High Throughput Secure MPC over Small Population in Hybrid Networks (Extended Abstract)

2020 ◽  
pp. 832-855
Author(s):  
Ashish Choudhury ◽  
Aditya Hegde
Keyword(s):  
High Throughput ◽  
Small Population ◽  
Hybrid Networks

scholarly journals A Flow Cytometry–Based Screening System for Directed Evolution of Proteases

2011 ◽  
Vol 16 (3) ◽  
pp. 285-294 ◽  
Author(s):  
Ran Tu ◽  
Ronny Martinez ◽  
Radivoje Prodanovic ◽  
Mathias Klein ◽  
Ulrich Schwaneberg
Keyword(s):  
Flow Cytometry ◽  
Directed Evolution ◽  
High Throughput ◽  
Double Emulsion ◽  
Catalytic Efficiency ◽  
Small Population ◽  
Subtilis Strain ◽  
Screening System ◽  
Screening Technology

Proteases are industrially important enzymes but often have to be improved for their catalytic efficiency and stabilities to suit applications. Flow cytometry screening technology based on in vitro compartmentalization in double emulsion had been developed and applied on directed evolution of paraoxonase and β-galactosidase. Further advancements of flow cytometry–based screening technologies will enable an ultra-high throughput of variants offering novel opportunities in directed enzyme evolution under high mutational loads. For the industrially important enzyme class of proteases, a first flow cytometry–based screening system for directed protease evolution has been developed based on an extracellular protease-deficient Bacillus subtilis strain (WB800N), a model protease (subtilisin Carlsberg), and a water-in-oil-in-water double-emulsion technology. B. subtilis WB800N cells are encapsulated in double emulsion with a fluorogenic substrate (rhodamine 110–containing peptide), allowing the screening of protease variants in femtoliter compartments at high throughput. The protease screening technology was validated by employing an epPCR mutant library with a high mutational load and screened for increased resistance toward the inhibitor antipain dihydrochloride. A variant (K127R, T237P, M239I, I269V, Y310F, I372V) with an improved relative resistance was isolated from a small population of active variants, validating the reported protease flow cytometry screening technology for increased inhibitor resistance.

Abstract 19641: Novel Microprinted Elisa Platform for High Throughput Screening of Protein Secretion Reveals a Comprehensive Strategy to Prevent Ischemia Reperfusion Induced Apoptosis

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Kshitiz Kz ◽  
David E Ellison ◽  
Junaid Afzal ◽  
Maimon E Hubbi ◽  
Segun Bernard ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Death ◽  
Stem Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Ischemia Reperfusion ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Small Population ◽  
Cardiac Differentiation

Introduction: A major challenge in using stem cells to treat myocardial infarction (MI) is the massive cell death post transplant. Many progenitor cell types limit disrepair post MI without significant cardiac differentiation, possibly by anti-apoptotic signals in their secretions. The signature of these secretions is however not well known. Methods: We created a novel high throughput microELISA platform by combining microprinting and microfluidics to precisely estimate secretions of a small population of cells over time. Screening the secretions in many stem cell types used previously to treat MI, i.e. BMSCs (Bone marrow derived stem cells), CDCs (Cardiosphere derived cells), and iPSC-CMs (induced pluripotent stem cell derived cardiomyocytes), we found a common preserved secretory signature of growth factors. Results: Using a high throughput screen of pro-apoptotic factors that prevent CDCs from peroxide induced cell death, we surprisingly found that these factors were identical to the preserved secretory signature. Using the constituents of the anti-apoptotic secretory signature in combination with ischemic and mechanical preconditioning in myocardium mimicking rigidity, we created a comprehensive cytoprotective cocktail to prevent CDCs from ischemia induced cell death. We tested the cocktail in a rat model of ischemia reperfusion, and found a stark reduction in CDC retention post injection. Figures: (A) MicroELISA schematic. Secretions from cells in left are detected by rows of microprinted capture antibodies; photograph in (C-D). Bioluminescence imaging of CDCs treated with comprehensive cocktail show significantly high cell retention vs untreated CDCs.

