Analysis of sunscreens and antibiotics in groundwater during the Covid-19 pandemic in the Riviera Maya, Mexico.

Tourism contributes to groundwater pollution, but quantifying its exact impact is challenging due to the presence of multiple pollution sources. However, the COVID-19 pandemic presented a unique opportunity to conduct a natural experiment and assess the influence of tourism on groundwater pollution. One such tourist destination is the Riviera Maya in Quintana Roo, Mexico (specifically Cancun). Here, water contamination occurs due to the addition of sunscreen and antibiotics during aquatic activities like swimming, as well as from sewage. In this study, water samples were collected during the pandemic and when tourists returned to the region. Samples were taken from sinkholes (cenotes), beaches, and wells then tested using liquid chromatography for antibiotics and active ingredients found in sunscreens. The data revealed that contamination levels from specific sunscreens and antibiotics persisted even when tourists were absent, indicating that local residents significantly contribute to groundwater pollution. However, upon the return of tourists, the diversity of sunscreen and antibiotics found increased, suggesting that tourists bring along various compounds from their home regions. During the initial stages of the pandemic, antibiotic concentrations were highest, primarily due to local residents incorrectly using antibiotics to combat COVID-19. Additionally, the research found that tourist sites had the greatest contribution to groundwater pollution, with sunscreen concentration increasing. Furthermore, installation of a wastewater treatment plant decreased overall groundwater pollution. These findings enhance our understanding of the pollution contributed by tourists in relation to other pollution sources.

Quantitation and localization of beta-blockers and SSRIs accumulation in fathead minnows by complementary mass spectrometry analyses.

Beta-blockers and selective serotonin reuptake inhibitors (SSRIs) enter the environment through the improper disposal of drugs in garbage, sinks, or toilets as well as via excretion after their intended use. Beta-blockers and SSRIs are resistant to biodegradation and highly water soluble, leading to limited removal capabilities by wastewater treatment plants. As a result, these compounds can easily enter the aquatic environment in a biologically active state, posing risks to the behavior, anatomy, and physiology of aquatic organisms through exposure. However, there is insufficient research on the effects of beta-blockers on aquatic vertebrates to have a complete understanding of the impact of beta-blocker pollution on aquatic organisms. Exposing fathead minnows at environmentally-relevant concentrations allows for an accurate representation of how beta-blockers and SSRIs affect and accumulate in aquatic vertebrates. Singular exposures to two beta-blockers and two selective serotonin reuptake inhibitors (SSRIs) at seven days modeled how pollution affects aquatic wildlife during acute exposure events. After exposure, the entire organism was analyzed quantitatively by liquid chromatography mass spectrometry and qualitatively by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). Imaging allowed for the determination of the part of the organism in which the drugs were accumulating. Results showed that after seven days of exposure there was accumulation of both the beta-blockers and SSRIs in the fathead minnows. This data represents the first application of LC-MS quantitation guided by MALDI-MSI in aquatic toxicology-type research in environmental analytical chemistry.

A Comparative Analysis of Methods for Quantitation of Sugars during the Corn-to-Ethanol Fermentation Process.

The quantitation of sugars, including glucose, the primary fermentable sugar; maltose (DP2); and maltotriose (DP3), is a standard procedure during the corn-to-ethanol fermentation process. The quantitation of glucose by the Megazyme Assay utilizing glucose oxidase and peroxidase enzymes (GOPOD) and UV-Vis detection, high-performance liquid chromatography with refractive index detection (HPLC-RID), and liquid chromatography mass spectrometry (LC-MS) with electrospray ionization (ESI) and selected ion monitoring (SIM) was studied. Three biological flask fermentation replicates were analyzed every 12 h beginning at 14 h of fermentation (T14) until near completion of fermentation (T62).The method comparison results for glucose quantitation showed that the LC-MS SIM analysis had the lowest limit of quantitation (LOQ) at 2 ppm and the widest dynamic range of 2.7 orders of magnitude. The HPLC-RID analysis had a linear dynamic range (LDR) of 1.5 orders of magnitude with an LOQ of 1500 ppm. The Megazyme GOPOD analysis had an LDR of 0.9 orders of magnitude with an LOQ of 120 ppm.The HPLC-RID method was ideal for glucose quantitation when it was present in high concentrations. In contrast, maltose and maltotriose components were found to be present in lower concentrations, such that simultaneous quantitation of the three analytes is difficult during fermentation. The LC-MS method was the only method able to quantify the concentration of glucose successfully and simultaneously with DP2 and DP3 in all the fermentation broth samples collected from T14 through T62 during the corn-to-ethanol fermentation process.

