Abstract: On seasonal time scales, vapor pressure deficit (VPD) is a known predictor of burned area in the southwestern United States (“the Southwest”). VPD increases with atmospheric warming due to the exponential relationship between temperature and saturation vapor pressure. Another control on VPD is specific humidity, such that increases in specific humidity can counteract temperature-driven increases in VPD. Unexpectedly, despite the increased capacity of a warmer atmosphere to hold water vapor, near-surface specific humidity decreased from 1970 to 2019 in much of the Southwest, particularly in spring, summer, and fall. Here, we identify declining near-surface humidity from 1970 to 2019 in the southwestern United States with both reanalysis and in situ station data. Focusing on the interior Southwest in the months preceding the summer forest fire season, we explain the decline in terms of changes in atmospheric circulation and moisture fluxes between the surface and the atmosphere. We find that an early spring decline in precipitation in the interior region induced a decline in soil moisture and evapotranspiration, drying the lower troposphere in summer. This prior season precipitation decline is in turn related to a trend toward a Northern Hemisphere stationary wave pattern. Finally, using fixed humidity scenarios and the observed exponential relationship between VPD and burned forest area, we estimate that with no increase in temperature at all, the humidity decline alone would still lead to nearly one-quarter of the observed VPD-induced increase in burned area over 1984–2019. Significance Statement: Burned forest area has increased significantly in the southwestern United States in recent decades, driven in part by an increase in atmospheric aridity [vapor pressure deficit (VPD)]. Increases in VPD can be caused by a combination of increasing temperature and decreasing specific humidity. As the atmosphere warms with climate change, its capacity to hold moisture increases. Despite this, there is a decrease in near-surface air humidity in the interior southwestern United States over 1970–2019, which during the summer is likely caused by a decline in early spring precipitation leading to limited soil moisture and evaporation in spring and summer. We estimate that this declining humidity alone, without an increase in temperature, would cause about one-quarter of the VPD-induced increase in burned forest area in this region over 1984–2019.

Abstract John Wesley Powell, in the nineteenth century, introduced the notion that the 100th meridian divides the North American continent into arid western regions and humid eastern regions. This concept remains firmly fixed in the national imagination. It is reexamined in terms of climate, hydrology, vegetation, land use, settlement, and the agricultural economy. It is shown there is a stark east–west gradient in aridity roughly at the 100th meridian that is well expressed in hydroclimate, soil moisture, and “potential vegetation.” The gradient arises from atmospheric circulations and moisture transports. In winter, the arid regions west of the 100th meridian are shielded from Pacific storm-related precipitation and are too far west to benefit from Atlantic storms. In summer, the southerly flow on the western flank of the North Atlantic subtropical high has a westerly component over the western plains, bringing air from the interior southwest, but it also brings air from the Gulf of Mexico over the eastern plains, generating a west–east moisture transport and precipitation gradient. The aridity gradient is realized in soil moisture and a west-to-east transition from shortgrass to tallgrass prairie. The gradient is sharp in terms of greater fractional coverage of developed land east of the 100th meridian than to the west. Farms are fewer but larger west of the meridian, reflective of lower land productivity. Wheat and corn cultivation preferentially occur west and east of the 100th meridian, respectively. The 100th meridian is a very real arid–humid divide in the physical climate and landscape, and this has exerted a powerful influence on human settlement and agricultural development.

Abstract: The cool season (November–March) of 2022–2023 was exceptional in the western United States (US), with the highest precipitation totals in ≥128 years in some areas. Recent precipitation extremes and expectations based on thermodynamics motivate us to evaluate the evidence for an anthropogenic intensification of western US cool‐season precipitation to date. Over cool seasons 1951–2023, trends in precipitation totals on the wettest cool‐season days were neutral or negative across the western US, and significantly negative in northern California and parts of the Pacific Northwest, counter to the expected net intensification effect from anthropogenic forcing. Multiple reanalysis data sets indicate a corresponding lack of increase in moisture transports into the western US, suggesting that atmospheric circulation trends over the North Pacific have counteracted the increases in atmospheric moisture expected from warming alone. The lack of precipitation intensification to date is generally consistent with climate model simulations. A large ensemble of 648 simulations from 35 climate models suggests it is too soon to detect anthropogenic intensification of precipitation across much of the western US. In California, the 35‐model median time of emergence for intensification of the wettest days is 2080 under a mid‐level emissions scenario. On the other hand, observed reductions of precipitation extremes in California and the Pacific Northwest are near the lower edge of the large ensemble of simulated trends, calling into question model representation of western US precipitation variability.

