Pathogenic Cav3.2 channel mutation in a child with primary generalized epilepsy.

Two paternally-inherited missense variants in CACNA1H were identified and characterized in a 6-year-old child with generalized epilepsy. Febrile and unprovoked seizures were present in this child. Both variants were expressed in cis or isolation using human recombinant Cav3.2 calcium channels in tsA-201 cells. Whole-cell patch-clamp recordings indicated that one variant (c.3844C > T; p.R1282W) caused a significant increase in current density consistent with a pathogenic gain-of-function phenotype; while the other cis-related variant (c.5294C > T; p.A1765V) had a benign profile

A Tree-Ring Record of Historical Fire Activity In a Piedmont Longleaf Pine (\u3ci\u3ePinus palustris\u3c/i\u3e Mill.) Woodland In North Carolina, USA

CO2 capture from industrial point source waste streams represents an important need for achieving the global goal of carbon-neutrality. Compared with conventional liquid sorbents, solid sorbents can exhibit several distinct advantages, including enhanced lifetime and reduced energy consumption for sorbent regeneration. Considering that reducing CO2 emission is a great challenge, reaching approximately 37 billion metric tons just in 2021, ideal sorbent solutions should not only exhibit a high capture performance but also enable large scale manufacturing using low-cost precursors and simple processes. In this work, we demonstrate the use of a commodity polymer, polystyrene-block-polyisoprene-block-polystyrene (SIS), as the starting material for preparing hierarchically porous, sulfur-doped carbons for CO2 capture. Particularly, the sulfonation-crosslinking reaction enables the formation of macropores in the polymer framework due to the release of gaseous byproducts. After carbonization and activation, the highly porous structure of SIS-derived carbons is successfully retained, while their surface area can reach up to 905 m2 g−1. These porous carbon sorbents exhibit excellent CO2 uptake performance, reaching sorption capacities of 3.8 mmol g−1 at 25 °C and 6.0 mmol g−1 at 0 °C, as well as a high selectivity up to 43 : 1 against N2 gas under ambient conditions. Overall, our work provides an industrially viable method for “template-free” fabrication of porous carbons from commodity polyolefin-based materials, which can be employed for reducing CO2 emission from industrial plants/sectors

Mycobacterium bovis: A model pathogen at the interface of livestock, wildlife, and humans

Complex and dynamic interactions involving domestic animals, wildlife, and humans create environments favorable to the emergence of new diseases, or reemergence of diseases in new host species. Today, reservoirs of Mycobacterium bovis, the causative agent of tuberculosis in animals, and sometimes humans, exist in a range of countries and wild animal populations. Free-ranging populations of white-tailed deer in the US, brushtail possum in New Zealand, badger in the Republic of Ireland and the United Kingdom, and wild boar in Spain exemplify established reservoirs of M. bovis. Establishment of these reservoirs is the result of factors such as spillover from livestock, translocation of wildlife, supplemental feeding of wildlife, and wildlife population densities beyond normal habitat carrying capacities. As many countries attempt to eradicate M. bovis from livestock, efforts are impeded by spillback from wildlife reservoirs. It will not be possible to eradicate this important zoonosis from livestock unless transmission between wildlife and domestic animals is halted. Such an endeavor will require a collaborative effort between agricultural, wildlife, environmental, and political interests.Peer Reviewe

Characterization of effector and memory T cell subsets in the immune response to bovine tuberculosis in cattle

Cultured IFN-γ ELISPOT assays are primarily a measure of central memory T cell (Tcm) responses with humans; however, this important subset of lymphocytes is poorly characterized in cattle. Vaccine-elicited cultured IFN-γ ELISPOT responses correlate with protection against bovine tuberculosis in cattle. However, whether this assay measures cattle Tcm responses or not is uncertain. The objective of the present study was to characterize the relative contribution of Tcm (CCR7+, CD62Lhi, CD45RO+), T effector memory (Tem, defined as: CCR7-, CD62Llow/int, CD45RO+), and T effector cells (CCR7-, CD62L-/low, CD45RO-), in the immune response to Mycobacterium bovis. Peripheral blood mononuclear cells (PBMC) from infected cattle were stimulated with a cocktail of M. bovis purified protein derivative, rTb10.4 and rAg85A for 13 days with periodic addition of fresh media and rIL-2. On day 13, cultured PBMC were re-stimulated with medium alone, rESAT-6:CFP10 or PPDb with fresh autologous adherent cells for antigen presentation. Cultured cells (13 days) or fresh PBMCs (ex vivo response) from the same calves were analyzed for IFN-γ production, proliferation, and CD4, CD45RO, CD62L, CD44, and CCR7 expression via flow cytometry after overnight stimulation. In response to mycobacterial antigens, ~75% of CD4+ IFN-γ+ cells in long-term cultures expressed a Tcm phenotype while less than 10% of the ex vivo response consisted of Tcm cells. Upon re-exposure to antigen, long-term cultured cells were highly proliferative, a distinctive characteristic of Tcm, and the predominant phenotype within the long-term cultures switched from Tcm to Tem. These findings suggest that proliferative responses of Tcm cells to some extent occurs simultaneously with reversion to effector phenotypes (mostly Tem). The present study characterizes Tcm cells of cattle and their participation in the response to M. bovis infection

Groundwater Flow and Exchange Across the Land Surface Explain Carbon Export Patterns in Continuous Permafrost Watersheds

