Phytoremediation of No. 2 Fuel Oil-Contaminated Soil

Phytoremediation has been implemented at an industrial site in Wisconsin to promote in situ remediation of No. 2 fuel oil-contaminated soil. The goal of the project is to utilize microbial-enhancing processes within the rhizosphere of trees to stimulate biodegradation of diesel range organics (DROs) within four contaminated hot spots at the site. Between 40 and 90% reductions in the concentrations of the DROs were observed over the course of a 24-week bench-scale bioventing study performed in 1994. In addition to a reduction in the concentration of DROs, the chromatograms for those analyses exhibited a relative decrease in the proportion of the more water soluble and available shorter chained or lower molecular weight DROs compared to their higher molecular weight counterparts in the fuel. In addition to a decrease in concentration, this observed change in the pattern of the chromatograms over time is consistent with biodegradation of DROs. An agronomic assessment performed in 1995 indicated that conditions were favorable for tree growth. Phytoremediation was implemented as a low-cost in situ alternative for remediation of the site. Willow trees were planted in the four hot spots in May 1996 and trees have exhibited fair to excellent growth in the first season.     

Changes in the organization of excitation-contraction coupling structures in failing human heart

Background

The cardiac myocyte t-tubular system ensures rapid, uniform cell activation and several experimental lines of evidence suggest changes in the t-tubular system and associated excitation-contraction coupling proteins may occur in heart failure.

Methods and Results

The organization of t-tubules, L-type calcium channels (DHPRs), ryanodine receptors (RyRs) and contractile machinery were examined in fixed ventricular tissue samples from both normal and failing hearts (idiopathic (non-ischemic) dilated cardiomyopathy) using high resolution fluorescent imaging. Wheat germ agglutinin (WGA), Na-Ca exchanger, DHPR and caveolin-3 labels revealed a shift from a predominantly transverse orientation to oblique and axial directions in failing myocytes. In failure, dilation of peripheral t-tubules occurred and a change in the extent of protein glycosylation was evident. There was no change in the fractional area occupied by myofilaments (labeled with phalloidin) but there was a small reduction in the number of RyR clusters per unit area. The general relationship between DHPRs and RyR was not changed and RyR labeling overlapped with 51±3% of DHPR labeling in normal hearts. In longitudinal (but not transverse) sections there was an ∼30% reduction in the degree of colocalization between DHPRs and RyRs as measured by Pearson''s correlation coefficient in failing hearts.

Conclusions

The results show that extensive remodelling of the t-tubular network and associated excitation-contraction coupling proteins occurs in failing human heart. These changes may contribute to abnormal calcium handling in heart failure. The general organization of the t-system and changes observed in failure samples have subtle differences to some animal models although the general direction of changes are generally similar.     

The Development of Purpose During Adolescence

While there has been much research on adults' abilities to detect deception, there have been very few studies examining both children's and adults' abilities to detect children's real, spontaneous lies. The present study asked both children and adults to make judgments of children's true and false reports. Participants (N = 156) were shown videotaped sessions of children who were either spontaneously lying or telling the truth. Results indicate that both children's and adults' accuracy for detecting children's true statements was below chance. However, older children were significantly better at detecting lies than both younger children and adults.     

Spontaneous autopolyploidy in the Acipenseriformes,with recommendations for management

Reviews in Fish Biology and Fisheries - Sturgeon and paddlefishes (Acipenseriformes) are cultured worldwide for commercial and conservation purposes. Over the past decade, we have learned many...     

Egg lipid and thiamine vary between early and late spawning lake sturgeon

Nutritional deficiency associated with reduced thiamine (vitamin B1) and reduced natural reproduction of salmonid species in the Great Lake Region is well established. The negative relationship between egg thiamine and lipid concentration to post-hatch larval growth and survival in teleost species, coupled with the limited research of egg thiamine in Acipenseriform species of conservation concern, including lake sturgeon, indicates that study of thiamine concentrations lake sturgeon eggs is warranted. Eggs were collected from females (N = 12) during the early and late portion of the spawning run in 2007 in a wild population from Black Lake, MI. Concentrations of thiamine, lipid and fatty acid concentration were measured along with female biological information (body size and egg size) and characteristics of larvae at hatch. Significant differences in egg thiamine concentrations were observed between early- and late-spawning females (mean ± SD: 2.36 nmol·g⁻¹ ± 1.09 vs. 0.73 ± 0.25 nmol·g⁻¹, W = 0.05, p < .01). No significant relationships were observed between female body size or egg size and egg lipid or thiamine concentration. Differences in lipid and thiamine concentrations were not predictive of larval body size or yolk sac volume at hatch. Total and phosphorylated thiamine were correlated with n-3 polyunsaturated fatty acids, suggesting that dietary items were likely partially responsible for provisioning of essential compounds. Given the negative effects of low egg thiamine concentration on larval survival in other fish species globally, results indicate that further research in areas of nutrient acquisition and thiamine effects on larval survival, natural recruitment, and hatchery feeding strategies is warranted for lake sturgeon.     

