Field Demonstration and Laboratory Evaluation of Foamed Asphalts - Muscatine County, HR-233, 1985

In view of the energy, environmental, and economic advantages of the foamed asphalt process using local aggregates in cold mixes and the promising results from Research Project HR-212, a 4.2-mile section of county road in Muscatine County was built with foamed asphalt and local aggregates during August-September 1983. Extensive laboratory evaluation was carried out on five plant mixes representing foamed mixes used in the nine test sections, a laboratory prepared foamed mix, and a laboratory prepared hot mix similar to Plant Mix 1. The foamed mixes were compacted, cured under 15 curing conditions and tested for bulk specific gravity, Marshall stability at 77° F and at 140° F, cured moisture content, resilient modulus and effects of moisture damage due to freeze-thaw cycles, water soaking, and vacuum saturation. In addition, four sets of 83 core samples were taken at 1 to 15 months and tested for moisture content, specific gravity, Marshall stability, and resilient modulus. In summary, the test road has performed satisfactorily for almost two years. The few early construction problems encountered were to be expected for experimental projects dealing with new materials and technologies. Overall results to date are encouraging and foamed asphalt mixes have proved to have the potential as a viable base material in areas where marginal aggregates are available. It is hoped and expected that performance evaluation of the test sections will be continued and that more foamed asphalt trial projects will be constructed and monitored so that experiences and findings from this project can be verified and mix design criteria can be gradually established. For future foamed asphalt projects it is recommended that anti-stripping additives, such as hydrated lime, be added in view of the potential moisture susceptibility of foamed mixes observed in the laboratory evaluation.

Lime or Chloride Treatment of Granular Base Course Materials, HR-99

Results are presented of triaxial testing of three crushed limestones to which either hydrated high-calcium lime, sodium chloride or calcium chloride had been added. Lime was added at rates of 1, 3, 10 and 16 percent, chlorides were added at 0.5 percent rate only. Speciments were compacted using vibratory compaction apparatus and were tested in triaxial compression using lateral pressures from 10 to 100 psi. Triaxial test results indicate that: (1) sodium chloride slightly decreased the angle of internal friction and increased cohesion, (2) calcium chloride slightly increased the angle of internal friction and decreased cohesion, and (3) lime had no appreciable effect on angle of internal friction but increased cohesion, decreased density and increased pore water pressure.

An SH2 domain-containing 5' inositolphosphatase inhibits insulin-induced GLUT4 translocation and growth factor-induced actin filament rearrangement.

Tyrosine kinase receptors lead to rapid activation of phosphatidylinositol 3-kinase (PI3 kinase) and the subsequent formation of phosphatidylinositides (PtdIns) 3,4-P2 and PtdIns 3,4, 5-P3, which are thought to be involved in signaling for glucose transporter GLUT4 translocation, cytoskeletal rearrangement, and DNA synthesis. However, the specific role of each of these PtdIns in insulin and growth factor signaling is still mainly unknown. Therefore, we assessed, in the current study, the effect of SH2-containing inositol phosphatase (SHIP) expression on these biological effects. SHIP is a 5' phosphatase that decreases the intracellular levels of PtdIns 3,4,5-P3. Expression of SHIP after nuclear microinjection in 3T3-L1 adipocytes inhibited insulin-induced GLUT4 translocation by 100 +/- 21% (mean +/- the standard error) at submaximal (3 ng/ml) and 64 +/- 5% at maximal (10 ng/ml) insulin concentrations (P < 0.05 and P < 0.001, respectively). A catalytically inactive mutant of SHIP had no effect on insulin-induced GLUT4 translocation. Furthermore, SHIP also abolished GLUT4 translocation induced by a membrane-targeted catalytic subunit of PI3 kinase. In addition, insulin-, insulin-like growth factor I (IGF-I)-, and platelet-derived growth factor-induced cytoskeletal rearrangement, i.e., membrane ruffling, was significantly inhibited (78 +/- 10, 64 +/- 3, and 62 +/- 5%, respectively; P < 0.05 for all) in 3T3-L1 adipocytes. In a rat fibroblast cell line overexpressing the human insulin receptor (HIRc-B), SHIP inhibited membrane ruffling induced by insulin and IGF-I by 76 +/- 3% (P < 0.001) and 68 +/- 5% (P < 0.005), respectively. However, growth factor-induced stress fiber breakdown was not affected by SHIP expression. Finally, SHIP decreased significantly growth factor-induced mitogen-activated protein kinase activation and DNA synthesis. Expression of the catalytically inactive mutant had no effect on these cellular responses. In summary, our results show that expression of SHIP inhibits insulin-induced GLUT4 translocation, growth factor-induced membrane ruffling, and DNA synthesis, indicating that PtdIns 3,4,5-P3 is the key phospholipid product mediating these biological actions.

