CHARACTERIZATION AND TREATMENT OF A NOVEL MOUSE MODEL OF TSC-ASSOCIATED AUTISM

Tuberous sclerosis complex (TSC) is a dominant tumor suppressor disorder caused by mutations in either TSC1 or TSC2. The proteins of these genes form a complex to inhibit the mammalian target of rapamycin complex 1 (mTORC1), which controls protein translation and cell growth. TSC causes substantial neuropathology, often leading to autism spectrum disorders (ASDs) in up to 60% of patients. The anatomic and neurophysiologic links between these two disorders are not well understood. However, both disorders share cerebellar abnormalities. Therefore, we have characterized a novel mouse model in which the Tsc2 gene was selectively deleted from cerebellar Purkinje cells (Tsc2f/-;Cre). These mice exhibit progressive Purkinje cell degeneration. Since loss of Purkinje cells is a well-reported postmortem finding in patients with ASD, we conducted a series of behavior tests to assess if Tsc2f/-;Cre mice displayed autistic-like deficits. Using the three chambered social choice assay, we found that Tsc2f/-;Cre mice showed behavioral deficits, exhibiting no preference between a stranger mouse and an inanimate object, or between a novel and a familiar mouse. Tsc2f/-;Cre mice also demonstrated increased repetitive behavior as assessed with marble burying activity. Altogether, these results demonstrate that loss of Tsc2 in Purkinje cells in a haploinsufficient background lead to behavioral deficits that are characteristic of human autism. Therefore, Purkinje cells loss and/or dysfunction may be an important link between TSC and ASD. Additionally, we have examined some of the cellular mechanisms resulting from mutations in Tsc2 leading to Purkinje cell death. Loss of Tsc2 led to upregulation of mTORC1 and increased cell size. As a consequence of increased protein synthesis, several cellular stress pathways were upregulated. Principally, these included altered calcium signaling, oxidative stress, and ER stress. Likely as a consequence of ER stress, there was also upregulation of ubiquitin and autophagy. Excitingly, treatment with an mTORC1 inhibitor, rapamycin attenuated mTORC1 activity and prevented Purkinje cell death by reducing of calcium signaling, the ER stress response, and ubiquitin. Remarkably, rapamycin treatment also reversed the social behavior deficits, thus providing a promising potential therapy for TSC-associated ASD.

Ethnic differences in food availability at home among 3--5 year old preschoolers attending Head Start centers in 2008

Background. The childhood obesity epidemic has disproportionately impacted the lives of low-income, minority preschoolers and their families. Research shows that parents play a major role as "gatekeepers" who control what food is brought into the home and as role models for dietary practices. Currently, there is limited research regarding ethnic differences in families of low-income preschoolers. ^ Objective. The objective of this study was to look at ethnic differences in food availability at home among the low-income families of Hispanic and African American preschoolers attending Head Start centers in Harris County, Texas. ^ Design/Subjects. Descriptive data on food availability at home between Hispanic and African American families were used and analyzed for this study. Parents or primary caregivers (n = 718) of children enrolled at Head Start Centers in Houston, Texas completed the Healthy Home Survey. ^ Methods. In the Healthy Home Survey, participants were asked to answer open-ended questions regarding various types of foods currently available at home, such as fresh, canned or jarred, dried and frozen fruits; fresh, canned or jarred, and frozen vegetables; salty snacks, sweet snacks, candy, and soda. Descriptive analyses were conducted to identify significant differences between Hispanics and African Americans via a paired t-test to compare the means of variables between the study groups and a Pearson's chi-square or Fischer's exact (if cell size was <5) test calculated for food availability (food types) between ethnicities to determine differences in distributions. ^ Results. Although both Hispanics and African Americans reported having all categories of food types at home, there were statistically significant differences between ethnic groups. Hispanics were more likely to have fresh fruits and vegetables at home than African Americans. At the same time, more African American families reported having canned or jarred fruits and canned green/leafy vegetables than Hispanics. More Hispanic families reported having diet, regular, and both diet and regular sodas available compared to African American families. However, high percentages of unhealthy foods (including snacks and candy) were reported by both ethnicities. ^ Conclusions. The findings presented in this study indicate the implicit ethnic differences that exist in the food availability among low-income families of Hispanic and African American preschoolers. Future research should investigate the associations between food availability and children's weight status by ethnicity to identify additional differences that may exist.^

