Spoken Communication

This Grants Collection uses the grant-supported open course Spoken Communication from Clayton State University: http://clayton.libguides.com/SpokenCommunication This Grants Collection for Spoken Communication was created under a Round Three ALG Textbook Transformation Grant. Affordable Learning Georgia Grants Collections are intended to provide faculty with the frameworks to quickly implement or revise the same materials as a Textbook Transformation Grants team, along with the aims and lessons learned from project teams during the implementation process. Documents are in .pdf format, with a separate .docx (Word) version available for download. Each collection contains the following materials: Linked Syllabus Initial Proposal Final Reporthttps://oer.galileo.usg.edu/communication-collections/1001/thumbnail.jp

Investigating the Effects of Tissue-Specific Extracellular Matrix on the Adipogenic and Osteogenic Differentiation of Human Adipose-Derived Stromal Cells Within Composite Hydrogel Scaffolds

© Copyright © 2019 Shridhar, Amsden, Gillies and Flynn. While it has been postulated that tissue-specific bioscaffolds derived from the extracellular matrix (ECM) can direct stem cell differentiation, systematic comparisons of multiple ECM sources are needed to more fully assess the benefits of incorporating tissue-specific ECM in stem cell culture and delivery platforms. To probe the effects of ECM sourced from decellularized adipose tissue (DAT) or decellularized trabecular bone (DTB) on the adipogenic and osteogenic differentiation of human adipose-derived stem/stromal cells (ASCs), a novel detergent-free decellularization protocol was developed for bovine trabecular bone that complemented our established detergent-free decellularization protocol for human adipose tissue and did not require specialized equipment or prolonged incubation times. Immunohistochemical and biochemical characterization revealed enhanced sulphated glycosaminoglycan content in the DTB, while the DAT contained higher levels of collagen IV, collagen VI and laminin. To generate platforms with similar structural and biomechanical properties to enable assessment of the compositional effects of the ECM on ASC differentiation, micronized DAT and DTB were encapsulated with human ASCs within methacrylated chondroitin sulfate (MCS) hydrogels through UV-initiated crosslinking. High ASC viability (\u3e90%) was observed over 14 days in culture. Adipogenic differentiation was enhanced in the MCS+DAT composites relative to the MCS+DTB composites and MCS controls after 14 days of culture in adipogenic medium. Osteogenic differentiation studies revealed a peak in alkaline phosphatase (ALP) enzyme activity at 7 days in the MCS+DTB group cultured in osteogenic medium, suggesting that the DTB had bioactive effects on osteogenic protein expression. Overall, the current study suggests that tissue-specific ECM sourced from DAT or DTB can act synergistically with soluble differentiation factors to enhance the lineage-specific differentiation of human ASCs within 3-D hydrogel systems

Techniques for the isolation of high-quality RNA from cells encapsulated in chitosan hydrogels

Extracting high-quality RNA from hydrogels containing polysaccharide components is challenging, as traditional RNA isolation techniques designed for cells and tissues can have limited yields and purity due to physiochemical interactions between the nucleic acids and the biomaterials. In this study, a comparative analysis of several different RNA isolation methods was performed on human adipose-derived stem cells photo-encapsulated within methacrylated glycol chitosan hydrogels. The results demonstrated that RNA isolation methods with cetyl trimethylammonium bromide (CTAB) buffer followed by purification with an RNeasy® mini kit resulted in low yields of RNA, except when the samples were preminced directly within the buffer. In addition, genomic DNA contamination during reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was observed in the hydrogels processed with the CTAB-based methods. Isolation methods using TRIzol® in combination with one of a Qiaex® gel extraction kit, an RNeasy® mini kit, or an extended solvent purification method extracted RNA suitable for gene amplification, with no evidence of genomic contamination. The latter two methods yielded the best results in terms of yield and amplification efficiency. Predigestion of the scaffolds with lysozyme was investigated as a possible means of enhancing RNA extraction from the polysaccharide gels, with no improvements observed in terms of the purity, yield, or amplification efficiency. Overall, this work highlights the application of a TRIzol®+extended solvent purification method for optimizing RNA extraction that can be applied to obtain reliable and accurate gene expression data in studies investigating cells seeded in chitosan-based scaffolds. © 2013 Mary Ann Liebert, Inc

The distinct roles of the nucleus and nucleus-cytoskeleton connections in three-dimensional cell migration

