Abstract only The accumulation of vascular smooth muscle (SMC)-like cells within the intima contributes significantly to intimal medial thickening (IMT) and vascular remodeling typical of ...arteriosclerotic disease. Light has emerged as a powerful tool to interrogate cells label-free and facilitates discriminant observations both in vitro and in vivo . The auto-fluorescence (AF) profile of individual cells isolated from arteriosclerotic vessels, captured on V-cup array and interrogated across five wavelengths using a novel Lab-on-a-Disc platform, was significantly increased at the 565 ± 20nm wavelength concomitant with a reduction in Myh11 expression, when compared to differentiated vascular smooth muscle (SMC) cells from control vessels. In vitro, TGF-β1 promoted myogenic differentiation of murine bone-marrow derived Sca1 + /CD44 + mesenchymal stem cells (MSC) and murine Sca1 + C3H 10T1/2 cells concomitant with enrichment of the specific SMC epigenetic histone mark, H3K4me2 at the Myh11 promoter, Myh11 promoter transactivation and increased SMC differentiation marker mRNA and protein expression. Myogenic differentiation resulted in a significant increase in the AF intensity across 565 ± 20nm wavelength, an effect not observed for TGF-β1 treated RAMOS human B lymphocytes but mimicked by Notch activation of resident Sca1 + multipotent vascular stem cells (MVSCs) with Jagged1 and inhibited following elastin and collagen III depletion, respectively. Moreover, the temporal increase in the AF intensity at 565 ± 20nm wavelength during myogenic differentiation was similar to the AF profile of dissociated cells from arteriosclerotic vessels at this same wavelength. These data suggest that an AF photonic fingerprint of stem cell-derived myogenic progeny in vitro mimics that of vascular cells ex vivo following injury.
Cellular activation and inflammation leading to endothelial dysfunction is associated with cardiovascular disease (CVD). We investigated whether a single cell label-free multi parameter optical ...interrogation system can detect endothelial cell and endothelial progenitor cell (EPC) activation in vitro and ex vivo, respectively. Cultured human endothelial cells were exposed to increasing concentrations of tumour necrosis factor alpha (TNF-α) or lipopolysaccharide (LPS) before endothelial activation was validated using fluorescence-activated cell sorting (FACS) analysis of inflammatory marker expression (PECAM-1, E-selectin and ICAM-1). A centrifugal microfluidic system and V-cup array was used to capture individual cells before optical measurement of light scattering, immunocytofluorescence, auto-fluorescence (AF) and cell morphology was determined. In vitro, TNF-α promoted specific changes to the refractive index and cell morphology of individual cells concomitant with enhanced photon activity of fluorescently labelled inflammatory markers and increased auto-fluorescence (AF) intensity at three different wavelengths, an effect blocked by inhibition of downstream signalling with Iκβ. Ex vivo, there was a significant increase in EPC number and AF intensity of individual EPCs from CVD patients concomitant with enhanced PECAM-1 expression when compared to normal controls. This novel label-free 'lab on a disc' (LoaD) platform can successfully detect endothelial activation in response to inflammatory stimuli in vitro and ex vivo.
This study describes the prevalence and patterns of searching for meaning in the aftermath of breast cancer and asks how the search relates to made meaning and emotional adjustment. Women (n=72) ...reported their level of searching for meaning, made meaning and negative affect at multiple time points in the first 18 months after breast cancer treatment. Over time, four search for meaning patterns emerged: continuous (44%), exiguous (28%), delayed (15%) and resolved (13%). Just over half of the participants reported having made meaning at early and late time points. A higher level of searching for meaning was unrelated to made meaning, but was associated with a higher level of negative affect in longitudinal analyses controlling for baseline levels. Women who engaged in an ongoing, unresolved search for meaning from baseline to follow-up also had a significantly higher level of negative affect at follow-up than women who infrequently or never engaged in a search for meaning over time. These analyses reveal that: a) there is great variability in the prevalence and pattern of searching for meaning in the aftermath of breast cancer, and b) searching for meaning may be both futile and distressing.