Analysis of beta-blocker bioconcentration in brown planaria (Girardia dorotocephala) and its effects on regeneration.

Production, distribution, and disposal of pharmaceutical products, including beta-blockers, have become a global issue. Beta-blockers are known to persist in the environment months after their release and may result in the disruption of the homeostatic system in non-target organisms. Here, we study the bioconcentration of three of the most commonly used beta-blockers and their effect on the regeneration of Girardia dorotocephala, a freshwater brown planarian. Acute toxicity tests determined LC50s for acebutolol, metoprolol, and propranolol to be 778 mg/L, 711 mg/L, and 111 mg/L, respectively. The quantification and analysis of beta-blocker bioconcentration during acute exposure were performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After 4 days of exposure to beta-blockers, the bioconcentration drastically decreased for all three beta-blockers at all exposure levels, suggesting that an effective mechanism to reduce uptake or excrete beta-blockers could be present. Additionally, Girardia dorotocephala were cut proximal to the head and the quality of regeneration was documented from each fragment daily. No significant difference was visually observed after 2 weeks of regeneration between the brown planarians placed in beta-blocker solution and those placed in control solution.

Estimation of cancer risks and benefits associated with a potential increased consumption of fruits and vegetables.

The current paper provides an analysis of the potential number of cancer cases that might be prevented if half the U.S. population increased its fruit and vegetable consumption by one serving each per day. This number is contrasted with an upper-bound estimate of concomitant cancer cases that might be theoretically attributed to the intake of pesticide residues arising from the same additional fruit and vegetable consumption. The cancer prevention estimates were derived using a published meta-analysis of nutritional epidemiology studies. The cancer risks were estimated using U.S. Environmental Protection Agency (EPA) methods, cancer potency estimates from rodent bioassays, and pesticide residue sampling data from the U.S. Department of Agriculture (USDA). The resulting estimates are that approximately 20,000 cancer cases per year could be prevented by increasing fruit and vegetable consumption, while up to 10 cancer cases per year could be caused by the added pesticide consumption. These estimates have significant uncertainties (e.g., potential residual confounding in the fruit and vegetable epidemiologic studies and reliance on rodent bioassays for cancer risk). However, the overwhelming difference between benefit and risk estimates provides confidence that consumers should not be concerned about cancer risks from consuming conventionally-grown fruits and vegetables.

Secondary ion mass spectrometry imaging of molecular distributions in cultured neurons and their processes: comparative analysis of sample preparation.

Neurons often exhibit a complex chemical distribution and topography; therefore, sample preparation protocols that preserve structures ranging from relatively large cell somata to small neurites and growth cones are important factors in secondary ion mass spectrometry (SIMS) imaging studies. Here, SIMS was used to investigate the subcellular localization of lipids and lipophilic species in neurons from Aplysia californica. Using individual neurons cultured on silicon wafers, we compared and optimized several SIMS sampling approaches. After an initial step to remove the high salt culturing media, formaldehyde, paraformaldehyde, and glycerol, and various combinations thereof, were tested for their ability to achieve cell stabilization during and after the removal of extracellular media. These treatments improved the preservation of cellular morphology as visualized with SIMS imaging. For analytes >250 Da, coating the cell surface with a 3.2 nm-thick gold layer increased the ion intensity; multiple analytes previously not observed or observed at low abundance were detected, including intact cholesterol and vitamin E molecular ions. However, once a sample was coated, many of the lower molecular mass (<200 Da) analyte signals were suppressed. The optimum approach depended on the analyte being studied; the approaches evaluated included rinsing with water and cell stabilization with glycerol and 4 % paraformaldehyde. The sample preparation methods described here enhance SIMS imaging of processes of individual cultured neurons over a broad mass range with enhanced image contrast.

Localized mandibular enlargement in end-stage renal disease: two case reports and a review of the literature.

Enlargement of the jaws is an infrequently reported complication of chronic kidney disease mineral and bone disorder (CKD-MBD). Two cases of localized mandibular swellings in young patients with histories of end-stage renal disease are discussed with a review of the literature. Although 17 of the first 19 cases that were reported exhibited diffuse enlargement, these reports increase the number of localized swellings to 8 and support the contention that localized expansion of the jaws as a manifestation of CKD-MBD is more common than originally recognized.

Stretched tissue mounting for MALDI mass spectrometry imaging.