Transition from hemifusion to pore opening is rate limiting for vacuole membrane fusion.

Fusion pore opening and expansion are considered the most energy-demanding steps in viral fusion. Whether this also applies to soluble N-ethyl-maleimide sensitive fusion protein attachment protein receptor (SNARE)- and Rab-dependent fusion events has been unknown. We have addressed the problem by characterizing the effects of lysophosphatidylcholine (LPC) and other late-stage inhibitors on lipid mixing and pore opening during vacuole fusion. LPC inhibits fusion by inducing positive curvature in the bilayer and changing its biophysical properties. The LPC block reversibly prevented formation of the hemifusion intermediate that allows lipid, but not content, mixing. Transition from hemifusion to pore opening was sensitive to guanosine-5'-(gamma-thio)triphosphate. It required the vacuolar adenosine triphosphatase V0 sector and coincided with its transformation. Pore opening was rate limiting for the reaction. As with viral fusion, opening the fusion pore may be the most energy-demanding step for intracellular, SNARE-dependent fusion reactions, suggesting that fundamental aspects of lipid mixing and pore opening are related for both systems.

Preservation of rhizobia by lyophilization with trehalose

The objective of this study was to determine if trehalose can effectively enhance the viability and storage lifetime of Rhizobium tropici and Rhizobium etli after lyophilization as compared to the traditional protective combination of sucrose and peptone. Two strains of bean-nodulating rhizobia were effectively preserved in the lyophilized state by 100 mM trehalose, and survived for at least 12 days, even when stored under non-ideal conditions. Trehalose provided better protection than the sucrose/peptone combination. When protective excipients were introduced inside the cells, strain CFN 42 was more sensitive to the type of additive used than strain CIAT 899, probably because CIAT 899 produces beta (1-2) glucan, which may have a protective effect. beta (1-2) glucan combined with sucrose protected better than trehalose against leakage from freeze-dried liposomes, but by itself it had no protective capability. There was a high level of unfrozen water associated with the hydrated glucan, in addition to a thermal event at approximately 70ºC, corresponding to an observed gel-sol transition in the glucan. These findings may shed light on the mechanism by which the glucan contributes to the protection of dried cells and liposomes, and may lead to further improvements in rhizobial inoculants.

Sequencing human-gibbon breakpoints of synteny reveals mosaic new insertions at rearrangement sites

The gibbon genome exhibits extensive karyotypic diversity with an increased rate of chromosomal rearrangements during evolution. In an effort to understand the mechanistic origin and implications of these rearrangement events, we sequenced 24 synteny breakpoint regions in the white-cheeked gibbon (Nomascus leucogenys, NLE) in the form of high-quality BAC insert sequences (4.2 Mbp). While there is a significant deficit of breakpoints in genes, we identified seven human gene structures involved in signaling pathways (DEPDC4, GNG10), phospholipid metabolism (ENPP5, PLSCR2), beta-oxidation (ECH1), cellular structure and transport (HEATR4), and transcription (ZNF461), that have been disrupted in the NLE gibbon lineage. Notably, only three of these genes show the expected evolutionary signatures of pseudogenization. Sequence analysis of the breakpoints suggested both nonclassical nonhomologous end-joining (NHEJ) and replication-based mechanisms of rearrangement. A substantial number (11/24) of human-NLE gibbon breakpoints showed new insertions of gibbon-specific repeats and mosaic structures formed from disparate sequences including segmental duplications, LINE, SINE, and LTR elements. Analysis of these sites provides a model for a replication-dependent repair mechanism for double-strand breaks (DSBs) at rearrangement sites and insights into the structure and formation of primate segmental duplications at sites of genomic rearrangements during evolution.

Effects of Cement on Moisture Migration in Concrete, Phase A, Laboratory Study of Moisture Migration in Hardened Cement Paste, HR-141, 1972

The Columbus Laboratories of Battelle Memorial Institute is currently conducting a study of the effect of cement on moisture migration in concrete as related to the problem of D-cracking of portland cement concrete pavements. The study began on December 31, 1970, and is planned as a 3-year program. The work plan, approved by the policy committee of the members of the Iowa, Kansas, and Missouri highway departments and the Federal Highway Administration, is composed of four parts. The first phase (A) of the investigation concerned the movement of moisture into and from hardened cement pastes and the dimensional changes accompanying the moisture changes. Small slab specimens of hardened neat cement pastes were prepared from 32 different cements which were prepared at the same water/cement ratio and hydrated to the same maturity factor.

Structure of intracellular mature vaccinia virus observed by cryoelectron microscopy.

