Application of the adverse outcome pathway framework to genotoxic modes of action

In May 2017, the Health and Environmental Sciences Institute's Genetic Toxicology Technical Committee hosted a workshop to discuss whether mode of action (MOA) investigation is enhanced through the application of the adverse outcome pathway (AOP) framework. As AOPs are a relatively new approach in genetic toxicology, this report describes how AOPs could be harnessed to advance MOA analysis of genotoxicity pathways using five example case studies. Each of these genetic toxicology AOPs proposed for further development includes the relevant molecular initiating events, key events, and adverse outcomes (AOs), identification and/or further development of the appropriate assays to link an agent to these events, and discussion regarding the biological plausibility of the proposed AOP. A key difference between these proposed genetic toxicology AOPs versus traditional AOPs is that the AO is a genetic toxicology endpoint of potential significance in risk characterization, in contrast to an adverse state of an organism or a population. The first two detailed case studies describe provisional AOPs for aurora kinase inhibition and tubulin binding, leading to the common AO of aneuploidy. The remaining three case studies highlight provisional AOPs that lead to chromosome breakage or mutation via indirect DNA interaction (inhibition of topoisomerase II, production of cellular reactive oxygen species, and inhibition of DNA synthesis). These case studies serve as starting points for genotoxicity AOPs that could ultimately be published and utilized by the broader toxicology community and illustrate the practical considerations and evidence required to formalize such AOPs so that they may be applied to genetic toxicity evaluation schemes. Environ. Mol. Mutagen. 61:114–134, 2020. © 2019 Wiley Periodicals, Inc.     

Learning a high-precision locomotor task in patients with Parkinson's disease.

We evaluated the acquisition and performance of a high-precision locomotor task in patients with Parkinson's disease (PD) and healthy subjects. All subjects walked on a treadmill and had to step repetitively as low as possible over an obstacle without touching it. During blocks 1 and 2, the subjects had full vision and received additional acoustic warning and feedback signals. During block 3, vision became restricted. Changes in foot clearance and the number of obstacle hits were evaluated. Initially, PD patients performed poorer and improved foot clearance slower. After task repetition, the groups performed similarly. Restricting vision deteriorated performance in both groups. The similar performance of PD patients after task repetition might indicate that adequate training could improve adaptive locomotor behavior in PD patients.     

Occupational exposure to hazardous substances and risk of cancer in the area of the mouth cavity, oropharynx, hypopharynx and larynx. A case-control study

A case-control study of squamous cell carcinoma of the upper aerodigestive tract conducted in Heidelberg and Giessen (FRG) provided information on occupational factors in 200 patients and 800 controls (adjusted to sex, age and area of living; 4:1 matched design). The number of subjects exposed to wood dusts, organic chemicals, coal products or to cement was significantly elevated in the tumour group. An increased risk for head and neck cancer was observed after exposition to wood dust (RR = 2,2), organic compounds (RR = 2,4), coal products (RR = 2,7) and especially to cement (RR = 4,4). The cancer risk due to cement exposition showed a positive correlation to the duration of exposition and remained significantly elevated after adjustment for alcohol and tobacco consumption.     

Ultrastructural changes in the blood-brain barrier after nimodipine treatment and induced hypertension

Fourty-four narcotized rats were split into two equal groups, one being treated with nimodipine and the other with a placebo. By use of norfenefrine the blood pressure was raised to values of 150 and 180 mm Hg within the limits of the autoregulation of brain perfusion and under continuous measurement. Fifteen minutes after application of the standard tracer, horseradish peroxidase, the animals were exsanguinated using a saline perfusion and then perfusion-fixed with Karnovsky's solution. After development of the peroxidase staining the brain sections were evaluated and then allocated to their respective groups. In brain tissues from the experimental group significantly more frequent perivascular accumulations of horseradish peroxidase reaction product were found (P less than 0.001). In electron micrographs it could be seen that the tight junctions were intact and that there was a neuroendothelial transport, with horseradish peroxidase-filled vesicles, in the endothelium, muscle cells, and brain parenchyma. These vesicles represent a medium of transport for all proteins of high molecular weight and can therefore result in brain edema. It is concluded that nimodipine damages the blood-brain barrier by disturbance of the autoregulation of the cerebral blood flow.     

Asymmetric space closure

In a vertically slotted 0.022 X 0.028 inch edgewise system, the use of a two-dimensional ribbon arch--0.022 X 0.016 inch from lateral incisor to lateral incisor and 0.016 X 0.022 inch from the canine to the molars--plus an uprighting spring on the canine maintained incisor position while unilaterally protracting a molar-premolar unit by means of sliding mechanics. Five degrees of labial crown torque was placed in the incisor portion of the wire and the uprighting spring exerted a force of 200 to 250 g. The intra-arch force applied to protract the molar--premolar unit was 300 to 350 g.     

Anleitung für eine vollständige Kehlkopfpräparation

Zusammenfassung Anhand von 9 Abbildungen wird eine Methode zur Kehlkopfpräparation beschrieben, bei der sämtliche Knorpel, Gelenke und Muskeln dargestellt werden. Nach Ablösung des Schildknorpels und des Hypopharynx werden die Mm. postici und arytaenoidei isoliert, die Cricoarytaenoidgelenke sowie die Arytaenoidknorpel präpariert und die Mm. laterales eingeschnitten. Ein Horizontalschnitt durch den Ringknorpel und die median-sagittale Spaltung des proximalen Fragmentes liefern schließlich den Zugang zu den inneren Kehlkopfweichteilen.     

Interaction of a plasminogen activator proteinase, LV-PA with human alpha2-macroglobulin.

Lachesis venom plasminogen activator (LV-PA) is a 33-kDa serine proteinase isolated from bushmaster (Lachesis muta muta) snake venom, which activates the fibrinolytic system in vitro. This study has examined the effect of the plasma proteinase inhibitor alpha2-macroglobulin (alpha2-M) towards LV-PA and compares it with the effect on tissue type plasminogen activator (t-PA). The proteolytic activity of LV-PA alone or previously incubated with human plasminogen (Plg) on the large molecular mass protein substrates, dimethylcasein (DMC) and fibrinogen (Fg) was completely inhibited by human alpha2-M. However, the synthetic peptides Tos-Gly-Pro-Lys-pNA and H-D-Pro-Phe-Arg-pNA (S-2302) were hydrolyzed with almost no reduction in rate. At pH 7.4 and 37 degrees C the proteinase (0.15 microM over 15 min) interacted with alpha2-M, and each mole of alpha2-M bound 2 mol of enzyme. Sodium dodecyl sulfate gel electrophoresis of reduced samples showed that the interaction of alpha2-M with either LV-PA or t-PA preincubated with Plg resulted in the formation of approximately 90 kDa fragments and high molecular mass complexes (Mr 180 kDa), generated by the incubation mixture (LV-PA or t-PA) and Plg. The data suggest that LV-PA is a direct-type PA and its fibrinolytic effect can be reduced by alpha2-M in vivo.     

An improved method for determining 89Sr and 90Sr in urine

A method has been developed for the rapid isolation and quantitation of 89Sr and 90Sr in urine samples. The radiostrontium is concentrated from the bulk urine sample by coprecipitation with calcium phosphate. The precipitate is then wet ashed with nitric acid, and a solution of the resulting residue in 2 M HNO3-0.5 M Al(NO3)3 is passed through an extraction chromatographic column containing a supported crown ether that preferentially retains strontium. Sorbed strontium may then be eluted from the column with either dilute HNO3 or water and counted via liquid scintillation. A new counting scheme that permits quantitation of both 89Sr and 90Sr on the same day the separation is performed is described.