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) combines information-rich chemical detection with spatial localization of analytes. For a given instrumental platform and analyte class, the data acquired can represent a compromise between analyte extraction and spatial information. Here, we introduce an improvement to the spatial resolution achievable with MALDI MSI conducted with standard mass spectrometric systems that also reduces analyte migration during matrix application. Tissue is placed directly on a stretchable membrane that, when stretched, fragments the tissue into micrometer-sized pieces. Scanning electron microscopy analysis shows that this process produces fairly homogeneous distributions of small tissue fragments separated and surrounded by areas of hydrophobic membrane surface. MALDI matrix is then applied by either a robotic microspotter or an artist's airbrush. Rat spinal cord samples imaged with an instrumental resolution of 50-250 µm demonstrate lipid distributions with a 5-fold high spatial resolution (a 25-fold increase in pixel density) after stretching compared to tissues that were not stretched.

The modified-bead stretched sample method: development and application to MALDI-MS imaging of protein localization in the spinal cord.

Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been used to create spatial distribution maps from lipids, peptides, and proteins in a variety of biological tissues. MALDI-MSI often involves trade-offs between the extent of analyte extraction and desired spatial resolution, compromises that can adversely affect detectability. For example, increasing the extraction time can lead to unwanted analyte spatial redistribution. With the stretched sample method (SSM), the extraction period can be extended, resulting in reduced analyte redistribution while suppressing detection of cationic salt adducts. The SSM involves thaw-mounting a thin tissue section onto a substrate of small glass beads embedded in Parafilm M and then stretching the membrane to fragment the tissue into thousands of bead-sized pieces. Here, we applied the SSM method to MALDI-MSI using rat spinal cord as a model. We used surface-modified beads coated with trypsin or chymotrypsin in order to facilitate controlled digestion and detection of proteins. The enzymatic reactions were maintained by repeatedly condensing water on the stretched sample surface. As a result, new peptides formed by tryptic or chymotryptic protein digestion were detected and identified using a combination of MALDI-MSI and offline liquid chromatography tandem mass spectrometric analysis. Localization of these peptides indicated the distribution of their proteins of origin, including myelin basic protein, actin beta, and tubulin alpha chain. Additionally, we used uncoated beads to create distribution maps of many endogenous lipids and small peptides. The extension of the SSM using modified beads resulted in the creation of mosaic bead surfaces where adjacent beads were coated with different enzymes or other reactive chemicals, permitting investigation of the distributions of a wider range of analytes in biological samples within a single experiment.

MALDI mass spectrometric imaging using the stretched sample method to reveal neuropeptide distributions in aplysia nervous tissue.

Neuropeptides are a diverse set of complex cell-cell signaling molecules that modulate behavior, learning, and memory. Their spatially heterogeneous distributions, large number of post-translational modifications, and wide range of physiologically active concentrations make their characterization challenging. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometric imaging is well-suited to characterizing and mapping neuropeptides in the central nervous system. Because matrix application can cause peptide migration within tissue samples, application parameters for MALDI typically represent a compromise between attaining the highest signal quality and preserving native spatial distributions. The stretched sample approach minimizes this trade-off by fragmenting the tissue section into thousands of spatially isolated islands, each approximately 40 mum in size. This inhibits analyte migration between the pieces and, at the same time, reduces analyte-salt adduct formation. Here, we present methodological improvements that enable the imaging of stretched tissues and reveal neuropeptide distributions in nervous tissue from Aplysia californica. The distributions of known neuropeptides are shown to correspond with previous immunohistochemical results, demonstrating that the stretched imaging method is well-suited for working with easily redistributed molecules and heterogeneous tissues and reduces adducts from physiological salts.

Chapter 13: Imaging of cells and tissues with mass spectrometry: adding chemical information to imaging.

Techniques that map the distribution of compounds in biological tissues can be invaluable in addressing a number of critical questions in biology and medicine. One of the newer methods, mass spectrometric imaging, has enabled investigation of spatial localization for a variety of compounds ranging from atomics to proteins. The ability of mass spectrometry to detect and differentiate a large number of unlabeled compounds makes the approach amenable to the study of complex biological tissues. This chapter focuses on recent advances in the instrumentation and sample preparation protocols that make mass spectrometric imaging of biological samples possible, including strategies for both tissue and single-cell imaging using the following mass spectrometric ionization methods: matrix-assisted laser desorption/ionization, secondary ion, electrospray, and desorption electrospray.