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The estuary of the Elwha River, on Washington’s Olympic Peninsula, has been degraded and simplified over the past century from sediment retention behind two large dams, levee construction, and channelization. With the removal of Elwha Dam and initiation of Glines Canyon Dam’s removal in fall 2011, sediment deposits will change the estuary and affect anadromous and nearshore marine fishes. Juvenile Chinook salmon commonly use estuaries and the river’s population is part of an Evolutionarily Significant Unit listed as Threatened under the U.S. Endangered Species Act. This study reports on monthly sampling in part of the river’s estuary from March 2007 through September 2011 to characterize the seasonal changes in relative abundance of yearlings and sub-yearlings, and size distributions prior to dam removal. Most (69 %) of the yearlings were caught in April, when this life history type was released from the hatchery, and to a lesser extent in May (28 %) and June (3 %). Yearlings caught in the estuary were smaller than those released from the hatchery (means: 153 mm ± 28 SD vs. 175 mm ± 5 SD), suggesting more rapid departure by larger fish. Sub-yearlings were much more abundant in the estuary, and were caught from March through November, increasing in mean fork length by 8.7 mm month-1. The hatchery-origin sub-yearlings were not marked externally and so were not distinguishable from natural origin fish. However, 39 % of the sub-yearlings were caught prior to June, when sub-yearlings were released from the hatchery, indicating substantial use of the estuary by natural-origin fish. Thus, even in a reduced state after a century of dam operation, the highly modified estuary was used over many months by juvenile Chinook salmon. The information on juvenile Chinook salmon prior to dam removal provides a basis for comparison to patterns in the future, when the anticipated increase in estuarine complexity may further enhance habitat use by juvenile Chinook salmon.

In addition to the downstream migration of smolts in spring, Coho Salmon Oncorhynchus kisutch also enter estuaries throughout the year but especially in the spring as fry and in the fall as parr. The removal of two large dams on the Elwha River, Washington, has increased the area accessible to salmon and is affecting many aspects of the system. For comparison with the postdam period, when the Elwha River estuary will likely expand in size and complexity, monthly sampling was conducted in the estuary during 2007–2011 to determine patterns of Coho Salmon presence and size prior to dam removal; Salt Creek, a nearby undammed stream, was also sampled to allow comparison of fish size and seasonal timing patterns. The spring smolt migration in the Elwha River included a large fraction of unmarked fish (primarily of natural origin) as well as marked fish from the Lower Elwha Klallam Tribe Fish Hatchery. Subyearlings entered both estuaries during much of the year, exhibiting a peak in September. Coho Salm...

The demand for trees for industrial application is growing steadily and not expected to plateau for at least two decades; consequently, the supply of wood must increase to meet this need. One method for increasing yield without compromising land requirements is to modify wood density. The current study uses a transcriptomic approach to identify the genes associated with wood density that are likely to be of value in Sitka spruce (Picea sitchensis (Bong.) Carr.) breeding programmes. Following extensive wood density analysis from a Sitka field grown clonal trial, three detailed microarray studies were conducted to compare the transcriptome of cambium and xylem tissue from contrasting density clonal lines. Twenty-five genes exhibited differential expression, with up to 50-fold differences in expression observed, in at least two of the three microarray experiments, and this was verified using real-time polymerase chain reaction. Identified genes included those involved in cell wall synthesis, transcriptional regulation and plant pathogen defence functional categories. A wide range of processes influence wood density, but this study has identified potential regulators in these pathways. Future studies can now use this information to understand natural variation in wood density, and manipulate the expression of these genes to improve timber quality and yield.

Tafazzin, a mitochondrial acyltransferase, plays an important role in cardiolipin side chain remodeling. Previous studies have shown that dysfunction of tafazzin reduces cardiolipin content, impairs mitochondrial function, and causes dilated cardiomyopathy in Barth syndrome. Reactive oxygen species (ROS) have been implicated in the development of cardiomyopathy and are also the obligated byproducts of mitochondria. We hypothesized that tafazzin knockdown increases ROS production from mitochondria, and a mitochondria-targeted antioxidant prevents tafazzin knockdown induced mitochondrial and cardiac dysfunction. We employed cardiac myocytes transduced with an adenovirus containing tafazzin shRNA as a model to investigate the effects of the mitochondrial antioxidant, mito-Tempo. Knocking down tafazzin decreased steady state levels of cardiolipin and increased mitochondrial ROS. Treatment of cardiac myocytes with mito-Tempo normalized tafazzin knockdown enhanced mitochondrial ROS production and cellular ATP decline. Mito-Tempo also significantly abrogated tafazzin knockdown induced cardiac hypertrophy, contractile dysfunction, and cell death. We conclude that mitochondria-targeted antioxidant prevents cardiac dysfunction induced by tafazzin gene knockdown in cardiac myocytes and suggest mito-Tempo as a potential therapeutic for Barth syndrome and other dilated cardiomyopathies resulting from mitochondrial oxidative stress.

