Effects of troponoids on mitochondrial function and cytotoxicity

The α-hydroxytropolones (αHT) are troponoid inhibitors of hepatitis B virus (HBV) replication that can target the HBV ribonuclease H (RNase H) with sub-micromolar efficacies. αHTs and related troponoids (tropones and tropolones) can be cytotoxic in cell lines as measured by MTS assays that assesses mitochondrial function. Earlier studies suggest that tropolones induce cytotoxicity through inhibition of mitochondrial respiration. Therefore, we screened 35 diverse troponoids for effects on mitochondrial function, mitochondrial:nuclear genome ratio, cytotoxicity, and reactive oxygen species (ROS) production. Troponoids as a class did not inhibit respiration or glycolysis, although the α-ketotropolone subclass did interfere with these processes. The troponoids had no impact on the mitochondrial DNA to nuclear DNA ratio after three days of compound exposure. Patterns of troponoid-induced cytotoxicity among three hepatic cell lines were similar for all compounds, but three potent HBV RNase H inhibitors were not cytotoxic in primary human hepatocytes. Tropolones and αHTs increased ROS production in cells at cytotoxic concentrations but had no effect at lower concentrations that efficiently inhibit HBV replication. Troponoid-mediated cytotoxicity was significantly decreased upon addition of the ROS scavenger N-acetylcysteine. These studies show that troponoids can increase ROS production at high concentrations within cell lines leading to cytotoxicity, but are not be cytotoxic in primary hepatocytes. Future development of αHTs as potential therapeutics against HBV may need to mitigate ROS production by altering compound design and/or by co-administration with ROS antagonists to ameliorate increased ROS levels.

The birth-and-death evolution of cytochrome P450 genes in bees

The birth-and-death model of multigene family evolution describes how families can expand by duplication and contract by gene deletion and formation of pseudogenes. The phylogenetic stability of a gene is thought to be related to the degree of functional importance. However, it is unclear how much evolution of a gene in a gene family is driven by adaptive versus neutral processes. The cytochrome P450s are one of the most diverse and well-studied multigene families, involved in both physiological and xenobiotic functions. Bees have a high toxin exposure due to their diet of nectar and pollen, as well as the resin-collecting behavior exhibited by some bees. Here, we describe the P450s of the orchid bee Euglossa dilemma. Orchid bees are a neotropical clade in which males form perfume bouquets used in courtship displays by collecting a diverse set of volatile compounds, resulting in high chemical compound exposure. We conducted phylogenetic and selection analyses across ten bee species encompassing three bee families. We do not find a relationship between the ecology of a bee species and its P450 repertoire. Our analyses reveal that P450 clades can be classified into stable and unstable clades, and that genes involved in xenobiotic metabolism are more likely to belong to unstable clades. Furthermore, we find that unstable clades are under more dynamic evolutionary pressures, with signals of adaptive evolution detected, suggesting that both gene duplication and positive selection driving sequence divergence have played a role in the diversification of bee P450s. Our works highlights the complexity of multigene family evolution which does not always follow generalized predictions.

Immunotoxicity of Per- and Polyfluoroalkyl Substances: Insights into Short-Chain PFAS Exposure

Novel per- and polyfluoroalkyl substances (PFAS) were recently identified in drinking water sources throughout North Carolina. These include the perfluoroether acids (PFEAs) perfluoro-2-methoxyacetic acid (PFMOAA), perfluoro-2-methoxypropanoic acid (PFMOPrA), and perfluoro-4-methoxybutanioc acid (PFMOBA). Little toxicological data exist for these PFEAs. Therefore, the present study described signs of toxicity and immunotoxicity following oral exposure. Adult male and female C57BL/6 mice were exposed once/day for 30 days to PFMOAA (0, 0.00025, 0.025, or 2.5 mg/kg), PFMOPrA, or PFMOBA (0, 0.5, 5, or 50 mg/kg). A dose of 7.5 mg/kg of perfluorooctanoic acid (PFOA) was used as a positive control. Terminal body weights, and absolute liver, spleen, or thymus weights did not differ by dose for any compound; exposure to 50 mg/kg of PFMOBA increased relative liver weights in males. Changes in splenic cellularity were observed in males exposed to PFMOPrA and decreased numbers of B and natural killer (NK) cells were observed in males and females exposed to PFMOBA. Exposure did not alter NK cell cytotoxicity or T cell-dependent antibody responses at doses administered. Our results indicate that these “understudied” PFAS have toxicological potential but require additional investigation across endpoints and species, including humans, to understand health effects via drinking water exposure.

Altered Toxicological Endpoints in Humans with Quaternary Ammonium Compound Exposure

Humans are extensively exposed to Quaternary Ammonium Compounds (QACs). QACs are ubiquitously used in medical settings, restaurants, and homes as cleaners and disinfectants. They are also used on food and in personal care products as preservatives. Despite their prevalence, nothing is known about the health effects associated with chronic low-level exposure. Chronic QAC toxicity was recently identified in mice and resulted in developmental and reproductive deficits and altered immune function. Cell based studies show that QACs increase inflammation, disrupt cholesterol synthesis, and decrease mitochondria function. If these studies translate to human toxicity, multiple physiological functions could be affected. QAC concentrations in humans have not been monitored previously. This study tested whether QAC concentrations could be detected in the blood of 43 random volunteers, and whether QAC concentrations were associated with markers of inflammation, mitochondrial function, and cholesterol synthesis in a dose dependent manner. QAC concentrations were detected in 80% of study participants, and were associated with decreased mitochondrial function and an increase in inflammatory cytokines in a dose dependent manner. Cholesterol synthesis pathway intermediaries were generally increased, indicating disruption in cholesterol homeostasis. This is the first study to demonstrate that chronic exposure to QACs results in measurable concentrations in human blood, and to also demonstrate significant correlations between QAC level and meaningful biomarkers related to health.

Occupational Lead Exposure and Brain Tumors: Systematic Review and Meta-Analysis

(1) Background: Due to inconsistencies in epidemiological findings, there has been uncertainty regarding the association of lead compounds with brain tumors. We performed a meta-analysis of published case-control and cohort studies exploring lead compound exposure and brain tumor risk. (2) Methods: We searched PubMed, Embase®, and Cochrane to find eligible studies. Eighteen studies were selected for assessment of occupational exposure to lead compound and brain tumor. Pooled estimates of odds ratios (ORs) were obtained using random effects models. We assessed the differences through subgroup analysis according to tumor type, study design, measurements of exposure, and tumor outcome. Statistical tests for publication bias, heterogeneity, and sensitivity analysis were applied. (3) Results: Our systematic review and meta-analysis showed a not significant association with lead exposure and risk of benign and malignant brain tumors (pooled OR = 1.11, 95% Confidence Interval (CI): 0.95–1.29). Including only malignant brain tumors, the risk of brain tumor was significantly increased (pooled OR = 1.13, 95% CI: 1.04–1.24). (4) Conclusions: This meta-analysis provides suggestive evidence for an association between lead compound exposure and brain tumor. In future studies, it will be necessary to identify the effect of lead compounds according to the types of brain tumor.