No abstract

Mucous cell hyperplasia and airway smooth muscle (ASM) hyperresponsiveness are hallmark features of inflammatory airway diseases, including asthma. Here, we show that the recently identified calciumactivated chloride channel (CaCC) TMEM16A is expressed in the adult airway surface epithelium and ASM. The epithelial expression is increased in asthmatics, particularly in secretory cells. Based on this and the proposed functions of CaCC, we hypothesized that TMEM16A inhibitors would negatively regulate both epithelial mucin secretion and ASM contraction. We used a high-throughput screen to identify small-molecule blockers of TMEM16A-CaCC channels. We show that inhibition of TMEM16A-CaCC significantly impairs mucus secretion in primary human airway surface epithelial cells. Furthermore, inhibition of TMEM16A-CaCC significantly reduces mouse and human ASM contraction in response to cholinergic agonists. TMEM16A-CaCC blockers, including those identified here, may positively impact multiple causes of asthma symptoms.A sthma is a significant cause of morbidity and mortality worldwide, and the prevalence of this disease is increasing among all age, sex, and racial groups. Characteristic features of asthma include inflammation, subepithelial fibrosis, hyperplasia of mucus-producing cells, accumulation of mucus within airway lumens, hyperplasia of airway smooth muscle (ASM), and ASM hyperresponsiveness. Together, these symptoms impair lung function by limiting the flow of gases to and from the alveoli in the distal lung.The current standard of care for asthma involves inhaled corticosteroids for the management of inflammation combined with long-acting agonists of β2-adrenergic receptors. Despite this treatment, lung function is not improved in 30-45% of asthmatic patients, and exacerbations continue to be a major problem (reviewed in ref. 1). Asthma can be divided into at least two distinct molecular phenotypes defined by the degree of Th2 inflammation (2, 3). Cytokines, including IL-4 and IL-13, promote airway epithelial mucous cell metaplasia, subepithelial fibrosis, and hyperplasia of smooth muscle in Th2-high asthmatics, and these patients generally show improved lung function with inhaled corticosteroid therapy. A greater understanding of this heterogeneity and the molecular and physiological events that lead to airway remodeling might lead to improved diagnosis and treatment.Calcium-activated chloride channels (CaCCs) have been ascribed numerous cellular functions (reviewed in refs. 4 and 5), among these are epithelial fluid secretion and smooth muscle contraction, both of which contribute to the progression and severity of asthma. Moreover, calcium-activated chloride currents in the airway epithelium are enhanced by the Th2 cytokines IL-4 and IL-13, as well as IFN-γ (6). For these reasons, CaCC is an attractive potential therapeutic target for asthma (7). However, the study of CaCC was impeded by lack of information about the gene(s) encoding this channel. It was only relatively recently that TMEM16A (transmembrane p...

TMEM16A forms calcium-activated chloride channels (CaCCs) that regulate physiological processes such as the secretions of airway epithelia and exocrine glands, the contraction of smooth muscles, and the excitability of neurons. Notwithstanding intense interest in the mechanism behind TMEM16A-CaCC calcium-dependent gating, comprehensive surveys to identify and characterize potential calcium sensors of this channel are still lacking. By aligning distantly related calcium-activated ion channels in the TMEM16 family and conducting systematic mutagenesis of all conserved acidic residues thought to be exposed to the cytoplasm, we identify four acidic amino acids as putative calcium-binding residues. Alterations of the charge, polarity, and size of amino acid side chains at these sites alter the ability of different divalent cations to activate the channel. Furthermore, TMEM16A mutant channels containing double cysteine substitutions at these residues are sensitive to the redox potential of the internal solution, providing evidence for their physical proximity and solvent accessibility.DOI: http://dx.doi.org/10.7554/eLife.02772.001

Effectiveness of vegetative buffer strips for herbicide retention from agricultural runoff was evaluated in a twoyear natural rainfall study. A source area of 0.41 ha (mainly Canisteo silty clay loam soil), having an average slope of 3%, was fall chisel-plowed, spring disked, and planted to corn. Three herbicides (atrazine, metolachlor, and cyanazine) were applied to the source area in each spring. Six vegetative buffer strips, 1.52 m wide ¥ 20.12 m long, were isolated with metal borders downslope of the source area in a well established bromegrass (Bromus inermis) waterway. These strips provided for three replications of two drainage to buffer area ratio treatments of 15:1 and 30:1. Herbicide retention was dependent on the antecedent moisture conditions of the strips. These retentions ranged from 11 to 100% for atrazine, 16 to 100% for metolachlor, and 8 to 100% for cyanazine. Herbicide retention by the buffer strips for the two treatments were not significantly different for the observed storm events. Herbicide concentrations in solution in outflow from the strips were less than the inflow concentrations for all the three herbicides. Infiltration was the key process for herbicide retention by the buffer strips, although there was some adsorption to in-place soil and/or vegetation. Metolachlor concentrations in sediment increased in outflow for the two treatments; however, the opposite was true for atrazine and cyanazine. Herbicide retention by sediment deposition in the strip represented about 5% of the total herbicide retention by the buffer strips. The buffer strips were found to have high percent sediment retention, ranging from 40 to 100%; thus, the strips would be more effective for retaining strongly adsorbed herbicides.

