Coronavirus envelope (E) proteins play an important, not fully understood role(s) in the virus life cycle. All E proteins have conserved cysteine residues located on the carboxy side of the long hydrophobic domain, suggesting functional significance. In this study, we confirmed that mouse hepatitis coronavirus A59 E protein is palmitoylated. To understand the role of the conserved residues and the necessity of palmitoylation, three cysteines at positions 40, 44, and 47 were changed singly and in various combinations to alanine. Double-and triple-mutant E proteins resulted in decreased virus-like particle output when coexpressed with the membrane (M) protein. Mutant E proteins were also studied in the context of a full-length infectious clone. Single-substitution viruses exhibited growth characteristics virtually identical to those of the wild-type virus, while the double-substitution mutations gave rise to viruses with less robust growth phenotypes indicated by smaller plaques and decreased virus yields. In contrast, replacement of all three cysteines resulted in crippled virus with significantly reduced yields. Triplemutant viruses did not exhibit impairment in entry. Mutant E proteins localized properly in infected cells. A comparison of intracellular and extracellular virus yields suggested that release is only slightly impaired. E protein lacking all three cysteines exhibited an increased rate of degradation compared to that of the wild-type protein, suggesting that palmitoylation is important for the stability of the protein. Altogether, the results indicate that the conserved cysteines and presumably palmitoylation are functionally important for virus production.Coronaviruses are medically important viruses that cause primarily respiratory and enteric infections in humans and a broad range of animals. The emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) and the recent identification of new human coronaviruses (HCoVs), HCoV-NL63 and HCoV-HKU1 (10, 21), have significantly increased the importance of understanding key interactions during virus assembly, since these interactions may provide insight into potential targets for antiviral and vaccine development.Coronaviruses are enveloped, positive-stranded RNA viruses that belong to the Coronaviridae family in the Nidovirales order. The virion envelope contains at least three proteins: membrane (M), spike (S), and envelope (E). The genomic RNA is encapsidated by the phosphorylated nucleocapsid. Coronaviruses assemble at and bud into membranes of the endoplasmic reticulum Golgi intermediate compartment (ERGIC) (22,46). The focus of this paper is the E protein. Coronavirus E proteins are small (76 to 109 amino acids) integral membrane proteins bearing long hydrophobic domains. The protein plays an important, but not fully understood role(s) in virus production (7,11,24,38). Virus-like particles (VLPs) are formed only when the E and M proteins are expressed (3,5,50). When E protein is expressed by itself, E protein-containing vesicles are relea...

In breast reconstructive procedures, adipose-derived stem cells (ASCs) that are present in clinical fat grafting isolates are considered to play the main role in improving wound healing. In patients following chemotherapy for breast cancer, poor soft tissue wound healing is a major problem. However, it is unclear if tamoxifen (TAM) as the most widely used hormonal therapeutic agent for breast cancer treatment, affects the ASCs and ultimately wound healing. This study evaluated whether TAM exposure to in vitro human ASCs modulate cellular functions. Human ASCs were isolated and treated with TAM at various concentrations. The effects of TAM on cell cycle, cell viability and proliferation rates of ASCs were examined by growth curves, MTT assay and BrdU incorporation, respectively. Annexin V and JC-1 Mitochondrial Membrane Potential assays were used to analyze ASC apoptosis rates. ASCs were cultured in derivative-specific differentiation media with or without TAM (5 uM) for 3 weeks. Adipogenic and osteogenic differentiation levels were measured by quantitative RT-PCR and histological staining. TAM has cytotoxic effects on human ASCs through apoptosis and inhibition of proliferation in dose-and time-dependent manners. TAM treatment significantly down-regulates the capacity of ASCs for adipogenic and osteogenic differentiation (p<0.05 vs. control), and inhibit the ability of the ASCs to subsequently formed cords in Matrigel. This study is the first findings to our knowledge that demonstrated that TAM inhibited ASC proliferation and multi-lineage ASC differentiation rates. These results may provide insight into the role of TAM with associated poor soft tissue wound healing and decreased fat graft survival in cancer patients receiving TAM.Autologous fat grafting is a versatile tool in reconstructive surgery, and is increasingly used for breast reconstruction.

Pretreatment of the scaffold with fibronectin offers a method to increase cell adhesion and delivery. ASCs maintain their immunophenotype and ability to differentiate while on SIS. Seeding freshly isolated SVF onto the scaffold demonstrated that minimally manipulated cells may be useful for perioperative surgical applications within the OR suite. We have shown that this model for a "living mesh" can be successfully used in abdominal wall reconstruction.

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Infrainguinal arterial occlusive disease can lead to potentially disabling and limb-threatening conditions. Revascularization may be indicated for claudication, rest pain, or tissue loss. Although endovascular interventions are becoming more prevalent, open surgeries such as endarterectomy and bypass are still needed and performed regularly. Open reconstruction has been associated with postoperative morbidity, both at the local and at the systemic levels. Local complications include surgical site infections (SSIs 0–5.3%), graft failure (12–60%), and amputation (5.7–27%), and more systemic issues include cardiac (2.6–18.4%), respiratory (2.5%), renal (4%), neurovascular (1.5%), and thromboembolic (0.2–1%) complications. While such outcomes present an additional challenge to the postoperative management of surgical patients, it may be possible to minimize their occurrence through careful risk stratification and preoperative assessment. Therefore, individualized selection of candidates for open repair requires weighing the need for intervention against the likelihood of adverse outcomes based on preoperative risk factors. This review provides an overview of open reconstruction, focusing on identifying the clinical indications for surgery and perioperative morbidity and mortality.

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services, Directorate for Information , 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302. Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. REPORT DATE (DD-MM-YYYY)October 27, 2010 14. ABSTRACT Strategic communication is an essential tool for achieving national objectives in peacetime and in war. The military, diplomatic and academic communities continue to struggle to define strategic communication. Four characteristics that differentiate strategic communications at the operational and strategic levels are: the extent to which the engagement is in reaction to a precipitating event or at the initiative of the communicator; the duration of the engagement; whether the target of the engagement is to change audience analysis or to change the audience's analytic framework; and whether the engagement implies reflexiveness, i.e., the need or potential for behavioral change by the communicator as well as the audience. Identifying characteristics that differentiate between strategic communication at the operational and strategic levels, and incorporating them into the analysis of problems and operational plans, will help operational planners to distinguish between the two levels, determine whether chosen objectives are more appropriately categorized as operational or strategic, select tactics appropriate to the operational environment and the chosen objective, and establish correct expectations for success of a communication plan. Discussion and Analysis 9 SUBJECT TERMSThe Strategic Level of Strategic Communication 9Strategic Communication and Public Diplomacy 12 Conclusion and Recommendations 18Endnotes 21Bibliography 23iii AbstractStrategic communication is an essential tool for achieving national objectives in peacetime and in war. The military, diplomatic and academic communities continue to struggle to define strategic communication. Four characteristics that differentiate strategic communications at the operational and strategic levels are: the extent to which the engagement is in reaction to a precipitating event or at the initiative of the communicator; the duration of the engagement; whether the target of the engagement is to change audience analysis or to change the audience's analytic framework; and whether the engagement implies reflexiveness, i.e., a need or potential for behaviora...

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