Chlorophyll content and temperature-dependent net photosynthesis and respiration rate in the lichen Cetraria aculeata

We studied polar and temperate samples of the lichen Cetraria aculeata to investigate whether genetical differences between photobionts are correlated with physiological properties of the lichen holobiont. Net photosynthesis and dark respiration (DR) at different temperatures (from 0 to 30 °C) and photon flux densities (from 0 to 1,200 ?mol/m**2/s) were studied for four populations of Cetraria aculeata. Samples were collected from maritime Antarctica, Svalbard, Germany and Spain, representing different climatic situations. Sequencing of the photobiont showed that the investigated samples fall in the polar and temperate clade described in Fernández-Mendoza et al. (2011, doi:10.1111/j.1365-294X.2010.04993.x). Lichens with photobionts from these clades differ in their temperature optimum for photosynthesis, maximal net photosynthesis, maximal DR and chlorophyll content. Maximal net photosynthesis was much lower in Antarctica and Svalbard than in Germany and Spain. The difference was smaller when rates were expressed by chlorophyll content. The same is true for the temperature optima of polar (11 °C) and temperate (15 and 17 °C) lichens. Our results indicate that lichen mycobionts may adapt or acclimate to local environmental conditions either by selecting algae from regional pools or by regulating algal cell numbers (chlorophyll content) within the thallus.

Global distributions of picophytoplankton abundance and biomass - Gridded data product (NetCDF) - Contribution to the MAREDAT World Ocean Atlas of Plankton Functional Types

The smallest marine phytoplankton, collectively termed picophytoplankton, have been routinely enumerated by flow cytometry since the late 1980s, during cruises throughout most of the world ocean. We compiled a database of 40,946 data points, with separate abundance entries for Prochlorococcus, Synechococcus and picoeukaryotes. We use average conversion factors for each of the three groups to convert the abundance data to carbon biomass. After gridding with 1° spacing, the database covers 2.4% of the ocean surface area, with the best data coverage in the North Atlantic, the South Pacific and North Indian basins. The average picophytoplankton biomass is 12 ± 22 µg C L-1 or 1.9 g C m-2. We estimate a total global picophytoplankton biomass, excluding N2-fixers, of 0.53 - 0.74 Pg C (17 - 39 % Prochlorococcus, 12 - 15 % Synechococcus and 49 - 69 % picoeukaryotes). Future efforts in this area of research should focus on reporting calibrated cell size, and collecting data in undersampled regions.

Vertical distribution of phytoplankton pigments in the equatorial Pacific

Abundance distribution and cellular characteristics of picophytoplankton were studied in two distinct regions of the equatorial Pacific: the western warm pool (0°, 167°E), where oligotrophic conditions prevail, and the equatorial upwelling at 150°W characterized by high-nutrient low-chlorophyll (HNLC) conditions. The study was done in September-October 1994 during abnormally warm conditions. Populations of Prochlorococcus, orange fluorescing Synechococcus and picoeukaryotes were enumerated by flow cytometry. Pigment concentrations were studied by spectrofluorometry. In the warm pool, Prochlorococcus were clearly the dominant organisms in terms of cell abundance, estimated carbon biomass and measured pigment concentration. Integrated concentrations of Prochlorococcus, Synechococcus and picoeukaryotes were 1.5x10**13, 1.3x10**11 and 1.5x10**11 cells/m**2, respectively. Integrated estimated carbon biomass of picophytoplankton was 1 g/m**2, and the respective contributions of each group to the biomass were 69, 3 and 28%. In the HNLC waters, Prochlorococcus cells were slightly less numerous than in the warm pool, whereas the other groups were several times more abundant (from 3 to 5 times). Abundance of Prochlorococcus, Synechococcus and picoeukaryotes were 1.2x10**13, 6.2x10**11 and 5.1x10**11 cells/m**2, respectively. The integrated biomass was 1.9 g C/m**2. Prochlorococcus was again the dominant group in terms of abundance and biomass (chlorophyll, carbon); the respective contributions of each group to the carbon biomass were 58, 7 and 35%. In the warm pool the total chlorophyll biomass was 28 mg/m**2, 57% of which was divinyl chlorophyll a. In the HNLC waters, the total chlorophyll biomass was 38 mg/m**2, 44% of which was divinyl chlorophyll a. Estimates of Prochlorococcus, Synechococcus and picoeukaryotes cell size were made in both hydrological conditions.