Growing interest in local food has sparked debate about the merits of attempting to reduce the distance food travels. One point of contention is the capacity of local agriculture to meet the food needs of local people. In hopes of informing this debate, this research presents a method for mapping potential foodsheds, land areas that could theoretically feed urban centers. The model was applied to New York State (NYS). Geographic information systems were used to estimate the spatial distribution of food production capacity relative to the food needs of NYS population centers. Optimization tools were then applied to allocate production potential to meet food needs in the minimum distance possible. Overall, the model showed that NYS could provide 34% of its total food needs within an average distance of just 49 km. However, the model did not allocate production potential evenly. Most NYS population centers could have the majority of their food needs sourced in-state, except for the greater New York City (NYC) area. Thus, the study presents a mixed review of the potential for local food systems to reduce the distance food travels. While small- to medium-sized cities of NYS could theoretically meet their food needs within distances two orders of magnitude smaller than the current American food system, NYC must draw on more distant food-producing resources. Nonetheless, the foodshed model provides a successful template for considering the geography of food production and food consumption simultaneously. Such a tool could be valuable for examining how cities might change their food procurement to curb greenhouse gas emissions and adapt to depletion of petroleum and other energy resources necessary for long-distance transport of food.

Agriculture faces a multitude of challenges in the 21st century, and new tools are needed to help determine how it should respond. Among these challenges is a need to reconcile how human food consumption patterns should change to both improve human nutrition and reduce agriculture's environmental footprint. A complete-diet framework is needed for better understanding how diet influences demand for a fundamental agricultural resource, land. We tested such a model, measuring the impact of fat and meat consumption on the land requirements of food production in New York State (NYS). Analysis was confined to this geographic area to simplify the modeling procedure and to examine the state's ability to reduce environmental impact by supplying food locally. Per capita land resource requirements were calculated with a spreadsheet model for 42 diets ranging from 0 to 381 g d−1 (0 to 12 oz d−1) of meat and eggs and 20 to 45% total calories from fat. Many of these diets meet national dietary recommendations. The potential human carrying capacity of the NYS land base was then derived, based on recent estimates of available agricultural land. A nearly fivefold difference (0.18–0.86 ha) in per capita land requirements was observed across the diets. Increasing meat in the diet increased per capita land requirements, while increasing total dietary fat increased the land requirements of low meat diets but reduced the land needed for high meat diets. Higher meat diets used a larger share of the available cropland suited only to pasture and perennial crops. Thus, only a threefold difference was observed for the potential number of people fed from the NYS land base (2.0–6.2 million). In addition, some high-fat vegetarian diets supported fewer people than lower fat diets containing 63–127 g d−1 of meat (approximately one- to two-thirds of the national average per capita consumption in the US). These results support the assertion that diet should be considered in its entirety when assessing environmental impact. To more completely understand how diet influences land requirements and potential carrying capacity, this model should be applied across a larger geographic area that encompasses a wider variety of climates and soil resources. To better understand the ability of a local region to supply more of its own food, the model should be moved into a geospatial framework.

SUMMARY Abnormalities in dendritic spines are manifestations of several neurodevelopmental and psychiatric diseases. TAOK2 is one of the genes in the 16p11.2 locus, copy number variations of which are associated with autism and schizophrenia. Here, we show that the kinase activity of the serine/threonine kinase encoded by TAOK2 is required for spine maturation. TAOK2 depletion results in unstable dendritic protrusions, mislocalized shaft-synapses, and loss of compartmentalization of NMDA receptor-mediated calcium influx. Using chemical-genetics and mass spectrometry, we identified several TAOK2 phosphorylation targets. We show that TAOK2 directly phosphorylates the cytoskeletal GTPase Septin7, at an evolutionary conserved residue. This phosphorylation induces translocation of Septin7 to the spine, where it associates with and stabilizes the scaffolding protein PSD95, promoting dendritic spine maturation. This study provides a mechanistic basis for postsynaptic stability and compartmentalization via TAOK2-Sept7 signaling, with implications toward understanding the potential role of TAOK2 in neurological deficits associated with the 16p11.2 region.

Strategies for environmental sustainability and global food security must account for dietary change. Using a biophysical simulation model we calculated human carrying capacity under ten diet scenarios. The scenarios included two reference diets based on actual consumption and eight "Healthy Diet" scenarios that complied with nutritional recommendations but varied in the level of meat content. We considered the U.S. agricultural land base and accounted for losses, processing conversions, livestock feed needs, suitability of land for crops or grazing, and land productivity. Annual per capita land requirements ranged from 0.13 to 1.08 ha person -1 year -1 across the ten diet scenarios. Carrying capacity varied from 402 to 807 million persons; 1.3 to 2.6 times the 2010 U.S. population. Carrying capacity was generally higher for scenarios with less meat and highest for the lacto-vegetarian diet. However, the carrying capacity of the vegan diet was lower than two of the healthy omnivore diet scenarios. Sensitivity analysis showed that carrying capacity estimates were highly influenced by starting assumptions about the proportion of cropland available for cultivated cropping. Population level dietary change can contribute substantially to meeting future food needs, though ongoing agricultural research and sustainable management practices are still needed to assure sufficient production levels.