Downregulation of PDGFRß Signaling Overcomes Crizotinib Resistance in a TYRO3 and ALK Mutated Neuroendocrine-Like Tumor

Patient-derived xenografts provide significant advantages over long-term passage cell lines when investigating efficacy of treatments for solid tumors. Our laboratory encountered a high-grade, metastatic, neuroendocrine-like tumor from a pediatric patient that presented with a unique genetic profile. In particular, mutations in TYRO3 and ALK were identified. We established a human patient-derived xenoline (PDX) of this tumor for use in the current study. We investigated the effect of crizotinib, a chemotherapeutic known to effectively target both TYRO3 and ALK mutations. Crizotinib effectively decreased viability, proliferation, growth, and the metastatic properties of the PDX tumor through downregulation of STAT3 signaling, but expression of PDGFRß was increased. Sunitinib is a small molecule inhibitor of PDGFRß and was studied in this PDX independently and in combination with crizotinib. Sunitinib alone decreased viability, proliferation, and growth in vitro and decreased tumor growth in vivo. In combination, sunitinib was able to overcome potential crizotinib-induced resistance through downregulation of ERK 1/2 activity and PDGFRß receptor expression; consequently, tumor growth was significantly decreased both in vitro and in vivo. Through the use of the PDX, it was possible to identify crizotinib as a less effective therapeutic for this tumor and suggest that targeting PDGFRß would be more effective. These findings may translate to other solid tumors that present with the same genetic mutations.     

Types and rates of sequence evolution at the high-molecular-weight glutenin locus in hexaploid wheat and its ancestral genomes

The Glu-1 locus, encoding the high-molecular-weight glutenin protein subunits, controls bread-making quality in hexaploid wheat (Triticum aestivum) and represents a recently evolved region unique to Triticeae genomes. To understand the molecular evolution of this locus region, three orthologous Glu-1 regions from the three subgenomes of a single hexaploid wheat species were sequenced, totaling 729 kb of sequence. Comparing each Glu-1 region with its corresponding homologous region from the D genome of diploid wheat, Aegilops tauschii, and the A and B genomes of tetraploid wheat, Triticum turgidum, revealed that, in addition to the conservation of microsynteny in the genic regions, sequences in the intergenic regions, composed of blocks of nested retroelements, are also generally conserved, although a few nonshared retroelements that differentiate the homologous Glu-1 regions were detected in each pair of the A and D genomes. Analysis of the indel frequency and the rate of nucleotide substitution, which represent the most frequent types of sequence changes in the Glu-1 regions, demonstrated that the two A genomes are significantly more divergent than the two B genomes, further supporting the hypothesis that hexaploid wheat may have more than one tetraploid ancestor.     

13C-Labelling of lipids to investigate microbial communities in the environment

The introduction of (13)C-labelled substrates to soils, sediments or cultures followed by (13)C analysis of phospholipid fatty acids (PLFAs) provides quantitative and chemotaxonomic information for the groups of microorganisms utilizing a given substrate. Gas chromatography-combustion-isotope ratio mass spectrometry has provided the high precision necessary to measure small isotopic changes (differences in the relative abundances of (13)C to (12)C expressed as delta(13)C values) for nanogram amounts of individual compounds, such as microbial PLFAs. This methodology constitutes a powerful new culture-independent method for investigating microbial communities in the environment. The information obtained is highly complementary to that obtained from gene-probe-based methods, and considerable possibilities exist to extend this methodology to include other biochemical components of microorganisms.     

Mycobacterium tuberculosis WhiB3 Maintains Redox Homeostasis by Regulating Virulence Lipid Anabolism to Modulate Macrophage Response

The metabolic events associated with maintaining redox homeostasis in Mycobacterium tuberculosis (Mtb) during infection are poorly understood. Here, we discovered a novel redox switching mechanism by which Mtb WhiB3 under defined oxidizing and reducing conditions differentially modulates the assimilation of propionate into the complex virulence polyketides polyacyltrehaloses (PAT), sulfolipids (SL-1), phthiocerol dimycocerosates (PDIM), and the storage lipid triacylglycerol (TAG) that is under control of the DosR/S/T dormancy system. We developed an in vivo radio-labeling technique and demonstrated for the first time the lipid profile changes of Mtb residing in macrophages, and identified WhiB3 as a physiological regulator of virulence lipid anabolism. Importantly, MtbΔwhiB3 shows enhanced growth on medium containing toxic levels of propionate, thereby implicating WhiB3 in detoxifying excess propionate. Strikingly, the accumulation of reducing equivalents in MtbΔwhiB3 isolated from macrophages suggests that WhiB3 maintains intracellular redox homeostasis upon infection, and that intrabacterial lipid anabolism functions as a reductant sink. MtbΔwhiB3 infected macrophages produce higher levels of pro- and anti-inflammatory cytokines, indicating that WhiB3-mediated regulation of lipids is required for controlling the innate immune response. Lastly, WhiB3 binds to pks2 and pks3 promoter DNA independent of the presence or redox state of its [4Fe-4S] cluster. Interestingly, reduction of the apo-WhiB3 Cys thiols abolished DNA binding, whereas oxidation stimulated DNA binding. These results confirmed that WhiB3 DNA binding is reversibly regulated by a thiol-disulfide redox switch. These results introduce a new paradigmatic mechanism that describes how WhiB3 facilitates metabolic switching to fatty acids by regulating Mtb lipid anabolism in response to oxido-reductive stress associated with infection, for maintaining redox balance. The link between the WhiB3 virulence pathway and DosR/S/T signaling pathway conceptually advances our understanding of the metabolic adaptation and redox-based signaling events exploited by Mtb to maintain long-term persistence.