aberrantly, and lose their differentiated phenotype1,2. While these two-dimensional cell culture studies have played a pivotal role in furthering our understanding of many biological processes, they do not emulate in vivo conditions.
In the body, cells reside in a complex extracellular matrix (ECM) consisting of a three-dimensional (3D) architecture that allows direct interactions with neighboring cells and matrix molecules through biochemical and mechanical cues. Cell–cell and cell–ECM interactions establish a 3D communication network that maintains the specificity and homeostasis of the tissue3. Therefore it is not surprising that fundamental differences in the microenvironment of 2D and 3D cell culture systems influence various cellular behaviors including the way in which cells attach, spread and grow, their morphology and polarity, gene and protein expression, viability, proliferation, motility, differentiation, response to stimuli, cellular metabolism, and overall function1,2,4-16.
The 3D ECM surrounding a cell performs several critical functions. To begin with, it provides a complex, nanoscale architecture of structural proteins such as collagen, laminin, fibronectin and glycosaminoglycans to establish the mechanical properties inherent in the cellular microenvironment17,18. Cells sense these mechanics through their cell surface integrins and bind to specific adhesion motifs present on the ECM proteins and glycosaminoglycans. Furthermore, the ECM is vital for sequestering soluble biomolecules and growth factors, and releasing these signaling molecules with spatial-temporal control to guide processes such as cell migration, matrix degradation and deposition17,18. Thus, to truly mimic the native ECM, it is necessary to develop 3D culture models that exhibit the mechanical and chemical properties of the ECM, not only at the initial stage of seeding the cells, but rather, in a dynamic and tunable manner as the cells grow and develop. To address the requirements of different cell types and applications in the body, a vast array of materials and fabrication techniques have been employed to develop 3D microenvironments with different physical, biological, and mechanical characteristics. Each model comes with its own set of advantages and limitations, and one distinct model is not suitable for all applications.
3D Culture systems
A. 3D Cultures using Hydrogels and Scaffolds
Hydrogels are comprised of networks of cross-linked polymer chains or complex protein or polysaccharide-like molecules of natural or synthetic origin. Due to their significant water content and range of tunable mechanical properties, hydrogels possess biophysical characteristics similar to natural tissue, and serve as highly effective matrices for 3D cell culture. Hydrogels can be used as stand-alone 3D matrices, as a coating, or as a method of encapsulating cells to be combined with other technologies, such as solid scaffolds, permeable supports, cellular microarrays, 3D printing or microfluidics devices. The morphology, growth and functionality of cells within the hydrogel depend on the presentation of biophysical and biochemical cues, as well as physical properties such as permeability and matrix stiffness.
Natural Hydrogels and Extracellular Matrices (ECMs)
Naturally derived hydrogels for cell culture are typically formed of proteins, polysaccharides and ECM components such as collagen, laminin, fibrin, hyaluronic acid, chitosan or Matrigelä (a reconstituted basement membrane extract from Engelbreth–Holm–Swarm mouse tumors containing a mixture of laminin, collagen IV, heparin sulfate proteoglycan, and nidogen/entactin19, Figure 1). Gels derived from natural sources are inherently biocompatible and bioactive20. They also promote many cellular functions due to the presence of various endogenous factors, which can be advantageous for supporting viability, proliferation, function and development of many cell types21.
However, natural hydrogels do present some disadvantages, including their isolation from animal-derived sources, and inherent batch-to-batch variability in composition. Also, they contain endogenous bioactive components such as growth factors that can be advantageous for creating some 3D models, but in other instances, can confound the specific cell behavior or response that is under investigation. For instance Matrigel activates pathways that control angiogenesis22,23, cancer cell motility24, and drug sensitivity25 while collagen type 1 alters the production of matrix metalloproteinases (MMPs), enzymes that degrade ECM components and allow tumor cell invasion26, cell sensitivity to anti-cancer drugs27, cell proliferation, and migration28,29.
To circumvent some of the issues associated with animal-derived biomaterials, matrices have been developed in organisms that are animal-free, or derived from recombinant nucleic acid technology. Hyaluronic acid (hyaluronan or HA) is an increasingly popular polysaccharide-based biologically derived matrix30-32. Most commercial grade HA is of bacterial origin, and characterized by high purity and homogeneous quality. These gels may be modified by the addition of ECM components for improving cell attachment and growth properties.
Synthetic Hydrogels and Extracellular Matrices
Synthetic hydrogels are an option for 3D cell culture applications when naturally derived biological matrices are unsuitable. Synthetic hydrogels are formed of purely non-natural molecules33 such as poly(ethylene glycol) (PEG)34, poly(vinyl alcohol)35, and poly(2-hydroxy ethyl methacrylate)36. They are biologically inert, but provide structural support for various cell types. They are also highly reproducible, allow for facile tuning of the mechanical properties, and are simple to process and manufacture.
Synthetic hydrogels can maintain the viability of encapsulated cells while allowing for ECM deposition as the hydrogel degrades37, thereby demonstrating that synthetic gels can function as 3D cell culture platforms in the absence of integrin-binding ligands. As an alternative approach inert synthetic hydrogels can be modified with appropriate biological components. An example of a synthetic hydrogel that is tunable is Corning PuraMatrix™ Peptide Hydrogel, a self-assembling, synthetic oligopeptide that exhibits nanometer scale fibers (Figure 2). Synthetic hydrogels can be customized with specific peptide sequences to improve cell attachment, cell homing, and other behaviors38 or the hydrogels can be supplemented with bioactive molecules (e.g., growth factors or ECM proteins). Furthermore, devices such as permeable supports (e.g., Transwell® inserts) can be incorporated into 3D culture models with hydrogel substrates to study the interaction between different cell types, soluble factors, and the culture environment.
Solid Scaffolds
Solid scaffolds for 3D cell culture are fabricated with a broad range of materials including metal, ceramics, glass, and polymers. In particular, polymers are a common choice for generating solid scaffolds of diverse size, varying structure, stiffness, porosity, and permeability39. A multitude of fabrication techniques are being utilized to generate solid scaffolds for 3D cell culture, including soft-lithography, electro-spinning, microarray, bio-printing, and many others. The major drawbacks of using solid scaffolds are limited scope for cell imaging and difficulties that are encountered when recovering cells from the matrix.
An important consideration for designing scaffolds for 3D cell culture is the scale and topography of the internal structures within the scaffold. In the body, the ECM provides an intricate nanoscale infrastructure to support cells, and presents an instructive background that governs their behavior40-44. Cells binding to scaffolds that exhibit microscale architectures usually flatten and spread out as they would if cultured on flat surfaces45. Even minute nanoscale level alterations in topography of the cell's environment can elicit diverse effects on cell behavior46. Apart from scale and structure, the material used for constructing the scaffold, the surface chemical properties, matrix stiffness, permeability, and mechanical forces can significantly impact cell adhesion, growth, and behavior47.
B. Spheroids
Cellular spheroids are simple 3D models that can be generated from a wide range of cell types, which form spheroids naturally due to the tendency of adherent cells to aggregate. Common examples of spheroids include embryoid bodies, mammospheres, tumor spheroids, hepatospheres, and neurospheres. In particular, adherent cells tend to aggregate under circumstances that impede adhesion to cell culture substrates. Common matrix-free methods employed for generating spheroids include the use of ultra-low attachment surfaces for cell culture, or by maintaining the cells as suspension cultures in media (e.g. hanging drop technology, rotary cultures, and bioreactors). Several cell types also form spheroids in 3D hydrogels, and to a limited extent, in some solid scaffolds depending on the structural and mechanical properties of the material. The overall size of the spheroids is limited to a few hundred micrometers, beyond which, necrosis ensues within the core of the spheroids48.
As cells spontaneously aggregate into spheroids they naturally mimic various aspects of solid tissues and are equipped with inherent gradients for efficient diffusion of oxygen and nutrients as well as the removal of metabolic wastes. These cellular aggregates can emulate avascular, solid tumor behavior more effectively than standard 2D environments because spheroids, much like tumors, usually contain a heterogeneous population of surface-exposed and deeply buried cells, proliferating and non-proliferating cells, and well-oxygenated and hypoxic cells49. Additionally, differentiation of pluripotent stem cells typically involves the formation of spherical structures called embryoid bodies, an important step for subsequent cell differentiation studies in vitro. Spheroids thus represent an especially good physiological 3D model for studying solid tumorigenesis and stem cell differentiation. In addition, spheroids can be readily analyzed by imaging using light, fluorescence, and confocal microscopies, which is an advantage over more complex 3D cell culture models. Furthermore, it is relatively simple to mass-produce uniformly sized 3D spheroids making them highly amenable for many in vitro high-throughput (HTP) and toxicity screening applications.
C. Organoids
Another major advancement in stem cell differentiation is the formation of self-organizing 3D mini "organs" known as organoids. Organoids can recapitulate histological details and provide functional representation of multiple cell types that are present within the native organs. Organoids representative of intestinal50,51, retinal52, pancreatic53, mammary54, colonic55, and cerebral tissues56 have been developed recently using 3D models comprised of Matrigel Matrix. Figure 3 illustrates a schematic of 3D culture conditions that enabled efficient expansion of dissociated mouse embryonic pancreatic progenitors53. Another example is shown in Figure 4, where Lancaster et al. generated cerebral organoids with structural similarity to brain tissue57 . While, in vitro culture of organoids is a major step toward elucidating the principles of organ development and the mechanisms responsible for genetic diseases, the consistency and predictability of obtaining the same structures varies from sample to sample57. Similar to spheroids, they are also limited in size by the development of necrotic cores, limiting their usefulness for regenerative approaches.
