C8- and C9-Alkylphenols and Ethoxylates: II. Assessment of Environmental Persistence and Bioaccumulation Potential

C8- and C9-alkylphenols and their ethoxylates (APE) are widely used commercial products mainly used in industrial applications, in the formulation of crop protection chemicals, and in industrial and household cleaners. Recent regulatory focus on these compounds has included an assessment of their potential to meet criteria for persistent, bioaccumulative, and toxic compounds (PBT). To fully evaluate either the relative persistence or bioaccumulation potential of any APE, degradation intermediates and metabolic by-products of these compounds should also be considered. To facilitate the evaluation of the ultimate fate of APE in the environment, a review of the degradation pathways and identification of degradation intermediates was performed (part I of a two-part series). In part II of this series, the relative persistence of APE as indicated by degradation half-lives was examined based on a review of abiotic and biological degradation data. To assess the bioaccumulation potential of APE, the relevant literature was also reviewed. The available data for C8- and C9-APE show that the commercial products and their degradation intermediates do not meet any national or international criteria for identifying these compounds as PBT substances.     

Degradation of organic contaminants found in organic waste

In recent years, great interest has arisen in recycling of the waste created by modern society. A common way of recycling the organic fraction is amendment on farmland. However, these wastes may contain possible hazardous components in small amounts, which may prevent their use in farming. The objective of our study has been to develop biological methods by which selected organic xenobiotic compounds can be biotransformed by anaerobic or aerobic treatment. Screening tests assessed the capability of various inocula to degrade two phthalates di-n-butylphthalate, and di(2-ethylhexyl)phthalate, five polycyclic aromatic hydrocarbons, linear alkylbenzene sulfonates and three nonylphenol ethoxylates under aerobic and anaerobic conditions. Under aerobic conditions, by selecting the appropriate inoculum most of the selected xenobiotics could be degraded. Aerobic degradation of di(2-ethylhexyl)phthalate was only possible with leachate from a landfill as inoculum. Anaerobic degradation of some of the compounds was also detected. Leachate showed capability of degrading phthalates, and anaerobic sludge showed potential for degrading, polycyclic aromatic hydrocarbons, linear alkylbenzene sulfonates and nonyl phenol ethoxylates. The results are promising as they indicate that a great potential for biological degradation is present, though the inoculum containing the microorganisms capable of transforming the recalcitrant xenobiotics has to be chosen carefully.     

Conserved Structural Chemistry for Incision Activity in Structurally Non-homologous Apurinic/Apyrimidinic Endonuclease APE1 and Endonuclease IV DNA Repair Enzymes

Non-coding apurinic/apyrimidinic (AP) sites in DNA form spontaneously and as DNA base excision repair intermediates are the most common toxic and mutagenic in vivo DNA lesion. For repair, AP sites must be processed by 5′ AP endonucleases in initial stages of base repair. Human APE1 and bacterial Nfo represent the two conserved 5′ AP endonuclease families in the biosphere; they both recognize AP sites and incise the phosphodiester backbone 5′ to the lesion, yet they lack similar structures and metal ion requirements. Here, we determined and analyzed crystal structures of a 2.4 Å resolution APE1-DNA product complex with Mg²⁺ and a 0.92 Å Nfo with three metal ions. Structural and biochemical comparisons of these two evolutionarily distinct enzymes characterize key APE1 catalytic residues that are potentially functionally similar to Nfo active site components, as further tested and supported by computational analyses. We observe a magnesium-water cluster in the APE1 active site, with only Glu-96 forming the direct protein coordination to the Mg²⁺. Despite differences in structure and metal requirements of APE1 and Nfo, comparison of their active site structures surprisingly reveals strong geometric conservation of the catalytic reaction, with APE1 catalytic side chains positioned analogously to Nfo metal positions, suggesting surprising functional equivalence between Nfo metal ions and APE1 residues. The finding that APE1 residues are positioned to substitute for Nfo metal ions is supported by the impact of mutations on activity. Collectively, the results illuminate the activities of residues, metal ions, and active site features for abasic site endonucleases.     

