A pan‐Arctic synthesis of CH4 and CO2 production from anoxic soil incubations

Permafrost thaw can alter the soil environment through changes in soil moisture, frequently resulting in soil saturation, a shift to anaerobic decomposition, and changes in the plant community. These changes, along with thawing of previously frozen organic material, can alter the form and magnitude of greenhouse gas production from permafrost ecosystems. We synthesized existing methane (CH₄) and carbon dioxide (CO₂) production measurements from anaerobic incubations of boreal and tundra soils from the geographic permafrost region to evaluate large‐scale controls of anaerobic CO₂ and CH₄ production and compare the relative importance of landscape‐level factors (e.g., vegetation type and landscape position), soil properties (e.g., pH, depth, and soil type), and soil environmental conditions (e.g., temperature and relative water table position). We found fivefold higher maximum CH₄ production per gram soil carbon from organic soils than mineral soils. Maximum CH₄ production from soils in the active layer (ground that thaws and refreezes annually) was nearly four times that of permafrost per gram soil carbon, and CH₄ production per gram soil carbon was two times greater from sites without permafrost than sites with permafrost. Maximum CH₄ and median anaerobic CO₂ production decreased with depth, while CO₂:CH₄ production increased with depth. Maximum CH₄ production was highest in soils with herbaceous vegetation and soils that were either consistently or periodically inundated. This synthesis identifies the need to consider biome, landscape position, and vascular/moss vegetation types when modeling CH₄ production in permafrost ecosystems and suggests the need for longer‐term anaerobic incubations to fully capture CH₄ dynamics. Our results demonstrate that as climate warms in arctic and boreal regions, rates of anaerobic CO₂ and CH₄ production will increase, not only as a result of increased temperature, but also from shifts in vegetation and increased ground saturation that will accompany permafrost thaw.     

Investigation of Radical and Cationic Cross‐Linking in High‐Efficiency,Low Band Gap Solar Cell Polymers

Dithienogermole‐co‐thieno[3,4‐c]pyrroledione (DTG‐TPD) polymers incorporating chemically cross‐linkable sidechains are reported and their properties compared to a parent polymer with simple octyl sidechains. Two cross‐linking groups and mechanisms are investigated, UV‐promoted radical cross‐linking of an alkyl bromide cross‐linker and acid‐promoted cationic cross‐linking of an oxetane cross‐linker. It is found that random copolymers with a 20% incorporation of the cross‐linker demonstrate a higher performance in bulk heterojunction solar cells than the parent polymer, while 100% cross‐linker incorporation results in deterioration in device efficiency. The use of 1,8‐diiodooctane (DIO) as a processing additive improves as‐cast solar cell performance, but is found to have a significant deleterious impact on solar cell efficiency after UV exposure. The instability to UV can be overcome by the use of an alternative additive, 1‐chloronapthalene, which also promotes high device efficiency. Cross‐linking of the polymer is investigated in the presence and absence of fullerene highlighting significant differences in behavior. Intractable films cannot be obtained by radical cross‐linking in the presence of fullerene, whereas cationic cross‐linking is successful.     

Maximum diastolic potential of human induced pluripotent stem cell-derived cardiomyocytes depends critically on I(Kr)

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) hold promise for therapeutic applications. To serve these functions, the hiPSC-CM must recapitulate the electrophysiologic properties of native adult cardiomyocytes. This study examines the electrophysiologic characteristics of hiPSC-CM between 11 and 121 days of maturity. Embryoid bodies (EBs) were generated from hiPS cell line reprogrammed with Oct4, Nanog, Lin28 and Sox2. Sharp microelectrodes were used to record action potentials (AP) from spontaneously beating clusters (BC) micro-dissected from the EBs (n = 103; 37°C) and to examine the response to 5 µM E-4031 (n = 21) or BaCl₂ (n = 22). Patch-clamp techniques were used to record I_Kr and I_K1 from cells enzymatically dissociated from BC (n = 49; 36°C). Spontaneous cycle length (CL) and AP characteristics varied widely among the 103 preparations. E-4031 (5 µM; n = 21) increased Bazett-corrected AP duration from 291.8±81.2 to 426.4±120.2 msec (p<0.001) and generated early afterdepolarizations in 8/21 preparations. In 13/21 BC, E-4031 rapidly depolarized the clusters leading to inexcitability. BaCl₂, at concentrations that selectively block I_K1 (50–100 µM), failed to depolarize the majority of clusters (13/22). Patch-clamp experiments revealed very low or negligible I_K1 in 53% (20/38) of the cells studied, but presence of I_Kr in all (11/11). Consistent with the electrophysiological data, RT-PCR and immunohistochemistry studies showed relatively poor mRNA and protein expression of I_K1 in the majority of cells, but robust expression of I_Kr. In contrast to recently reported studies, our data point to major deficiencies of hiPSC-CM, with remarkable diversity of electrophysiologic phenotypes as well as pharmacologic responsiveness among beating clusters and cells up to 121 days post-differentiation (dpd). The vast majority have a maximum diastolic potential that depends critically on I_Kr due to the absence of I_K1. Thus, efforts should be directed at producing more specialized and mature hiPSC-CM for future therapeutic applications.     