D. Perfusion bioreactors
One of the major challenges of creating 3D cultures in vitro is the limited mass transfer distances required for nutritional supply and removal. Bioreactors address this challenge by controlling the physiological environment surrounding 3D constructs (i.e. nutrient supply, oxygen tension, waste removal, mechanical inputs, pH, and temperature). Bioreactors therefore are essential when larger tissue constructs are required and also to sustain tissues in vitro for extended periods of time, as increased transport through perfusion helps to avoid necrotic centers. In particular, bioreactors can be used to sustain long term cultures where dynamic changes can be monitored58. Additionally bioreactors control the size and shape of 3D cultures, and promote uniform cell distributions59-64. Bioreactors come in many different formats including: (direct) perfusion bioreactors65, (indirect) hollow fiber perfusion bioreactors66, perfusion bioreactors with mechanical stimulation67, rotating wall vessel bioreactors68, spinner flasks69, and orbital shakers70.
E. Other ways to address limited mass transfer
Another physiologically relevant solution to address limited mass transfer in large tissue constructs is to recreate the tissue vasculature (which provides and removes nutrients in vivo) with interconnected, vascularizable channels 71. As an artificial substitute to support high-density cell growth, 'pseudovascularization' through hollow fiber membrane bioreactors has also shown promising results66,72-74. The hollow fibers act as the blood vessels transferring culture medium throughout the tissue engineered constructs while the bulk can be seeded with extracellular matrix or other scaffold materials and cells.
Recent improvements in microscale engineering techniques, such as fabrication of microfluidic devices and microarrays have the potential to address nutrient supply issues by creating micro scale models. Microfluidic devices manipulate small volumes (10-9 to 10-6 L) to generate and precisely control dynamic fluid flow and spatio-temporal gradients, as well as deliver nutrients and other chemical cues to cells in a controlled manner75. Microfluidic networks have been incorporated directly within cell embedded hydrogels, as well as several other matrices, to enable efficient convective transport of nutrients and other soluble cues throughout the 3D scaffolds76. Microarrays consist of a solid support wherein small volumes of different biomolecules and cells can be positioned in defined locations, allowing multiplexed investigation of living cells and their responses to stimuli. Use of these technologies can improve cell viability and functions. Moreover, microfluidic technologies can increase throughput and significantly reduce the cost of culture due to low reagent volume requirements.
F. Cell considerations
Different types of cells can be incorporated in 3D models resulting in altered outcomes. There are three main classifications of cells that can be used: stem cells, primary cells, and immortalized cell lines. Stem cells are undifferentiated cells that can be stimulated to differentiate down different lineages. Stem cells are an attractive option since they self-renew in culture and can be used to create patient specific models. There are many types of stem cells which are defined by their stage of development. The different types include: adult stem cells (often from bone marrow or adipose tissue), fetal stem cells, cord blood stems, embryonic stem cells, or induced pluripotent stem cells. Primary cells on the other hand are obtained directly from a specific tissue and therefore are already committed to a specific lineage. They have a limited lifetime in culture and thus are more limited in numbers, and they eventually undergo senescence and stop proliferating. Immortalized cells in contrast have been altered to proliferate and evade cellular senescence. Gene delivery systems are also an option for modulating a specific phenotype in tissue engineered models.
In the body, communication between different cell types is critical to maintaining tissue homeostasis and specificity. Enhanced differentiation and survival is accomplished by co-culturing appropriate cell types77-81. Furthermore, co-cultures lead to increased ECM deposition82,83, improved functional outcome measurements84, proper morphology83,84, and vascular structures81 not achieved in monocultures. Besides mimicking normal physiology better, 3D co-culture models have been useful for studying complex interactions between cancer cells and other cell types by elucidating their contribution to tumor growth, vascularization and metastasis.
Unique cellular considerations for organs that interface with air have resulted in cultures of multiple cell types at an air-liquid interface, including lung85, epidermis86, cornea87, and inner cheek models88.
Culturing cells at an air-liquid interface using permeable supports has enabled the development ofin vitro3D models for drug discovery, including modeling of the lung epithelium. Using cells from diseased donors, these cellular models are used for studying a variety of pulmonary disorders.
Representative Results
Figure 1. In vitro developed endometrial glands display epithelial apicobasal polarity. Double immunostaining of either in vitro developed glands cultured for 8 days in BIE medium (A–E) or cryostat sections obtained from mouse endometrium (F–I). A and F: Cytokeratin (green) and vimentin (red). B and G: Laminin (green) and phalloidin (red). C and I: GM130 (green) and phalloidin (red). D and H: ZO-1 (green) and E-cadherin (red). E: ZO-1 (green) and b-catenin (red). White scale bar = 20 mm. BIE medium = basal medium supplemented with 5ng/mL EGF and 1:100 dilution of ITS supplement and 3% fresh Matrigel™ Matrix; EGF = epidermal growth factor; ITS = insulin transferrin selenium (Eritja et al., 2010)89.
Figure 1 description - Glandular epithelial cell organization, signaling, and secretion more closely resemble the properties observed in vivo when cultured in a 3D environment, in contrast to the behavior that occurs on 2D surfaces 10,11. A 3D in vitro model for endometrial glands is shown in Figure 1. In this model, endometrial epithelial cells were cultured in medium containing Matrigel™ Matrix for 7-8 days followed by immunofluorescence staining of various polarity markers. Marker expression analysis confirmed that the glandular structures were composed exclusively of cells of epithelial origin (1A and 1F). Furthermore, spheroids representing endometrial glands displayed correct apical-basolateral polarity (1B and 1G), positioning of Golgi apparatus (1C and 1I), adherent junctions (ID and IH), and tight junctions (IE).
Figure 2. Proliferation and differentiation in 3D scaffold. (A + B) Transmission light picture of proliferating cells in PuraMatrix with 0.25 % laminin (PML) and differentiating cells in PML 0.25% (C + D). Scanning electron microscope picture of proliferating cells and differentiated cells in PML 0.25%. (B + D) Upon induction of differentiation one observes the development of a dense 3 dimensional network of processes. (E) Immunocytochemistry for bIII-tubulin and TH of uninduced cells in PML 0.25% and (F) Cells after 7 days of differentiation revealed a dense network of bIII-tubulin positive cells. TH+ cells were found to possess processes, but without building up a dense network (Ortinau et al. 2010)90.
Figure 2 description – In this study, Ortinau and colleagues used a synthetic hydrogel, PuraMatrix™ supplemented with laminin (PML 0.25%) for neuronal differentiation of human neural progenitor cells90. Using transmission light microscopy (FIGURE 2A + B) and scanning electron microscopy (FIGURE 2C + D) the researchers showed that neural progenitor cells can develop into a dense network of neuronal processes (FIGURE 2B + D). Furthermore, immunocytochemistry revealed that after 7 days of differentiation, the neuronal precursor cells began to express neuronal markers (bIII-tubulin and tyrosine hydroxylase (TH)) (FIGURE 2E + F). These observations demonstrate how functionalization with appropriate biological factors can convert inert synthetic matrices into useful in vitro models for supporting differentiation of stem and progenitor cells.
Fig. 3. Organoids recapitulate progenitor expansion and organized differentiation.
(A-E) Immunohistochemistry on sections of 7-day organoids showing that (A) all cells (DRAQ5, red nuclei) are epithelial (E-cadherin) and many proliferate [phospho-histone H3 (pHH3)] and (B, C) retain pancreatic markers PDX1, SOX9 and HNF1B. (C) Cells polarize and form tubes lined by mucin 1. (D) Exocrine differentiation (amylase) is seen at the periphery. (E) Endocrine differentiation (insulin) is detected in the center. The section in B is close to that in A, and the section in D is close to that in C. (F) Experimental scheme to test endocrine differentiation after back- transplantation of cells grown in organoid in a pancreatic niche. WT, wild type. (G-J) The cells that were first grown in vitro integrate into the host epithelium (white in G and green in H- J). Some remain progenitors (G; HNF1B), some become acinar (H; amylase) or ductal (I; DBA) and others become endocrine (glucagon or insulin). Insets are magnifications of the dashed boxes. Scale bars: 50 μm (Greggio et al., 2013)53.
Figure 3 description – Figure 3 illustrates a schematic of 3D culture conditions that enabled efficient expansion of dissociated mouse embryonic pancreatic progenitors53. By manipulating the medium composition, researchers generated either hollow spheres, which are mainly composed of pancreatic progenitors, or complex organoids that spontaneously undergo pancreatic morphogenesis and differentiation.
Figure 4 | Description of cerebral organoid culture system. A, Schematic of the culture system used to generate cerebral organoids. Example images of each stage are shown. bFGF, basic fibroblast growth factor; hES, human embryonic stem cell; hPSCs, human pluripotent stem cells; RA, retinoic acid. B, Neuroepithelial tissues generated using this approach (left) exhibited large fluid-filled cavities and typical apical localization of the neural N-cadherin (arrow). These tissues were larger and more continuous than tissues grown in stationary suspension without Matrigel (right). C, Sectioning and immunohistochemistry revealed complex morphology with heterogeneous regions containing neural progenitors (SOX2, red) and neurons (TUJ1, green) (arrow). D, Low-magnification bright-field images revealing fluid-filled cavities reminiscent of ventricles (white arrow) and retina tissue, as indicated by retinal pigmented epithelium (black arrow). Scale bars, 200 mm (Lancaster et al., 2013)57.
Figure 4 description – Lancaster et al. generated cerebral organoids with remarkable structural similarity to brain tissue57. These cerebral organoids grew larger than typical cellular spheroids and survived for weeks to months.
