Microhomology-mediated end joining is the principal mediator of double-strand break repair during mitochondrial DNA lesions

Mitochondrial DNA (mtDNA) deletions are associated with various mitochondrial disorders. The deletions identified in humans are flanked by short, directly repeated mitochondrial DNA sequences; however, the mechanism of such DNA rearrangements has yet to be elucidated. In contrast to nuclear DNA (nDNA), mtDNA is more exposed to oxidative damage, which may result in double-strand breaks (DSBs). Although DSB repair in nDNA is well studied, repair mechanisms in mitochondria are not characterized. In the present study, we investigate the mechanisms of DSB repair in mitochondria using in vitro and ex vivo assays. Whereas classical NHEJ (C-NHEJ) is undetectable, microhomology-mediated alternative NHEJ efficiently repairs DSBs in mitochondria. Of interest, robust microhomology-mediated end joining (MMEJ) was observed with DNA substrates bearing 5-, 8-, 10-, 13-, 16-, 19-, and 22-nt microhomology. Furthermore, MMEJ efficiency was enhanced with an increase in the length of homology. Western blotting, immunoprecipitation, and protein inhibition assays suggest the involvement of CtIP, FEN1, MRE11, and PARP1 in mitochondrial MMEJ. Knock-down studies, in conjunction with other experiments, demonstrated that DNA ligase III, but not ligase IV or ligase I, is primarily responsible for the final sealing of DSBs during mitochondrial MMEJ. These observations highlight the central role of MMEJ in maintenance of mammalian mitochondrial genome integrity and is likely relevant for deletions observed in many human mitochondrial disorders.

Low Spring Constant Regulates P-Selectin-Psgl-1 Bond Rupture

Forced dissociation of selectin-ligand bonds is crucial to such biological processes as leukocyte recruitment, thrombosis formation, and tumor metastasis. Although the bond rupture has been well known at high loading rate r(f) (>= 10(2) pN/s), defined as the product of spring constant k and retract velocity v, how the low r(f) (< 10(2) pN/s) or the low k regulates the bond dissociation remains unclear. Here an optical trap assay was used to quantify the bond rupture at r(f) <= 20 pN/s with low k (similar to 10(-3)-10(-2) pN/nm) when P-selectin and P-selectin glycoprotein ligand 1 (PSGL-1) were respectively coupled onto two glass microbeads. Our data indicated that the bond rupture force f retained the similar values when r(f) increased up to 20 pN/s. It was also found that f varied with different combinations of k and v even at the same r(f). The most probable force, f

Superelastic And Spring Properties Of Si3N4 Microcoils

Silicon nitride with helical structure was prepared on a large scale by CVD. On the microscale, these coiled Si3N4 ceramics still possess superelasticity and can recover their original shapes after cyclic loadings without noticeable deformations. These results suggest helical microcoils could have potential in microdevices for MEMS, motors, electromagnets, generators, and related equipment.

Influence of shelf-break fronts on shellfish and fish stocks off Argentina [Contribution to ICES Annual Science Conference, Reykjavik, Iceland, 27 Sep- 04 Oct 1996]

The general circulation pattern in the western boundary of the SW Atlantic is dominated by the opposite flows of Malvinas (-Falkland)and Brazil Current. In the Confluence region both currents separate from the continental slope and flow offshore creating an area of strong contracts and complex dynamics. The shelf-break fronts off Argentina mark the transition between shelf waters of mixed origin and nutrient rich Malvinas waters. Two areas deserve special attention due to the steep gradients introduced by the outflow of important sources of continental waters: the Rio de la Plata and the Magellan Strait to the north and south of the study area. Characteristics of the front is the high primary and secondary production, and the presence of important invertebrate and fish stocks that concentrate along the front to feed or spawn. The area comprises nearly 30 o/o (333 million U$S in 1995)of all Argentine catches of fish and squid. Resources in the area, beyond the EEZ limits, support international fisheries mainly of Russia, Poland and Spain. (Document contains 15 pages & figs)

Lithology and palynology of cave floor sediment cores from Wakulla Spring, Wakulla County, Florida

Five short bottom sediment cores taken in Wakulla Spring Wakulla County, Florida, were described lithologically and sampled for palynological study. Four of the cores were recoveredfrom sediments at the spring cave entrance (130 feet water depth). One core was taken in a fossil vertebrate bone bed, 280 feet distance into the main spring cave at a water depth of 240 feet. Sediments in the cores are composed of alternating intervals of quartz sand and calcilitite, containing freshwater diatoms, freshwater mollusk shells and plant remains. The predominant pollen present in all cores consists of a periporate variety typical of the herb families Chenopodiaceae and Amaranthaceae. Arboreal flora, typical of the area surrounding the spring today, represent a very low percentage of thle pollen assemblage in the cores. Clustered Chenopod-Amaranth type pollen observed in one core suggest minimal transport prior to deposition, and indicate that the bottom sediments in the cave may be essentially In situ. An absence of exotic flora suggests a Quaternary age for the sediments. (PDF contains 11 pages.)

Nutrient Limitation of Periphyton in a Spring-Fed, Coastal Stream in Florida, USA.

