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A Manual Kinetic Assay in a Fixed Yeast Model for Drug Discovery

Received: 12 May 2019     Accepted: 10 June 2019     Published: 24 June 2019
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Abstract

Manual cell counting assays are often considered as a gold standard in some applications because of direct observation of results. Here a manual kinetic cell counting method is used to evaluate the efficacy of reagents to unclump cells in a fixed yeast (Saccharomyces cerevisae) model. Clumped cells are more dangerous than single cells in many human venues such as cancer spread, thrombocytic blockages and biofilm infectivity. In this study percentage of single cells are analyzed over time in the presence or absence of specific reagents and the number of cell clumps and cells per clump are also assessed. The results show that when the percentage of single cells increases, the number of clumps and cells per clump decrease, helping to validate the assay. These findings suggest that this assay can be a gold standard for evaluating the effects of specific reagents on cell unclumping in a model system that can be used in drug discovery investigations. This study is part of a program that won a U.S. Presidential Award for science mentoring presented by President Obama at the White House. The experiments are easily accomplished by undergraduate students and can be done by research classes without background in complex science methodology.

Published in American Journal of Applied Scientific Research (Volume 5, Issue 1)
DOI 10.11648/j.ajasr.20190501.15
Page(s) 28-34
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2019. Published by Science Publishing Group

Keywords

Cell Unclumping Kinetic Assay, Fixed Yeast Model, Drug Discovery

References
[1] G. Zem, B. Cooperman, F. Bahri, A. Mahjoubi, N. Warner, L. Malekian, R. Mirebrahimian, M. Pistalu, M. Patel, E. Choi, T. Baronian, O. Gilani, A. Cardenas, A. Hambarsoomian, D. Gomez, F. Gallgos, J. Holmes, V. Vahdati, L. Jorshari, P. Grigorian, K. Ohanessian, E. Baum, G. Majarian, K. Aldzhyan, H. Manasyan, N. Allatakakhsh, S. Oppenheimer Reliability of yeast unclumping assay, a model for testing potentially clinically useful reagents FASEB J, 29 (2015), abstract 925.3.
[2] G. Zem, A. Chimayan, V. Aleksanyan, J. Gordon, F. Gomez, A. Seyedroubari, J. Chang, T. Botello, N. Tan, D. Arefin, D. Tobar, A. Khachekian, L. Gama, E. Durodola, J. Batty, C. Plascencia, L. Barillas, A. Roverud, S. Kreuz, L. Sarkisyan, F. Lee, J. Munoz, L. Reque, V. Abed, L. Kinog, S. B. Oppenheimer A kinetic assay for non-automated drug screening FASEB J, 32 (2018), abstract 113.
[3] G. Zem, V. Nahapetyan, K. Crocker, D. Tobar, S. Delos Santos, D. Nazarian, J. Hovannes, G. Beltran, R. Dubin, L. Reque, P. Singh, L. sarkisyan, B. Cardona, G. Bachinela, S. B. Oppenheimer Validation of a manual cell counting assay for assessing the kinetics of cell unclumping reagents in drug discovery FASEB J, 33 (2019), abstract 19.
[4] National Science Foundation Excellence Awards in Science and Engineering Program: Presidential Awards for Excellence in Science, Mathematics and Engineering Mentoring (EASE: PAESMEM). Solicitation 16-534. 2019.
[5] OSTP and NSF to honor 140 individuals and organizations with highest US award for teachers and mentors. News Release 18-043, June 25, 2018.
[6] Presidential Awards for Excellence in Science, Mathematics and Engineering Mentoring, NSF Home Page, 2019.
[7] NSF Proposal 07316333, March 6, 2007, Individual-Steven Oppenheimer Mentor, Researcher, Teacher.
[8] Herstein, O. Steven Oppenheimer CSUN Magazine, no. 67 (2016), p. 20.
[9] P. C. Zeidler-Erdely, J. M. Antonini, T. G. Meighan, S-H. Young, T. J. Eve, M. A. Hammer, A. Erdely Comparison of cell counting methods in rodent pulmonary toxicity studies: automated and manual protocols and considerations for experimental design Inhal Toxicol, 28 (2016), pp. 410-420.
[10] V. M. Navarro, S. L. Walker, O. Badali, L. L. Ngo, S. Weerasinghe, M. Barajas, G. Zem, S. B. Oppenheimer Analysis of surface properties of fixed and live cells using derivatized agarose beads Acta Histochem, 104 (2002), pp. 99-106.
[11] K. Germanovich, E. Alessandra Femiaq, C. Y. Cheng, N. Dovlatova, M. Cattaneo Effects of pH and concentration of sodium citrate anticoagulant on platelet aggregation measured by light transmission aggregometry induced by adenosine diphosphate Platelets, 29 (2018), pp. 21-26.
[12] N. Aceto, A. Bardia, D. T. Miyamoto, M. C. Donandson, B. S. Wittner, J. A. Spencer, M. Yu, A. Pely, A. Engstrom, H. Zhu, B. W. Brannigan, R. Kapur, S. L. Stott, T. Shioda, S. Ramaswamy, D. T. Ting, C. P. Lin, M. Toner, D. A. Haber, S. Maheswaran Circulating tumor cell clusters are oligoclonal precursors of breast cancer metastasis Cell, 28 (2014), pp1110-1122.
[13] S. H. Au, B. D. Storey, J. C. Moore, Q. Tang, Y-L. Chen, S. Javaid, A. F. Sarioglu, R. Sullivan, M. W. Madden, R. O’Keefe, D. A. Haber, S. Maheswaran, D. M. Langenau, S. L. Stott, M. Toner Clusters of circulating tumor cells traverse capillary-sized vessels Proc Natl Acad Sci US, 113 (2016), pp. 4947-4952.
[14] D. Cadena-Herrera, J. E. Esparza-De Lara, N. D. Ramirez-Ibanez, C. A. Lopez-Marales, N. O. Perez, L. F. Flores-Ortiz, E. Medina-Rivero Validation of three viable cell counting methods: manual, semi-automated, and automated Biotechnology Rep, 7 (2015), pp. 9-16.
[15] Kolodkin-Gal, D. Romero, S. Cao, J. Clardy, R. Kolter, R. Losick D-amino acids trigger biofilm disassembly Science 30, (2010), pp. 627-629.
Cite This Article
  • APA Style