XRCC1 interactions with base excision repair DNA intermediates

Abasic (AP) sites in DNA arise either spontaneously, or through glycosylase-catalyzed excision of damaged bases. Their removal by the base excision repair (BER) pathway avoids their mutagenic and cytotoxic consequences. XRCC1 coordinates and facilitates single-strand break (SSB) repair and BER in mammalian cells. We report that XRCC1, through its NTD and BRCT1 domains, has affinity for several DNA intermediates in BER. As shown by its capacity to form a covalent complex via Schiff base, XRCC1 binds AP sites. APE1 suppresses binding of XRCC1 to unincised AP sites however, affinity was higher when the DNA carried an AP-lyase- or APE1-incised AP site. The AP site binding capacity of XRCC1 is enhanced by the presence of strand interruptions in the opposite strand. Binding of XRCC1 to BER DNA intermediates could play an important role to warrant the accurate repair of damaged bases, AP sites or SSBs, in particular in the context of clustered DNA damage.     

Slow base excision by human alkyladenine DNA glycosylase limits the rate of formation of AP sites and AP endonuclease 1 does not stimulate base excision

The base excision repair pathway removes damaged DNA bases and resynthesizes DNA to replace the damage. Human alkyladenine DNA glycosylase (AAG) is one of several damage-specific DNA glycosylases that recognizes and excises damaged DNA bases. AAG removes primarily damaged adenine residues. Human AP endonuclease 1 (APE1) recognizes AP sites produced by DNA glycosylases and incises the phophodiester bond 5' to the damaged site. The repair process is completed by a DNA polymerase and DNA ligase. If not tightly coordinated, base excision repair could generate intermediates that are more deleterious to the cell than the initial DNA damage. The kinetics of AAG-catalyzed excision of two damaged bases, hypoxanthine and 1,N6-ethenoadenine, were measured in the presence and absence of APE1 to investigate the mechanism by which the base excision activity of AAG is coordinated with the AP incision activity of APE1. 1,N6-ethenoadenine is excised significantly slower than hypoxanthine and the rate of excision is not affected by APE1. The excision of hypoxanthine is inhibited to a small degree by accumulated product, and APE1 stimulates multiple turnovers by alleviating product inhibition. These results show that APE1 does not significantly affect the kinetics of base excision by AAG. It is likely that slow excision by AAG limits the rate of AP site formation in vivo such that AP sites are not created faster than can be processed by APE1.     

Complete degradation of xenobiotic surfactants by consortia of aerobic microorganisms

Linear alkylbenzene sulphonates are primarily attacked via a hydroxylation of the alkyl chain from the methyl group followed by β-oxidation. The alkyl chain is metabolized by pure cultures to give sulphophenyl carboxylates which accumulate in the medium. In mixed culture, other microorganisms are capable of degrading sulphophenyl carboxylates. Formation of ethylene glycol monosulphates as major products of alkyl ethoxy sulphates demonstrates that the ether bonds are cleaved. The bacteria involved in growing on the alkyl chain are unable to utilize the hydrophilic moiety. This hydrophilic moiety, in turn, is degraded by other microorganisms. The degradation of alkylphenol ethoxylates and highly branched alcohol ethoxylates proceeds by shortening the polyoxyethylene chain leaving the hydrophobic part of the molecule. The biodegradation of linear alcohol ethoxylates and ethoxylated fatty amines is initiated by a central cleavage or ω-oxidation. Subsequent oxidation of the alkyl chains results in the production of polyethylene glycols and secondary ethoxylated amines. Both polar moieties are metabolized by other microorganisms. Degradation of alkyltrimethylammonium salts and alkylamines is initiated by a cleavage of the C _alkyl -N bond. The central fission leads to the formation of alkanals which are readily converted by β-oxidation. The alkyl chain-utilizing bacteria are not able to degrade the methylamines. The methylamines, in turn, are subject to biodegradation by methylotrophs. The limited metabolic capacities of pure cultures of microorganisms utilizing surfactants point to the requirement of consortia to degrade surfactants completely. Complete degradation of surfactants is accomplished by mixed cultures of microorganisms constructed on the basis of synergistic and commensalistic relationships. However, degradation of a surfactant by one member of a commensalistic consortium may lead to the production of toxic or non-toxic metabolites. Waste water treatment without the build up of such metabolites can be achieved in plants operated with sludge retention times that are suitable for maintaining all microorganisms of the consortium. In contrast, in natural ecosystems the introduction of a surfactant may result in a transient formation of a metabolite.     