High resolution bathymetric compilation for Potter Cove, WAP, Antarctica, with links to data in ArcGIS format

The bathymetry raster with a resolution of 5 m x 5 m was processed from unpublished single beam data from the Argentine Antarctica Institute (IAA, 2010) and multibeam data from the United Kingdom Hydrographic Office (UKHO, 2012) with a cell size of 5 m x 5 m. A coastline digitized from a satellite image (DigitalGlobe, 2014) supplemented the interpolation process. The 'Topo to Raster' tool in ArcMap 10.3 was used to merge the three data sets, while the coastline represented the 0-m-contour to the interpolation process ('contour type option').

Global distributions of diatoms abundance, biovolume and biomass - Gridded data product (NetCDF) - Contribution to the MAREDAT World Ocean Atlas of Plankton Functional Types

This study is a first effort to compile the largest possible body of data available from different plankton databases as well as from individual published or unpublished datasets regarding diatom distribution in the world ocean. The data obtained originate from time series studies as well as spatial studies. This effort is supported by the Marine Ecosystem Data (MAREDAT) project, which aims at building consistent data sets for the main PFTs (Plankton Functional Types) in order to help validate biogeochemical ocean models by using converted C biomass from abundance data. Diatom abundance data were obtained from various research programs with the associated geolocation and date of collection, as well as with a taxonomic information ranging from group down to species. Minimum, maximum and average cell size information were mined from the literature for each taxonomic entry, and all abundance data were subsequently converted to biovolume and C biomass using the same methodology.

Seasonal changes in labile organic matter composition on the continental shelf and bathyal sediments of the Cretan Sea

Bacterial abundance, biomass and cell size were studied in the oligotrophic sediments of the Cretan Sea (Eastern Mediterranean), in order to investigate their response to the seasonal varying organic matter (OM) inputs. Sediment samples were collected on a seasonal basis along a transect of seven stations (ranging from 40 to 1570 m depth) using a multiple-corer. Bacterial parameters were related to changes in chloroplastic pigment equivalents (CPE), the biochemical composition (proteins, lipids, carbohydrates) of the sedimentary organic matter and the OM flux measured at a fixed station over the deep basin (1570 m depth). The sediments of the Cretan Sea represent a nutrient depleted ecosystem characterised by a poor quality organic matter. All sedimentary organic compounds were found to vary seasonally, and changes were more evident on the continental shelf than in deeper sediments. Bacterial abundance and biomass in the sediments of the Cretan Sea (ranging from 1.02 to 4.59 * 10**8 cells/g equivalent to 8.7 and 38.7 µgC/g) were quite high and their distribution appeared to be closely related to the input of fresh organic material. Bacterial abundance and biomass were sensitive to changes in nutrient availability, which also controls the average cell size and the frequency of dividing cells. Bacterial abundance increased up to 3-fold between August '94 and February '95 in response to the increased amount of sedimentary proteins and CPE, indicating that benthic bacteria were constrained more by changes in quality rather than the quantity of the sedimentary organic material. Bacterial responses to the food inputs were clearly detectable down to 10 cm depth. The distribution of labile organic compounds in the sediments appeared to influence the vertical patterns of bacterial abundance and biomass. Cell size decreased significantly with water depth. Bacterial abundance and biomass were characterised by clear seasonal changes in response to seasonal OM pulses. The strong coupling between protein flux and bacterial biomass together with the strong bacterial dominance over the total biomass suggest that the major part of the carbon flow was channelled through the bacteria and the benthic microbial loop.

