Spatial variability in tree-ring carbon isotope discrimination in response to local drought across the entire loblolly pine natural range.

Considering the temporal responses of carbon isotope discrimination (Δ13C) to local water availability in the spatial analysis of Δ13C is essential for evaluating the contribution of environmental and genetic facets of plant Δ13C. Using tree-ring Δ13C from years with contrasting water availability at 76 locations across the natural range of loblolly pine, we decomposed site-level Δ13C signals to maximum Δ13C in well-watered conditions (Δ13Cmax) and isotopic drought sensitivity (m) as a change in Δ13C per unit change of Palmer's Drought Severity Index (PDSI). Site water status, especially the tree lifetime average PDSI, was the primary factor affecting Δ13Cmax. The strong spatial correlation exhibited by m was related to both genetic and environmental factors. The long-term average water availability during the period relevant to trees as indicated by lifetime average PDSI correlated with Δ13Cmax, suggesting acclimation in tree gas-exchange traits, independent of incident water availability. The positive correlation between lifetime average PDSI and m indicated that loblolly pines were more sensitive to drought at mesic than xeric sites. The m was found to relate to a plant's stomatal control and may be employed as a genetic indicator of efficient water use strategies. Partitioning Δ13C to Δ13Cmax and m provided a new angle for understanding sources of variation in plant Δ13C, with several fundamental and applied implications.

Ecosystem carbon density and allocation across a chronosequence of longleaf pine forests.

Forests can partially offset greenhouse gas emissions and contribute to climate change mitigation, mainly through increases in live biomass. We quantified carbon (C) density in 20 managed longleaf pine (Pinus palustris Mill.) forests ranging in age from 5 to 118 years located across the southeastern United States and estimated above- and belowground C trajectories. Ecosystem C stock (all pools including soil C) and aboveground live tree C increased nonlinearly with stand age and the modeled asymptotic maxima were 168 Mg C/ha and 80 Mg C/ha, respectively. Accumulation of ecosystem C with stand age was driven mainly by increases in aboveground live tree C, which ranged from <1 Mg C/ha to 74 Mg C/ha and comprised <1% to 39% of ecosystem C. Live root C (sum of below-stump C, ground penetrating radar measurement of lateral root C, and live fine root C) increased with stand age and represented 4-22% of ecosystem C. Soil C was related to site index, but not to stand age, and made up 39-92% of ecosystem C. Live understory C, forest floor C, downed dead wood C, and standing dead wood C were small fractions of ecosystem C in these frequently burned stands. Stand age and site index accounted for 76% of the variation in ecosystem C among stands. The mean root-to-shoot ratio calculated as the average across all stands (excluding the grass-stage stand) was 0.54 (standard deviation of 0.19) and higher than reports for other conifers. Long-term accumulation of live tree C, combined with the larger role of belowground accumulation of lateral root C than in other forest types, indicates a role of longleaf pine forests in providing disturbance-resistant C storage that can balance the more rapid C accumulation and C removal associated with more intensively managed forests. Although other managed southern pine systems sequester more C over the short-term, we suggest that longleaf pine forests can play a meaningful role in regional forest C management.

Circulating fibrocytes stabilize blood vessels during angiogenesis in a paracrine manner.

Accumulating evidence supports that circulating fibrocytes play important roles in angiogenesis. However, the specific role of fibrocytes in angiogenesis and the underlying mechanisms remain unclear. In this study, we found that fibrocytes stabilized newly formed blood vessels in a mouse wound-healing model by inhibiting angiogenesis during the proliferative phase and inhibiting blood vessel regression during the remodeling phase. Fibrocytes also inhibited angiogenesis in a Matrigel mouse model. In vitro study showed that fibrocytes inhibited both the apoptosis and proliferation of vascular endothelial cells (VECs) in a permeable support (Transwell) co-culture system. In a three-dimensional collagen gel, fibrocytes stabilized the VEC tubes by decreasing VEC tube density on stimulation with growth factors and preventing VEC tube regression on withdrawal of growth factors. Further mechanistic investigation revealed that fibrocytes expressed many prosurvival factors that are responsible for the prosurvival effect of fibrocytes on VECs and blood vessels. Fibrocytes also expressed angiogenesis inhibitors, including thrombospondin-1 (THBS1). THBS1 knockdown partially blocked the fibrocyte-induced inhibition of VEC proliferation in the Transwell co-culture system and recovered the fibrocyte-induced decrease of VEC tube density in collagen gel. Purified fibrocytes transfected with THBS1 siRNA partially recovered the fibrocyte-induced inhibition of angiogenesis in both the wound-healing and Matrigel models. In conclusion, our findings reveal that fibrocytes stabilize blood vessels via prosurvival factors and anti-angiogenic factors, including THBS1.

Improved function and growth of pancreatic cells in a three-dimensional bioreactor environment.

Methods of three-dimensional (3D) cell culture have made significant progress in recent years due to a better understanding of cell to cell interactions and the cell's interface with their surrounding environment. We hypothesized that a microgravity 3D culture system would improve upon the growth and function of a pancreatic progenitor cell population. We developed a rotating wall vessel bioreactor and established a culture system using a pancreatic cell line. Cells in the bioreactors showed robust proliferation, enhanced transcriptional signaling, and improved translation of pancreatic genes compared with two-dimensional static culture. Cells also gained the ability to respond to glucose stimulation, which was not observed in the control cultures. These findings suggest that a 3D microgravity bioreactor environment mimics the niche of the pancreas yielding a cell source with potential for cell-based therapy in the treatment of diabetes.

