Differential Downregulation of ß1-Adrenergic Receptor Signaling in the Heart.

BACKGROUND: Chronic sympathetic stimulation drives desensitization and downregulation of ß1 adrenergic receptor (ß1AR) in heart failure. We aim to explore the differential downregulation subcellular pools of ß1AR signaling in the heart. METHODS AND RESULTS: We applied chronic infusion of isoproterenol to induced cardiomyopathy in male C57BL/6J mice. We applied confocal and proximity ligation assay to examine ß1AR association with L-type calcium channel, ryanodine receptor 2, and SERCA2a ((Sarco)endoplasmic reticulum calcium ATPase 2a) and Förster resonance energy transfer-based biosensors to probe subcellular ß1AR-PKA (protein kinase A) signaling in ventricular myocytes. Chronic infusion of isoproterenol led to reduced ß1AR protein levels, receptor association with L-type calcium channel and ryanodine receptor 2 measured by proximity ligation (puncta/cell, 29.65 saline versus 14.17 isoproterenol, P<0.05), and receptor-induced PKA signaling at the plasma membrane (Förster resonance energy transfer, 28.9% saline versus 1.9% isoproterenol, P<0.05) and ryanodine receptor 2 complex (Förster resonance energy transfer, 30.2% saline versus 10.6% isoproterenol, P<0.05). However, the ß1AR association with SERCA2a was enhanced (puncta/cell, 51.4 saline versus 87.5 isoproterenol, P<0.05), and the receptor signal was minimally affected. The isoproterenol-infused hearts displayed decreased PDE4D (phosphodiesterase 4D) and PDE3A and increased PDE2A, PDE4A, and PDE4B protein levels. We observed a reduced role of PDE4 and enhanced roles of PDE2 and PDE3 on the ß1AR-PKA activity at the ryanodine receptor 2 complexes and myocyte shortening. Despite the enhanced ß1AR association with SERCA2a, the endogenous norepinephrine-induced signaling was reduced at the SERCA2a complexes. Inhibiting monoamine oxidase A rescued the norepinephrine-induced PKA signaling at the SERCA2a and myocyte shortening. CONCLUSIONS: This study reveals distinct mechanisms for the downregulation of subcellular ß1AR signaling in the heart under chronic adrenergic stimulation.

Towards Whole Health Toxicology: In-Silico Prediction of Diseases Sensitive to Multi-Chemical Exposures.

Chemical exposures from diverse sources merge on a limited number of molecular pathways described as toxicity pathways. Changes in the same set of molecular pathways in different cell and tissue types may generate seemingly unrelated health conditions. Today, no approaches are available to predict in an unbiased way sensitivities of different disease states and their combinations to multi-chemical exposures across the exposome. We propose an inductive in-silico workflow where sensitivities of genes to chemical exposures are identified based on the overlap of existing genomic datasets, and data on sensitivities of individual genes is further used to sequentially derive predictions on sensitivities of molecular pathways, disease states, and groups of disease states (syndromes). Our analysis predicts that conditions representing the most significant public health problems are among the most sensitive to cumulative chemical exposures. These conditions include six leading types of cancer in the world (prostatic, breast, stomach, lung, colorectal neoplasms, and hepatocellular carcinoma), obesity, type 2 diabetes, non-alcoholic fatty liver disease, autistic disorder, Alzheimer's disease, hypertension, heart failure, brain and myocardial ischemia, and myocardial infarction. Overall, our predictions suggest that environmental risk factors may be underestimated for the most significant public health problems.

Effects of diluted Concord grape juice laced with sodium chloride and selected boron-containing compounds on attraction, consumption, crop muscle contractions, and mortality of adult Drosophila suzukii Matsumura (Diptera: Drosophilidae).

BACKGROUND: In recent years, there has been interest in low-cost, reduced-risk materials that could be used for attract-and-kill of the invasive pest, spotted-wing Drosophila, Drosophila suzukii. This pest causes heavy economic damage to soft-skinned fruits in many countries. In this study, we evaluated physiological and behavioral effects of adding either borax, boric acid, or sodium chloride to diluted Concord grape juice (DGJ), a material that is attractive to adult D. suzukii. RESULTS: Results showed that the addition of borax, sodium chloride and boric acid did not significantly affect the response of adult D. suzukii, relative to DGJ alone. Increases in concentrations (to 5% and 10%) of borax, sodium chloride and boric acid were correlated with decreased ingestion of materials. Mortality of males and females was almost 100% with lower concentrations (1% and 5%) of borax and boric acid within 72 h. The higher concentrations of sodium chloride (5% and 10%) resulted in 100% mortality of both sexes within 72 h. There was no significant effect of chemicals on the number of crop contractions of flies when fed for 4 h. CONCLUSIONS: This study suggests that some substances such as boric acid and borax may act as toxicants without influencing the behavioral response of D. suzukii. © 2021 Society of Chemical Industry.