Mutation of the mitochondrial protein tafazzin causes dilated cardiomyopathy in Barth syndrome. Previous studies have shown that tafazzin knockdown promotes hypertrophy of neonatal cardiac myocytes. The current investigation was designed to show whether tafazzin knockdown affects cardiac fibroblast proliferation and collagen secretion, which contribute to fibrosis in dilated cardiomyopathy. In primary cultures of neonatal ventricular fibroblasts (NVFs) transduced with a tafazzin short hairpin RNA adenovirus, tafazzin knockdown increased production of reactive oxygen species and activation of mitogen-activated protein kinases and induced protein and DNA synthesis via cell cycle regulators. It also reduced intracellular ATP, activated AMPK, and caused multinucleation, hypertrophy, and enhanced collagen secretion. We concluded that tafazzin knockdown interrupts the NVF cell cycle and this in turn may contribute to fibrosis and dilated cardiomyopathy in Barth syndrome.

Base excision repair (BER) is necessary for removal of DNA damage that has occurred from spontaneous decomposition, alkylation and oxidation. Defects in BER have been associated with cancer predisposition, neurodegeneration and premature aging. BER is initiated by a damage-specific DNA glycosylase that excises the damaged base, creating an apurinic/apyrimidinic (AP) site. The AP site is recognized by the multifunctional AP endonuclease 1 (APE1), which cleaves the phosphodiester backbone, producing a DNA strand break. DNA polymerase beta incorporates the correct nucleotide, and BER is completed via ligation by a protein complex of Ligase III and XRCC1. Like many BER proteins, APE1 is essential for survival, as deletion of both alleles in mice leads to embryonic lethality. We are pursuing the hypothesis that more mild reductions in APE1 activity will contribute to disease risk and development. To address this issue, we have engaged two strategies: (1) identification and characterization of APE1 missense mutations and (2) design and characterization of genetically-altered mutant human cell lines. In our first study, we have characterized eight APE1 variants found within the population or identified in endometrial cancer. Our studies reveal that while most of these proteins are normal for protein stability, biochemical activities, and intracellular localization, the endometrial cancer-associated variant Arg237Cys has decreased 3′-functions and AP-DNA binding. In our second study, we have created a heterozygous knockout HCT116 cell line, in which we have deleted one allele of APE1. The heterozygous knockout cells show a loss in AP endonuclease activity and are hypersensitive to DNA damaging drugs. Current cellular strategies are determining the essential nature of APE1 activity in HCT116 and other cell lines Citation Format: Jennifer Illuzzi, Nicole A. Harris, Brittney A. Manvilla, Daemyung Kim, Mengxia Li, Alexander C. Drohat, David M. Wilson. Deciphering the role of APE1 protein variants in disease etiology. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2388. doi:10.1158/1538-7445.AM2014-2388

Apurinic/apyrimidinic endonuclease 1 (APE1) is the predominant AP site repair enzyme in mammals. APE1 also maintains 3′–5′ exonuclease and 3′-repair activities, and regulates transcription factor DNA binding through its REF-1 function. Since complete or severe APE1 deficiency leads to embryonic lethality and cell death, it has been hypothesized that APE1 protein variants with slightly impaired function will contribute to disease etiology. Our data indicate that except for the endometrial cancer-associated APE1 variant R237C, the polymorphic variants Q51H, I64V and D148E, the rare population variants G241R, P311S and A317V, and the tumor-associated variant P112L exhibit normal thermodynamic stability of protein folding; abasic endonuclease, 3′–5′ exonuclease and REF-1 activities; coordination during the early steps of base excision repair; and intracellular distribution when expressed exogenously in HeLa cells. The R237C mutant displayed reduced AP-DNA complex stability, 3′–5′ exonuclease activity and 3′-damage processing. Re-sequencing of the exonic regions of APE1 uncovered no novel amino acid substitutions in the 60 cancer cell lines of the NCI-60 panel, or in HeLa or T98G cancer cell lines; only the common D148E and Q51H variants were observed. Our results indicate that APE1 missense mutations are seemingly rare and that the cancer-associated R237C variant may represent a reduced-function susceptibility allele.

Background Concern over land use for non-food bioenergy crops requires breeding programmes that focus on producing biomass on the minimum amount of land that is economically-viable. To achieve this, the maximum potential yield per hectare is a key target for improvement. For long lived tree species, such as poplar, this requires an understanding of the traits that contribute to biomass production and their genetic control. An important aspect of this for long lived plants is an understanding of genetic interactions at different developmental stages, i.e. how genes or genetic regions impact on yield over time. Results QTL mapping identified regions of genetic control for biomass yield. We mapped consistent QTL across multiple coppice cycles and identified five robust QTL hotspots on linkage groups III, IV, X, XIV and XIX, calling these 'Poplar Biomass Loci' (PBL 1–5). In total 20% of the variation in final harvest biomass yield was explained by mapped QTL. We also investigated the genetic correlations between yield related traits to identify 'early diagnostic' indicators of yield showing that early biomass was a reasonable predictor of coppice yield and that leaf size, cell number and stem and sylleptic branch number were also valuable traits. Conclusion These findings provide insight into the genetic control of biomass production and correlation to 'early diagnostic' traits determining yield in poplar SRC for bioenergy. QTL hotspots serve as useful targets for directed breeding for improved biomass productivity that may also be relevant across additional poplar hybrids.