Intracellular mature vaccinia virus, also called intracellular naked virus, and its core envelope have been observed in their native, unfixed, unstained, hydrated states by cryoelectron microscopy of vitrified samples. The virion appears as a smooth rounded rectangle of ca. 350 by 270 nm. The core seems homogeneous and is surrounded by a 30-nm-thick surface domain delimited by membranes. We show that surface tubules and most likely also the characteristic dumbbell-shaped core with the lateral bodies which are generally observed in negatively stained or conventionally embedded samples are preparation artifacts.

Actin cables and the exocyst form two independent morphogenesis pathways in the fission yeast.

Cell morphogenesis depends on polarized exocytosis. One widely held model posits that long-range transport and exocyst-dependent tethering of exocytic vesicles at the plasma membrane sequentially drive this process. Here, we describe that disruption of either actin-based long-range transport and microtubules or the exocyst did not abolish polarized growth in rod-shaped fission yeast cells. However, disruption of both actin cables and exocyst led to isotropic growth. Exocytic vesicles localized to cell tips in single mutants but were dispersed in double mutants. In contrast, a marker for active Cdc42, a major polarity landmark, localized to discreet cortical sites even in double mutants. Localization and photobleaching studies show that the exocyst subunits Sec6 and Sec8 localize to cell tips largely independently of the actin cytoskeleton, but in a cdc42 and phospholipid phosphatidylinositol 4,5-bisphosphate (PIP₂)-dependent manner. Thus in fission yeast long-range cytoskeletal transport and PIP₂-dependent exocyst represent parallel morphogenetic modules downstream of Cdc42, raising the possibility of similar mechanisms in other cell types.

Evaluation of the Consolid System of Soil Stabilization, MLR-86-10, 1987

The Consolid System by American Consolid Inc. is a three product system that, according to product literature, "enables any soil, found anywhere, to be upgraded to achieve better characteristics necessary in improving road life and quality". Consolid was evaluated along with mixes of cement-fly ash and hydrated lime on two soils. The soils were an A-2-4(0) with zero plasticity index and an A-7-8(18) with a 31 plasticity index. American Consolid Inc. recommended an application rate of 0.10% Consolid 444 and 1.00% Conservex by dry soil weight. The application rate chosen for cement-fly ash was 5% cement and 15% fly ash and for hydrated lime it was 6.5%. Testing involved triaxial testing of specimens after water soaking, unconfined compressive strength of specimens before and after water soaking, and freeze and thaw testing of specimens after water soaking. All specimens were compacted to standard proctor at optimum moisture. The cement-fly ash treated mixes had the highest strength and durability followed by the hydrated lime treated mixes.

Fly Ash Soil Stabilization for Non-Uniform Subgrade Soils, Volume I: Engineering Properties and Construction Guidelines, TR-461, 2005

Soil treated with self-cementing fly ash is increasingly being used in Iowa to stabilize fine-grained pavement subgrades, but without a complete understanding of the short- and long-term behavior. To develop a broader understanding of fly ash engineering properties, mixtures of five different soil types, ranging from ML to CH, and several different fly ash sources (including hydrated and conditioned fly ashes) were evaluated. Results show that soil compaction characteristics, compressive strength, wet/dry durability, freeze/thaw durability, hydration characteristics, rate of strength gain, and plasticity characteristics are all affected by the addition of fly ash. Specifically, Iowa selfcementing fly ashes are effective at stabilizing fine-grained Iowa soils for earthwork and paving operations; fly ash increases compacted dry density and reduces the optimum moisture content; strength gain in soil-fly ash mixtures depends on cure time and temperature, compaction energy, and compaction delay; sulfur contents can form expansive minerals in soil–fly ash mixtures, which severely reduces the long-term strength and durability; fly ash increases the California bearing ratio of fine-grained soil–fly ash effectively dries wet soils and provides an initial rapid strength gain; fly ash decreases swell potential of expansive soils; soil-fly ash mixtures cured below freezing temperatures and then soaked in water are highly susceptible to slaking and strength loss; soil stabilized with fly ash exhibits increased freeze-thaw durability; soil strength can be increased with the addition of hydrated fly ash and conditioned fly ash, but at higher rates and not as effectively as self-cementing fly ash. Based on the results of this study, three proposed specifications were developed for the use of self-cementing fly ash, hydrated fly ash, and conditioned fly ash. The specifications describe laboratory evaluation, field placement, moisture conditioning, compaction, quality control testing procedures, and basis of payment.