Construction of physiologically relevant 3D tissues is a multifaceted research initiative where the resulting tissues can be used for many applications. Since 3D cell culture systems provide a more accurate environment for cell growth, tissue morphogenesis, and stem cell differentiation they have potential to be used for modeling tissue development, disease progression and for implantation in situ for large tissue defects. 3D cell culture models are also suited to bridge the gap between conventional 2D preclinical models for drug and toxicity screening and in vivo clinical studies in humans. Improved 3D preclinical models will better recapitulate pathobiological processes underlying diseases of specific tissues and organs in humans, and also more accurately predict physiological responses to therapeutic compounds.
Great strides have already been achieved over the past few decades in this field, yet there remain many practical limitations before 3D cell culture models can be widely implemented in many applications. To begin with, many of the 3D models are novel, and data generated using these methods need to be validated against established in vivo responses for calibration. Additionally, the outcome of 3D assays can be highly variable owing to a lack of standardized protocols and the use of heterogeneous, and sometimes multiple, cell populations. Nondestructive data analysis techniques (secreted factors, morphological changes, and/or non-invasive functional markers) are therefore essential for tracking the same 3D construct over culture. Also, some 3D matrices have limited permeability or poor diffusion dynamics and therefore must incorporate perfusion systems to maintain viability and function. Furthermore, cell recovery and visualization can be restricted in certain 3D scaffolds. Another major limitation is that many of the 3D culture models cannot be easily automated or scaled for high throughput screening.
These limitations however, are being overcome with emerging technologies and newer 3D in vitro assays. In vitro 3D cell culture models have progressed in supporting cell growth, tissue morphogenesis, stem cell differentiation, disease modeling, drug discovery, and toxicity testing. It is also evident that different 3D models with varying characteristics are required to meet the needs of specific cell types or applications. Combining newer technologies such as microarrays on a chip, microfluidics, or bio-printing with biologically relevant matrices could help to scale-up and drive some of these complex 3D cell culture models to highly predictive and relevant drug screening assays and toxicity tests, as well as provide novel systems for regenerative research.
No conflicts of interest declared.
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- Oragui, E., Nannaparaju, M., & Khan, W. S. The role of bioreactors in tissue engineering for musculoskeletal applications. The open orthopaedics journal. 5 Suppl 2, 267-270, doi:10.2174/1874325001105010267 (2011).
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- Mueller, D. et al. In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor. Journal of tissue engineering and regenerative medicine. 5, e207-218, doi:10.1002/term.418 (2011).
- Seidel, J. O. et al. Long-term culture of tissue engineered cartilage in a perfused chamber with mechanical stimulation. Biorheology. 41, 445-458 (2004).
- Lin, H. J., O'Shaughnessy, T. J., Kelly, J., & Ma, W. Neural stem cell differentiation in a cell-collagen-bioreactor culture system. Brain research. Developmental brain research. 153, 163-173, doi:10.1016/j.devbrainres.2004.08.010 (2004).
- Bellas, E., Marra, K., & Kaplan, D. L. P. Sustainable 3D tissue model of human adipose tissue. Tissue engineering. Part C, Methods., doi:10.1089/ten.TEC.2012.0620 (2013).
- Farrell, M. J. et al. Functional properties of bone marrow-derived MSC-based engineered cartilage are unstable with very long-term in vitro culture. Journal of biomechanics, doi:10.1016/j.jbiomech.2013.10.030 (2013).
- Tiruvannamalai-Annamalai, R., Armant, D. R., & Matthew, H. W. A glycosaminoglycan based, modular tissue scaffold system for rapid assembly of perfusable, high cell density, engineered tissues. PloS one. 9, e84287, doi:10.1371/journal.pone.0084287 (2014).
- Chesnick, I. E. et al. Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR microspectroscopy. Bone. 40, 904-912, doi:10.1016/j.bone.2006.10.020 (2007).
- Bettahalli, N. M. et al. Integration of hollow fiber membranes improves nutrient supply in three-dimensional tissue constructs. Acta biomaterialia. 7, 3312-3324, doi:10.1016/j.actbio.2011.06.012 (2011).
- Ellis, M. J., & Chaudhuri, J. B. Poly(lactic-co-glycolic acid) hollow fibre membranes for use as a tissue engineering scaffold. Biotechnology and bioengineering. 96, 177-187, doi:10.1002/bit.21093 (2007).
- Huh, D., Hamilton, G. A., & Ingber, D. E. From 3D cell culture to organs-on-chips. Trends in cell biology. 21, 745-754, doi:10.1016/j.tcb.2011.09.005 (2011).
- Whitesides, G. M. The origins and the future of microfluidics. Nature. 442, 368-373, doi:10.1038/nature05058 (2006).
- Ikuta, S., Sekino, N., Hara, T., Saito, Y., & Chida, K. Mouse epidermal keratinocytes in three-dimensional organotypic coculture with dermal fibroblasts form a stratified sheet resembling skin. Bioscience, biotechnology, and biochemistry. 70, 2669-2675 (2006).
- Futai, N., Gu, W., Song, J. W., & Takayama, S. Handheld recirculation system and customized media for microfluidic cell culture. Lab on a chip. 6, 149-154, doi:10.1039/b510901a (2006).
- Majumdar, D., Gao, Y., Li, D., & Webb, D. J. Co-culture of neurons and glia in a novel microfluidic platform. Journal of neuroscience methods. 196, 38-44, doi:10.1016/j.jneumeth.2010.12.024 (2011).
- Gingras, M., Beaulieu, M. M., Gagnon, V., Durham, H. D., & Berthod, F. In vitro study of axonal migration and myelination of motor neurons in a three-dimensional tissue-engineered model. Glia. 56, 354-364, doi:10.1002/glia.20617 (2008).
- Hofmann, A. et al. The effect of human osteoblasts on proliferation and neo-vessel formation of human umbilical vein endothelial cells in a long-term 3D co-culture on polyurethane scaffolds. Biomaterials. 29, 4217-4226, doi:10.1016/j.biomaterials.2008.07.024 (2008).
- Hayden, R. S., Quinn, K. P., Alonzo, C. A., Georgakoudi, I., & Kaplan, D. L. Quantitative characterization of mineralized silk film remodeling during long-term osteoblast-osteoclast co-culture. Biomaterials., doi:10.1016/j.biomaterials.2014.01.034 (2014).
- Wang, P. C., & Takezawa, T. Reconstruction of renal glomerular tissue using collagen vitrigel scaffold. Journal of bioscience and bioengineering. 99, 529-540, doi:10.1263/jbb.99.529 (2005).
- Hussain, A., Collins, G., Yip, D., & Cho, C. H. Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds. Biotechnology and bioengineering. 110, 637-647, doi:10.1002/bit.24727 (2013).
- Choe, M. M., Tomei, A. A., & Swartz, M. A. Physiological 3D tissue model of the airway wall and mucosa. Nature protocols. 1, 357-362, doi:10.1038/nprot.2006.54 (2006).
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- Liu, X. Y. et al. In vitro tissue engineering of lamellar cornea using human amniotic epithelial cells and rabbit cornea stroma. International journal of ophthalmology. 6, 425-429, doi:10.3980/j.issn.2222-3959.2013.04.03 (2013).
- Bhargava, S., Chapple, C. R., Bullock, A. J., Layton, C., & MacNeil, S. Tissue-engineered buccal mucosa for substitution urethroplasty. BJU international. 93, 807-811, doi:10.1111/j.1464-410X.2003.04723.x (2004).
- Eritja, N. et al. A novel three-dimensional culture system of polarized epithelial cells to study endometrial carcinogenesis. The American journal of pathology. 176, 2722-2731, doi:10.2353/ajpath.2010.090974 (2010).
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text:
Фанаты DropBox самостоятельно начали писать версию под Windows Mobile.
Программа - клиент для работы с сервисом DropBox.com
Вход под своим логином
Просмотр папок и файлов в DropBox
Скачивание файлов
Скачивание и отправка по e-mail
Загрузка на DropBox
Фотографирование и отправка фото
Удаление файлов из Dropbox
DroppedBoxx v0.4.2
DroppedBoxx v0.5
DroppedBoxx v0.5.4
DroppedBoxx v0.7.1
DroppedBoxx v.1.4 (edict)
DroppedBoxx v1.5
Системные требования: Нет данных
Необходим .NetCF не ниже: Нет данных
Тип установки: Нет данных
Автор/Разработчик: d1k_is
Оф. сайт/Источник:
Язык: ENG
Разрешения Lite/Free: показать/скрыть
История изменений: показать/скрыть
- Few bug fixes
- Note: there was a problem with the Marketplace version of this update which causes the update to fail.
* Added the ability to open a file with the default application after its been downloaded
* Fixes Login bug (oauth_timestamp) caused by API changes
* Fixed bug where application adds a newline to the end of uploaded files (makes .cab installers corrupt)
* Fixed bug where subfolders were being deleted unexpectedly
* Fixed bug where selecting a file then creating a folder turns the file into a folder and creates a folder inside there
* Fixed bug in email attach (now working again)
* Added setting for camera Resolution (VGA, 1MP, 2MP or 3MP) (Subject to suggestions)
* Added labels to the buttons on the homescreen (other menus to follow if feedback is good)
* Upped the camera resolution to 2MP (will work on getting the devices max or allowing user to set this)
* Added background uploading to the camera-to-box feature (bigger images take longer )
* Added application data usage to the info screen (resets when you close the app)
* Fixed bug: Error handling files bigger than 2, 147, 483, 647 bytes
* Fixed Icons looking terrible (now they only look average)
* Login screen now has an average looking background!!
* Changed the "Remember Me" function to store the tokens instead of user info (Dropbox prefer) Also this now acts as an auto login. (Everyone will have to login again, sorry :P)
* Added an "Exit" (door) button to the Sync Folders screen to Exit the app instead of having to logout then exit (Requested)
* Info/About screen added with DroppedBoxx version number and Dropbox account usage (Quota, used, shared. Note: The actual usage is the used + shared) Will work on a graph or progress bar thing for this
* Added the settings screen (at the moment all it does is let you "Forget Me" to stop the auto login/remembering password)
* more but i forgot...
* Fixed bug in the modal popups (now the backgrounds stay as the page you were on)
* Added Copy/Paste style Moving and Copying of files around dropbox
* Added Remember Me functionality to Login (Soon to come, auto logins)
* Fixed bug where downloading non-text based files caused corruption, Also changed improved the process managing these files (helps performance)
* Added functionality to Create/Delete Folders from Dropbox (Add opens up a modal box to enter folder name)
* Added new Sync folder icon
* Temp folder has been moved to app data (anyone with a "DroppedBoxx" folder in the device root please delete)
* Temp folder now gets cleared on exit
* Backend stuff for AccountInfo/Sync Folders stuff (coming soon)
* Updated the Link for all the sites who just hot-linked to my download...
* Login Button is now Green!
* Some wording/spelling changes
* Added an Icon!
* Put in some error handling...
* Added a keyboard button to Login to activate the SIP keyboard (will try to do this for any inputs in the future)
* Fixed a few issues with the File Upload/Download screens on WVGA
PRE v0.4:
* Fixed uploading files to dropbox root
* Fixed the installer to put files in the correct place
* Fixed bug where selecting a file then camera upload deleted the file, created a folder with the files name and uploaded the photo there (weird...)
* Compressed the cab installer (now half the size)
* Fixed file upload, YAY!