Effects of chitosan and erythritol on labellar taste neuron activity, proboscis extension reflex, daily food intake, and mortality of male and female spotted-winged drosophila, Drosophila suzukii.

In recent years, there has been interest in reduced-risk materials with insecticidal properties for the invasive pest spotted-wing drosophila, Drosophila suzukii. Here, we compared the peripheral sensitivity (via the tip-recording technique, used to monitor the neural activity of gustatory receptor neurons [GRNs]) and palatability (via the Proboscis Extension Reflex [PER]) of chitosan, a polysaccharide derived from chitin, with that of erythritol, a sugar alcohol, to male and female D. suzukii. Because in some insect species it has previously been shown that chitosan has some insecticidal properties, then treatment effects on mortality rates of male and female D. suzukii were quantified. Physiological recordings from the l-type labellar sensilla showed that erythritol evoked responses from one GRN, while chitosan elicited spiked activity from a second one. The first PER bioassay revealed that the level of response to erythritol increased significantly for males and females as the concentrations increased, and the effect of fly sex was non-significant. The second PER bioassay compared the male and female response to chitosan and erythritol each at 0.125, 0.25, 0.5, 1, and 2% concentrations. The overall female PER to erythritol was significantly greater than that exhibited by males, and no differences were noted between sexes when chitosan was evaluated. These results indicate that chitosan alone can elicit PER responses in adult D. suzukii. In the third experiment, chitosan was toxic to D. suzukii. When combined with sucrose (2%), chitosan elicited high levels (80-100%) of mortality of adult D. suzukii within 3 days, particularly in males. The presence of erythritol did not seem to increase the toxic effect of chitosan.

Unbiased approach for the identification of molecular mechanisms sensitive to chemical exposures.

Targeted methods that dominated toxicological research until recently did not allow for screening of all molecular changes involved in toxic response. Therefore, it is difficult to infer if all major mechanisms of toxicity have already been discovered, or if some of them are still overlooked. We used data on 591,084 unique chemical-gene interactions to identify genes and molecular pathways most sensitive to chemical exposures. The list of identified pathways did not change significantly when analyses were done on different subsets of data with non-overlapping lists of chemical compounds indicative that our dataset is saturated enough to provide unbiased results. One of the most important findings of this study is that almost every known molecular mechanism may be affected by chemical exposures. Predictably, xenobiotic metabolism pathways, and mechanisms of cellular response to stress and damage were among the most sensitive. Additionally, we identified highly sensitive molecular pathways, which are not widely recognized as major targets of toxicants, including lipid metabolism pathways, longevity regulation cascade, and cytokine-mediated signaling. These mechanisms are relevant to significant public health problems, such as aging, cancer, metabolic and autoimmune disease. Thus, public health field will benefit from future focus of toxicological research on identified sensitive mechanisms.

Data on chemical-gene interactions and biological categories enriched with genes sensitive to chemical exposures.

A dataset of chemical-gene interactions was created by extracting data from the Comparative Toxicogenomics Database (CTD) with the following filtering criteria: data was extracted only from experiments that used human, rat, or mouse cells/tissues and used high-throughput approaches for gene expression analysis. Genes not present in genomes of all three species were filtered out. The resulting dataset included 591,084 chemical-gene interaction. All chemical compounds in the database were annotated for their major uses. For every gene in the database number of chemical-gene interactions was calculated and used as a metric of gene sensitivity to a variety of chemical exposures. The lists of genes with corresponding numbers of chemical-gene interactions were used in gene-set enrichment analysis (GSEA) to identify potential sensitivity to chemical exposures of molecular pathways in Hallmark, KEGG and Reactome collections. Thus, data presented here represent unbiased and searchable datasets of sensitivity of genes and molecular pathways to a broad range of chemical exposures. As such the data can be used for a diverse range of toxicological and regulatory applications. Approach for the identification of molecular mechanisms sensitive to chemical exposures may inform regulatory toxicology about best toxicity testing strategies. Analysis of sensitivity of genes and molecular pathways to chemical exposures based on these datasets was published in Chemosphere (Suvorov et al., 2021) [1].