Groundwater flow regimes in the seasonally thawed soils in areas of continuous permafrost are relatively unknown despite their potential role in delivering water, carbon, and nutrients to streams. Using numerical groundwater flow models informed by observations from a headwater catchment in arctic Alaska, United States, we identify several mechanisms that result in substantial surface‐subsurface water exchanges across the land surface during downslope transport and create a primary control on dissolved organic carbon loading to streams and rivers. The models indicate that surface water flowing downslope has a substantial groundwater component due to rapid surface‐subsurface exchanges across a range of hydrologic states, from unsaturated to flooded. Field‐based measurements corroborate the high groundwater contributions, and river dissolved organic carbon concentrations are similar to that of groundwater across large discharge ranges. The persistence of these groundwater contributions in arctic watersheds will influence carbon export to rivers as thaw depth increases in a warmer climate

Groundwater Flow and Exchange Across the Land Surface Explain Carbon Export Patterns in Continuous Permafrost Watersheds

Groundwater flow regimes in the seasonally thawed soils in areas of continuous permafrost are relatively unknown despite their potential role in delivering water, carbon, and nutrients to streams. Using numerical groundwater flow models informed by observations from a headwater catchment in arctic Alaska, United States, we identify several mechanisms that result in substantial surface‐subsurface water exchanges across the land surface during downslope transport and create a primary control on dissolved organic carbon loading to streams and rivers. The models indicate that surface water flowing downslope has a substantial groundwater component due to rapid surface‐subsurface exchanges across a range of hydrologic states, from unsaturated to flooded. Field‐based measurements corroborate the high groundwater contributions, and river dissolved organic carbon concentrations are similar to that of groundwater across large discharge ranges. The persistence of these groundwater contributions in arctic watersheds will influence carbon export to rivers as thaw depth increases in a warmer climate

Exposure of Soil Microbial Communities to Chromium and Arsenic Alters Their Diversity and Structure

Extensive use of chromium (Cr) and arsenic (As) based preservatives from the leather tanning industry in Pakistan has had a deleterious effect on the soils surrounding production facilities. Bacteria have been shown to be an active component in the geochemical cycling of both Cr and As, but it is unknown how these compounds affect microbial community composition or the prevalence and form of metal resistance. Therefore, we sought to understand the effects that long-term exposure to As and Cr had on the diversity and structure of soil microbial communities. Soils from three spatially isolated tanning facilities in the Punjab province of Pakistan were analyzed. The structure, diversity and abundance of microbial 16S rRNA genes were highly influenced by the concentration and presence of hexavalent chromium (Cr (VI)) and arsenic. When compared to control soils, contaminated soils were dominated by Proteobacteria while Actinobacteria and Acidobacteria (which are generally abundant in pristine soils) were minor components of the bacterial community. Shifts in community composition were significant and revealed that Cr (VI)-containing soils were more similar to each other than to As contaminated soils lacking Cr (VI). Diversity of the arsenic resistance genes, arsB and ACR3 were also determined. Results showed that ACR3 becomes less diverse as arsenic concentrations increase with a single OTU dominating at the highest concentration. Chronic exposure to either Cr or As not only alters the composition of the soil bacterial community in general, but affects the arsenic resistant individuals in different ways

Initial Thrust Measurements of Marshall's Ion-ioN Thruster

Electronegative ion thrusters are a variation of tradition gridded ion thruster technology differentiated by the production and acceleration of both positive and negative ions. Benefits of electronegative ion thrusters include the elimination of lifetime-limiting cathodes from the thruster architecture and the ability to generate appreciable thrust from both charge species. Following the continued development of electronegative ion thruster technology as exhibited by the PEGASES (Plasma Propulsion with Electronegative GASES) thruster, direct thrust measurements are required to push interest in electronegative ion thruster technology forward. For this work, direct thrust measurements of the MINT (Marshall's Ion-ioN Thruster) will be taken on a hanging pendulum thrust stand for propellant mixtures of Sulfur Hexafluoride and Argon at volumetric flow rates of 5-25 sccm at radio frequency power levels of 100-600 watts at a radio frequency of 13.56 MHz. Acceleration grid operation is operated using a square waveform bias of +/-300 volts at a frequency of 25 kHz

Using a Robust and Sensitive GFP-Based cGMP Sensor for Real-Time Imaging in Intact Caenorhabditis elegans.

cGMP plays a role in sensory signaling and plasticity by regulating ion channels, phosphodiesterases, and kinases. Studies that primarily used genetic and biochemical tools suggest that cGMP is spatiotemporally regulated in multiple sensory modalities. FRET- and GFP-based cGMP sensors were developed to visualize cGMP in primary cell culture and Caenorhabditis elegans to corroborate these findings. While a FRET-based sensor has been used in an intact animal to visualize cGMP, the requirement of a multiple emission system limits its ability to be used on its own as well as with other fluorophores. Here, we demonstrate that a C. elegans codon-optimized version of the cpEGFP-based cGMP sensor FlincG3 can be used to visualize rapidly changing cGMP levels in living, behaving C. elegans We coexpressed FlincG3 with the blue-light-activated guanylyl cyclases BeCyclOp and bPGC in body wall muscles, and found that the rate of change in FlincG3 fluorescence correlated with the rate of cGMP production by each cyclase. Furthermore, we show that FlincG3 responds to cultivation temperature, NaCl concentration changes, and sodium dodecyl sulfate in the sensory neurons AFD, ASEL/R, and PHB, respectively. Intriguingly, FlincG3 fluorescence in ASEL and ASER decreased in response to a NaCl concentration upstep and downstep, respectively, which is opposite in sign to the coexpressed calcium sensor jRGECO1a and previously published calcium recordings. These results illustrate that FlincG3 can be used to report rapidly changing cGMP levels in an intact animal, and that the reporter can potentially reveal unexpected spatiotemporal landscapes of cGMP in response to stimuli