scholarly journals A ROBOTIC SYSTEM FOR HIGH-THROUGHPUT AUTOMATED LIFESPAN ANALYSIS IN C. ELEGANS

2019 ◽  
Vol 3 (Supplement_1) ◽  
pp. S102-S102
Author(s):  
Ben Blue ◽  
Elena Vayndorf ◽  
Matt Kaeberlein ◽  
Jason Pitt
Keyword(s):  
High Throughput ◽  
Digital Camera ◽  
Nematode Species ◽  
Time Lapse ◽  
Small Population ◽  
Robotic System ◽  
Model Organisms ◽  
C Elegans ◽  
Age Related ◽  
Short Video

Abstract Over the past decade, the identification of potential genetic and pharmacological modifiers of lifespan and age-related pathologies in C. elegans and other model organisms has yielded fruitful leads for follow-up investigation. A major limitation of such studies, however, is that they are often time-consuming and labor-intensive. The advent of affordable high-quality digital cameras, robotics systems, and 3D printers, as well as the decreasing costs of image storage and processing have allowed us to automate data capture and analysis at an unprecedented scale. To this end, our group developed a tool consisting of an unbiased, high-throughput, automated robotic system to perform genetic and pharmacological quantification of lifespan and health measures in C. elegans and related nematode species. The WormBot utilizes industry-standard, commercially available robotics components to position a digital camera over individual wells of standard 12-well culture plates, containing a small population of C. elegans per well. A high-resolution image is captured of each plate every 10 minutes throughout the course of the experiment. Our software processes the images for stabilization, compiles them into a time-lapse series for each well, and quantifies survival and mobility (paralysis) with minimal input. In addition, a short video is captured of each well once each day, to allow for quantitative analyses of activity and coordinated movement. We will describe this technology and present applications to screen genetic and pharmacological libraries in aging and age-related disease.

scholarly journals A Cytotoxic Three-Dimensional-Spheroid, High-Throughput Assay Using Patient-Derived Glioma Stem Cells

2018 ◽  
Vol 23 (8) ◽  
pp. 842-849 ◽  
Author(s):  
Victor Quereda ◽  
Shurong Hou ◽  
Franck Madoux ◽  
Louis Scampavia ◽  
Timothy P. Spicer ◽  
...  
Keyword(s):  
Stem Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Three Dimensional ◽  
Treatment Resistance ◽  
Small Population ◽  
Molecular Probes ◽  
Glioma Stem Cells

Glioblastoma (GBM) is the most aggressive primary brain cancer with an average survival time after diagnosis of only 12–14 months, with few (<5%) long-term survivors. A growing body of work suggests that GBMs contain a small population of glioma stem cells (GSCs) that are thought to be major contributors to treatment resistance and disease relapse. Identifying compounds that modulate GSC proliferation would provide highly valuable molecular probes of GSC-directed signaling. However, targeting GSCs pharmacologically has been challenging. Patient-derived GSCs can be cultured as neurospheres, and in vivo these cells functionally recapitulate the heterogeneity of the original tumor. Using patient-derived GSC-enriched cultures, we have developed a 1536-well spheroid-based proliferation assay and completed a pilot screen, testing ~3300 compounds comprising approved drugs. This cytotoxic and automation-friendly assay yielded a signal-to-background (S/B) ratio of 161.3 ± 7.5 and Z′ of 0.77 ± 0.02, demonstrating its robustness. Importantly, compounds were identified with anti-GSC activity, demonstrating the applicability of this assay for large-scale high-throughput screening (HTS).

154: Integration of TPSA and High-Throughput Mass Spectrometry Data Improves Prostate Cancer Prediction

2007 ◽  
Vol 177 (4S) ◽  
pp. 52-53
Author(s):  
Stefano Ongarello ◽  
Eberhard Steiner ◽  
Regina Achleitner ◽  
Isabel Feuerstein ◽  
Birgit Stenzel ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Mass Spectrometry ◽  
High Throughput ◽  
Mass Spectrometry Data ◽  
Cancer Prediction
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