The use of a centrifuge-free RABiT-II system for high-throughput micronucleus analysis

ABSTRACT The cytokinesis-block micronucleus (CBMN) assay is considered to be the most suitable biodosimetry method for automation. Previously, we automated this assay on a commercial robotic biotech high-throughput system (RABiT-II) adopting both a traditional and an accelerated micronucleus protocol, using centrifugation steps for both lymphocyte harvesting and washing, after whole blood culturing. Here we describe further development of our accelerated CBMN assay protocol for use on high-throughput/high content screening (HTS/HCS) robotic systems without a centrifuge. This opens the way for implementation of the CBMN assay on a wider range of commercial automated HTS/HCS systems and thus increases the potential capacity for dose estimates following a mass-casualty radiological event.

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The use of a centrifuge-free RABiT-II system for high-throughput micronucleus analysis

10.1101/627596 ◽

2019 ◽

Author(s):

Mikhail Repin ◽

Sergey Pampou ◽

David J. Brenner ◽

Guy Garty

Keyword(s):

High Throughput ◽

Whole Blood ◽

Mass Casualty ◽

Robotic Systems ◽

High Content Screening ◽

Potential Capacity ◽

Cbmn Assay ◽

Further Development ◽

Assay Protocol ◽

The Way

AbstractThe cytokinesis-block micronucleus (CBMN) assay is considered as the most suitable biodosimetry method for automation. Previously, we automated this assay on a commercial robotic biotech high-throughput system (RABiT-II) adopting both a traditional and an accelerated micronucleus protocol, both using centrifugation steps for lymphocyte harvesting and washing, after whole blood culturing. Here we describe further development of our accelerated CBMN assay protocol for using on High Throughput/High Content Screening (HTS/HCS) robotic systems without a centrifuge. This opens the way for implementation of the CBMN assay on a wider range of commercial automated HTS/HCS systems and thus increases the potential capacity of dose estimates following a mass-casualty radiological event.

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New Imidazole Inhibitors of Mycobacterial FtsZ: the Way from High-Throughput Molecular Screening in Grid up to in vitro Verification

Science and innovation ◽

10.15407/scine12.03.043 ◽

2016 ◽

Vol 12 (3) ◽

pp. 43-55 ◽

Cited By ~ 4

Author(s):

P.A. Karpov ◽

◽

O.M. Demchuk ◽

V.M. Britsun ◽

D.I. Lytvyn ◽

...

Keyword(s):

High Throughput ◽

Molecular Screening ◽

The Way

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Whole blood gene expression within days after total-body irradiation predicts long term survival in Gottingen minipigs

Scientific Reports ◽

10.1038/s41598-021-95120-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sunita Chopra ◽

Maria Moroni ◽

Jaleal Sanjak ◽

Laurel MacMillan ◽

Bernadette Hritzo ◽

...

Keyword(s):

Gene Expression ◽

Total Body Irradiation ◽

Whole Blood ◽

Radiation Effects ◽

Expression Profiles ◽

Mass Casualty ◽

Machine Learning Algorithms ◽

Term Survival ◽

Blood Gene Expression ◽

Total Body

AbstractGottingen minipigs mirror the physiological radiation response observed in humans and hence make an ideal candidate model for studying radiation biodosimetry for both limited-sized and mass casualty incidents. We examined the whole blood gene expression profiles starting one day after total-body irradiation with increasing doses of gamma-rays. The minipigs were monitored for up to 45 days or time to euthanasia necessitated by radiation effects. We successfully identified dose- and time-agnostic (over a 1–7 day period after radiation), survival-predictive gene expression signatures derived using machine-learning algorithms with high sensitivity and specificity. These survival-predictive signatures fare better than an optimally performing dose-differentiating signature or blood cellular profiles. These findings suggest that prediction of survival is a much more useful parameter for making triage, resource-utilization and treatment decisions in a resource-constrained environment compared to predictions of total dose received. It should hopefully be possible to build such classifiers for humans in the future.

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In vitro effects of temperature on red blood cell deformability and membrane stability in human and various vertebrate species

Clinical Hemorheology and Microcirculation ◽

10.3233/ch-211118 ◽

2021 ◽

pp. 1-10

Author(s):

Adam Attila Matrai ◽

Gabor Varga ◽

Bence Tanczos ◽

Barbara Barath ◽

Adam Varga ◽

...