Comparison of photomixotrophic and heterotrophic callus and suspension cultures of Pinus elliottii. 1. Photosynthetic properties and ultrastructural evidence for coexistence of starch granules and secondary metabolites

Photomixotrophic callus and suspension cultures of salsh pine (Pinus elliottii var. elliottii Engelm.) have been established. Callus tissues contained up to 2.76 g chlorophyll mg^-1 dry wt and suspensions 2.98 g chlorophyll mg^-1 dry wt. Maximum photosynthetic oxygen evolution was 25–32 mol O₂ h^-1 mg^-1 chlorophyll for callus and 35–39 mol O₂h^-1 mg^-1 chlorophyll for suspension, respectively. Photomixotrophic callus was friable with a high moisture content during early and exponential growth, but evolved into a compact and dense tissue during the latter stage of growth. Compact photomixotrophic callus accumulated and deposited secondary metabolites in the central vacuole and developed large starch granules in the chloroplasts. Secondary metabolites were not observed in photomixotrophic suspensions or in heterotrophic calli and suspensions. Photomixotrophic callus contained numerous mitochondria closely associated with well-developed chloroplasts containing 2–6 thylakoids per granum. Heterotrophic callus was characterized by a poorly developed cytoplasm and cup-shaped mitochondria.     

Effect of polyethylene glycol-electrolyte lavage solution on intestinal microflora.

The bacterial flora of colonic aspirates taken after oral administration of a polyethylene glycol-electrolyte lavage solution to patients undergoing colonoscopy was analyzed by standard bacteriological methods and DNA hybridization techniques. The data were compared with those for normal feces collected from the same patients the day before treatment. The results indicate that the bacterial composition of feces differs from individual to individual but is not altered by the oral intake of a polyethylene glycol-electrolyte lavage solution. This pilot study demonstrates that routine colonoscopy aspirates taken after standard colonoscopy preparation accurately reflect the microflora of feces and therefore may be used in large-scale epidemiological studies.     

Developmental and wound-, cold-, desiccation-, ultraviolet-B-stress-induced modulations in the expression of the petunia zinc finger transcription factor gene ZPT2-2

The ZPT2-2 gene belongs to the EPF gene family in petunia (Petunia hybrida), which encodes proteins with TFIIIA-type zinc-finger DNA-binding motifs. To elucidate a possible function for ZPT2-2, we analyzed its pattern of expression in relation to different developmental and physiological stress signals. The activity of the ZPT2-2 promoter was analyzed using a firefly luciferase (LUC) reporter gene, allowing for continuous measurements of transgene activity in planta. We show that ZPT2-2::LUC is active in all plant tissues, but is strongly modulated in cotyledons upon germination, in leaves in response to desiccation, cold treatment, wounding, or ultraviolet-B light, and in petal tissue in response to pollination of the stigma. Analysis of mRNA levels indicated that the modulations in ZPT2-2::LUC expression reflect modulations in endogenous ZPT2-2 gene expression. The change in ZPT2-2::LUC activity by cold treatment, wounding, desiccation, and ultraviolet-B light suggest that the phytohormones ethylene and jasmonic acid are involved in regulating the expression of ZPT2-2. Although up-regulation of expression of ZPT2-2 can be blocked by inhibitors of ethylene perception, expression in plants is not induced by exogenously applied ethylene. The application of jasmonic acid does result in an up-regulation of gene activity and, thus, ZPT2-2 may play a role in the realization of the jasmonic acid hormonal responses in petunia.     

High-resolution comprehensive radiation hybrid maps of the porcine chromosomes 2p and 9p compared with the human chromosome 11

We are constructing high-resolution, chromosomal 'test' maps for the entire pig genome using a 12,000-rad WG-RH panel (IMNpRH2(12,000-rad))to provide a scaffold for the rapid assembly of the porcine genome sequence. Here we present an initial, comparative map of human chromosome (HSA) 11 with pig chromosomes (SSC) 2p and 9p. Two sets of RH mapping vectors were used to construct the RH framework (FW) maps for SSC2p and SSC9p. One set of 590 markers, including 131 microsatellites (MSs), 364 genes/ESTs, and 95 BAC end sequences (BESs) was typed on the IMNpRH2(12,000-rad) panel. A second set of 271 markers (28 MSs, 138 genes/ESTs, and 105 BESs) was typed on the IMpRH(7,000-rad) panel. The two data sets were merged into a single data-set of 655 markers of which 206 markers were typed on both panels. Two large linkage groups of 72 and 194 markers were assigned to SSC2p, and two linkage groups of 84 and 168 markers to SSC9p at a two-point LOD score of 10. A total of 126 and 114 FW markers were ordered with a likelihood ratio of 1000:1 to the SSC2p and SSC9p RH(12,000-rad) FW maps, respectively, with an accumulated map distance of 4046.5 cR(12,000 )and 1355.2 cR(7,000 )for SSC2p, and 4244.1 cR(12,000) and 1802.5 cR(7,000) for SSC9p. The kb/cR ratio in the IMNpRH2(12,000-rad) FW maps was 15.8 for SSC2p, and 15.4 for SSC9p, while the ratio in the IMpRH(7,000-rad) FW maps was 47.1 and 36.3, respectively, or an approximately 3.0-fold increase in map resolution in the IMNpRH(12,000-rad) panel over the IMpRH(7,000-rad) panel. The integrated IMNpRH(12,000-rad) andIMpRH(7,000-rad) maps as well as the genetic and BAC FPC maps provide an inclusive comparative map between SSC2p, SSC9p and HSA11 to close potential gaps between contigs prior to sequencing, and to identify regions where potential problems may arise in sequence assembly.