subset: commoncrawl


text:
The Chinese Space Science and Technology Corporation has presented a transport unmanned aircraft, which is currently the largest in the world.
For the development of the aircraft, the engineers took as a basis the so-called corncobber Yun-5B, which was improved by new innovative developments and equipped with an unmanned control system.
Unmanned aircraft "Feyhun-98", whose name in Chinese means "Flying Swan" has already made the first flight tests. For the first time the ship flew from the landfill in the city of Baotou.
Engineers designed this drone special for massive cargo. Its maximum take-off weight is 2.5 tons, and directly on board the Flying Swan can transport one and a half tons of cargo.
For take-off and landing, the "Feyhun-98" requires only 150 meters of runway, so the ship will cope with the transportation of goods in almost any terrain.
It is reported that the aircraft is designed to fly at a maximum altitude of 4.5 km. The cruising speed of the aircraft is 180 km / h. At one refueling unmanned transport aircraft can fly 1,200 kilometers.
Earlier we reported that at a conference of the American Institute of Aeronautics and Astronautics, the American company Boeing presented the concept of a future hypersonic passenger aircraft. The airliner will be able to carry passengers at a speed exceeding the speed of sound about five times (5M). The airline noted that such an aircraft would allow getting from New York to Tokyo in about 2:00. The aircraft provided a revolutionary fuselage extremely streamlined shape.
The liner of the future received a nose that looked like a needle. According to experts, such solutions will minimize the acoustic impact arising from the transition of the aircraft to supersonic speed. Specialists of the publication New Atlas note that the design of the liner may indicate the choice of cryogenic hydrogen fuel.
Something similar will appear on the market in about 20-30 years, provided that Boeing is seriously engaged in the development of such a machine. The results obtained in the framework of the research can be applied both in civil and military spheres.


text:
On the day of the opening of German Spring, one of the most illustrious Estonian jazz pianists Kristjan Randalu, who was nominated for a Grammy award in 2006, came to Jazzkaar festival with his international ensemble and special project Limes Occidentalis.
All the musicians of the project have their own personality and are leaders of bands themselves, but Randalu successfully created a true symbiosis between them. Randalu has lived in Germany, England and America and has performed in various configurations in many prestigious concert halls in the world. The French guitarist Nguyên Lê has been a technology innovator in the guitar since the early 80s and his country of origin is Vietnam. The Syrian clarinetist Kinan Azmeh'i considered a virtuoso in his field, is currently based in New-York. The bassist Petros Klampanis from Greece has made a name for himself in New York and is also living in the United States now. Bodek Janke is a Polish-born drummer currently living in Germany, and whose musical collaboration with Kristjan Randalu has been going on for twenty-five years. He possesses a percussions arsenal from different countries and is, like every musician of the project, very enthusiastic about world music.
Emigration is definitely the central theme and the common thread of this ensemble, where travelling abroad is for every musician synonym of open-mindedness and personal enrichment.
They performed a series of long and brilliant deep songs, where every song appeared as being a piece of the puzzle, a chapter of the story. The story about crossing, about the current migration of thousands of people, fleeing their countries for numerous reasons. The songs "Crossing" and "Into the flow", among others, are a nod to the current refugees crossing, redrawing the perilous journey of those whose future is uncertain.
Taking inspiration from negative things of the world to do something good, here is their musical aim and aspiration. Like a travel into the darkest recesses of the humanity, but with a note of hope, an ode to life, a touch of peace.
KRISTJAN RANDALU LIMES OCCIDENTALIS (Estonia-Germany-France-Greece-Syria)
26th of April (6pm) at Vaba Lava (Tallinn)
Kristjan Randalu – piano
Nguyên Lê – guitar
Kinan Azmeh – clarinet
Petros Klampanis – double bass
Bodek Janke – drums, percussion
Check out the photos from the concert


text:
UP: In ICU After Snake Bite, Minor Girl Allegedly Raped By Hospital Staffer & 4 Others
A minor girl was allegedly gang-raped in a hospital in Uttar Pradesh's Bareilly on Saturday, November 3. Reportedly, she was admitted to the hospital's ICU ward due to a poisonous snake bite, reported NDTV. The police have already arrested one person and are on the lookout for four others.
Although NDTV reports that she was the only person in ICU, Hindustan Times has reported that even though there were other people in the ICU, the perpetrators closed the curtains from all sides. The accused gave sedatives to the victim and blindfolded her before performing the cruel act.
When the victim's relatives came to visit her the next day, she could not speak and tried informing about the incident in sign language. She explained and informed everyone about the incident when she was shifted to the general ward.
"A case has been registered and an investigation is underway," senior police officer A Singh told news agency ANI. The police have seized the CCTV footage of the ICU. The girl reportedly told her grandmother that the men in uniform came to her at night and forcefully gave her an injection. When she refused they gagged her, tied up her hands and raped her.
Not the first instance
Two weeks ago, a ward boy and a medical student were arrested for raping a woman who was admitted to a hospital in UP's Bagpat. The 17-year-old nursing student was raped when her sister, who was looking after her, went out for tea.
The accused hatched a plan and took the victim to the examination room under the false pretext of running some tests on her and then drugged and raped her.
The Logical Indian condemns the heinous crime and hopes that police catch all the culprits as soon as possible.
Also Read: UP: 27-Year Old Gang-Rape Survivor Sets Herself, Son Ablaze After Police Allegedly Refused To Register Complaint