Background

The role of connective tissue growth factor (CTGF/CCN2) in pathological angiogenesis in the retina is unknown.

Results

CTGF/CCN2 stimulates retinal neovascularization through transactivation of p53 target genes such as matrix metalloproteinase (MMP)-2.

Conclusion

CTGF/CCN2 effects on abnormal vessel formation in the retina are mediated by p53 and MMP-2.

Significance

CTGF/CCN2 and its downstream effectors are potential targets in the development of new antiangiogenic treatments. Pathological angiogenesis in the retina is driven by dysregulation of hypoxia-driven stimuli that coordinate physiological vessel growth. How the various components of the neovascularization signaling network are integrated to yield pathological changes has not been defined. Connective tissue growth factor (CTGF/CCN2) is an inducible matricellular protein that plays a major role in fibroproliferative disorders. Here, we show that CTGF/CCN2 was dynamically expressed in the developing murine retinal vasculature and was abnormally increased and localized within neovascular tufts in the mouse eye with oxygen-induced retinopathy. Consistent with its propitious vascular localization, ectopic expression of the CTGF/CCN2 gene further accelerated neovascularization, whereas lentivirus-mediated loss-of-function or -expression of CTGF/CCN2 harnessed ischemia-induced neovessel outgrowth in oxygen-induced retinopathy mice. The neovascular effects of CTGF/CCN2 were mediated, at least in part, through increased expression and activity of matrix metalloproteinase (MMP)-2, which drives vascular remodeling through degradation of matrix and non matrix proteins, migration and invasion of endothelial cells, and formation of new vascular patterns. In cultured cells, CTGF/CCN2 activated the MMP-2 promoter through increased expression and tethering of the p53 transcription factor to a highly conserved p53-binding sequence within the MMP-2 promoter. Concordantly, the neovascular effects of CTGF/CCN2 were suppressed by p53 inhibition that culminated in reduced enrichment of the MMP-2 promoter with p53 and decreased MMP-2 gene expression. Our data identified new gene targets and downstream effectors of CTGF/CCN2 and provided the rational basis for targeting the p53 pathway to curtail the effects of CTGF/CCN2 on neovessel formation associated with ischemic retinopathy.

Background

Human prostate basal cells expressing alpha-6 integrin (CD49f(Hi)) and/or CD44 form prostaspheres in vitro. This functional trait is often correlated with stem/progenitor (S/P) activity, including the ability to self-renew and induce differentiated tubules in vivo. Antigenic profiles that distinguish tubule-initiating prostate stem cells (SCs) from progenitor cells (PCs) and mature luminal cells (LCs) with less regenerative potential are unknown.

Methodology/principle findings

Prostasphere assays and RT-PCR analysis was performed following FACS separation of total benign prostate cells based upon combinations of Epcam, CD44, and/or CD49f expression. Epithelial cell fractions were isolated, including Epcam(+)CD44(+) and Epcam+CD44+CD49f(Hi) basal cells that formed abundant spheres. When non-sphere-forming Epcam(+)CD44(-) cells were fractionated based upon CD49f expression, a distinct subpopulation (Epcam(+)CD44(-)CD49f(Hi)) was identified that possessed a basal profile similar to Epcam(+)CD44(+)CD49f(Hi) sphere-forming cells (p63(+)AR(Lo)PSA(-)). Evaluation of tubule induction capability of fractionated cells was performed, in vivo, via a fully humanized prostate tissue regeneration assay. Non-sphere-forming Epcam(+)CD44(-) cells induced significantly more prostate tubular structures than Epcam(+)CD44(+) sphere-forming cells. Further fractionation based upon CD49f co-expression identified Epcam(+)CD44(-)CD49f(Hi) (non-sphere-forming) basal cells with significantly increased tubule induction activity compared to Epcam(+)CD44(-)CD49f(Lo) (true) luminal cells.