Northeast Greenland - Digital Bathymetric Compilation, with links to DBM in ArcGIS format

A new digital bathymetric model (DBM) for the Northeast Greenland (NEG) continental shelf (74°N - 81°N) is presented. The DBM has a grid cell size of 250 m × 250 m and incorporates bathymetric data from 30 multibeam cruises, more than 20 single-beam cruises and first reflector depths from industrial seismic lines. The new DBM substantially improves the bathymetry compared to older models. The DBM not only allows a better delineation of previously known seafloor morphology but, in addition, reveals the presence of previously unmapped morphological features including glacially derived troughs, fjords, grounding-zone wedges, and lateral moraines. These submarine landforms are used to infer the past extent and ice-flow dynamics of the Greenland Ice Sheet during the last full-glacial period of the Quaternary and subsequent ice retreat across the continental shelf. The DBM reveals cross-shelf bathymetric troughs that may enable the inflow of warm Atlantic water masses across the shelf, driving enhanced basal melting of the marine-terminating outlet glaciers draining the ice sheet to the coast in Northeast Greenland. Knolls, sinks, and hummocky seafloor on the middle shelf are also suggested to be related to salt diapirism. North-south-orientated elongate depressions are identified that probably relate to ice-marginal processes in combination with erosion caused by the East Greenland Current. A single guyot-like peak has been discovered and is interpreted to have been produced during a volcanic event approximately 55 Ma ago.

Culture experiments to evaluate CO2 effects on growth and morphology of the coccolithophore Pleurochrysis cf. pseudoroscoffensis

In the framework of the Italian project CO2 Monitor, two culture experiments were carried out in vertical closed photobioreactors with Pleurochrysis cf. pseudoroscoffensis Gayral & Fresnel 1983, a coccolithophore isolated from the Gulf of Trieste (North Adriatic Sea). The aim of this study was to investigate the effects induced by pH variations due to CO2 emissions on its growth and morphology. Two experiments were carried out with two different CO2 concentrations (1 and 2%). Growth and cell size in light microscopy, morphology and coccolith size in scanning electron microscopy, particulate nitrogen (PN) and particulate inorganic and organic carbon (PIC and POC) content of the coccolithophore were investigated during the light and dark phases. Dissolved inorganic nutrient (nitrate and phosphate) concentrations and pH of the medium and the presence of heterotrophic prokaryotes (HP) were monitored as well.

Global Ocean Particulate Organic Carbon flux merged with satellite parameters

The efficiency of the biological pump of carbon to the deep ocean depends largely on the biologically mediated export of carbon from the surface ocean and its remineralization with depth. Global satellite studies have primarily focused on chlorophyll concentration and net primary production (NPP) to understand the role of phytoplankton in these processes. Recent satellite retrievals of phytoplankton composition now allow for the size of phytoplankton cells to be considered. Here, we improve understanding of phytoplankton size structure impacts on particle export, remineralization and transfer. Particulate organic carbon (POC) flux observations from sediment traps and 234Th are compiled across the global ocean. Annual climatologies of NPP, percent microplankton, and POC flux at four time series locations and within biogeochemical provinces are constructed, and sinking velocities are calculated to align surface variables with POC flux at depth. Parameters that characterize POC flux vs. depth (export flux ratio, labile fraction, remineralization length scale) are then fit to the aligned dataset. Times of the year dominated by different size compositions are identified and fit separately in regions of the ocean where phytoplankton cell size showed enough dynamic range over the annual cycle. Considering all data together, our findings support the paradigm of high export flux but low transfer efficiency in more productive regions and vice versa for oligotrophic regions. However, when parsing by dominant size class, we find periods dominated by small cells to have both greater export flux and lower transfer efficiency than periods when large cells comprise a greater proportion of the phytoplankton community.