text:
...My 4 Boys and Me...
Wednesday, February 24, 2016
Tyty takes on Basketball!
On January 9th we laughed and enjoyed watching Ty's very first basketball game! We knew this day was coming, we just didn't know it would be so soon! To no surprise he did great...he sure is lucky to have his older brothers to watch!
Coach Keith talking strategy. Just don't take 16 steps without dribbling the ball at least once.
Scott, Ben Lee and Tyty getting geared up for their first game. Watch out for these three!
always ready on defense!
Posted by
Caroline Armstrong
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text:
|I guess this is my Spooky Halloween Post
|Author's Note: These mechanics were originally written up for fortified keeps as a way to have a greater amount of interactions. Settlements have a wide variety of things for characters to do, such as carouse or research, but isolated shelters don't have so many options so this chart was made
Dreadful Foreboding Mechanics
I believe that these fortified keeps should be mechanically distinct from other settlements not only by their limited supplies but further in their sense of isolation. I think a way to mechanically reinforce the sense of these outposts as bastions of light encroached by darkness is to emphasize their dwindling resources and increasing social pressures. The following mechanics could easily be expanded to other settlements in order to play up the "grimdarkness" or "horribleness/terribleness" of a setting.
These settlements are isolated and each week spent there should feel more oppressive and unwelcoming. The following is a table with ways to mechanically reinforce that, each week roll a 1d12 and consult the Impending Doom Table below. Unless abated, the dooms continue on week after week. For those with a desire of a gross amount of bookkeeping, they may roll this for every single settlement on their hex map and create a living world.
Impending Doom Table
|Menace: Roll a wandering monster encounter appropriate to the terrain of the hex where the settlement is located. The monster encountered begins to harass the settlement. That week and every week further roll a 1d6 as it corresponds to the Modified Wilderness Anecdote Table. The roll corresponds to what is encountered by a person living in the settlement at night.
|Xenophobia: Those foreign to the settlement must succeed on a reaction roll in order to enter the settlement. Further those traveling alone within the settlement have a 1 in 6 chance of encountering hostile people and must roll a reaction roll.
|Hysteria: Anyone staying within the settlement cannot lose trauma points and have a 1 in 6 chance of gaining a point of trauma per week. This applies to every major figure in the settlement, not just the characters.
|Famish: Rations run low and their price doubles. If this is rolled twice there is a 1 in 6 chance of there only being 2d20 rations left in the entire settlement and panic ensues. Every time after the second that this is rolled, the chance for only 2d20 rations exist increases by 1 in 6.
|Rot: Though it is a safe place pestilence endures within. Any disease cured instead simply abates for the duration of the character's stay in the settlement. Further any disease deals an additional 1 point of damage.
|Ruin: The structure of the settlement is eroding and collapsing. In any great confrontation, there is only a 1 in 6 chance of it staying upright and not collapsing. Structures pick the climatically appropriate time to demolish themselves in vast spectacles.
|Woe: Despair falls on the settlement like rain. Characters resting within the settlement only have a 1 in 6 chance of regaining HP.
|Harrowing: Great tragedy strikes a random important figure in the settlement and there is a 1 in 6 chance that they die.
Modified Wilderness Anecdote Table
|Lair, the monster moves into the darkness within the settlement, (rolling an additional time on this table instead of just once every following week)
|Spoor, person sights the monster around the outskirts of the settlement
|Tracks, person finds signs of something moving about at night
|Traces 1 of something, as normally encountered
|Traces 2 of something, as normally encountered
|Monster encounter, roll a reaction check and treat the person appropriately after
Procedurally Generated Antagonist MechanicsThe table above works with the Polite Lands Encounter Tables in a rather unique way. While Fortified Keeps and Villages may be harassed by strange and horrible monsters found out in the wilderness, Towns and Cities will be mainly menaced by human beings. This means that in cities and towns the conflicts will be with people who have specific interests and desires, look at the table below.
Kind (1d8)
|Traveling Merchant
|Caravan of 1d6 Merchants and 2d10 Guards
|1d4 Lumberjacks
|1d4 Hunters
|1d6 Farmers
|1d6 Peasents
|1d3 Members of the Nobility
|1d3 Members of the Priesthood
Each of these presents a unique interaction with the Modified Wilderness Anecdote Table and honestly can be used to generate a variety of conflicts in each city in your setting.


text:
Apple blog reviews, tutorials, news and info. About everything iPhone, iPad, Apple Watch, Mac and iOS.
Unique Alarm Clock | Weather
Developer: Smart Solutions Desenvolvimento de Sistemas Ltda Epp
Language: English, Portuguese, Spanish
Size: 17.8 MB
Price:Free for a limited time
Requirements:Requires iOS 6.0 or later. Compatible with iPhone, iPad, and iPod touch. This app is optimized for iPhone 5.
In brief, WakeApp Weather is an intelligent alarm clock that base on the your choice of weathers. As a result, you can set up your alarms base on your ideal climate such as Clear, Cloudy, Drizzle, Rain, Snow or All for your activity. Furthermore, you can decide the places and select the cities and the temperature for your activity. On the other hand, you also can share your activities through Facebook and Twitter.
-You will only be woken up if the weather is ideal for your activity, according to what you set up on the alarm.
-Your hours of sleep will be optimized!!!
-You can now plan your trips and count on Wakeapp Weather to only wake you up if the weather is ideal on your city of destination.
-It is now possible to only wake up early in the weekends if the weather is ideal for outdoor activities, such as going to the beach.
-It is now possible to be woken up a bit earlier to go to work on the days that are raining or snowing.
-By using this app, you can plan your activities as a group, you just need to share your alarms on your social network accounts.
-Activity: choose from more than 20 activities. In case you don't find the one you want you can always edit the name of the activities offered.
-Climate conditions: from Clear, Cloudy, Drizzle, Rain and Snow, you can choose up to three options, or select the option "All".
-Select the days of the week and hour.
-Location: Choose the location you want or use the default current location (GPS).
-Save your alarm!!!
Download Link
Click Here To Download
Page Link : WakeApp Weather
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text:
Christmas came right on time for some fortunate Don jazzy fans as he gave out money ranging from 50k – 200k.
It began when a woman answered a prayer by another fan,asking God to give him N200K before the 2017 runs out.
The Mavin boss who says he is dropping a song with Iyanya titled "Credit" at that point requested that fans send their bank details and he sent cash to the 1st group of fans who reacted.
See screenshots beneath…


text:
Search maps
Search Tags
Cambodia Laos Vietnam
2011 Cambodia, Laos and Vietnam, colour relief, major rivers and lakes. [11-123a]
Cambodia, Laos and Vietnam
Colour elevation map of Cambodia, Laos, Vietnam and eastern Thailand showing international borders, main rivers and major cities including the country capitals. [00-137]
Region: Asia - Southeast
The Islands of the Caribbean and surrounding countries showing island groups and major cities. [09-141]
Country: Antigua and Barbuda , Barbados , Cuba , Dominica , Dominican Republic , Guadeloupe
Central America & the Caribbean
Countries of Central America and the Caribbean showing capital cities. [00-451]
Region: Americas
Country: Belize , Costa Rica , Cuba , Dominican Republic , El Salvador , Guatemala
Elevation map of Colombia, South America showing department borders and main towns with location inset. [00-214]
Country: Colombia
Galapagos Islands base
Galapagos Islands showing island names, main towns and roads - 2012. PNG versions of the base map can be downloaded with or without elevation shading.
Country: Ecuador
Hanoi Central
Selected tourist destination around Hoam Kiem Lake, Central Hanoi.
Country: Vietnam
Myanmar (Burma) to Australia
Southeast Asian countries.
Region: Asia - Southeast
North Vietnam
Popular tourist destinations in northern Vietnam.
Country: Vietnam


text:
New fire truck for Wellington Voluntary Rural Fire Force
25 September 2019
Wellington Voluntary Rural Fire Force has received the first of Fire and Emergency New Zealand's new model fire appliances designed for fighting vegetation fires.
"The brigade is really excited about our new truck," says Rural Controller Colin Robson.
"It has more modern technology to fight wildfires and respond to emergencies, meaning our crews can do their job and serve our communities more efficiently."
The new model of fire truck can seat six people and has improved firefighting capabilities and safety features for the crew. These include; larger water capacity, four-wheel drive, increased locker volume, a stepped rear deck area, and better lighting for working in low visibility conditions.
"It's the first new model of medium rural fire appliance since Fire and Emergency was formed in 2017. Before the amalgamation, rural fire brigades had a variety of smaller firefighting trucks. This is the first time they've had a truck this big, with this up to date technology.
"In fact, this is the first brand new truck our brigade has received in over 20 years," says Colin Robson. "Our previous truck was a former military tanker made in 1985."
Twenty-two of these trucks will be put into service across the country over the next year, along with 16 single cab versions designed to seat a crew of three.
Members of Wellington Voluntary Rural Fire Force in their new truck
For more information contact:
Media | Fire and Emergency New Zealand
Email: email@example.com, Mobile: 027 591 8837


