Elevated susceptibility to exogenous seizure triggers and impaired interneuron excitability in a mouse model of Leigh syndrome epilepsy.

Mutations in the NADH dehydrogenase (ubiquinone reductase) iron­sulfur protein 4 (NDUFS4) gene, which encodes for a key structural subunit of the OXFOS complex I (CI), lead to the most common form of mitochondrial disease in children known as Leigh syndrome (LS). As in other mitochondrial diseases, epileptic seizures constitute one of the most significant clinical features of LS. These seizures are often very difficult to treat and are a sign of poor disease prognosis. Mice with whole-body Ndufs4 KO are a well-validated model of LS; they exhibit epilepsy and several other clinical features of LS. We have previously shown that mice with Ndufs4 KO in only GABAergic interneurons (Gad2-Ndufs4-KO) reproduce the severe epilepsy phenotype observed in the global KO mice. This observation indicated that these mice represent an excellent model of LS epilepsy isolated from other clinical manifestations of the disease. To further characterize this epilepsy phenotype, we investigated seizure susceptibility to selected exogenous seizure triggers in Gad2-Ndufs4-KO mice. Then, using electrophysiology, imaging, and immunohistochemistry, we studied the cellular, physiological, and neuroanatomical consequences of Ndufs4 KO in GABAergic interneurons. Homozygous KO of Ndufs4 in GABAergic interneurons leads to a prominent susceptibility to exogenous seizure triggers, impaired interneuron excitability and interneuron loss. Finally, we found that the hippocampus and cortex participate in the generation of seizure activity in Gad2-Ndufs4-KO mice. These findings further define the LS epilepsy phenotype and provide important insights into the cellular mechanisms underlying epilepsy in LS and other mitochondrial diseases.

Too Many Cooks: Bayesian Inference for Coordinating Multi-Agent Collaboration.

Collaboration requires agents to coordinate their behavior on the fly, sometimes cooperating to solve a single task together and other times dividing it up into sub-tasks to work on in parallel. Underlying the human ability to collaborate is theory-of-mind (ToM), the ability to infer the hidden mental states that drive others to act. Here, we develop Bayesian Delegation, a decentralized multi-agent learning mechanism with these abilities. Bayesian Delegation enables agents to rapidly infer the hidden intentions of others by inverse planning. We test Bayesian Delegation in a suite of multi-agent Markov decision processes inspired by cooking problems. On these tasks, agents with Bayesian Delegation coordinate both their high-level plans (e.g., what sub-task they should work on) and their low-level actions (e.g., avoiding getting in each other's way). When matched with partners that act using the same algorithm, Bayesian Delegation outperforms alternatives. Bayesian Delegation is also a capable ad hoc collaborator and successfully coordinates with other agent types even in the absence of prior experience. Finally, in a behavioral experiment, we show that Bayesian Delegation makes inferences similar to human observers about the intent of others. Together, these results argue for the centrality of ToM for successful decentralized multi-agent collaboration.

Caveolae-Mediated Activation of Mechanosensitive Chloride Channels in Pulmonary Veins Triggers Atrial Arrhythmogenesis.

Background Atrial fibrillation often occurs in the setting of hypertension and associated atrial dilation with pathologically increased cardiomyocyte stretch. In the setting of atrial dilation, mechanoelectric feedback has been linked to the development of ectopic beats that trigger paroxysmal atrial fibrillation mainly originating from pulmonary veins (PVs). However, the precise mechanisms remain poorly understood. Methods and Results We identify mechanosensitive, swelling-activated chloride ion channels (ICl,swell) as a crucial component of the caveolar mechanosensitive complex in rat and human cardiomyocytes. In vitro optical mapping of rat PV, single rat PV, and human cardiomyocyte patch clamp studies showed that stretch-induced activation of ICl,swell leads to membrane depolarization and decreased action potential amplitude, which trigger conduction discontinuities and both ectopic and reentrant activities within the PV. Reverse transcription quantitative polymerase chain reaction, immunofluorescence, and coimmunoprecipitation studies showed that ICl,swell likely consists of at least 2 components produced by mechanosensitive ClC-3 (chloride channel-3) and SWELL1 (also known as LRRC8A [leucine rich repeat containing protein 8A]) chloride channels, which form a macromolecular complex with caveolar scaffolding protein Cav3 (caveolin 3). Downregulation of Cav3 protein expression and disruption of caveolae structures during chronic hypertension in spontaneously hypertensive rats facilitates activation of ICl,swell and increases PV sensitivity to stretch 10- to 50-fold, promoting the development of atrial fibrillation. Conclusions Our findings identify caveolae-mediated activation of mechanosensitive ICl,swell as a critical cause of PV ectopic beats that can initiate atrial arrhythmias including atrial fibrillation. This mechanism is exacerbated in the setting of chronically elevated blood pressures.

Development and pilot testing of a text message vaccine reminder system for use during an influenza pandemic.