Effects of high CO2 levels on the ecophysiology of the diatom Thalassiosira weissflogii differ depending on the iron nutritional status

Iron availability in seawater, namely the concentration of dissolved inorganic iron ([Fe']), is affected by changes in pH. Such changes in the availability of iron should be taken into account when investigating the effects of ocean acidification on phytoplankton ecophysiology because iron plays a key role in phytoplankton metabolism. However, changes in iron availability in response to changes in ocean acidity are difficult to quantify specifically using natural seawater because these factors change simultaneously. In the present study, the availability of iron and carbonate chemistry were manipulated individually and simultaneously in the laboratory to examine the effect of each factor on phytoplankton ecophysiology. The effects of various pCO2 conditions (390, 600, and 800 µatm) on the growth, cell size, and elemental stoichiometry (carbon [C], nitrogen [N], phosphorus [P], and silicon [Si]) of the diatom Thalassiosira weissflogii under high iron ([Fe'] = 240 pmol/l) and low iron ([Fe'] = 24 pmol/l) conditions were investigated. Cell volume decreased with increasing pCO2, whereas intracellular C, N, and P concentrations increased with increasing pCO2 only under high iron conditions. Si:C, Si:N, and Si:P ratios decreased with increasing pCO2. It reflects higher production of net C, N, and P with no corresponding change in net Si production under high pCO2 and high iron conditions. In contrast, significant linear relationships between measured parameters and pCO2 were rarely detected under low iron conditions. We conclude that the increasing CO2 levels could affect on the biogeochemical cycling of bioelements selectively under the iron-replete conditions in the coastal ecosystems.

Seawater carbonate chemistry, POC, PIC, TPC, SPM, N, TEP and growth rate during experiments with coccolithophores Emiliania huxleyi (AC472), Calcidiscus leptoporus (AC370) and Syracosphaera pulchra (AC418) during experiments, 2011

The response of three coccolithophores (Emiliania huxleyi, Calcidiscus leptoporus and Syracosphaera pulchra) to elevated partial pressure (pCO2) of carbon dioxide was investigated in batch cultures. For the first time, we also report on the response of the non calcifying (haploid) life stage of these three species. The growth rate, cell size, inorganic (PIC) and organic carbon (POC) of both life stages were measured at two different pCO2 (400and 760 ppm) and their organic and inorganic carbon production calculated. The two lifestages within the same species generally exhibited a similar response to elevated pCO2, theresponse of the haploid stage being often more pronounced than that of the diploid stage. Thegrowth rate was consistently higher at higher pCO2 but the response of other processes varied among species. The calcification rate of C. leptoporus and of S. pulchra did not change at elevated pCO2 while increased in E. huxleyi. The POC production as well as the cell size of both life stages of S. pulchra and of the haploid stage of E. huxleyi markedly decreased at elevated pCO2. It remained unaltered in the diploid stage of E. huxleyi and C. leptoporus and increased in the haploid stage of the latter. The PIC:POC ratio increased in E. huxleyi and was constant in C. leptoporus and S. pulchra. These results suggest that the non-calcifying stage, is more responsive than the calcifying stage and that the most versatile genera will proliferate in a more acidic ocean rather than all coccolithophores will decline.

Experiment: Influence of CO2 and nitrogen limitation on the coccolith volume of Emiliania huxleyi (Haptophyta)

Coccolithophores, a key phytoplankton group, are one of the most studied organisms regarding their physiological response to ocean acidification/carbonation. The biogenic production of calcareous coccoliths has made coccolithophores a promising group for paleoceanographic research aiming to reconstruct past environmental conditions. Recently, geochemical and morphological analyses of fossil coccoliths have gained increased interest in regard to changes in seawater carbonate chemistry. The cosmopolitan coccolithophore Emiliania huxleyi (Lohm.) Hay and Mohler was cultured over a range of pCO2 levels in controlled laboratory experiments under nutrient replete and nitrogen limited conditions. Measurements of photosynthesis and calcification revealed, as previously published, an increase in particulate organic carbon production and a moderate decrease in calcification from ambient to elevated pCO2. The enhancement in particulate organic carbon production was accompanied by an increase in cell diameter. Changes in coccolith volume were best correlated with the coccosphere/cell diameter and no significant correlation was found between the coccolith volume and the particulate inorganic carbon production. The conducted experiments revealed that the coccolith volume of E. huxleyi is variable with aquatic CO2 concentration but its sensitivity is rather small in comparison with its sensitivity to nitrogen limitation. Comparing coccolith morphological and geometrical parameters like volume, mass and size to physiological parameters under controlled laboratory conditions is an important step to understand variations in fossil coccolith geometry.