Transcriptomic and behavioral analyses reveal unique target tissues and molecular pathways associated with embryonic exposure to low level glyphosate and metal mixtures in zebrafish.

Investigation of developmental molecular events following exposure to environmentally relevant agrochemical mixtures is critical to predicting their potential long-term ecological and human health risks. Here, we sought to uncover transcriptomic changes during zebrafish (Danio rerio) embryonic development following exposure to glyphosate and co-exposure to metals. Glyphosate is widely used globally with an allowable drinking water limit of 700 ppb. We examined effects of glyphosate (10 ppb) alone and when co-exposed to a metal mixture containing low levels of arsenic (4 ppb), lead (5 ppb), cadmium (2 ppb), and vanadium (15 ppb). This mixture was derived based on behavioral and morphological toxicity findings and environmentally relevant concentrations found in agricultural regions where glyphosate and metals are ubiquitously present. Gene expression patterns coupled to a single-cell transcriptomic dataset revealed that developmental exposure (28-72 h post fertilization) to glyphosate dysregulates expression of developmental genes specific to the central nervous system. Subsequent studies indicated significant suppression of larval zebrafish movement with 10 ppb glyphosate exposure. Studies with glyphosate + metals mixture and metals mixture alone showed unique developmental transcriptomic patterns and behavioral changes compared to glyphosate exposure alone. However, some outcomes (e.g., changes in expression of genes involved in epigenetic regulation and extracellular matrix patterning) were common across all three exposures compared to the control. Notably, glyphosate + metals co-exposure distinctly suppresses lysosomal transcripts and targets renal developmental genes. While further studies are required to uncover the precise nature of the interactions between glyphosate and metals, our study shows that glyphosate at very low levels is a behavioral and neurotoxicant that changes when metals are present. Given this herbicide affects distinctive physiological processes, including renal development and lysosomal dysregulation when co-exposed with metals, we conclude that environmental cation levels should be considered in glyphosate toxicity and risk assessment.

Urinary 1-hydroxypyrene in pregnant women in a Northeastern U.S. city: socioeconomic disparity and contributions from air pollution sources.

BACKGROUND: Maternal exposure to polycyclic aromatic hydrocarbons (PAHs), ubiquitous constituents of air pollution, has been associated with adverse birth outcomes. Yet it remains unclear whether and how socioeconomic status (SES) affects gestational PAH exposure. OBJECTIVE: To examine whether there are socioeconomic disparities in PAHs exposure among pregnant women from Rochester, NY, and if so, to what extent disproportionate proximity to air pollution sources, measured by residential distance to transportation-related sources, contributed to the exposure disparity. METHODS: We measured 1-hydroxypyrene concentrations in 726 urine samples collected from 305 pregnant women up to three samples throughout pregnancy. Residential distances to transportation-related sources were calculated based on participants' home addresses. We used linear mixed-effects models with random intercepts of participants to examine associations between 1-hydroxypyrene, SES indicators, and distance to transportation-related sources. We used structural equation modelling to assess to what extent distance to transportation-related sources contributes to the socioeconomic disparity in 1-hydroxypyrene concentrations. RESULTS: Reduced household income and maternal education level were both significant SES predictors of 1-hydroxypyrene concentrations, after the adjustment for other maternal demographic characteristics. Each interquartile range (IQR) increases in residential proximity to the airport (from 14.3 to 6.0 km), the railroad yard (from 22.3 to 6.0 km), and annual average daily traffic within 300 m (from 3796 to 99,933 vehicles/year) were associated with 15.0% (95%CI: 7.0-22.2%), 15.4% (95%CI: 6.5-23.5%), and 13.6% (95%CI: 4.7-23.3%) increases in 1-hydroxypyrene concentrations, respectively. Proximity to these sources jointly explained 10% (95%CI: 1.6-18.4%) of the 1-hydroxypyrene concentration change associated with decreases in SES as a latent variable defined by both household income and education level. IMPACT STATEMENT: Our findings suggest that efforts to address disproportionate residential proximity to transportation-related sources may reduce the socioeconomic disparity in PAH exposure.

Concurrent behavioral and electrophysiological longitudinal recordings for in vivo assessment of aging.

