Differential induction of apoptosis in undifferentiated and differentiated HL-60 cells by DNA topoisomerase I and II inhibitors

The effects of monocytic/macrophage and granulocytic differentiation induced by phorbol myristate acetate (TPA) and all-trans retinoic acid, respectively, were tested on the induction of apoptosis in human promyelocytic leukemia HL-60 cells treated with topoisomerase I and II inhibitors. Using a filter-binding assay, we observed a strong inhibition of DNA fragmentation induced by 3- and 24-hour continuous exposure to camptothecin, VP-16, VM-26, and m-AMSA in TPA- differentiated cells. The inhibition of the typical internucleosomal DNA fragmentation was confirmed by agarose gel electrophoresis. By contrast, drug-induced DNA fragmentation was not inhibited in retinoic acid-differentiated cells, and apoptosis occurred in these cells after 4 to 5 days in the absence of drug treatment. The TPA inhibitory effect was maximal after 24 hours of treatment and was correlated with differentiation, because phorbol dibutyrate ester was active, whereas 4- alpha-TPA, a nontumor promoter that does not induce differentiation, was not active. Using alkaline elution, we observed that TPA and retinoic acid differentiation were associated with changes in topoisomerase-mediated DNA breaks that were not correlated with their differential effects on drug-induced DNA fragmentation. Moreover, TPA also inhibited DNA fragmentation induced by vinblastine, cycloheximide, calphostin C, and x-rays. Using a cell-free system, we observed that DNA fragmentation was not inhibited in nuclei from TPA-differentiated cells. Rather, inhibition of apoptosis seemed to take place in the cytoplasm. We conclude that phenotypic changes associated with TPA- induced differentiation include inactivation of a cytoplasmic activity that can induce DNA fragmentation associated with apoptosis.     

Prediction of ARA/PPI Drug-Drug Interactions at the Drug Discovery and Development Interface

Advances in understanding of human disease have prompted the U.S. Food and Drug Administration to classify certain molecules as “break-through therapies,” providing an accelerated review that may potentially enhance the quality of patient lives. With this designation come compressed timelines to develop drug products, which are not only suitable for clinic trials but can also be approved and brought to the market rapidly. Early risk identification for decreased oral absorption due to drug-drug interactions with proton pump inhibitors (PPIs) or acid-reducing agents (ARAs) is paramount to an effective drug product development strategy. An early ARA/PPI drug-drug interaction (DDI) risk identification strategy has been developed using physiologically based absorption modeling that readily integrates ADMET predictor generated in silico estimates or measured in vitro solubility, permeability, and ionization constants. Observed or predicted pH-solubility profile data along with pKas and drug dosing parameters were used to calculate a fraction of drug absorbed ratio in absence and presence of ARAs/PPIs. An integrated physiologically based pharmacokinetic absorption model using GastroPlus? with pKa values fitted to measured pH-solubility profile data along with measured permeability data correctly identified the observed ARA/PPI DDI for 78% (16/22) of the clinical studies. Formulation strategies for compounds with an anticipated pH-mediated DDI risk are presented.     

Alteration of Root Growth by Lettuce,Wheat, and Soybean in Response to Wear Debris from Automotive Brake Pads

Brakes from motor vehicles release brake pad wear debris (BPWD) with increased concentrations of heavy metals. Germination and root-elongation assays with lettuce, wheat, and soybean were used to provide an initial evaluation of the phytotoxicity of either a water extract of BPWD or BPWD particulates. In terms of germination, the only effect observed was that lettuce germination decreased significantly in the BPWD particulate treatment. Lettuce and wheat showed decreased root length and root-elongation rate in the presence of the BPWD particulates, whereas lettuce produced a significantly greater number of lateral roots in response to BPWD extract. There was no significant effect of either BPWD treatment on soybean root elongation or lateral roots. Treatment with BPWD extracts or particulates caused significant alterations in the bending pattern of the plant roots. These initial results suggest that BPWD may have effects on the early growth and development of plants.     

Time‐ and concentration‐dependent genomic responses of the rat airway to inhaled nickel sulfate

While insoluble nickel subsulfide (Ni₃S₂) was carcinogenic in the lung in a 2‐year rat bioassay, soluble nickel sulfate hexahydrate (NiSO_4*6H₂O) was not. To investigate whether differences in the cellular responses to these two nickel compounds could underlie their differential activities, we conducted parallel studies to determine the gene expression changes in micro‐dissected lung distal airway cells from Fischer 344 rats following inhalation of the two compounds for one and four weeks (6 hr per day, 5 days per week). The results of the Ni₃S₂ study have been reported previously; this paper reports the results for NiSO₄ and provides a comparative analysis. The cellular responses to NiSO₄ were highly similar to those previously reported for Ni₃S₂, and a set of genes was identified whose expression could be used as biomarkers for comparing cellular nickel effects from in vitro or in vivo studies with soluble NiSO₄ and particulate Ni₃S₂. Evaluation of the genomic concentration‐responses for the two compounds suggests that the highest inhaled concentration in the tumor bioassay for NiSO₄, which was limited by toxicity, may not have achieved the Ni concentrations at which tumors were observed in the Ni₃S₂ bioassay. However, several key differences in the immune responses to NiSO₄ and Ni₃S₂ were identified that may result from the differential intracellular disposition of Ni from NiSO₄ entering the cell as an ion rather than as a slowly soluble Ni₃S₂ particle. These differences may also contribute to the observation of tumors in the bioassay for Ni₃S₂ but not NiSO₄. Environ. Mol. Mutagen. 58:607–618, 2017. © 2017 The Authors Environmental and Molecular Mutagenesis published by Wiley Periodicals, Inc. on behalf of Environmental Mutagen Society