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Additional file 2: Supplementary Figure 2. MFI measurement. Positive expression of THY1 and MSC surface markers in SF subsets was identified by comparing fluorescence-minus-one control. THY1 intensity was calculated as MFI of THY1+ SF population. Dash line indicated negative populations, while solid line indicated positive populations.

Additional file 6: Supplementary Figure 6. Expression of THY1 and MSC surface markers in the individual subset from OA and RA. The MFI of THY1 (A) and other MSC surface markers (B) was evaluated with flow cytometry. Data was shown in each subset from OA and RA. OA and RA samples are plotted as white circles and white squares, respectively. Data were analyzed by Holm-Sidak���s multiple comparison test for comparing each subset from the same samples (*p

Additional file 7: Supplementary Figure 7. Osteogenic potentials of SF subsets from OA and RA. A. Quantification of the staining. Cells per donor were cultured in 3 dishes and the results of 10 donors (OA:7, RA:3) for alizarin red and 11 donors for ALP were plotted (OA:10, RA:1). B. Gene expression levels by qPCR. The expression levels of the genes were plotted as a ratio to the expression level of 18S of undifferentiated SF as a control. Cells per donor were cultured in 3 dishes and the results of 10 donors (OA:9, RA:1) for RUNX2 and ALPL, and 9 donors (OA:8, RA:1) for OCN. OA and RA samples are plotted as white circles and white squares, respectively. The average data of each subset are plotted as black squares. Data were analyzed by Dunn���s multiple comparison test for comparing each subset from the same patients (*p

Additional file 5: Supplementary Figure 5. Cell survival/proliferation and MSC surface marker expression by THY1 knockdown. WST-8 assay. Cell survival/proliferation by THY1 knockdown was assessed using WST-8 assay. Data were analyzed by Wilcoxon���s test for comparing each subset from the same samples (*p

Extracellular vesicles (EVs) play an important role in intercellular communication and regulation of cells, especially in the immune system where EVs can participate in antigen presentation and may have adjuvant effects. We aimed to identify small molecule compounds that can increase EV release and thereby enhance the immunogenicity of vaccines. We utilized a THP-1 reporter cell line engineered to release EV-associated tetraspanin (CD63)-Turbo-luciferase to quantitatively measure EVs released in culture supernatants as a readout of a high throughput screen (HTS) of 27,895 compounds. In parallel, the cytotoxicity of the compounds was evaluated by PrestoBlue dye assay. For screening immunostimulatory potency, we performed two additional independent HTS on the same compound library using NF-κB and interferon-stimulated response element THP-1 reporter cell lines. Hit compounds were then identified in each of the 3 HTS’s, using a “Top X″ and a Gaussian Mixture Model approach to rule out false positive compounds and to increase the sensitivity of the hit selection. Thus, 644 compounds were selected as hits which were further evaluated for induction of IL-12 in murine bone-marrow derived dendritic cells (mBMDCs) and for effects of cell viability. The resulting 130 hits were then assessed from a medicinal chemistry perspective to remove compounds with functional group liabilities. Finally, 80 compounds were evaluated as vaccine adjuvants in vivo using ovalbumin as a model antigen. We analyzed 18 compounds with adjuvant activity for their ability to induce the expression of co-stimulatory molecules on mBMDCs. The full complement of data was then used to cluster the compounds into 4 distinct biological activity profiles. These compounds were also evaluated for quantitation of EV release and spider plot overlays were generated to compare the activity profiles of compounds within each cluster. This tiered screening process identified two compounds that belong to the 4-thieno-2-thiopyrimidine scaffold with identical screening profiles supporting data reproducibility and validating the overall screening process. Correlation patterns in the adjuvanticity data suggested a role for CD63 and NF-κB pathways in potentiating antigen-specific antibody production. Thus, our three independent cell-based HTS campaigns led to identification of immunostimulatory compounds that release EVs and have adjuvant activity.

Additional file 9: Supplementary Figure 9. Adipogenic differentiation markers of SF subsets from OA and RA. Cells per donor were cultured in 3 dishes and the results of 7 donors (OA:3, RA:4). The target genes were normalized with 18S of undifferentiated SF as a control. OA and RA samples are plotted as white circles and white squares, respectively. The average data of each subset are plotted as black squares. Data were analyzed by Dunn���s multiple comparison test for comparing each subset from the same samples.