Electrophysiological and behavioral alterations, including sleep and cognitive impairments, are critical components of age-related decline and neurodegenerative diseases. In preclinical investigation, many refined techniques are employed to probe these phenotypes, but they are often conducted separately. Herein, we provide a protocol for one-time surgical implantation of EMG wires in the nuchal muscle and a skull-surface EEG headcap in mice, capable of 9-to-12-month recording longevity. All data acquisitions are wireless, making them compatible with simultaneous EEG recording coupled to multiple behavioral tasks, as we demonstrate with locomotion/sleep staging during home-cage video assessments, cognitive testing in the Barnes maze, and sleep disruption. Time-course EEG and EMG data can be accurately mapped to the behavioral phenotype and synchronized with neuronal frequencies for movement and the location to target in the Barnes maze. We discuss critical steps for optimizing headcap surgery and alternative approaches, including increasing the number of EEG channels or utilizing depth electrodes with the system. Combining electrophysiological and behavioral measurements in preclinical models of aging and neurodegeneration has great potential for improving mechanistic and therapeutic assessments and determining early markers of brain disorders.

Defining drinking water metal contaminant mixture risk by coupling zebrafish behavioral analysis with citizen science.

Contaminated drinking water is an important public health consideration in New England where well water is often found to contain arsenic and other metals such as cadmium, lead, and uranium. Chronic or high level exposure to these metals have been associated with multiple acute and chronic diseases, including cancers and impaired neurological development. While individual metal levels are often regulated, adverse health effects of metal mixtures, especially at concentrations considered safe for human consumption remain unclear. Here, we utilized a multivariate analysis that examined behavioral outcomes in the zebrafish model as a function of multiple metal chemical constituents of 92 drinking well water samples, collected in Maine and New Hampshire. To collect these samples, a citizen science approach was used, that engaged local teachers, students, and scientific partners. Our analysis of 4016 metal-mixture combinations shows that changes in zebrafish behavior are highly mixture dependent, and indicate that certain combinations of metals, especially those containing arsenic, cadmium, lead, and uranium, even at levels considered safe in drinking water, are significant drivers of behavioral toxicity. Our data emphasize the need to consider low-level chemical mixture effects and provide a framework for a more in-depth analysis of drinking water samples. We also provide evidence for the efficacy of utilizing citizen science in research, as the broader impact of this work is to empower local communities to advocate for improving their own water quality.

The Role of the Anthropologist in the Application of NamUs.

The National Missing and Unidentified Person System (NamUs) provides a web-based platform where forensic scientists, law enforcement investigators, and the public can share information related to missing person and unidentified decedent cases across the United States. Through this secure online database, forensic anthropologists, along with medical examiners, coroners, law enforcement officers, family members of missing persons, and concerned citizens, can access varying levels of information on a common platform to assist in the resolution of these cases. Conventional wisdom and experience allows that an individual who is known within the community and has been dead a very short time is more likely to be identified than someone who is badly decomposed and/or skeletal. For others, even when there has been an extended postmortem interval, circumstances, associated evidence, and location of the remains are just some of the elements that direct the investigation and help establish the identity of the decedent. When the remains are degraded by factors such as extended postmortem interval, mechanical disruption, scattering, or burning, however, the identification becomes much more difficult. In the first week of May 2016, there were 10 767 open cases of unidentified remains in NamUs. Of these, 7257 were classified as unrecognizable by a variety of aforementioned factors. By the very nature of their work, anthropologists can develop key pieces of information from decomposed and/or skeletal elements and NamUs can provide the technology to consolidate and share their findings as well as those of all subject matter experts involved with the investigation.

Treating burn-associated joint contracture: results of an inpatient rehabilitation stretching protocol.

The prevention and treatment of scar contracture is critical after significant burn injuries to avoid functional impairment. However, descriptions of specific contracture treatment interventions and outcomes are limited. Our objective is to provide detailed information and range of motion outcomes regarding the use of an intensive stretching protocol for burn-associated contracture. As part of a quality improvement measure, all patients admitted to inpatient rehabilitation with burn injury were treated with at least 1 hour of daily stretching by experienced therapists and were tracked with standardized range of motion measurements. Eighty-eight joint contractures were treated across nine patients for up to 4 weeks. The average weekly improvement in range of motion was 8.2 degrees (95% confidence interval [CI], 6.5-9.9). The largest gains were seen in the first week of treatment with an average improvement of 11.2 degrees (95% CI, 8.7-13.6). Eighteen digit contractures were treated across four patients. The average weekly improvement in flexion was 7.2 mm (95% CI, 5.2-9.1) again with larger gains in the first week of treatment-12.8 mm (95% CI, 10.3-15.4). Thumb opposition improved across five patients in the first week with an average improvement of 1.4 on the opposition scale (95% CI, 0.4-2.5). Intensive stretching by experienced therapists yielded significant improvements in joint range of motion for patients with burn-associated joint contracture. Defining specific burn contracture interventions remains a key goal in advancing burn rehabilitation in the future.