In the project titled Hierarchy of Anti‐Clumping Salts in these proceedings, in 2525 separate trials by 45 independent investigators, a hierarchy of best to worst yeast unclumping agents was developed using 7 salts related to magnesium sulfate a known anti‐clumping agent. Here we test the long term reliability of this assay in 1404 trials by 29 mostly new investigators done 6 months after the original study. We focused on sodium sulfate, the best unclumper in the previous study. Using the same procedures described in the Hierarchy study, we found that after 60 min, when comparing the mean of percentage of single cells in experimentals vs controls, +20.0% single cells were found in the experimental samples. This is identical to the percent found a half year earlier, confirming the reliability of this assay. Here 18 of 20 groups of data with sodium sulfate, in 415 trials had a p value of less than 0.05 when comparing experimentals and controls. Other reagents were also examined here (magnesium carbonate and sodium bicarbonate) that were not effective unclumping agents. Identification of unclumping agents can be useful in anti‐thrombocytic, anti‐cancer, anti‐infection, anti‐biofilm as well as in industrial applications (support: Sidney Stern Memorial Trust, NIH RISE, NSF Presidential Award 0731633 and university funding sources).

We are identifying reagents that unclump cells in a model yeast system (Saccharomyces cerevisiae) for potential uses in clinical and other applications. Here 7 salts (0.1M concentration range) and no salt controls were stirred with fixed yeast at 20 min intervals over a 60 min time course in 1 ml distilled water droplets on glass microscope slides in 2545 trials by 45 independent investigators counting percentages of unclumped yeast vs clumps at each time point. The 7 salts were chosen because of their relationship to a known anti‐clumping agent magnesium sulfate. The following hierarchy of best to worst unclumping (compared with controls) was established at the 60 min time. Average percent single cells vs controls of all the trials appears in parentheses: sodium sulfate (+20.0), magnesium sulfate (+17.6), potassium sulfate (+9.9), magnesium carbonate (‐1.6), magnesium chloride (‐7.3), calcium carbonate (‐8.4), calcium sulfate (‐13.0). Individual groups of values varied from +59% to ‐41%. T tests were performed to compare results of experimentals vs controls. These results might help identify reagents that can be useful in unclumping cells in the bloodstream such as in anti‐thrombocytic and anti‐cancer applications, in reducing infectivity and biofilm development as well as in industrial applications (support: Sidney Stern Memorial Trust, NIH RISE, NSF Presidential Award 0731633 and Cal State).

MgS04 suppresses platelet aggregation (Brit J Hematol 119: 1033, 2002; Circulation 105: 1970, 2002; Magnes Res 18: 7, 2005; J Matern Fetal Neonat Med: 478, 2012). Intracellular metabolic mechanisms have been proposed to cause this suppression. Here we show, using a simple, inexpensive quantitative assay (FASEB J 27: abs 341, 2013) that MgSO4 disaggregates fixed cells in a yeast (Saccharomyces cerevisiae) model. In 1715 separate experiments by 35 independent investigators, MgSO4 (3mg/slide droplet) increased percent of fixed single cells by 60 min by an average of 17% to as much as 33% (with p values often less than 0.05 compared to controls without reagent), while alpha methyl glucose, D‐glucose and D‐mannitol decreased percent single cells by an average of 15%. It remains to be determined if MgSO4 can unclump fixed platelets, implicating a cell surface mechanism in its antithrombocytic activity. MgSO4 may be a useful anticlumping agent in many applications (Science 341: 186, 2013). Anti‐infection, anti‐biofilm, and anti‐cancer reagents could be identified using this simple, inexpensive, quantitative assay system (Supported by university incentives and grants from NIH RISE, SCORE, Sidney Stern Memorial Trust, Joseph Drown Fdn and NSF Presidential Award 0731633).