Developmental Manganese Exposure Causes Lasting Attention Deficits Accompanied by Dysregulation of mTOR Signaling and Catecholaminergic Gene Expression in Brain Prefrontal Cortex.

Elevated manganese (Mn) exposure is associated with attentional deficits in children, and is an environmental risk factor for attention deficit hyperactivity disorder (ADHD). We have shown that developmental Mn exposure causes lasting attention and sensorimotor deficits in a rat model of early childhood Mn exposure, and that these deficits are associated with a hypofunctioning catecholaminergic system in the prefrontal cortex (PFC), though the mechanistic basis for these deficits is not well understood. To address this, male Long-Evans rats were exposed orally to Mn (50 mg/kg/d) over PND 1-21 and attentional function was assessed in adulthood using the 5-Choice Serial Reaction Time Task. Targeted catecholaminergic system and epigenetic gene expression, followed by unbiased differential DNA methylation and gene regulation expression transcriptomics in the PFC, were performed in young adult littermates. Results show that developmental Mn exposure causes lasting focused attention deficits that are associated with reduced gene expression of tyrosine hydroxylase, dopamine transporter, and DNA methyltransferase 3a. Further, developmental Mn exposure causes broader lasting methylation and gene expression dysregulation associated with epigenetic regulation, inflammation, cell development, and hypofunctioning catecholaminergic neuronal systems. Pathway enrichment analyses uncovered mTOR and Wnt signaling pathway genes as significant transcriptomic regulators of the Mn altered transcriptome, and Western blot of total, C1 and C2 phospho-mTOR confirmed mTOR pathway dysregulation. Our findings deepen our understanding of the mechanistic basis of how developmental Mn exposure leads to lasting catecholaminergic dysfunction and attention deficits, which may aid future therapeutic interventions of environmental exposure associated disorders. Significance Statement: Attention deficit hyperactivity disorder (ADHD) is associated with environmental risk factors, including exposure to neurotoxic agents. Here we used a rodent model of developmental manganese (Mn) exposure producing lasting attention deficits to show broad epigenetic and gene expression changes in the prefrontal cortex, and to identify disrupted mTOR and Wnt signaling pathways as a novel mechanism for how developmental Mn exposure may induce lasting attention and catecholaminergic system impairments. Importantly, our findings establish early development as a critical period of susceptibility to lasting deficits in attentional function caused by elevated environmental toxicant exposure. Given that environmental health threats disproportionately impact communities of color and low socioeconomic status, our findings can aid future studies to assess therapeutic interventions for vulnerable populations.

Chemotherapeutic agents doxorubicin and epirubicin induce a procoagulant phenotype on endothelial cells and blood monocytes.

BACKGROUND: Although chemotherapy is associated with an increased risk of thrombosis, the pathogenic mechanisms by which chemotherapeutic agents exert prothrombotic effects are unclear. OBJECTIVES: In this study we explored the possibility that chemotherapeutic agents doxorubicin, epirubicin, 5-fluorouracil and methotrexate induce a procoagulant phenotype on vascular endothelial cells and/or on blood monocytes. METHODS: Thrombin generation was measured in defibrinated plasma exposed to chemotherapy-treated human umbilical vein endothelial cells (HUVECs). Tissue factor activity assays were performed on chemotherapy-treated HUVECs and blood monocytes. The effects of chemotherapy drugs on phosphatidylserine exposure and the protein C pathway were also measured. RESULTS: Exposure of defibrinated plasma to either doxorubicin- or epirubicin-treated HUVECs resulted in an increase in plasma thrombin generation. The procoagulant activity of doxorubicin- and epirubicin-treated HUVECs reflects an increase in tissue factor activity and phosphatidylserine exposure. Doxorubicin-mediated increase in tissue factor activity is related to increased levels of phosphatidylserine rather than to protein disulfide isomerase activity, and is likely to involve reactive oxygen species generation. Unlike doxorubicin, epirubicin does not have an impact on the protein C anticoagulant pathway. Interestingly, neither methotrextate nor 5-fluorouracil altered endothelial or monocyte hemostatic properties. CONCLUSIONS: These studies suggest that doxorubicin and epirubicin have the greatest 'prothrombotic potential' by virtue of their ability to alter endothelial and monocyte hemostatic properties.

Longitudinal age-related changes in motor activities and spatial orientation in CD-1 mice.

Female CD-1 mice were evaluated on three occasions over a nineteen month span in tests of exploration, motor coordination, and spatial orientation in a water maze. Aging decreased motor activity and exploration of specific environmental stimuli found in a hole-board and in a T-maze. Age-related deficits were also found in three motor coordination tasks (inclined grid, coat-hanger, and round bridge) and during retention but not acquisition of the hidden platform version of the water maze task. Performance on some motor coordination tests was linearly correlated with either motor activity or exploration, implying the existence of similar neurobiological pathways responsible for these age-related changes.

The effects of pentobarbital on spatial learning, motor coordination, and exploration.

Mice injected with either 8, 16 or 32 mg/kg of pentobarbital were as efficient as control subjects in learning and recalling the location of a submerged platform in a water maze. The highest dose of pentobarbital decreased fall latencies in the coat-hanger test of motor coordination. Exploratory activity was not affected by these doses of pentobarbital. The absence of a deficit in spatial learning and in exploratory activity occurred even at a dose sufficient to cause a deficit in motor coordination. These results stand in contrast to previous findings indicating spatial deficits in rats injected with benzodiazepines.

Effects of sodium azide on motor activity, motor coordination, and learning.

Because of the proposed importance of cytochrome oxidase in some neurological disorders, an inhibitor of this enzyme was evaluated in a battery of tests measuring exploration, motor coordination, and learning. Mice injected with sodium azide (6 or 12 mg/kg) were slower to initiate a response in a T maze and had less rears in a small chamber than mice injected with placebo. Drugged mice did not alternate spontaneously even at a minimal retention interval (0 min), but were not impaired in water maze spatial and visual discrimination learning tasks. No group differences emerged in terms of horizontal motor activity and its habituation, number of grooming episodes, and motor coordination. These results indicate that azide-induced slowing of motor activity is situation-specific and is accompanied by abnormalities in choice behavior in a T maze.

CHIME-Net, the Connecticut Health Information Network: a pilot study.

CHIME-Net is a state-wide community health information network project which uses a frame-relay approach to interfacility and internet connectivity. This is a collaborative effort among competitive institutions, which embraces technologies new to the health care industry. The experiences of implementation of the CHIME-Net pilot project are presented as a first milestone for the state-wide effort.

A frame-relay approach for a state-wide health information network.

An approach for a frame-relay implementation is described which is intended to establish connectivity between the health care providers in the state of Connecticut with a gateway to the internet. While other health care networking efforts have based the interconnectivity efforts on direct connections to the internet for each institution, our design takes a more cost effective approach by establishing a private health care network with a single entry point to the internet. This will not only provide the advantages of internet connections to all participating providers, but it will also isolate intrastate patient care traffic from the internet, reserving internet traffic for those information needs not available within the statewide network. In addition to the network solution, an extensive user support infrastructure is also presented.