ALBERT

Description: We applied multiple geochemical tracers ( 87 Sr/ 86 Sr, [Sr], 13 C, and 18 O) to waters and carbonates of the lower Colorado River system to evaluate its paleohydrology over the past 12 Ma. Modern springs in Grand Canyon reflect mixing of deeply derived (endogenic) fluids with meteoric (epigenic) recharge. Travertine (〈1 Ma) and speleothems (2–4 Ma) yield 87 Sr/ 86 Sr and 13 C and 18 O values that overlap with associated water values, providing justification for use of carbonates as a proxy for the waters from which they were deposited. The Hualapai Limestone (12–6 Ma) and Bouse Formation (5.6–4.8 Ma) record paleohydrology immediately prior to and during integration of the Colorado River. The Hualapai Limestone was deposited from 12 Ma (new ash age) to 6 Ma; carbonates thicken eastward to ~210 m toward the Grand Wash fault, suggesting that deposition was synchronous with fault slip. A fanning-dip geometry is suggested by correlation of ashes between subbasins using tephrochronology. New detrital-zircon ages are consistent with the "Muddy Creek constraint," which posits that Grand Wash Trough was internally drained prior to 6 Ma, with limited or no Colorado Plateau detritus, and that Grand Wash basin was sedimentologically distinct from Gregg and Temple basins until after 6 Ma. New isotopic data from Hualapai Limestone of Grand Wash basin show values and ranges of 87 Sr/ 86 Sr, 13 C, and 18 O that are similar to Grand Canyon springs and travertines, suggesting a long-lived spring-fed lake/marsh system sourced from western Colorado Plateau groundwater. Progressive up-section decrease in 87 Sr/ 86 Sr and 13 C and increase in 18 O in the uppermost 50 m of the Hualapai Limestone indicate an increase in meteoric water relative to endogenic inputs, which we interpret to record progressively increased input of high-elevation Colorado Plateau groundwater from ca. 8 to 6 Ma. Grand Wash, Hualapai, Gregg, and Temple basins, although potentially connected by groundwater, were hydrochemically distinct basins before ca. 6 Ma. The 87 Sr/ 86 Sr, 13 C, and 18 O chemostratigraphic trends are compatible with a model for downward integration of Hualapai basins by groundwater sapping and lake spillover. The Bouse Limestone (5.6–4.8 Ma) was also deposited in several hydrochemically distinct basins separated by bedrock divides. Northern Bouse basins (Cottonwood, Mojave, Havasu) have carbonate chemistry that is nonmarine. The 87 Sr/ 86 Sr data suggest that water in these basins was derived from mixing of high- 87 Sr/ 86 Sr Lake Hualapai waters with lower- 87 Sr/ 86 Sr, first-arriving "Colorado River" waters. Covariation trends of 13 C and 18 O suggest that newly integrated Grand Wash, Gregg, and Temple basin waters were integrated downward to the Cottonwood and Mojave basins at ca. 5–6 Ma. Southern, potentially younger Bouse basins are distinct hydrochemically from each other, which suggests incomplete mixing during continued downward integration of internally drained basins. Bouse carbonates display a southward trend toward less radiogenic 87 Sr/ 86 Sr values, higher [Sr], and heavier 18 O that we attribute to an increased proportion of Colorado River water through time plus increased evaporation from north to south. The 13 C and 18 O trends suggest alternating closed and open systems in progressively lower (southern) basins. We interpret existing data to permit the interpretation that the southernmost Blythe basin may have had intermittent mixing with marine water based on 13 C and 18 O covariation trends, sedimentology, and paleontology. [Sr] versus 87 Sr/ 86 Sr modeling suggests that southern Blythe basin 87 Sr/ 86 Sr values of ~0.710–0.711 could be produced by 25%–75% seawater mixed with river water (depending on [Sr] assumptions) in a delta–marine estuary system. We suggest several refinements to the "lake fill-and-spill" downward integration model for the Colorado River: (1) Lake Hualapai was fed by western Colorado Plateau groundwater from 12 to 8 Ma; (2) high-elevation Colorado Plateau groundwater was progressively introduced to Lake Hualapai from ca. 8 to 6 Ma; (3) Colorado River water arrived at ca. 5–6 Ma; and (4) the combined inputs led to downward integration by a combination of groundwater sapping and sequential lake spillover that first delivered Colorado Plateau water and detritus to the Salton Trough at ca. 5.3 Ma. We propose that the groundwater sapping mechanism strongly influenced lake evolution of the Hualapai and Bouse Limestones and that groundwater flow from the Colorado Plateau to Grand Wash Trough led to Colorado River integration.