A hallmark of subclinical atherosclerosis is the accumulation of vascular smooth muscle cell (SMC)-like cells leading to intimal thickening and lesion formation. While medial SMCs contribute to ...vascular lesions, the involvement of resident vascular stem cells (vSCs) remains unclear. We evaluated single cell photonics as a discriminator of cell phenotype in vitro before the presence of vSC within vascular lesions was assessed ex vivo using supervised machine learning and further validated using lineage tracing analysis
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Using a novel lab-on-a-Disk(Load) platform, label-free single cell photonic emissions from normal and injured vessels ex vivo were interrogated and compared to freshly isolated aortic SMCs, cultured Movas SMCs, macrophages, B-cells, S100β
+
mVSc, bone marrow derived mesenchymal stem cells (MSC) and their respective myogenic progeny across five broadband light wavelengths (λ465 - λ670 ± 20 nm). We found that profiles were of sufficient coverage, specificity, and quality to clearly distinguish medial SMCs from different vascular beds (carotid vs aorta), discriminate normal carotid medial SMCs from lesional SMC-like cells ex vivo following flow restriction
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and identify SMC differentiation of a series of multipotent stem cells following treatment with transforming growth factor beta 1 (TGF- β1), the Notch ligand Jagged1, and Sonic Hedgehog using multivariate analysis, in part, due to photonic emissions from enhanced collagen III and elastin expression. Supervised machine learning supported genetic lineage tracing analysis of S100β
+
vSCs and identified the presence of S100β
+
vSC-derived myogenic progeny within vascular lesions. We conclude disease-relevant photonic signatures may have predictive value for vascular disease.
Graphical abstract
This paper describes a novel, smart-grid technique for isolating, sorting and capturing cancer cells based on size and deformability by lateral displacement. In stopped-flow, centrifugal ...sedimentation mode, the bioparticles are sizeselectively displaced along the dynamically spaced grid of the microstructures in a deterministic fashion. Following displacement, particles are captured in claw-like structures for onward processing. This displacement method was first demonstrated for sorting seven bead populations with diameters between 5 μm and 30 μm from a mixture. Then HeLa and melanoma cells spiked in buffer and whole blood were isolated and captured. Finally, we succeeded to sort normal and fixed HeLa cells according to their deformability.
There is growing evidence that many women with breast cancer engage in a meaning-making process. Much of the research is descriptive and cross-sectional, and as such provides little insight into the ...progression of the meaning-making process over time. Further, many psychological interventions are based on several clinical assumptions about the frequency, trajectory, and importance of meaning-making relative to adjustment to cancer. With these clinical assumptions in mind, this correlational, longitudinal analysis investigated: (a) the prevalence and patterns of searching for meaning (SFM) and made meaning (MM) among a group of women recovering from breast cancer surgery, (b) concurrent and longitudinal associations between the two meaning variables, and (c) associations between the meaning variables and negative affect (NA). Seventy-two women treated for breast cancer completed the negative affect subscale of the Positive and Negative Affect Scale (PANAS) at three time points over an 18-month period. The SFM and MM variables were measured at Times 1 and 2. Analysis of the data was conducted using SPSS statistical procedures, including analysis of covariance (ANCOVA). A small majority of women reported an active SFM at both Time 1 and Time 2 with a significant minority reporting a lower frequency SFM at both time points. Four patterns of SFM emerged to illustrate the trajectory of the meaning search over time. Just over half of the participants reported MM at both time points. Chi-Square analysis revealed no significant concurrent or longitudinal associations between SFM and MM. ANCOVA analysis revealed a significant association between SFM and Time 2 NA. Further analysis revealed that women who engaged in an ongoing, unresolved SFM had significantly higher Time 2 mean NA scores than women who infrequently or never searched for meaning. These analyses revealed several important findings: (1) not all women reported SFM, (2) some women reported MM without ever having engaged in an active SFM, (3) for some women, making meaning did not signal an end to their search, (4) for some women, the search for meaning may be futile. Results are discussed within the context of the aforementioned clinical assumptions. Implications and suggestions for future research are proposed.
The American Diabetes Association and the European Association for the Study of Diabetes convened a panel to update the prior position statements, published in 2012 and 2015, on the management of ...type 2 diabetes in adults. A systematic evaluation of the literature since 2014 informed new recommendations. These include additional focus on lifestyle management and diabetes self-management education and support. For those with obesity, efforts targeting weight loss, including lifestyle, medication, and surgical interventions, are recommended. With regards to medication management, for patients with clinical cardiovascular disease, a sodium-glucose cotransporter 2 (SGLT2) inhibitor or a glucagon-like peptide 1 (GLP-1) receptor agonist with proven cardiovascular benefit is recommended. For patients with chronic kidney disease or clinical heart failure and atherosclerotic cardiovascular disease, an SGLT2 inhibitor with proven benefit is recommended. GLP-1 receptor agonists are generally recommended as the first injectable medication.
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