A meta-analysis framework to assess the role of units in describing nanoparticle toxicity.

Despite ample research on nanoparticles, their environmental toxicity is still debatable. The lack of consensus is due in part to the challenge of comparing studies because of variability in parameters like test organism, test medium, and duration of experiment. However, the unit used to compare the toxicology of nanoparticles is one variable that experimentalists can control. Traditionally, mass per volume is the most common unit used to make comparisons, but there is growing evidence that alternative units such as surface area per volume or particles per volume may provide a better and more mechanistic measure of toxicity. Herein, we propose and test a meta-analytic framework to study the effect of units on nanotoxicology using data from the NanoE-Tox database, a freely available database containing 1518 toxicology values from 224 published articles of which 42 records met our basic inclusion criteria. These data were augmented with more recent data published over the past five years as archived by the Web of Science citation index. An additional 27 records from 1676 papers met the inclusion criteria and were also included in the analysis. The meta-analysis framework measures the degree of heterogeneity for each of three units (grams/L, particles/L, surface area/L) grouped by the type of test organism, particle chemistry, and manner in which a nanoparticle's size was measured (e.g., nominal particle size reported by the manufacturer vs. measurement of size for particles suspended in the liquid medium used in a subsequent toxicity experiment). The result of the meta-analysis reveals that surface area per volume reduces the heterogeneity in the Ag crustacean subgroup when nanoparticle size was measured in the test medium, and the ZnO crustacean subgroup when nanoparticle size was measured out the test medium and may therefore be a more appropriate estimate of the toxicity of soluble nanoparticles. No subgroups in our analysis showed a reduction in heterogeneity for particles per volume in either soluble or insoluble nanoparticles. The lack of conclusion on insoluble nanoparticles was not due to a limitation of our meta-analysis but rather highlights a critical deficiency in the primary literature. The majority of published studies fail to report common measures of error that are essential for further analysis (i.e. error of the measured nanoparticle size and/or interoperable error of the measured half-maximal concentration of the toxic endpoint). If future nanotoxicity studies report such error, as they should, then the framework of our meta-analysis could be used more broadly to provide a simple, statistically rigorous way to assess the role of units on the toxicity of nanoparticles.

Fibrinogen binding is affected by amino acid substitutions in C-terminal repeat region of fibronectin binding protein A.

Fibronectin-binding protein A (FnBPA), a protein displayed on the outer surface of Staphylococcus aureus, has a structured A-domain that binds fibrinogen (Fg) and a disordered repeat-region that binds fibronectin (Fn). Amino acid substitutions in Fn-binding repeats (FnBRs) have previously been linked to cardiovascular infection in humans. Here we used microtiter and atomic force microscopy (AFM) to investigate adhesion by variants of full-length FnBPA covalently anchored in the outer cell wall of Lactococcus lactis, a Gram-positive surrogate that otherwise lacks adhesins to mammalian ligands. Fn adhesion increased in five of seven FnBPA variants under static conditions. The bond targeting Fn increased its strength with load under mechanical dissociation. Substitutions extended bond lifetime (1/koff) up to 2.1 times for FnBPA-Fn. Weaker adhesion was observed for Fg in all FnBPA variants tested with microtiter. However, mechanical dissociation with AFM showed significantly increased tensile strength for Fg interacting with the E652D/H782Q variant. This is consistent with a force-induced mechanism and suggests that the dock, lock, and latch (DLL) mechanism is favored for Fg-binding under mechanical stress. Collectively, these experiments reveal that FnBPA exhibits bimodal, ligand-dependent adhesive behavior. Amino acid substitutions in the repeat-region of FnBPA impact binding to both ligands. This was unexpected for Fg since all variants have the same A-domain sequence, and the Fg-binding site is distant from the repeat region. This indicates that FnBRs may fold back on the A-domain in a way that impacts the DLL binding mechanism for Fg.