Aerobic MTBE biodegradation: an examination of past studies, current challenges and future research directions

With the current practice of amending gasoline with up to 15% by volume MTBE, the contamination of groundwater by MTBE has become widespread. As a result, the bioremediation of MTBE-impacted aquifers has become an active area of research. A review of the current literature on the aerobic biodegradation of MTBE reveals that a number of cultures from diverse environments can either partially degrade or completely mineralize MTBE. MTBE is either utilized as a sole carbon and energy source or is degraded cometabolically by cultures grown on alkanes. Reported degradation rates range from 0.3 to 50 mg MTBE/g cells/h while growth rates (0.01–0.05 g MTBE/g cells/d) and cellular yields (0.1–0.2 g cells/g MTBE) are generally low. Studies on the mechanisms of MTBE degradation indicate that a monooxygenase enzyme cleaves the ether bond yielding tert-butyl alcohol (TBA) and formaldehyde as the dominant detectable intermediates. TBA is further degraded to 2-methyl-2-hydroxy-1-propanol, 2-hydroxyisobutyric acid, 2-propanol, acetone, hydroxyacteone and eventually, carbon dioxide. The majority of these intermediates are also common to mammalian MTBE metabolism. Laboratory studies on the degradation of MTBE in the presence of gasoline aromatics reveal that while degradation rates of other gasoline components are generally not inhibited by MTBE, MTBE degradation could be inhibited in the presence of more easily biodegradable compounds. Controlled field studies are clearly needed to elucidate MTBE degradation potential in co-contaminant plumes. Based on the reviewed studies, it is likely that a bioremediation strategy involving direct metabolism, cometabolism, bioaugmentation, or some combination thereof, could be applied as a feasible and cost-effective treatment method for MTBE contamination.     

A Weight of Evidence Analysis of the Chronic Ecotoxicity of Nonylphenol Ethoxylates,Nonylphenol Ether Carboxylates,and Nonylphenol

Nonylphenol ethoxylates (NPE) are widely used surfactant compounds with many domestic and industrial applications. Due to the nature of their use in down-the-drain applications, spent NPE are discharged to septic systems or to wastewater treatment plants. Biodegradation of parent material during treatment is often incomplete, leading to release of NPE and their degradation intermediates into the environment. Considerable aquatic toxicity research has occurred on NPE and particularly on nonylphenol (NP). Available data were subjected to a quality review and all studies of acceptable quality were used in a weight of evidence hazard assessment. Data for NP were further analyzed using a Species Sensitivities Distribution (SSD) approach. About 90 chronic values are available (ChVs, geometric mean of the no-observed effect concentration and lowest-observed effect concentration for each endpoint reported), which may be reduced to average ChVs for each tested species. Higher mole NPE (NPE ≥ 9) had ChVs ranging from 900 to 14,100 μg/L, ChVs for the low mole nonylphenol ether carboxylate (NPEC1) ranged from 3200 to 12,000 μg/L, ChVs for lower mole NPE (NPE1,2) ranged from 11 to 500 μg/L, and ChVs for NP ranged from 5 to 3500 μg/L. Using the SSD analysis for NP with higher quality study results, the 10th percentile chronic effect value is 5.7 μg/L, which supports the draft USEPA criteria on NP of 5.9 μg/L.     

Triclosan susceptibility and co-metabolism--a comparison for three aerobic pollutant-degrading bacteria

The antimicrobial agent triclosan is an emerging and persistent environmental pollutant. This study evaluated the susceptibility and biodegradation potential of triclosan by three bacterial strains (Sphingomonas wittichii RW1, Burkholderia xenovorans LB400 and Sphingomonas sp. PH-07) that are able to degrade aromatic pollutants (dibenzofuran, biphenyl and diphenyl ether, respectively) with structural similarities to triclosan. These strains showed less susceptibility to triclosan when grown in complex and mineral salts media. Biodegradation experiments revealed that only strain PH-07 was able to catabolize triclosan to intermediates that included hydroxylated compounds (monohydroxy-triclosan, and dihydroxy-triclosan) and the ether bond cleavage products (4-chlorophenol and 2,4-dichlorophenol), indicating that the initial dihydroxylation occurred on both aromatic rings of triclosan. Additional growth inhibition tests demonstrated that the main intermediate, 2,4-dichlorophenol, was less toxic to strain PH-07 than was triclosan. Our results indicate that ether bond cleavage might be the primary mechanism of avoiding triclosan toxicity by this strain.     