Description: Cleft lip and/or palate (CL/P) are common structural birth defects in humans. We used exome sequencing to study a patient with bilateral CL/P and identified a single nucleotide deletion in the patient and her similarly affected son—c.546_546delG, predicting p.Gln183Arg fs *57 in the Distal-less 4 ( DLX4 ) gene. The sequence variant was absent from databases, predicted to be deleterious and was verified by Sanger sequencing. In mammals, there are three Dlx homeobox clusters with closely located gene pairs ( Dlx1 / Dlx2 , Dlx3 / Dlx4 , Dlx5 / Dlx6 ). In situ hybridization showed that Dlx4 was expressed in the mesenchyme of the murine palatal shelves at E12.5, prior to palate closure. Wild-type human DLX4 , but not mutant DLX4 _c.546delG, could activate two murine Dlx conserved regulatory elements, implying that the mutation caused haploinsufficiency. We showed that reduced DLX4 expression after short interfering RNA treatment in a human cell line resulted in significant up-regulation of DLX3 , DLX5 and DLX6 , with reduced expression of DLX2 and significant up-regulation of BMP4 , although the increased BMP4 expression was demonstrated only in HeLa cells. We used antisense morpholino oligonucleotides to target the orthologous Danio rerio gene, dlx4b , and found reduced cranial size and abnormal cartilaginous elements. We sequenced DLX4 in 155 patients with non-syndromic CL/P and CP, but observed no sequence variants. From the published literature, Dlx1 / Dlx2 double homozygous null mice and Dlx5 homozygous null mice both have clefts of the secondary palate. This first finding of a DLX4 mutation in a family with CL/P establishes DLX4 as a potential cause of human clefts.

Description: The impact of cigarette smoking can persist for extended periods following smoking cessation and may involve epigenetic reprogramming. Changes in DNA methylation associated with smoking may help to identify molecular pathways that contribute to the latency between exposure and disease onset. Cross-sectional cohort data from subjects in the International COPD Genetics Network ( n = 1085) and the Boston Early-Onset COPD study ( n = 369) were analyzed as the discovery and replication cohorts, respectively. Genome-wide methylation data on 27 578 CpG sites in 14 475 genes were obtained on DNA from peripheral blood leukocytes using the Illumina HumanMethylation27K Beadchip in both cohorts. We identified 15 sites significantly associated with current smoking, 2 sites associated with cumulative smoke exposure, and, within the subset of former smokers, 3 sites associated with time since quitting cigarettes. Two loci, factor II receptor-like 3 ( F2RL3 ) and G-protein-coupled receptor 15 ( GPR15 ), were significantly associated in all three analyses and were validated by pyrosequencing. These findings (i) identify a novel locus ( GPR15 ) associated with cigarette smoking and (ii) suggest the existence of dynamic, site-specific methylation changes in response to smoking which may contribute to the extended risks associated with cigarette smoking that persist after cessation.

Description: Spatio-temporal variability in fluvial incision rates in bedrock channels provides data regarding uplift and denudation histories of landscapes. The longitudinal profile of the Gunnison River (Colorado), tributary to the Colorado River, contains a prominent knickzone with 800 m of relief across it within the Black Canyon of the Gunnison. Average bedrock incision rates over the last 0.64 Ma surrounding the knickpoint vary from 150 m/Ma (downstream) to 400–550 m/Ma (within) to 90–95 m/Ma (upstream), suggesting it is a transient feature. Lava Creek B ash constrains strath terraces along a paleoprofile of the river. An isochron cosmogenic burial date in the paleo–Bostwick River of 870 ± 220 ka is consistent with the presence of 0.64 Ma Lava Creek B ash in locally derived, stratigraphically younger sediment. With 350 m of incision since deposition, we determine an incision rate of 400–550 m/Ma, reflecting incision through resistant basement rock at 2–3 times regional incision rates. Such contrast is attributed to a wave of transient incision, potentially initiated by downstream base-level fall during abandonment of Unaweep Canyon at ca. 1 Ma. Rate extrapolation indicates that the ~700 m depth of Black Canyon has been eroded since 1.3–1.75 Ma. The Black Canyon knickpoint overlies a strong gradient between low-velocity mantle under the Colorado Rockies and higher-velocity mantle of the Colorado Plateau. We interpret recent reorganization and transient incision of both the Gunnison River and upper Colorado River systems to be a response to mantle-driven epeirogenic uplift of the southern Rockies in the last 10 Ma.

Description: The lower Pliocene Bouse Formation in the lower Colorado River Valley (southwestern USA) consists of basal marl and dense tufa overlain by siltstone and fine sandstone. It is locally overlain by and interbedded with sands derived from the Colorado River. We briefly review 87 Sr/ 86 Sr analyses of Bouse carbonates and shells and carbonate and gypsum of similar age east of Las Vegas that indicate that all of these strata are isotopically similar to modern Colorado River water. We also review and add new data that are consistent with a step in Bouse Formation maximum elevations from 330 m south of Topock Gorge to 555 m to the north. New geochemical data from glass shards in a volcanic ash bed within the Bouse Formation, and from an ash bed within similar deposits in Bristol Basin west of the Colorado River Valley, indicate correlation of the two ash beds and coeval submergence of both areas. The tuff bed is identified as the 4.83 Ma Lawlor Tuff derived from the San Francisco Bay region. We conclude, as have some others, that the Bouse Formation was deposited in lakes produced by first-arriving Colorado River water that entered closed basins inherited from Basin and Range extension, and estimate that first arrival of river water occurred ca. 4.9 Ma. If this interpretation is correct, addition of Bristol Basin to the Blythe Basin inundation area means that river discharge was sufficient to fill and spill a lake with an area of ~10,000 km 2 . For spillover to occur, evaporation rates must have been significantly less in early Pliocene time than modern rates of ~2–4 m/yr, and/or Colorado River discharge was significantly greater than the current ~15 km 3 /yr. In this lacustrine interpretation, evaporation rates were sufficient to concentrate salts to levels that were hospitable to some marine organisms presumably introduced by birds.