    Vahe Nahapetyan, Shiela Delos Santos, Kristel Joy Crocker, Dayana Tobar, Dante Nazarian, et al. (2019). A Manual Kinetic Assay in a Fixed Yeast Model for Drug Discovery. American Journal of Applied Scientific Research, 5(1), 28-34. https://doi.org/10.11648/j.ajasr.20190501.15

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    ACS Style

    Vahe Nahapetyan; Shiela Delos Santos; Kristel Joy Crocker; Dayana Tobar; Dante Nazarian, et al. A Manual Kinetic Assay in a Fixed Yeast Model for Drug Discovery. Am. J. Appl. Sci. Res. 2019, 5(1), 28-34. doi: 10.11648/j.ajasr.20190501.15

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    AMA Style

    Vahe Nahapetyan, Shiela Delos Santos, Kristel Joy Crocker, Dayana Tobar, Dante Nazarian, et al. A Manual Kinetic Assay in a Fixed Yeast Model for Drug Discovery. Am J Appl Sci Res. 2019;5(1):28-34. doi: 10.11648/j.ajasr.20190501.15

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  • @article{10.11648/j.ajasr.20190501.15,
      author = {Vahe Nahapetyan and Shiela Delos Santos and Kristel Joy Crocker and Dayana Tobar and Dante Nazarian and Hovannes Chirishyan and Gerard Beltran and Rachel Dubin and Leticia Reque and Prabkiran Singh and Brandie Cardona and Graciel Royce Bachinela and Lillyt Sarkisyan and Gregory Zem and Steven Oppenheimer},
      title = {A Manual Kinetic Assay in a Fixed Yeast Model for Drug Discovery},
      journal = {American Journal of Applied Scientific Research},
      volume = {5},
      number = {1},
      pages = {28-34},
      doi = {10.11648/j.ajasr.20190501.15},
      url = {https://doi.org/10.11648/j.ajasr.20190501.15},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajasr.20190501.15},
      abstract = {Manual cell counting assays are often considered as a gold standard in some applications because of direct observation of results. Here a manual kinetic cell counting method is used to evaluate the efficacy of reagents to unclump cells in a fixed yeast (Saccharomyces cerevisae) model. Clumped cells are more dangerous than single cells in many human venues such as cancer spread, thrombocytic blockages and biofilm infectivity. In this study percentage of single cells are analyzed over time in the presence or absence of specific reagents and the number of cell clumps and cells per clump are also assessed. The results show that when the percentage of single cells increases, the number of clumps and cells per clump decrease, helping to validate the assay. These findings suggest that this assay can be a gold standard for evaluating the effects of specific reagents on cell unclumping in a model system that can be used in drug discovery investigations. This study is part of a program that won a U.S. Presidential Award for science mentoring presented by President Obama at the White House. The experiments are easily accomplished by undergraduate students and can be done by research classes without background in complex science methodology.},
     year = {2019}
    }
    

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  • TY  - JOUR
    T1  - A Manual Kinetic Assay in a Fixed Yeast Model for Drug Discovery
    AU  - Vahe Nahapetyan
    AU  - Shiela Delos Santos
    AU  - Kristel Joy Crocker
    AU  - Dayana Tobar
    AU  - Dante Nazarian
    AU  - Hovannes Chirishyan
    AU  - Gerard Beltran
    AU  - Rachel Dubin
    AU  - Leticia Reque
    AU  - Prabkiran Singh
    AU  - Brandie Cardona
    AU  - Graciel Royce Bachinela
    AU  - Lillyt Sarkisyan
    AU  - Gregory Zem
    AU  - Steven Oppenheimer
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    PY  - 2019
    N1  - https://doi.org/10.11648/j.ajasr.20190501.15
    DO  - 10.11648/j.ajasr.20190501.15
    T2  - American Journal of Applied Scientific Research
    JF  - American Journal of Applied Scientific Research
    JO  - American Journal of Applied Scientific Research
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    EP  - 34
    PB  - Science Publishing Group
    SN  - 2471-9730
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    AB  - Manual cell counting assays are often considered as a gold standard in some applications because of direct observation of results. Here a manual kinetic cell counting method is used to evaluate the efficacy of reagents to unclump cells in a fixed yeast (Saccharomyces cerevisae) model. Clumped cells are more dangerous than single cells in many human venues such as cancer spread, thrombocytic blockages and biofilm infectivity. In this study percentage of single cells are analyzed over time in the presence or absence of specific reagents and the number of cell clumps and cells per clump are also assessed. The results show that when the percentage of single cells increases, the number of clumps and cells per clump decrease, helping to validate the assay. These findings suggest that this assay can be a gold standard for evaluating the effects of specific reagents on cell unclumping in a model system that can be used in drug discovery investigations. This study is part of a program that won a U.S. Presidential Award for science mentoring presented by President Obama at the White House. The experiments are easily accomplished by undergraduate students and can be done by research classes without background in complex science methodology.
    VL  - 5
    IS  - 1
    ER  - 

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Author Information
  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

  • Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, United States

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