Isolation and characterization of Halomonas sp. strain C2SS100, a hydrocarbon-degrading bacterium under hypersaline conditions

Aims:  To isolate and characterize an efficient hydrocarbon-degrading bacterium under hypersaline conditions, from a Tunisian off-shore oil field.
Methods and Results:  Production water collected from 'Sercina' petroleum reservoir, located near the Kerkennah island, Tunisia, was used for the screening of halotolerant or halophilic bacteria able to degrade crude oil. Bacterial strain C2SS100 was isolated after enrichment on crude oil, in the presence of 100 g l⁻¹ NaCl and at 37°C. This strain was aerobic, Gram-negative, rod-shaped, motile, oxidase + and catalase +. Phenotypic characters and phylogenetic analysis based on the 16S rRNA gene of the isolate C2SS100 showed that it was related to members of the Halomonas genus. The degradation of several compounds present in crude oil was confirmed by GC–MS analysis. The use of refined petroleum products such as diesel fuel and lubricating oil as sole carbon source, under the same conditions of temperature and salinity, showed that significant amounts of these heterogenic compounds could be degraded. Strain C2SS100 was able to degrade hexadecane (C₁₆). During growth on hexadecane, cells surface hydrophobicity and emulsifying activity increased indicating the production of biosurfactant by strain C2SS100.
Conclusions:  A halotolerant bacterial strain Halomonas sp. C2SS100 was isolated from production water of an oil field, after enrichment on crude oil. This strain is able to degrade hydrocarbons efficiently. The mode of hydrocarbon uptake is realized by the production of a biosurfactant which enhances the solubility of hydrocarbons and renders them more accessible for biodegradation.
Significance and Impact of the Study:  The biodegradation potential of the Halomonas sp. strain C2SS100 gives it an advantage for possibly application on bioremediation of water, hydrocarbon-contaminated sites under high-salinity level.     

环境激素类有机污染物的微生物降解和药物类化合物的残留问题

合成有机物在环境中的残留和危害已不仅仅局限于其毒性、富集、致畸和致突变,同时还能干扰包括人类在内的生物的内分泌调节作用.近年来发达国家已开始逐渐有了环境方面的条例,限制和控制这类化合物在水及食物链中的含量.现已清楚地知道,部分除草剂和杀虫剂(如阿特拉津、DDT),塑料的添加增塑剂均有内分泌激素活性,从而对生物的正常生长发育造成不良的影响.而这些化合物不但广泛存在于环境中,在特定的环境中其含量更是非常之高.以增塑剂邻苯二甲酸和邻苯二甲酸二甲酯为例,它们在填埋渗出液中的含量可高达10g·L^-1.在我们研究这类化合物的微生物降解时发现,从活性污泥和红树林中富集到的好氧微生物能将这类化合物完全矿化,且反应速度很快.同时也发现,在降解邻苯二甲酸二甲酯时,单一的纯菌不能完全降解这类化合物,而二种或三种组合的纯菌可以在一周内将500mg·L^-1的底物完全矿化.我们已分离、鉴定出中间产物,建立起了降解途径.研究的结果证实,邻苯二甲酸二甲酯类环境激素是能够在排放前通过微生物的作用达到完全矿化的.另一方面,药物类化合物的残留问题也是一个逐渐显现出的环境问题,这方面的研究应引起更多的关注和重视.     

Catabolism of arylglycerol-beta-aryl ethers lignin model compounds by Pseudomonas cepacia 122

Pseudomonas cepacia 122 can grow on several lignin model compounds including the arylglycerol-beta-aryl ethers guaiacylglycerol-beta-coniferyl ether and guaiacylglycerol-beta-guaiacyl ether. Non-phenolic lignin model compounds are not degraded by this bacterium. The enzyme system catalyzing guaiacylglycerol-beta-guaiacyl ether dissimilation in Pseudomonas cepacia 122 is inducible and repressed by glucose. Guaiacylglycerol and guaiacylglycerol-beta-guaiacyl ether were identified as intermediates in guaiacylglycerol-beta-coniferyl ether catabolism. Guaiacol, guaiacoxyethanol, vanillin and vanillic acid were identified as intermediates of guaiacylglycerol-beta-guaiacyl ether breakdown indicating that a C alpha-C beta splitting mechanism is involved in the degradation of aryl-alkyl ethers by this bacterium.     

