Characteristics and Value of Directed Algorithms in High Content Screening

Ghosh, Rahul; Lapets, Oleg; Haskins, Jeffrey R.

doi:10.1385/1-59745-217-3:63

Cited by 8 publications

(9 citation statements)

References 20 publications

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“…VEGF stimulated complete and well-formed networking of capillary tubes while Suramin completely impeded tube formation. Angiogenic Index (AI) is the measure of the percentage of the image area covered by tubes multiplied by 10 [22]. PA at 3.5, 7 and 14 µM significantly reduced the AI compared to untreated control.…”

Section: Resultsmentioning

confidence: 99%

In Vitro and In Vivo Anti-Angiogenic Activities of Panduratin A

et al. 2012

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BackgroundTargeting angiogenesis has emerged as an attractive and promising strategy in anti-cancer therapeutic development. The present study investigates the anti-angiogenic potential of Panduratin A (PA), a natural chalcone isolated from Boesenbergia rotunda by using both in vitro and in vivo assays.Methodology/Principal FindingsPA exerted selective cytotoxicity on human umbilical vein endothelial cells (HUVECs) with IC50 value of 6.91±0.85 µM when compared to human normal fibroblast and normal liver epithelial cells. Assessment of the growth kinetics by cell impedance-based Real-Time Cell Analyzer showed that PA induced both cytotoxic and cytostatic effects on HUVECs, depending on the concentration used. Results also showed that PA suppressed VEGF-induced survival and proliferation of HUVECs. Furthermore, endothelial cell migration, invasion, and morphogenesis or tube formation demonstrated significant time- and dose-dependent inhibition by PA. PA also suppressed matrix metalloproteinase-2 (MMP-2) secretion and attenuated its activation to intermediate and active MMP-2. In addition, PA suppressed F-actin stress fiber formation to prevent migration of the endothelial cells. More importantly, anti-angiogenic potential of PA was also evidenced in two in vivo models. PA inhibited neo-vessels formation in murine Matrigel plugs, and angiogenesis in zebrafish embryos.Conclusions/SignificanceTaken together, our study demonstrated the distinctive anti-angiogenic properties of PA, both in vitro and in vivo. This report thus reveals another biological activity of PA in addition to its reported anti-inflammatory and anti-cancer activities, suggestive of PA’s potential for development as an anti-angiogenic agent for cancer therapy.

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Section: Resultsmentioning

confidence: 99%

In Vitro and In Vivo Anti-Angiogenic Activities of Panduratin A

et al. 2012

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“…Images of fluorescently labeled cells were analyzed by an algorithm that identified the angiogenic tubes formed by the association and clustering of the endothelial cells and quantitatively measured the properties of the identified tubes such as the number of tubes, tube lengths, tube areas, number of nodal branch points, and the angiogenic index. The angiogenic index is defined as the percentage of image area covered by tubes multiplied by 10 (Ghosh et al 2007; Grove and Ghosh 2006). Both compounds were as effective as vascular endothelial cell growth factor (VEGF) in increasing the angiogenic index above the value for untreated control cells in both cell types (Figures 1 and 2).…”

Section: Resultsmentioning

confidence: 99%

“…Images of fluorescently labeled cells were collected with a Thermo Scientific Cellomics ArrayScan HCS Reader (Thermo Fisher Scientific, Pittsburgh, PA, USA) and analyzed by an automated algorithm that identified the tubes formed by the association and clustering of the endothelial cells. This algorithm provided quantitative measurements of tube properties such as the number of tubes, tube lengths, tube areas, number of nodal branch points, and the angiogenic index—defined as the percentage of image area covered by tubes multiplied by 10 (Ghosh et al 2007; Grove and Ghosh 2006). Each experiment was repeated 3 times, and each treatment was probed in at least six wells.…”

Section: Methodsmentioning

confidence: 99%

Angiogenic Potential of 3-Nitro-4-Hydroxy Benzene Arsonic Acid (Roxarsone)

Basu

Ghosh

Grove

et al. 2008

Environ Health Perspect

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BackgroundRoxarsone (3-nitro-4-hydroxy benzene arsonic acid) is an arsenic compound widely used in the poultry industry as a feed additive to prevent coccidiosis, stimulate growth, and to improve tissue pigmentation. Little is known about the potential human health effects from roxarsone released into the environment from chicken waste or from residual compound in chicken products.ObjectiveThe growth potentiation and enhanced tissue pigmentation suggest that low levels of roxarsone exposure may have an angiogenic potential similar to that of inorganic arsenite (AsIII). The goal of this investigation was to test the hypothesis described above using cultured human aortic and lung microvascular endothelial cells in high-content imaging tube-forming assays and begin developing a molecular level understanding of the process.MethodsWe used a three-dimensional Matrigel assay for probing angiogenesis in cultured human endothelial cells, and a polymerase chain reaction (PCR) array to probe the gene changes as a function of roxarsone or AsIII treatment. In addition, we used Western blot analysis for changes in protein concentration and activation.ResultsRoxarsone was found to exhibit a higher angiogenic index than AsIII at lower concentrations. Increased endothelial nitric oxide synthase (eNOS) activity was observed for roxarsone but not for AsIII-induced angiogenesis. However, AsIII caused more rapid and pronounced phosphorylation of eNOS. Quantitative PCR array on select genes revealed that the two compounds have different and often opposite effects on angiogenic gene expression.ConclusionsThe results demonstrate that roxarsone and AsIII promote angiogenic phenotype in human endothelial cells through distinctly different signaling mechanisms.

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“…The phagocytic index of each strain may be compared to that of the control strain and other strains using a one-way ANOVA and the Kruskal-Wallis test. For medium-or highthroughput screens the final optimized assay should also be assessed to be certain it has the required power (Ghosh, Lapets, & Haskins, 2007;Zhang, Chung, & Oldenburg, 1999).…”

Section: Statistical Analysesmentioning

confidence: 99%

An Automated Assay to Measure Phagocytosis of Cryptococcus neoformans

2019

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Cryptococcus neoformans is an opportunistic fungal pathogen that causes meningoencephalitis, which kills 200,000 individuals worldwide each year. It is ubiquitous in the environment and is first inhaled into the lungs of the host, where it is taken up by phagocytes. The interaction of these fungal cells with host phagocytes, therefore, is a critical step in the pathogenesis of this disease. One characteristic of this initial step in host–pathogen interactions is the avidity with which fungal cells are taken up by phagocytes, described by the phagocytic index. In this chapter, we detail a high‐throughput method of directly assessing the phagocytic index of fungal cells using an imaging‐based paradigm. By automating image collection and processing, this method permits rapid assessment of this critical host interaction. © 2019 by John Wiley & Sons, Inc.

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Characteristics and Value of Directed Algorithms in High Content Screening

Cited by 8 publications

References 20 publications

In Vitro and In Vivo Anti-Angiogenic Activities of Panduratin A

In Vitro and In Vivo Anti-Angiogenic Activities of Panduratin A

Angiogenic Potential of 3-Nitro-4-Hydroxy Benzene Arsonic Acid (Roxarsone)

An Automated Assay to Measure Phagocytosis of Cryptococcus neoformans

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