In an influenza pandemic, two vaccine doses administered 21 days apart may be needed for individuals of all ages to achieve seroprotection. Achieving dose-series completion can be challenging even for routinely recommended vaccines. To prepare for a two-dose influenza pandemic vaccine campaign and promote dose-series completion and correct dosing intervals, CDC and its partners developed a text message-based vaccine reminder system to remind persons who receive a first dose of pandemic influenza vaccine to receive the second dose. Taking advantage of the high prevalence of cell phones in the United States, the system sent second-dose text message reminders and hyperlinks to educational information. The system was pilot tested from November 2015 to April 2016 among graduate public health students enrolled at four United States universities. Universities were selected based on convenience, and each university used a different recruitment method. Among 59 volunteers who pilot tested the system and completed a survey, 57 (92%) felt the system would be helpful during a pandemic. Forty (68%) respondents felt the information included in the messages was informative. Volunteers recommended including actionable ways to stay healthy during a pandemic, though specific suggestions varied. With further development, text reminder systems could be used to promote adherence to a two-dose regimen in a future pandemic, although audience-specific messaging and other complementary systems will likely be needed. Public and private partners can adapt and implement this tool in conjunction with their routine patient information systems to improve dose-series completion and ensure optimal protection during an influenza pandemic.

Green Tea, Intermittent Sprinting Exercise, and Fat Oxidation.

Fat oxidation has been shown to increase after short term green tea extract (GTE) ingestion and after one bout of intermittent sprinting exercise (ISE). Whether combining the two will result in greater fat oxidation after ISE is undetermined. The aim of the current study was to investigate the combined effect of short term GTE and a single session of ISE upon post-exercise fat oxidation. Fourteen women consumed three GTE or placebo capsules the day before and one capsule 90 min before a 20-min ISE cycling protocol followed by 1 h of resting recovery. Fat oxidation was calculated using indirect calorimetry. There was a significant increase in fat oxidation post-exercise compared to at rest in the placebo condition (p < 0.01). After GTE ingestion, however, at rest and post-exercise, fat oxidation was significantly greater (p < 0.05) than that after placebo. Plasma glycerol levels at rest and 15 min during post-exercise were significantly higher (p < 0.05) after GTE consumption compared to placebo. Compared to placebo, plasma catecholamines increased significantly after GTE consumption and 20 min after ISE (p < 0.05). Acute GTE ingestion significantly increased fat oxidation under resting and post-exercise conditions when compared to placebo.

Expression of lymphocyte homing receptors alpha4beta7 and MAdCAM-l in young and old rats.

The elderly constitute the most rapidly growing subpopulation in the United States. This age group represents a significant burden on the healthcare system due, in part, to increases in morbidity and mortality associated with an increase in the incidence of intestinal infectious diseases. Our previous studies suggest that impaired homing of IgA immunoblasts from the Peyer's patches to the intestinal lamina propria contributes to the diminished intestinal immune response in the elderly. The present study employs flow cytometry and quantitative immunohistochemistry to assess age-related changes in the numbers of peripheral blood mononuclear cells expressing the homing integrin alpha4beta7 and vascular endothelial cells in the intestine expressing its specific receptor, the address in MAdCAM-1, in inbred Fischer 344 rats. The proportion of alpha4beta7-positive mononuclear cells in young rats is significantly greater than that measured in the blood of senescent animals. Although the density of intestinal lamina propria blood vessels with MAdCAM-1-positive endothelium was greater in young adult rats in comparison to old animals, this difference achieved only borderline statistical significance. This is the first study to examine the expression of these two critical lymphocyte homing molecules as a function of age.

A self-contained 48-well fatty acid oxidation assay.

The modulation of fatty acid metabolism and especially the stimulation of fatty acid oxidation in liver or skeletal muscle are attractive therapeutic approaches for the treatment of obesity and the associated insulin resistance. However, current beta-oxidation assays are run in very low throughput, which represents an obstacle for drug discovery in this area. Here we describe results for a 48-well beta-oxidation assay using a new instrument design. A connecting chamber links two adjacent wells to form an experimental unit, in which one well contains the beta-oxidation reaction and the other captures CO(2). The experimental units are sealed from each other and from the outside to prevent release of radioactivity from the labeled substrate. CO(2) capture in this instrument is linear with time and over the relevant experimental range of substrate concentration. Cellular viability is maintained in the sealed environment, and cells show the expected responses to modulators of beta-oxidation, such as the AMP kinase activator 5-aminoimidazole carboxamide riboside. Data are presented for different lipid substrates and cell lines. The increased throughput of this procedure compared with previously described methods should facilitate the evaluation of compounds that modulate fatty acid metabolism.

gp120-independent HIV infection of cells derived from the female reproductive tract, brain, and colon.

The infection of CD4 cells may have significant involvement in the transmission and long-term persistency of HIV. Using HIV clones carrying the enhanced green fluorescent protein (EGFP), we infected epithelial and glioneuronal cell lines derived from the female reproductive tract, brain, colon, and intestine. HIV infection was quantified by counting EGFP-positive cells. Infection was quantified in cell lines from the female reproductive tract, brain tissue, and colon tissue (0.36%-3.15%). Virus replicated in the infected cells and the progeny virus were infectious for CD4 cells, HeLa-CD4, and CEM T lymphocytes. Furthermore, we found that infection of these epithelial and brain cell lines is independent of gp120. The results from the infection of CD4 epithelial cells suggest that HIV can traverse epithelial cell layers by infecting them through a gp120-independent mechanism. Infection of glial and neuronal cell lines suggests that HIV infection of these cells is a probable mechanism for HIV pathogenicity in the brain and a possible cause for persistent infection in patients.