Conclusions/significance

Our data delineates antigenic profiles that functionally distinguish human prostate epithelial subpopulations, including putative SCs that display superior tubule initiation capability and induce differentiated ductal/acini structures, sphere-forming PCs with relatively decreased tubule initiation activity, and terminally differentiated LCs that lack both sphere-forming and tubule-initiation activity. The results clearly demonstrate that sphere-forming ability is not predictive of tubule-initiation activity. The subpopulations identified are of interest because they may play distinct roles as cells of origin in the development of prostatic diseases, including cancer.

We pursued the hypothesis that specific glycans can be used to distinguish breast cancer stem cells (CSCs) and influence their function. Comparison of CSCs and non-CSCs from multiple breast cancer models revealed that CSCs are distinguished by expression of α2,3 sialylated core2 O-linked glycans. We identified a lectin, SLBR-N, which binds to O-linked α2,3 sialic acids, that was able to enrich for CSCs in vitro and in vivo. This O-glycan is expressed on CD44 and promotes its interaction with hyaluronic acid, facilitating CD44 signaling and CSC properties. In contrast, FUT3, which contributes to sialyl Lewis X (sLeX) production, is preferentially expressed in the non-CSC population, and it antagonizes CSC function. Collectively, our data indicate that SLBR-N can be more efficient at enriching for CSCs than CD44 itself because its use avoids the issues of CD44 splicing and glycan status. These data also reveal how differential glycosylation influences CSC fate.

Background

Human fetal prostate buds appear in the 10th gestational week as solid cords, which branch and form lumens in response to androgen 1. Previous in vivo analysis of prostate epithelia isolated from benign prostatectomy specimens indicated that Epcam⁺ CD44⁻ CD49f(Hi) basal cells possess efficient tubule initiation capability relative to other subpopulations 2. Stromal interactions and branching morphogenesis displayed by adult tubule-initiating cells (TIC) are reminiscent of fetal prostate development. In the current study, we evaluated in vivo tubule initiation by human fetal prostate cells and determined expression profiles of fetal and adult epithelial subpopulations in an effort to identify pathways used by TIC.

Methods

Immunostaining and FACS analysis based on Epcam, CD44, and CD49f expression demonstrated the majority (99.9%) of fetal prostate epithelial cells (FC) were Epcam⁺ CD44⁻ with variable levels of CD49f expression. Fetal populations isolated via cell sorting were implanted into immunocompromised mice. Total RNA isolation from Epcam⁺ CD44⁻ CD49f(Hi) FC, adult Epcam⁺ CD44⁻ CD49f(Hi) TIC, Epcam⁺ CD44⁺ CD49f(Hi) basal cells (BC), and Epcam⁺ CD44⁻ CD49f(Lo) luminal cells (LC) was performed, followed by microarray analysis of 19 samples using the Affymetrix Gene Chip Human U133 Plus 2.0 Array. Data was analyzed using Partek Genomics Suite Version 6.4. Genes selected showed >2-fold difference in expression and P < 5.00E-2. Results were validated with RT-PCR.

Results

Grafts retrieved from Epcam⁺ CD44⁻ fetal cell implants displayed tubule formation with differentiation into basal and luminal compartments, while only stromal outgrowths were recovered from Epcam- fetal cell implants. Hierarchical clustering revealed four distinct groups determined by antigenic profile (TIC, BC, LC) and developmental stage (FC). TIC and BC displayed basal gene expression profiles, while LC expressed secretory genes. FC had a unique profile with the most similarities to adult TIC. Functional, network, and canonical pathway identification using Ingenuity Pathway Analysis Version 7.6 compiled genes with the highest differential expression (TIC relative to BC or LC). Many of these genes were found to be significantly associated with prostate tumorigenesis.

Conclusions

Our results demonstrate clustering gene expression profiles of FC and adult TIC. Pathways associated with TIC are known to be deregulated in cancer, suggesting a cell-of-origin role for TIC versus re-emergence of pathways common to these cells in tumorigenesis.