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Python Fax – Resubmit a Transaction
This sample demonstrates how to resubmit a fax with Python. Resubmitting a fax transaction is useful if you want to resend a previously-submitted Python fax (either to the same number or to a different one), without having to upload the file or to provide all the parameters again.
The following Python fax snippet uses the ReSendFax method to resubmit a fax transaction. You are required to specify the ID of the fax that needs to be resubmitted and the destination fax number, which can be a new number or the same number you used in the last transaction.
A simple script resend a fax using the InterFAX
ReSendFax SOAP API call.
import osa
client = osa.Client("https://ws.interfax.net/dfs.asmx?WSDL")
print('Testing ReSendFax...')
result = client.service.ReSendFax( Username='', Password='', TransactionID=614416600, FaxNumber='') # Enter the TransactionId of the fax to re-send here.
# Also, enter the destination fax number here.
print(' ReSendFax returned with code {0}'.format(result))


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Systematic Review is a research method that aims to locate and summarize all available evidence for a specific question in order to guide decisions and practices (Definition from the Cochrane Handbook for Systematic Reviews of Interventions). It will help you gather knowledge about a particular field of study, identify recommendations for a future research, establish the context of a research topic or problem, and even identify the main methodologies and research techniques used in a particular research topic or field.
Process of Systematic Reviews
What we librarians can support you through your systematic review project?
- Provide feedback on your systematic review project protocol, especially help specify your research question
- Identify sources, design search strategies, and provide search results
- Write up a method section describing search strategies for your report
- Help you learn EndNote citation software for managing references
To get started, please complete the below request form or contact Marina Zhang at firstname.lastname@example.com with questions.


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Tony Seba, author of 'Clean Disruption' gives this brilliant talk at the AltCars Expo and Conference, September 19, 2014. Watch the first 1:35 and you won't want to stop because you'll realize the predictions made below are all going to come true. And I bet they come to fruition much sooner than 2030!
The keynote is titled "Clean Disruption: 100% electric transportation and 100% solar power by 2030″.
The book 'Clean Disruption' and this keynote assert that by 2030:
• All new mass-market vehicles will be electric.
• All of these vehicles will be autonomous (self-driving).
• Up to 80% of parking spaces and highways will be redundant.
• Taxis as we know them will be obsolete.
• The concept of car ownership will be obsolete.
• Oil will be obsolete
• All new energy will be provided by solar (and wind)
More information at
(Source: YouTube)


text: KATPILOT Report post Posted June 19, 2017 As I write this Patty is half way across the pond heading back to Maui to start loading the pod to ship the restaurant and her house to Scottsdale. She made it through her daughters graduation which I was blessed with the chance to see. I met her daughter for the first time. Ron had paid for her college education and I know it hurt Patty for him not to be there to see but I think he was and I bet he did see. He was proud as heck. I am certain of that. Now comes the next hard part which is the dismantling of home and business. I know this will be a difficult time as well but by mid August it and she will be calling this place home. Of course I didn't tell her much about the heat. It's supposed to reach 120 by Wednesday but who can tell right? In four weeks we will be married on a beach in Maui and that is like a dream come true...........a dream neither of us had, wanted, or expected but it happened just the same. I know all of this happened for a reason, a reason bigger than ourselves. 1 3 Quote Share this post Link to post Share on other sites


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Bluewater Productions sent Major Spoilers a sneak peek of titles arriving in stores this week.
Fame: Drake – the Mario Gully "Batman" homage cover
Print and Digital
Writer: Marc Shapiro
artist: Mike J.C.
He made a name for himself as the wheelchair bound Jimmy Brooks on "Degrassi: the Next Generation" but when Aubrey Graham went by his middle name, Drake, he rocketed to stardom. Now Bluewater Comics shares the story of how Drake went from Canadian teen-melodrama to the hip hop star that worked with Lil Wayne's stable of musicians with the comic "Fame: Drake."
Special cover by famed artist Mario Gully.
Mis-Adventures of Adam West #9
In Print and Digital
writer: Leon McKenzie and Edward Gross
artist: Tsubasa Yozora
Adam West finds himself in a world where reality is…warped at best. He ends up in his classic crime fighting costume, trying to save Generic City from its own costumed "hero," Fleischer, who doesn't seem to have a clue what's going on. As Adam discovers, with Fleischer that's par for the course!
Steve Harvey Presents: Roopster Roux #2
In Print and Digital
writer: Ryan Burton
artist: Erik Thompson
Comedian Steve Harvey and crimestopper Roopster Roux once again bring justice to their town by fighting a new threat-Victor Vain. Convinced the world would be a better place if everyone looked exactly like him, Vain has turned Woodville citizens into the spitting image of himself! It's up to Steve Harvey and Roopster Roux to stop Vain and his sidekick, Cheesecake, and return Woodville to normal.
Insane Jane: collected edition
In Print and Digital
writer: Zach Hunchar
artist: Mendoza
Take youthful energy. Add in some over-active imagination. Mix in some powdered donuts. Sprinkle it with delusional disorder and you get … Insane Jane! Will Jane save the day? Will Jane destroy the world? Will she remember to take her medication?
Vincent Price Presents collected editon: Dr. Phibes
In Print and Digital
writer: Mel Smith, Paul Birch
artist: Various
This one shot graphic novel featuring Vincent Price tells another gothic adventure about one of the characters that Price made famous, Dr. Phibes. Investigating a bad actor's death, Inspector Trout joins Thanatos, a mysterious man involved with Dr. Phibes' past, into a journey to London's sewers where he finds what happened to the master of bizarre murder. As well as Dr. Phibes is back in another new adventure. As the new world prepares to ring out the old millennium, Inspector Trout arrives in New York City to be greeted by a series of macabre murders and the knowledge that his arch-nemesis, Dr. Anton Phibes is behind them!
Tribute: Cory: Cory Monteith
Digital only
writer: C.W. Cooke, P.R. McCormack
artist: Beniamino Bradi
In this comic book novella developed with the Official Cory Monteith fan club the " Montourage". The comic book showcase the talent of a life that was cut to short. Tribute: Cory Monteith shows the superstar from his beginnings growing up to his success in Glee. A special forward was written by Ravital Zabarsky from the " Montourage".
FAME: The Cast of Glee the graphic novel
In Print and Digital
writer: C.W. Cooke, P.R. McCormack
artist: Beniamino Bradi
From the sold-out Fame series comes the story all Gleeks have been waiting for: the biography of the cast and creator of Glee. From creator Ryan Murphy and star Lea Michele to fan favorites Heather Morris and Darren Criss, Fame takes a look at how all of these men and women became part of a global phenomenon. From the hit FOX series, it's your turn to be a Gleek. You know you want to!
You can download these titles on Wowio, Comixology, DriveThru Comics, Google Play, My Digital Comics, Overdrive, Iverse, PanelFly, iTunes, Kindle, Wowio, Nook, Kobo and wherever eBooks are sold.
About Bluewater Productions
Bluewater Productions Inc. is one of the top independent production studios of comic books, young adult books and graphic novels. Its extensive catalog of titles includes the bestsellers "10th Muse" and "The Legend of Isis" "Bluewater publishes comic books in partnership with entertainment icon William Shatner ("Tear Chronicles"), legendary filmmaker Ray Harryhausen ("Wrath of the Titans," "Sinbad: Rogue of Mars," "Jason and the Argonauts," et al) and celebrated actor Vincent Price ("Vincent Price Presents"), Additionally, Bluewater publishes a highly successful line of biographical comics under the titles "Female Force" and "Political Power." Bluewater aims to unite cutting-edge art and engaging stories produced by its stable of the publishing industry's top artists and writers.


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Using RDP support to implement a sustainable model of agriculture that combines the production of high quality products and environmental protection. Winning initiative of the 2019 Rural Inspiration Awards in the 'Environment & Climate Action' category.
Projects & Practice
Total results: 3.
A project that demonstrates how agri-environment support can significantly help the protection of the environment while also increasing the profitability of a farm.
A family farm invested in a biogas plant to produce renewable energy while also purchasing a new tomato harvester to improve the farm's overall income.