There is strong evidence to suggest that ground-water nitrate concentrations have increased in recent years and further increases are expected along portions of the central Gulf coast of Florida. Much of the nitrate enriched groundwater is discharged into surface waters through numerous freshwater springs that are characteristic of the area and the potential for eutrophication of their receiving waters is a legitimate concern. To test the potential effects of elevated nutrient concentrations on the periphyton community an in situ nutrient addition experiment was conducted in the spring-fed Chassahowitzka River, FL, USA, during the summer of 1999. Plastic tubes housing arrays of glass microscope slides were suspended in the stream. Periphyton colonizing the microscope slides was subjected to artificial increases in nitrogen, phosphorus or a combination of both. Slides from each tube were collected at 3- to 4- day intervals and the periphyton communities were measured for chlorophyll concentration. The addition of approximately 10 μg/L of phosphate above ambient concentrations significantly increased the amount of periphyton on artificial substrates relative to controls; the addition of approximately 100 μg/L of nitrate above ambient concentrations did not. The findings from this experiment implicated phosphorus, rather than nitrogen, as the nutrient that potentially limits periphyton growth in this system.(PDF contains 4 pages.)

Land Use and Spring Protection

(1 poster)

Long-Term Monitoring of Ecological Conditions In Gray’s Reef National Marine Sanctuary: Comparison of soft-bottom benthic assemblages and contaminant levels in sediments and biota in Spring 2000 and 2005

As part of an ongoing program of benthic sampling and related assessments of sediment quality at Gray’s Reef National Marine Sanctuary (GRNMS) off the coast of Georgia, a survey of soft-bottom benthic habitats was conducted in spring 2005 to characterize condition of macroinfaunal assemblages and levels of chemical contaminants in sediments and biota relative to a baseline survey carried out in spring 2000. Distribution and abundance of macrobenthos were related foremost to sediment type (median particle size, % gravel), which in turn varied according to bottom-habitat mesoscale features (e.g., association with live bottom versus flat or rippled sand areas). Overall abundance and diversity of soft-bottom benthic communities were similar between the two years, though dominance patterns and relative abundances of component species were less repeatable. Seasonal summer pulses of a few taxa (e.g., the bivalve Ervilia sp. A) observed in 2000 were not observed in 2005. Concentrations of chemical contaminants in sediments and biota, though detectable in both years, were consistently at low, background levels and no exceedances of sediment probable bioeffect levels or FDA action levels for edible fish or shellfish were observed. Near-bottom dissolved oxygen levels and organic-matter content of sediments also have remained within normal ranges. Highly diverse benthic assemblages were found in both years, supporting the premise that GRNMS serves as an important reservoir of marine biodiversity. A total of 353 taxa (219 identified to species) were collected during the spring 2005 survey. Cumulatively, 588 taxa (371 identified to species) have been recorded in the sanctuary from surveys in 2000, 2001, 2002, and 2005. Species Accumulation Curves indicate that the theoretical maximum should be in excess of 600 species. Results of this study will be of value in advancing strategic science and management goals for GRNMS, including characterization and long-term monitoring of sanctuary resources and processes, as well as supporting evolving interests in ecosystem-based management of the surrounding South Atlantic Bight (SAB) ecosystem. (PDF contains 46 pages)

The ecological basis of fishery yield of the Puerto Rico-Virgin Islands Insular Shelf: 1987 Assessment

A literature review was conducted to locate information on the flow of energy from primary producers to the fishery stocks of the Puerto Rican-Virgin Islands insular shelf. This report uses site-specific information to describe the major ecological subsystems, or habitats, of the region, to identify the more common species and the subsystems in which they occur, to quantify productivity and biomass, and to outline trophic relationships. Discussions on each topic and subsystem vary in substance and detail, being limited by the availability and accessibility of information. (PDF contains 189 pages) Seven distinct subsystems are described: mangrove estuary, seagrass bed, coral reef, algal plain, sand/mud bottom, shelf break, and overlying pelagic. Over 50 tables provide lists of species found in each habitat on various surveys dating back to 1956. Estimates of density, relative abundance, and productivity are provided when possible. We evaluated whether sufficient information exists to support an analysis of the energy basis of fishery production in the area, beginning with the design and development of an ecosystem model. Data needs in three categories - species lists, biomass, and trophic relations - were examined for each subsystem and for each of three species groups - primary producers, invertebrates, and fish. We concluded that adequate data, sufficient for modeling purposes, are available in 16 (25%) of 64 categories; limited data, those requiring greater extrapolation, are available in 35 (55%) categories; and no data are available in 13 (20%) categories. The best-studied subsystems are seagrass beds and coral reefs, with at least limited data in all categories. Invertebrates, the intermediate link in the food web between primary producers and fishes, are the least quantified group in the region. Primary production and fishes, however, are relatively well-studied, providing sufficient data to support an ecosystem-level analysis and to initiate a modeling effort.

Generating cluster submodels from a multistage stochastic mixed integer optimization model using break stage

We present a scheme to generate clusters submodels with stage ordering from a (symmetric or a nonsymmetric one) multistage stochastic mixed integer optimization model using break stage. We consider a stochastic model in compact representation and MPS format with a known scenario tree. The cluster submodels are built by storing first the 0-1 the variables, stage by stage, and then the continuous ones, also stage by stage. A C++ experimental code has been implemented for reordering the stochastic model as well as the cluster decomposition after the relaxation of the non-anticipativiy constraints until the so-called breakstage. The computational experience shows better performance of the stage ordering in terms of elapsed time in a randomly generated testbed of multistage stochastic mixed integer problems.