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Whole Blood ◽

Mechanical Stability ◽

Cell Deformability ◽

Blood Samples ◽

Effects Of Temperature ◽

The Way ◽

Rbc Deformability

BACKGROUND: The effects of temperature on micro-rheological variables have not been completely revealed yet. OBJECTIVE: To investigate micro-rheological effects of heat treatment in human, rat, dog, and porcine blood samples. METHODS: Red blood cell (RBC) - buffer suspensions were prepared and immersed in a 37, 40, and 43°C heat-controlled water bath for 10 minutes. Deformability, as well as mechanical stability of RBCs were measured in ektacytometer. These tests were also examined in whole blood samples at various temperatures, gradually between 37 and 45°C in the ektacytometer. RESULTS: RBC deformability significantly worsened in the samples treated at 40 and 43°C degrees, more expressed in human, porcine, rat, and in smaller degree in canine samples. The way of heating (incubation vs. ektacytometer temperation) and the composition of the sample (RBC-PBS suspension or whole blood) resulted in the different magnitude of RBC deformability deterioration. Heating affected RBC membrane (mechanical) stability, showing controversial alterations. CONCLUSION: Significant changes occur in RBC deformability by increasing temperature, showing inter-species differences. The magnitude of alterations is depending on the way of heating and the composition of the sample. The results may contribute to better understanding the micro-rheological deterioration in hyperthermia or fever.

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Molecular replacement and high-throughput structure determination

Zeitschrift für Kristallographie - Crystalline Materials ◽

10.1524/zkri.217.12.715.20653 ◽

2002 ◽

Vol 217 (12) ◽

Author(s):

J. Navaza ◽

P. M. Alzari

Keyword(s):

High Throughput ◽

Structure Determination ◽

Solution Space ◽

Molecular Replacement ◽

Fast Rotation ◽

The Way

AbstractWe describe the philosophy of the MR method as implemented in the AMoRe package. Fast rotation and translation functions are first used to obtain a meaningful sampling of solution space, whose elements are subsequently assessed by using more robust criteria. The introduction of fast and accurate algorithms for screening a large number of possible solutions opened the way to automation, thus bringing MR methods to the realm of high-throughput structure determination. Selected examples are discussed to illustrate specific aspects of the method.

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A Novel Cell-Based, High-Content Assay for Phosphorylation of Lats2 by Aurora A

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057111413923 ◽

2011 ◽

Vol 16 (8) ◽

pp. 925-931 ◽

Cited By ~ 7

Author(s):

Amy Emery ◽

David A. Sorrell ◽

Stacy Lawrence ◽

Emma Easthope ◽

Mark Stockdale ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Kinase Activity ◽

High Content Screening ◽

Aurora A Kinase ◽

Aurora A ◽

Assay Performance ◽

A Cell ◽

Significant Addition ◽

Specific Inhibitors

Aurora A kinase is a key regulator of mitosis, which is upregulated in several human cancers, making it a potential target for anticancer therapeutics. Consequently, robust medium- to high-throughput cell-based assays to measure Aurora A kinase activity are critical for the development of small-molecule inhibitors. Here the authors compare measurement of the phosphorylation of two Aurora A substrates previously used in high-content screening Aurora A assays, Aurora A itself and TACC3, with a novel substrate Lats2. Using antibodies directed against phosphorylated forms of Aurora A (pThr288), P-TACC3 (pSer558), and P-Lats2 (pSer83), the authors investigate their suitability in parallel for development of a cell-based assay using several reference Aurora inhibitors: MLN8054, VX680, and AZD1152-HQPA. They validate a combined assay of target-specific phosphorylation of Lats2 at the centrosome and an increase in mitotic index as a measure of Aurora A activity. The assay is both sensitive and robust and has acceptable assay performance for high-throughput screening or potency estimation from concentration–response assays. It has the advantage that it can be carried out using a commercially available monoclonal antibody against phospho-Lats2 and the widely available Cellomics ArrayScan HCS reader and thus represents a significant addition to the tools available for the identification of Aurora A specific inhibitors.

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Interrogating the Founding Gestures of the New Materialism

Cultural Studies Review ◽

10.5130/csr.v22i2.4461 ◽

2016 ◽

Vol 22 (2) ◽

Cited By ~ 6

Author(s):

Dennis Bruining

Keyword(s):

New Materialism ◽

The Past ◽

The World ◽

Further Development ◽

Selection Of ◽

The Way

In this article, I aim to further thinking in the broadly ‘new materialist’ field by insisting it attends to some ubiquitous assumptions. More specifically, I critically interrogate what Sara Ahmed has termed ‘the founding gestures of the “new materialism”’. These founding rhetorical gestures revolve around a perceived neglect of the matter of materiality in ‘postmodernism’ and ‘poststructuralism’ and are meant to pave the way for new materialism’s own conception of matter-in/of-the-world. I argue in this article that an engagement with the postmodern critique of language as constitutive, as well as the poststructuralist critique of pure self-presence, does not warrant these founding gestures to be so uncritically rehearsed. Moreover, I demonstrate that texts which rely on these gestures, or at least the ones I discuss in this article, are not only founded on a misrepresentation of postmodern and poststructuralist thought, but are also guilty of repeating the perceived mistakes of which they are critical, such as upholding the language/matter dichotomy. I discuss a small selection of texts that make use of those popular rhetorical gestures to juxtapose the past that is invoked with a more nuanced reading of that past. My contention is that if ‘the founding gestures of the “new materialism”’ are not addressed, the complexity of the postmodern and poststructuralist positions continues to be obscured, with damaging consequences for the further development of the emerging field of new materialism, as well as our understanding of cultural theory’s past.

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