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- Description
One of our favourites walk with amazing views and a magic forest. An open ridge walk to Ivinghoe Beacon, gentle forests, and a classic village.
This route - following the Ridgeway along the Chiltern escarpment to Ivinghoe Beacon - is exhilarating, offering downland scenery as fine as anything on the South Downs. From the Beacon itself, it seems as if you can see half of England on a fine day.
Photos event:
Video event:
Transport & Meeting PointWe'll meet at Euston Station from 9:00 am and leave the meeting point at 9:15 am for the 9:24 am train.
Planned arrival time back into London of 7:00 pm (we have a group train ticket and you are committed for the day until then).
FoodWe will stop during the hike for a snack lunch so please bring food and drink. We will visit a pub after the hike where we can get refreshments.
CostThe trip is £26 per person which includes the guided hike and return train travel from the group departure station listed at the top of this event.
Price:• Members: £19
• No Members: £26
Under 18's must be accompanied at all times by a parent/s
Between 13 and 17 years old: £19
All events are limited to twenty-five assistants.
Do not forget to click Going! and send a copy of the payment with the full name and contact telephone number of the attendees. You can send this data through our Facebook, by email email@example.com or by whatsapp 07535295739.
Por any questions or more info ask to Maria: 07535 295 739 Whatsapp.
What to Bring• Suitable hiking clothes & hiking footwear (ideally hiking boots) - click here http://www.hikinginlondon.co.uk Waterproof jacket, trousers and footwear are recommended.
• Water (at least 1 and ideally 2 litres)
• Packed lunch/picnic
• Money for drinks and refreshments after the hike
• That's it! We will take care of train tickets, logistics on the day etc, so that you can just turn up and have a great day :-)
Safety and Terms and ConditionsPayments are non-refundable and non-transferable but we have a process to release your space for resale to another member. Please follow the instructions in our FAQ page http://www.senderismoenlondres.co.uk/frequent-questions.html and email us with your Pay Pal "invoice ID" immediately after you change your RSVP to "NO".
By signing up to this event you agree to the Terms and Conditions at the following link: http://www.senderismoenlondres.co.uk/club/club-rules.html
© Hiking in London Ltd 2020 in collaboration with Saturday Walkers Club.
Errors and Omissions Excepted
- Reviews
You may also be interested in the following product(s)
Adult full year - 12 months£45.00
Junior full year - 12 months
Regular Price: £30.00
Special Price £19.00


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3,400 year old figurine
one Israeli child is making headlines around the world after it discovered he found a precious ancient artefact.
According to media reports Friday,Ori Greenhunt spotted a figurine poking out of the earth when he was climbing a mound with friends at Tel Rehov archaeological site and carefully carried it home.
The small artefact is thought to be 3,400 years old and may be the image of the fertility goddess Astarte, or simply a curvaceous lady, reports The Daily Mail.
The boy came across a stone that had shifted and saw an image of a person covered with soil, according to the country's Antiquities Authority.
'Ori returned home with the impressive figurine and the excitement was great,' his mother Moriya Greenhut said.
'We explained to him this is an ancient artefact and that archaeological finds belong to the state,' she added.
The family, from the communal settlement of Tel Te'omin in the Beit Sheʽan Valley, gave the figurine to the Israel Antiquities Authority, which said the image of the woman was made by pressing soft clay into a mould.
Amihai Mazar, professor emeritus at Hebrew University and expedition director of the archaeological excavations at Tel Rehov said the figurine may represent an ordinary women, or that of the fertility goddess Astarte.
'Some researchers think the figure depicted here is that of a real flesh and blood woman, and others view her as the fertility goddess Astarte, known from Canaanite sources and from the Bible.'
'It is highly likely that the term trafim mentioned in the Bible indeed refers to figurines of this kind.'
Astarte was worshiped from the Bronze Age through classical antiquity and was connected with fertility, sexuality and war.
Her symbols were the lion, horse, sphinx, dove and a star within a circle indicating the planet Venus. She is usually depicted naked.
He continued: 'Evidently the figurine belonged to one of the residents of the city of Rehov, which was then ruled by the central government of the Egyptian pharaohs.'
Dr Mazar said: 'The figurine 'is typical of the Canaanite culture of the 15th-13th centuries BC.'
Ori has been awarded a certificate of appreciation for his good citizenship by the Israel Antiquities Authority at his school.
His teacher, Esther Ledell, said: 'It was an amazing occasion.
'The archaeologists entered the class during a Torah lesson, just when we were learning about Rahel stealing her father's household gods.
'I explained that the household gods were statues that were used in idol worship, and all of a sudden I realise that these very same idols are here in the classroom!'


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The window to the world and the gateway to the soul- eyes are a vital organ of the body which need constant care. Being surrounded by screens everywhere from phones, laptops, desktops to even on traffic signals and roads, the human eye to subject to electronic radiations for long hours every day. This leads to a negative effect on the eyes, and eventually requires a person to wear lenses or spectacles for vision correction.
Being in a corporate world, this is a common phenomenon and it is quite usual for people to ignore their eye health. To sensitize employees regarding the importance of the same, a health talk on taking care of eyes was organized for the employees of CapitalVia on Saturday, August 10, 2019 by Centre for Sight Eye Hospital.
The talk was conducted by Dr. Saurabh Maheshwari in which he explained general eye problems, their solutions, advance treatment in eye care etc. through a presentation and compilation of illustrative videos.
The presentation and video highlighted among the corporate employees the need to blink eyes at frequent intervals, to drink enough water, and to minimize use of mobile phones, to help eyes remain lubricated and to wear spectacles regularly for those whose eyes needed refractive correction.
He highlighted many other eye problems which are faced by people at different ages, including myopia, hypermetropia, glaucoma, dry eye syndrome, cataract etc. He also spoke about two procedures using which the refractive correction can be made in eyes – namely LASIK and SMILE. He explained the procedure of LASIK in detail along with a video.
"LASIK surgery requires a flap to be created over the cornea using a machine, following which the laser treatment is done to correct the vision. This is followed by placing the flap back on the cornea, the entire procedure taking not more than 30 minutes," Dr. Maheshwari said.
He also highlighted the necessary basic requirements for a person to be eligible for the surgery.
He also explained the second procedure i.e. SMILE which includes removal of a 'lenticule' from the cornea to improve vision. Both the procedures were explained with demonstrative videos which piqued interest of the attendees.
This was followed by an eye check up camp for the employees in which more than 150 employees got their vision tested. The new technology and impressive success rates of the new-age procedures has resulted in more than 25 employees showing interest in opting for the LASIK surgery procedure for vision correction.
All the employees who attended the camp also got coupons for eye check up at the Centre for Sight hospital. The hospital also promised to open its doors on a Sunday – which is usually an off day – specially for CapitalVia employees, whose timing would be announced in the days preceding such arrangement. A team of four experts from the hospital conducted the camp which went on till 6pm.
The eye check up camp successfully drew the attention of all CVians towards their eye health, spreading the message of taking care of their eyes and avoiding excessive use of screens which burden the eyes. Lastly, for those who were diagnosed with a refractive correction in their eyes were urged to start wearing spectacles and take care of their eyes.
Data is updated on 7th per month. It gets verified every quarter by independent auditor.
Data presented here is taken from company's inception
Importance given to satisfactory resolution as per prescribed TAT


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I got a good question at one of my Second Saturday seminars the other day. A woman asked, "What's a reconciliation clause? Do I even want one? I don't plan on reconciling!" She went on to say that a reconciliation clause wasn't included when she and her husband met with a mediator, but then that the draft of her separation agreement from our office included one. I was a little taken aback by her tone, to tell you the truth, because she sounded a little accusatory. Like we should have … [Read more...]
6 Big Misconceptions in Family Law
We all know a lot about how the law works because we watch TV. Right? Well, no—actually, it's quite the opposite. We don't know very much about the law, but most of us do know just enough to be truly dangerous. There are courtroom dramas right and left, and what most people really know is just enough to set them up for frustration and disappointment later on. There are lots of misconceptions when it comes to family law cases. Family law is, in a lot of ways, very, very different from other … [Read more...]


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Between-seasons jacket KEIKO BIKERJACKET suitable for all military men who value style and comfort of sportswear. In order to maintain the unique look, the jacket is equipped with epaulets, fastened with press stud, upstanding collar with warm puller, and special stitching at the waist, which also give the garment a motorcycle character. To ensure full functionality, this jacket is equipped with 1 inside pocket for wallet and 4 external pockets (2 chest with zipper, 2 for hands secured by flap with snaps).
Presented model of between-seasos jacket is made of cotton fibers that ensure adequate breathability. The material protects your body from overheating and excessive sweating, while protecting against the wind in the cooler spring weather.Due to the technique of coloring of the material (Vintage Washed), the color of the jacket may be changed when worn or after washing. The material can also dye.
|Total Length (cm/in)
|Chest Circumference (cm/in)
|Sleeve Lenght - from armpit to cuff (cm/in)
|66 cm (26")
|104 cm (41")
|51 cm (20")
|68 cm (26,5")
|109 cm (43")
|52 cm (20,5")
|69 cm (27")
|114 cm (45")
|52 cm (20,5")
|70 cm (27,5")
|120 cm (47")
|53 cm (21")
|71,5 cm (28")
|123 cm (48,5")
|53,5 cm (21")
Guarantee - Warranty is granted by the seller for a period of 12 months.
Users reviews


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Honda and Red Bull Group have signed an agreement whereby Honda will enter into a partnership with Red Bull Racing to supply it with power units for two years from the start of the 2019 FIA Formula One World Championship.
This will be in addition to its current agreement with Red Bull Scuderia Toro Rosso.
The two Red Bull-owned teams will run identical specification Honda Power Units.
Takahiro Hachigo, president and representative director of Honda Motor, said: "Having established a good relationship with Scuderia Toro Rosso, we have decided to extend our Formula One involvement to the other team in the Red Bull family, Aston Martin Red Bull Racing, as from the 2019 season. Having two teams means we can access twice as much data as previously. [Read more…] about Honda to supply Formula One power units to Red Bull Racing


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Tapnell Farm's award winning accommodation includes Safari Tents, charming stone cottages, wooden lodges and pods and an impressive Manor House. All enjoy panoramic views, comfortable beds, outdoor cooking and most enjoy wood burning hot tubs!
Wedding guests can all stay onsite, just a short walk from the reception venue making this the ideal location. Cosy Eco Pods make the ideal option for the happy couple on their wedding night.