Additional file 1: Supplementary Figure 1. Gating strategy. Gating strategy of freshly isolated cells from OA and RA synovial tissues using flow cytometry. Cells were stained with CD45, CD235a, CD31, THY1 and CD34. Individual subsets were analyzed for the expression of MSC surface markers and were sorted as CD34-THY1-, CD34-THY1+ and CD34+THY1+, respectively.

Additional file 3: Supplementary Figure 3. Expression of MSC surface markers. Expression levels of MSC surface markers (CD73, CD271, CD54, CD29 and CD44) on freshly isolated SF subsets were re-analyzed using microarray analysis previously reported ( https://www.ncbi.nlm.nih.gov/geo/geo2r/?acc=GSE109450&platform=GPL18573 ).

Additional file 4: Supplementary Figure 4. Expression of MSC surface markers after expansion. The MFI of MSC surface markers (THY1, CD34 and CD73) was evaluated with flow cytometry. OA and RA samples are plotted as white circles and white squares, respectively. The average data of each subset are plotted as black squares. Data were analyzed by Holm-Sidak���s multiple comparison test for comparing each subset from the same samples (*p

Additional file 8: Supplementary Figure 8. Chondrogenic potentials of SF subsets from OA and RA. A. Histology of the chondrocyte pellets. Chondrocyte matrix was stained as red by safranin O staining and measured the ratio of stained area (A). Cells per donor were cultured in 3 pellets and the results of 18 donors (OA: 10, RA:8). B. qPCR of chondrogenic differentiation marker. The relative expression levels of ACAN in differentiated cells. Cells per donor were cultured in 3 pellets and the results of 12 donors (OA:6, RA:6). The target genes were normalized with 18S of undifferentiated SF as a control. OA and RA samples are plotted as white circles and white squares, respectively. The average data of each subset are plotted as black squares. Data were analyzed by Dunn���s multiple comparison test for comparing each subset from the same samples (*p

In the original framework of Professors Acemoglu and Robinson, the government is unable to oppress the revolution once it is brought about. However, actual civil wars are unpredictable. With this notion, I introduce uncertainty depending on military expenditures of the government. Then an interesting argument follows: if the likelihood of successful oppression is sufficiently larger than a certain level of destruction rate and there are cheap-but-effective devices such as biochemicals, citizens in a dictatorial country may have a trade-off between economic prosperity and domestic military threats.

This paper provides a proof of Alchian and Allen’s thesis—superior good is relatively more consumed in distant market—with some restrictions on the utility function. Comparing to the proof by Borcherding and Silberberg (1978), those assumptions may restrict the model in some sense but we will find the Alchian-Allen theorem is independent of the third law of Hicks (Hicks 1946, pp. 309-311). The theorem is given by natural characteristics of relative demand curves.

This paper explores conditions for which Alchian-Allen theorem–the finer quality product is relatively more purchased than the lower one in regions where additional costs are imposed–holds in models of various but finite qualities. In this paper, “finer qualities raise ratios to lower ones in response to transportation costs” defines the various qualities version of Alchian-Allen theorem. Then we find Alchian-Allen theorem holds if and only if consumptions of neighbor qualities are not so different and weighted averages of compensated elasticities by inverse of prices are decreasing in qualities. In order to provide plausible intuitions, some sufficiency conditions are also suggested.

Relative efficiencies of antebellum slave farms are suggested by many empirical studies. This paper considers a theoretical aspect of those results using a repeated principal-agent problem. Within its theoretical analysis, with relevance to profitability of slave farms, it will be shown that when inter-temporal punishments are necessary and when they can perform efficiently in production. Applying those theoretical results, some empirical studies on relative profitability and relative efficiencies are discussed. In the empirical study, relative efficiencies of each farm scale—free farms, task farms, and gang farms—are estimated region by region by a stochastic profit frontier model.

This paper reports on a novel plastic-molded total-reflection type polygonal laser beam deflector. The present deflector is fabricated by the optical resin molding which is suitable for low cost mass production, because expensive and time consuming aluminum reflection film deposition process has been eliminated. A bar-code reader that employs the present laser beam deflector shows readout performance comparable to that of a bar-code reader that employs the conventional aluminum deflector. The condition of no deterioration of the field depth of the resolving power for the bar-code is that, waviness on a deflector surface Wmax is to be less than 10 nm.© (1993) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.