Degradation of 4,4'-dichlorobiphenyl, 3,3',4,4'-tetrachlorobiphenyl, and 2,2',4,4',5,5'-hexachlorobiphenyl by the white rot fungus Phanerochaete chrysosporium.

The white rot fungus Phanerochaete chrysosporium has demonstrated abilities to degrade many xenobiotic chemicals. In this study, the degradation of three model polychlorinated biphenyl (PCB) congeners (4,4'-dichlorobiphenyl [DCB], 3,3',4,4'-tetrachlorobiphenyl, and 2,2',4,4',5,5'-hexachlorobiphenyl) by P. chrysosporium in liquid culture was examined. After 28 days of incubation, 14C partitioning analysis indicated extensive degradation of DCB, including 11% mineralization. In contrast, there was negligible mineralization of the tetrachloro- or hexachlorobiphenyl and little evidence for any significant metabolism. With all of the model PCBs, a large fraction of the 14C was determined to be biomass bound. Results from a time course study done with 4,4'-[14C]DCB to examine 14C partitioning dynamics indicated that the biomass-bound 14C was likely attributable to nonspecific adsorption of the PCBs to the fungal hyphae. In a subsequent isotope trapping experiment, 4-chlorobenzoic acid and 4-chlorobenzyl alcohol were identified as metabolites produced from 4,4'-[14C]DCB. To the best of our knowledge, this the first report describing intermediates formed by P. chrysosporium during PCB degradation. Results from these experiments suggested similarities between P. chrysosporium and bacterial systems in terms of effects of congener chlorination degree and pattern on PCB metabolism and intermediates characteristic of the PCB degradation process.     

Ethers Illume Sodium‐Based Battery Chemistry: Uniqueness,Surprise, and Challenges

Sodium‐ion batteries (SIBs) have the potential to be practically applied in large‐scale energy storage markets. The rapid progress of SIBs research is primarily focused on electrodes, while electrolytes attract less attention. Indeed, the improvement of electrode performance is arguably correlated with the electrolyte optimization. In conventional lithium‐ion batteries (LIBs), ether‐based electrolytes are historically less practical owing to the insufficient passivation of both anodes and cathodes. As an important class of aprotic electrolytes, ethers have revived with the emerging lithium‐sulfur and lithium‐oxygen batteries in recent years, and are even booming in the wave of SIBs. Ether‐based electrolytes are unique to enabling these new battery chemistries in terms of producing stable ternary graphite intercalation compounds, modifying anode solid electrolyte interphases, reducing the solubility of intermediates, and decreasing polarization. Better still, ether‐based electrolytes are compatible with specific inorganic cathodes and could catalyze the assembly of full SIBs prototypes. This Research News article aims to summarize the recent critical reports on ether‐based electrolytes in sodium‐based batteries, to unveil the uniqueness of ether‐based electrolytes to advancing diverse electrode materials, and to shed light on the viability and challenges of ether‐based electrolytes in future sodium‐based battery chemistries.     

Microbial degradation of sulfur, nitrogen and oxygen heterocycles

Sulfur (S), nitrogen (N) and oxygen (O) heterocycles are among the most potent environmental pollutants. Microbial degradation of these pollutants is attracting more and more attention because such bioprocesses are environmentally friendly. The biotechnological potential of these processes is being investigated, for example, to achieve better sulfur removal by immobilized biocatalysts with magnetite nanoparticles or by solvent-tolerant bacteria, and to obtain valuable intermediates from these heterocycles. Other recent advances have demonstrated the mechanisms of angular dioxygenation of nitrogen heterocycles by microbes. However, these technologies are not yet available for large-scale applications so future research must investigate proper modifications for industrial applications of these processes. This review focuses on recent progress in understanding how microbes degrade S, N and O heterocycles.     

Apurinic/apyrimidinic endonuclease (APE/REF-1) haploinsufficient mice display tissue-specific differences in DNA polymerase beta-dependent base excision repair