Cells often migrate in vivo in an extracellular matrix that is intrinsically three-dimensional (3D) and the role of actin filament architecture in 3D cell migration is less well understood. Here we show that, while recently identified linkers of nucleoskeleton to cytoskeleton (LINC) complexes play a minimal role in conventional 2D migration, they play a critical role in regulating the organization of a subset of actin filament bundles – the perinuclear actin cap - connected to the nucleus through Nesprin2giant and Nesprin3 in cells in 3D collagen I matrix. Actin cap fibers prolong the nucleus and mediate the formation of pseudopodial protrusions, which drive matrix traction and 3D cell migration. Disruption of LINC complexes disorganizes the actin cap, which impairs 3D cell migration. A simple mechanical model explains why LINC complexes and the perinuclear actin cap are essential in 3D migration by providing mechanical support to the formation of pseudopodial protrusions

Adipose-Derived Stem Cells in a Resilient In Situ Forming Hydrogel Modulate Macrophage Phenotype

Injectable hydrogels have the potential to enhance stem cell-based therapies by improving cell localization, retention, and survival after transplantation. The inflammatory response to both the hydrogel and the encapsulated cells is a critical aspect of this strategy, with macrophages being highly involved in the process of hydrogel remodeling, angiogenesis, and tissue regeneration. As a step toward the development of a cell-based strategy for therapeutic angiogenesis, this work compared the intramuscular injection of allogeneic rat adipose-derived stem/stromal cells (rASCs) in an in situ gelling hydrogel with the injection of the hydrogel alone and rASCs in saline in an immunocompetent rat model by immunohistochemical analysis over 4 weeks. rASCs delivered in the hydrogel were retained intramuscularly at significantly higher densities as compared with cells delivered in saline. The encapsulated rASCs modulated the inflammatory response, promoting CD68(+) macrophage recruitment, with the majority of infiltrating cells expressing the M1 marker CCR7, as well as a higher fraction of CD163(+) M2c macrophages surrounding the hydrogel. Furthermore, rASCs reduced the initial expression of inducible nitric oxide synthase and promoted arginase-1 expression in the infiltrating macrophages over time, consistent with a shift toward a more proregenerative phenotype. Coincident with the enhanced macrophage infiltration, significantly more CD31(+) lumens were observed surrounding and within the hydrogels with rASCs at 2 and 4 weeks as compared with the hydrogels alone. Overall, these results are a promising indication that encapsulated rASCs can have immunomodulatory effects and may enhance angiogenic processes after intramuscular injection, promoting a regenerative macrophage response and blood vessel formation

Design of tissue-specific extracellular matrix composite hydrogels for adipose-derived stem/stromal cell (ASC) delivery

© 2019 Omnipress - All rights reserved. Statement of Purpose: There is a compelling need to develop hydrogel cell delivery systems that incorporate tissue specific ECM as a cell instructive component, to enhance survival and direct cell function. Building from our previous work supporting the potential of composite bioscaffolds incorporating decellularized adipose tissue (DAT) within methacrylated chondroitin sulphate (MCS) hydrogels,1,2 the current study focused on investigating the effects of incorporating tissue-specific ECM sourced from DAT or decellularized trabecular bone (DTB) on encapsulated human ASCs. We hypothesized that incorporating tissue-specific ECM would provide an inductive microenvironment that would promote lineage-specific ASC differentiation

XML-based tactical chat (XTC): requirements, capabilities and preliminary progress

The motivation for pursuing XML-based tactical chat includes the great potential of this technology and fixing limitations of current chat programs. XTC capabilities have the potential to completely upgrade and restructure all tactical military communications. The current tools for military chat include VRC, Yahoo, MSN, AIM, ICQ, and NKO. None of these provides the full functionality or interoperability needed in a joint environment. Moreover, if a nonproprietary chat protocol is developed, it can lead to a decision-support environment in which data, text, audio, and video can be logged, evaluated and managed, all in a Web environment where no additional specialized software or hardware is needed. Chat technology challenges for the military fit into three areas: tactical, technical and administrative. Tactically, there are many ways chat can be used, but effective practices are not yet defined in procedures or doctrine. Joint forces use a myriad of chat programs that don't interoperate and are usually proprietary. Technically many chat programs are barred by firewalls and lack a robust interface to allow logging and searching past chats, From an administrative prospective, plain-test chat has no structure. Scheduling and controlling who attends or converses remains undefined. Within DoD there is no standard for how, when, and by whom chats ought to be conducted. Possible approaches to these problems include adopting a proprietary chat system or customizing an open-source implementation. Proprietary solutions are costly, do not interoperate well, and are too inflexible for a technology that is evolving rapidly. Open-source software can provide a solution that is adaptable, extensible, quick to implement, straightforward to maintain, and relatively inexpensive. This report provides a preliminary assessment of XML-based tactical chat (XTC) using an open-source, open-standards solution. Promising initial results demonstrate that an XML document can be sent from an XHTML page in a Web browser to an off-the-shelf Jabber client via a Web serve. Further, available server and client implementation can enable a research and development plan for rapid development. further work on XTC as part of the Extensible Modeling and Simulation Framework (XNSF) is justified and needed.Approved for public release; distribution is unlimited