MAPK/ERK and Wnt/ß-Catenin pathways are synergistically involved in proliferation of Sca-1 positive hepatic progenitor cells.

Hepatic progenitor cells (HPCs) persist in adulthood and have the potential to play a major role in regenerating diseased liver. However, the signaling pathways that both directly and indirectly regulate HPCs' self-renewal and differentiation remain elusive. Previously, we identified a bipotent, stem cell antigen-1 (Sca-1) positive HPC population from naïve adult liver tissue. In the present study, we aimed to investigate the involvement of various signaling pathways in Sca-1(+) HPC proliferation. Epidermal growth factor (EGF) supplementation shows a significant increase in Sca-1(+) HPC proliferation and colony formation while stimulating phosphorylation of ERK1/2 and activating the induction of Cyclin D1. There were no demonstrable effects of EGF on Akt. The MEK inhibitor, PD0325901, inhibits proliferation and ERK1/2 phosphorylation while also suppressing the expression of Cyclin D1. In addition, activation of either IL-6/STAT3 or Wnt/ß-Catenin pathway did not independently support cell proliferation and colony formation of HPCs. The Wnt/ß-Catenin pathway can cooperate with EGF to significantly promote HPC colony formation ratio and maintain long-term HPC in vitro. The data indicates that the MAPK/ERK pathway is both essential and critical for HPC proliferation, and the Wnt signaling pathway is not sufficient, while it works synergistically with the MAPK/ERK signaling pathway to promote HPC proliferation.

Enrichment of a bipotent hepatic progenitor cell from naïve adult liver tissue.

BACKGROUND/AIM: Recent interest in the liver stem cell field has led to the identification and characterization of several hepatic progenitor cell populations from fetal and adult tissues. We isolated a hepatic progenitor cell from naïve adult liver and the current studies focus on differentiation and growth. RESULTS: A Sca-1(+) hepatic progenitor cell was identified within the liver parenchyma. This cell expresses numerous liver related genes and transcription found in the developing and/or adult liver. It is located in the peri-portal region and expresses markers associated with undifferentiated hepatic cell populations, mature hepatocytes and biliary cells which distinguish it from the Sca-1(-) fraction. CONCLUSION: This hepatic progenitor cell from uninjured liver has features of both hepatocytic and biliary populations and demonstrates proliferative potential. Further studies will focus on sca-HPC subsets and conditions that regulate differentiation towards hepatic or biliary lineages.

Influence of irrigation and fertilization on transpiration and hydraulic properties of Populus deltoides.

Long-term hydraulic acclimation to resource availability was explored in 3-year-old Populus deltoides Bartr. ex Marsh. clones by examining transpiration, leaf-specific hydraulic conductance (G(L)), canopy stomatal conductance (G(S)) and leaf to sapwood area ratio (A(L):A(S)) in response to irrigation (13 and 551 mm year(-1) in addition to ambient precipitation) and fertilization (0 and 120 kg N ha(-1) year(-1)). Sap flow was measured continuously over one growing season with thermal dissipation probes. Fertilization had a greater effect on growth and hydraulic properties than irrigation, and fertilization effects were independent of irrigation treatment. Transpiration on a ground area basis (E) ranged between 0.3 and 1.8 mm day(-1), and increased 66% and 90% in response to irrigation and fertilization, respectively. Increases in G(L), G(S) at a reference vapor pressure deficit of 1 kPa, and transpiration per unit leaf area in response to increases in resource availability were associated with reductions in A(L):A(S) and consequently a minimal change in the water potential gradient from soil to leaf. Irrigation and fertilization increased leaf area index similarly, from an average 1.16 in control stands to 1.45, but sapwood area was increased from 4.0 to 6.3 m(2) ha(-1) by irrigation and from 3.7 to 6.7 m(2) ha(-1) by fertilization. The balance between leaf area and sapwood area was important in understanding long-term hydraulic acclimation to resource availability and mechanisms controlling maximum productivity in Populus deltoides.

Scaling ozone effects from seedlings to forest trees.

Biospheric ozone has become a widely distributed air pollutant, and a growing body of research indicates that ozone impacts forest health and productivity. Ozone effects are mediated by the ozone concentration present in the external environment and the movement of ozone into the leaf via the stoma. The cumulative dose received by the plant is, in the simplest terms, a function of ambient ozone concentration and stomatal conductance to water vapor. This relationship is important in understanding ozone flux into the leaf and subsequent ozone response in plants. Here, current progress in understanding ozone uptake in juvenile and mature trees is examined. Through an analysis of two long-term case studies, the significant uncertainty in assessing ozone effects on forests is pinpointed to be the scaling of ozone sensitivity from controlled seedling studies to large forest trees. A rigorous statistical and monitoring approach, which includes ozone uptake as a cause variable, may provide the missing information on processes that are known to be important to risk assessment of ozone impacts on forest trees. Contents Summary 21 I. Introduction 22 1. Background 22 2. Characterization of ozone exposure 22 3. The need for scaling 23 II. Scaling from seedling to tree, evidence from a Quercus rubra case study 24 1. Study background 24 2. Facilities and measurements 24 3. Ozone exposure dynamics 24 4. Above-ground processes 25 5. Below-ground processes 26 6. A process modelling exercise 27 7. Conclusions 28 III. Scaling from chamber to forest, evidence from a field case study 29 1. Study background 29 2. Field sites and measurements 30 3. Ozone exposure dynamics 30 4. Stomatal conductance and ozone uptake in forest trees 30 5. Conclusions 32 IV. Evidence from a scaling exercise 33 V. Concluding remarks 36 Acknowledgements 37 References 37.