Apurinic/apyrimidinic (AP) endonuclease (APE) is a multifunctional protein possessing both DNA repair and redox regulatory activities. In base excision repair (BER), APE is responsible for processing spontaneous, chemical, or monofunctional DNA glycosylase-initiated AP sites via its 5'-endonuclease activity and 3'-"end-trimming" activity when processing residues produced as a consequence of bifunctional DNA glycosylases. In this study, we have fully characterized a mammalian model of APE haploinsufficiency by using a mouse containing a heterozygous gene-targeted deletion of the APE gene (Apex(+/-)). Our data indicate that Apex(+/-) mice are indeed APE-haploinsufficient, as exhibited by a 40-50% reduction (p < 0.05) in APE mRNA, protein, and 5'-endonuclease activity in all tissues studied. Based on gene dosage, we expected to see a concomitant reduction in BER activity; however, by using an in vitro G:U mismatch BER assay, we observed tissue-specific alterations in monofunctional glycosylase-initiated BER activity, e.g. liver (35% decrease, p < 0.05), testes (55% increase, p < 0.05), and brain (no significant difference). The observed changes in BER activity correlated tightly with changes in DNA polymerase beta and AP site DNA binding levels. We propose a mechanism of BER that may be influenced by the redox regulatory activity of APE, and we suggest that reduced APE may render a cell/tissue more susceptible to dysregulation of the polymerase beta-dependent BER response to cellular stress.     

Complete degradation of xenobiotic surfactants by consortia of aerobic microorganisms

Linear alkylbenzene sulphonates are primarily attacked via a hydroxylation of the alkyl chain from the methyl group followed by -oxidation. The alkyl chain is metabolized by pure cultures to give sulphophenyl carboxylates which accumulate in the medium. In mixed culture, other microorganisms are capable of degrading sulphophenyl carboxylates. Formation of ethylene glycol monosulphates as major products of alkyl ethoxy sulphates demonstrates that the ether bonds are cleaved. The bacteria involved in growing on the alkyl chain are unable to utilize the hydrophilic moiety. This hydrophilic moiety, in turn, is degraded by other microorganisms.The degradation of alkylphenol ethoxylates and highly branched alcohol ethoxylates proceeds by shortening the polyoxyethylene chain leaving the hydrophobic part of the molecule. The biodegradation of linear alcohol ethoxylates and ethoxylated fatty amines is initiated by a central cleavage or -oxidation. Subsequent oxidation of the alkyl chains results in the production of polyethylene glycols and secondary ethoxylated amines. Both polar moieties are metabolized by other microorganisms. Degradation of alkyltrimethylammonium salts and alkylamines is initiated by a cleavage of the C _alkyl -N bond. The central fission leads to the formation of alkanals which are readily converted by -oxidation. The alkyl chain-utilizing bacteria are not able to degrade the methylamines. The methylamines, in turn, are subject to biodegradation by methylotrophs.The limited metabolic capacities of pure cultures of microorganisms utilizing surfactants point to the requirement of consortia to degrade surfactants completely. Complete degradation of surfactants is accomplished by mixed cultures of microorganisms constructed on the basis of synergistic and commensalistic relationships. However, degradation of a surfactant by one member of a commensalistic consortium may lead to the production of toxic or non-toxic metabolites. Waste water treatment without the build up of such metabolites can be achieved in plants operated with sludge retention times that are suitable for maintaining all microorganisms of the consortium. In contrast, in natural ecosystems the introduction of a surfactant may result in a transient formation of a metabolite.     

Isolation of a small molecule inhibitor of DNA base excision repair

The base excision repair (BER) pathway is essential for the removal of DNA bases damaged by alkylation or oxidation. A key step in BER is the processing of an apurinic/apyrimidinic (AP) site intermediate by an AP endonuclease. The major AP endonuclease in human cells (APE1, also termed HAP1 and Ref-1) accounts for >95% of the total AP endonuclease activity, and is essential for the protection of cells against the toxic effects of several classes of DNA damaging agents. Moreover, APE1 overexpression has been linked to radio- and chemo-resistance in human tumors. Using a newly developed high-throughput screen, several chemical inhibitors of APE1 have been isolated. Amongst these, CRT0044876 was identified as a potent and selective APE1 inhibitor. CRT0044876 inhibits the AP endonuclease, 3'-phosphodiesterase and 3'-phosphatase activities of APE1 at low micromolar concentrations, and is a specific inhibitor of the exonuclease III family of enzymes to which APE1 belongs. At non-cytotoxic concentrations, CRT0044876 potentiates the cytotoxicity of several DNA base-targeting compounds. This enhancement of cytotoxicity is associated with an accumulation of unrepaired AP sites. In silico modeling studies suggest that CRT0044876 binds to the active site of APE1. These studies provide both a novel reagent for probing APE1 function in human cells, and a rational basis for the development of APE1-targeting drugs for antitumor therapy.