Fly Ash Soil Stabilization for Non-Uniform Subgrade Soils, Volume II: Influence of Subgrade Non-Uniformity on PCC Pavement Performance, TR-461, 2005

To provide insight into subgrade non-uniformity and its effects on pavement performance, this study investigated the influence of non-uniform subgrade support on pavement responses (stress and deflection) that affect pavement performance. Several reconstructed PCC pavement projects in Iowa were studied to document and evaluate the influence of subgrade/subbase non-uniformity on pavement performance. In situ field tests were performed at 12 sites to determine the subgrade/subbase engineering properties and develop a database of engineering parameter values for statistical and numerical analysis. Results of stiffness, moisture and density, strength, and soil classification were used to determine the spatial variability of a given property. Natural subgrade soils, fly ash-stabilized subgrade, reclaimed hydrated fly ash subbase, and granular subbase were studied. The influence of the spatial variability of subgrade/subbase on pavement performance was then evaluated by modeling the elastic properties of the pavement and subgrade using the ISLAB2000 finite element analysis program. A major conclusion from this study is that non-uniform subgrade/subbase stiffness increases localized deflections and causes principal stress concentrations in the pavement, which can lead to fatigue cracking and other types of pavement distresses. Field data show that hydrated fly ash, self-cementing fly ash-stabilized subgrade, and granular subbases exhibit lower variability than natural subgrade soils. Pavement life should be increased through the use of more uniform subgrade support. Subgrade/subbase construction in the future should consider uniformity as a key to long-term pavement performance.

Red blood cell-derived microparticles isolated from blood units initiate and propagate thrombin generation.

BACKGROUND: Red blood cell-derived microparticles (RMPs) are small phospholipid vesicles shed from RBCs in blood units, where they accumulate during storage. Because microparticles are bioactive, it could be suggested that RMPs are mediators of posttransfusion complications or, on the contrary, constitute a potential hemostatic agent. STUDY DESIGN AND METHODS: This study was performed to establish the impact on coagulation of RMPs isolated from blood units. Using calibrated automated thrombography, we investigated whether RMPs affect thrombin generation (TG) in plasma. RESULTS: We found that RMPs were not only able to increase TG in plasma in the presence of a low exogenous tissue factor (TF) concentration, but also to initiate TG in plasma in absence of exogenous TF. TG induced by RMPs in the absence of exogenous TF was neither affected by the presence of blocking anti-TF nor by the absence of Factor (F)VII. It was significantly reduced in plasma deficient in FVIII or F IX and abolished in FII-, FV-, FX-, or FXI-deficient plasma. TG was also totally abolished when anti-XI 01A6 was added in the sample. Finally, neither Western blotting, flow cytometry, nor immunogold labeling allowed the detection of traces of TF antigen. In addition, RMPs did not comprise polyphosphate, an important modulator of coagulation. CONCLUSIONS: Taken together, our data show that RMPs have FXI-dependent procoagulant properties and are able to initiate and propagate TG. The anionic surface of RMPs might be the site of FXI-mediated TG amplification and intrinsic tenase and prothrombinase complex assembly.

Changes in the isoflavone profile and in the chemical composition of tempeh during processing and refrigeration

The objective of this work was to analyze changes in the isoflavone profile, determined by high performance liquid chromatography, at different processing stages and after refrigeration of tempeh. For tempeh production, clean soybean grains from cultivars BR 36 (low isoflavone content) and IAS 5 (high) were dehulled, and the separated cotyledons were hydrated and then cooked in boiling water for 30 min. Spores of the fungus Rhizopus microsporus var. oligosporus were inoculated in the cooked and cooled cotyledons, and incubated at 32ºC for 6, 12, 18, and 24 hours in perforated polypropylene bags, for fermentation. The resulting tempeh was stored at 4ºC for 6, 12, 18, and 24 hours. After 24-hour fermentation, isoflavone glucosides were 50% reduced, and the aglycone forms in the tempeh from both cultivars was increased. The malonyl forms reduced 83% after cooking. Less than 24 hours of refrigeration did not affect the isoflavone profile of tempeh from either cultivar, which is a good indicator of its quality. The tempeh maintains the high and low isoflavone content of the cultivars, which indicates that cultivar differences in this trait should be considered when processing tempeh.

Microbial communities in Cerrado soils under native vegetation subjected to prescribed fire and under pasture

The objective of this work was to evaluate the effects of fire regimes and vegetation cover on the structure and dynamics of soil microbial communities, through phospholipid fatty acid (PLFA) analysis. Comparisons were made between native areas with different woody covers ("cerrado stricto sensu" and "campo sujo"), under different fire regimes, and a 20-year-old active palisadegrass pasture in the Central Plateau of Brazil. Microbial biomass was higher in the native plots than in the pasture, and the highest monthly values were observed during the rainy season in the native plots. No significant differences were observed between fire regimes or between communities from the two native vegetation types. However, the principal component (PC) analysis separated the microbial communities by vegetation cover (native x pasture) and season (wet x dry), accounting for 45.8% (PC1 and PC3) and 25.6% (PC2 and PC3), respectively, of the total PLFA variability. Changes in land cover and seasonal rainfall in Cerrado ecosystems have significant effects on the total density of soil microorganisms and on the abundance of microbial groups, especially Gram-negative and Gram-positive bacteria.