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Western Canada Select crude's discount to US benchmark West Texas Intermediate oil narrowed on Monday to the tightest since July. NY time, data compiled by Bloomberg show. The move also found support among her political rivals.
Alex Pourbaix, chief executive officer of oil-sands producer Cenovus, which may have to shut in about 35,000 barrels a day of production, said the industry is not happy to have the government so involved in the industry but realized that the curtailment plan was the only way to avoid "disaster".
Above: Price of Central Alberta blend of oil.
A new threat looms for Canada's largest oil producing province, even as it imposes mandatory output cuts to ease a glut that has driven down crude prices.
Oil prices were pressured by a weekly report from the American Petroleum Institute (API) that said US crude inventories rose by 5.4 million barrels in the week to November 30, to 448 million barrels, in a sign that USA oil markets are in a growing glut. The crisis has caused some producers to reduce production on their own, slash dividends and delay next year's drilling plans. Construction is underway on Enbridge Inc's Line 3 pipeline replacement from Alberta to the United States, with the project expected to be in service by the end of 2019.
Official U.S. government oil production and inventory data is due later on Wednesday. Further reductions in the curtailment are expected in the fall and winter as additional rail capacity comes online.
Federal Natural Resources Minister Amarjeet Sohi said Monday he is asking the National Energy Board to make sure Canada's oil pipelines are being used as efficiently as possible. The province estimates 25 producers will have to impose cuts.
How much production would be affected if Saskatchewan did implement cuts in a similar manner to Alberta?
The first 10,000 bpd for each producer will be excluded from the mandatory cuts, meant to avoid negatively impacting small producers.
Health care fell 1.8 per cent as shares of cannabis producer Aphria Inc. plunged almost 28 per cent after short-sellers called the Canadian company a "black hole" and alleged that its recent global acquisitions totalling $280-million were "largely worthless". The move is meant to combat steep discounts now placed on Alberta oil. Hopefully, the implementation of the decision will be effective, since the impact of curtailment on small and medium oil producers, which have already been hammered by this government's regulatory changes, could be drastic.
About the only dissenting voice has come from Canada's integrated oil companies, whose refineries have been benefiting from the cheaper feedstock.
As of December 3, Husky Energy, Saskatchewan's largest heavy oil producer, had the most drilling rigs working in the country, at 16. U.S. refiners have turned to rail to ship incremental crude barrels to the U.S. Gulf Coast to take advantage of discounted prices. The system would require a total of about 80 locomotives and more than 7,000 cars.
"I think those two factors combined are providing a little bit of lift for oil prices", said Craig Fehr, Canadian markets strategist for Edward Jones.
Discounts for upgraded synthetic oil improved to US$13.50 per barrel Monday afternoon from US$18.50 on Friday and Edmonton-priced light oil differentials fell to US$15.25 from US$23.00. In October, WCS' discount to futures dropped to $50 a barrel.


text: Beautiful blonde Cherry Kiss is enjoying sucking her own toes on her bed when Dean comes to have a piece of the action. He gently starts licking her feet and sucking her toes. Cherry proceeds to stroke his cock with her feet while he fingers her sweet pussy. She then goes down and swallows his thick cock until it is properly lubricated. Dean goes on by drilling her cooch and kissing her foot at the same time. Cherry moans in pleasure from this deep pounding. She goes back to sucking and gives him a deep blowjob until he cums all over her pretty face. To top this off this naughty babe goes to have a warm pee in her bath tub and have a little taste off of her fingers.


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John Bolton to Meet with Turks to Discuss Announced U.S. Withdrawal from Syria by Karen DeYoung – Washington Post
Turkey wants the United States to disarm Syrian Kurdish forces it has trained and supplied for the fight against the Islamic State, and to provide air and logistical support for Turkish troops and allied Syrian opposition forces who plan to pick up the anti-militant battle after a U.S. withdrawal.
The list of Turkey's "concerns and expectations" will be conveyed next week to White House national security adviser John Bolton when he visits Ankara to explain the still-unspecified U.S. plan for a withdrawal from Syria, according to Turkish officials.
Bolton's trip comes as Secretary of State Mike Pompeo is also heading to the region to try to persuade Arab partners and allies that "the United States is not leaving the Middle East. Despite reports to the contrary and false narratives surrounding the Syria decision, we are not going anywhere," a senior administration official said Friday in a briefing for reporters…


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Serving the Cleveland and Chattanooga areas
Bradley Awning Aluminum & Window Co of Cleveland, TN gives you outstanding patio covers that generate curb appeal as well as protect you from the elements up above.
You can select from canvas or aluminum for your new patio cover and you're guaranteed quality with durability.
Patio and deck covers
Screened-in rooms or glassed-in enclosures
Limitless design options
Living rooms
Reading rooms
Full panel screens
Swing screen doors
Sliding screen doors
Screened-in rooms


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Today, OmniVirt is expanding its platform to provide a monetization solution for Virtual Reality Inventory. While previous announcements and case studies from OmniVirt have called out work with The New York Times, Time Inc, AOL, and many other traditional publishers to power 360° Video advertising for web inventory, with this announcement OmniVirt's platform can be used to monetize a publisher's Virtual Reality inventory as well.
While publishers have invested into VR, the industry has lacked technology to connect web and VR inventory together, creating limited scale and monetization for VR providers. Now, both traditional publishers and VR developers can monetize their VR inventory and deliver campaigns that scale for brands.
See our demos below:
- For publishers, OmniVirt supports ad serving into VR apps: Intro Video, Demo Video
- For developers, OmniVirt provides monetization for VR apps: Intro Video
For VR app developers, OmniVirt provides monetization for VR apps (iOS, Android, Unity, and more):
For publishers, OmniVirt supports ad integration with ad servers (DFP, Freewheel, and more):
Demo – Ad integration with existing ad servers:
To date, OmniVirt has delivered over 500M 360° VR ad impressions, supporting high profile campaigns for clients like Toyota, Infiniti, Samsung, Netflix, Chevron, Universal Pictures and Johnnie Walker. Now these campaigns can scale into VR inventory as well.
OmniVirt's mission is to make it easy for publishers and advertisers to launch and monetize their VR experiences and campaigns. Now, with one platform both traditional publisher's with a VR app and emerging VR developer's looking to monetize their experience with native VR formats can get started with 3 simple steps:
- Sign Up for an OmniVirt Account at www.omnivirt.com
- Integrate the OmniVirt SDK and Set Ad Campaigns Live
- See revenues report at our earning page
If interested in learning more about OmniVirt and recent updates to its Monetization Platform, please reach out at [email protected]
About OmniVirt
OmniVirt is 360 Video and Virtual Reality advertising platform founded by former Google and YouTube employees. The company has received funding from top venture capitalists including Andreessen Horowitz, Greycroft, BDMI, Horizon Media, First Round Capital and many others. OmniVirt's technology distributes 360° VR across top publisher websites like The New York Times, AOL, Time Inc, Gannet, Wall Street Journal, Vice Media and more. These experiences are customizable and interactive, with detailed performance reporting showcasing where audiences are engaging in the immersive experience. The company has powered multiple high profile campaigns for clients like Toyota, Infiniti, Samsung, Chevron, Universal Pictures and Johnnie Walker.


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Attention: For textbook, access codes and supplements are not guaranteed with used items.
Arrives in 3-7 Business Days
In this book master dulcimer player Madeline MacNeil teaches beginning players tuning, scales, strumming, fingering, chord studies, alternate tunings, use of the capo, and how to use the noter. She endorses both playing by ear and learning to read standard notation. In twelve intensive lessons, she explains everything from the basics to some very complex concepts in easily understood language, making this a particularly valuable book for the beginning instrumentalist. The lessons and songs are written in both standard notation and tablature. This package also includes the companion DVD in which MacNeil demonstrates the techniques and songs taught in the book. CD also included.


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- 2 frozen Maine lobster tails (12 ounces each), thawed
- 2 small sweet peppers
- Salt and Freshly Ground black pepper
- 1/2 cup mayonnaise
- 1/4 cup minced green onion
- Shredded romaine lettuce
- 4 soft rolls, such as large hot dog buns or French roll
Grill or boil lobster tails until cooked through. Remove meat from shells and cut into 1/2-inch dice. Roast peppers on grill until charred. Peel, remove seeds and mince or puree. Combine mayonnaise, celery, green onion and peppers in a bowl. Add the lobster and stir to blend. Season the mixture to taste with salt and pepper. To serve: Lay the lettuce on each bun and spoon the lobster filling over. Cut into quarters.
Source:Lobster Sliders with Roasted Red Pepper Sauce Recipe