Successful Medical Management of Recalcitrant Fusarium solani Keratitis: Molecular Identification and Susceptibility Patterns

Fungal keratitis is a rare but sight-threatening infection of the cornea that may be caused by several fungal pathogens. A delay in diagnosis and inadequate treatment may even lead to loss of the affected eye. Fungal keratitis is often misdiagnosed as bacterial keratitis because isolation and identification of the fungal pathogen is difficult and requires experience, and fungal growth in culture requires time. In this report, a 14-year-old boy with recalcitrant Fusarium solani keratitis, unresponsive to initial therapy, is presented. CLSI M38-A2 in vitro antifungal susceptibility tests demonstrated that only amphotericin B (0.5 μg/ml) had potent activity against F. solani; however, fluconazole (>64 μg/ml), itraconazole (>16 μg/ml), voriconazole (8 μg/ml), and posaconazole (>16 μg/ml) had high minimum inhibitory concentrations. In addition, caspofungin (>16 μg/ml) and anidulafungin (>16 μg/ml) exhibited high minimum effective concentrations. Repeated intrastromal voriconazole injections, topical voriconazole, and caspofungin combined with systemic antifungal agents improved of the corneal lesion with a significant increase in visual acuity. Intrastromal voriconazole injection may be used as an adjunctive treatment method for recalcitrant fungal keratitis with no prominent complications. The intrastromal route could be an effective route of administration of antifungal agents, especially for F. solani keratitis, as in this case. A combination of various antifungal agents administered by different routes prevented loss of the eye.     

Limitations of Time-Resolved Fluorescence Suggested by Molecular Simulations: Assessing the Dynamics of T?cell Receptor Binding Loops

Time-resolved fluorescence anisotropy (TRFA) has a rich history in evaluating protein dynamics. Yet as often employed, TRFA assumes that the motional properties of a covalently tethered fluorescent probe accurately portray the motional properties of the protein backbone at the probe attachment site. In an extensive survey using TRFA to study the dynamics of the binding loops of a αβ T cell receptor, we observed multiple discrepancies between the TRFA data and previously published results that led us to question this assumption. We thus simulated several of the experimentally probed systems using a protocol that permitted accurate determination of probe and protein time correlation functions. We found excellent agreement in the decays of the experimental and simulated correlation functions. However, the motional properties of the probe were poorly correlated with those of the backbone of both the labeled and unlabeled protein. Our results warrant caution in the interpretation of TRFA data and suggest further studies to ascertain the extent to which probe dynamics reflect those of the protein backbone. Meanwhile, the agreement between experiment and computation validates the use of molecular dynamics simulations as an accurate tool for exploring the molecular motion of T cell receptors and their binding loops.     

The interaction between propagule pressure,habitat suitability and density‐dependent reproduction in species invasion

Seedling recruitment limitations create a demographic bottleneck that largely determines the viability and structure of plant populations and communities, and pose a core restriction on the colonization of novel habitat. We use a shade‐tolerant, invasive grass, Microstegium vimineum, to examine the interplay between seed and establishment limitations – phenomena that together determine recruitment success but usually are investigated individually. We add increasing amounts of seed to microhabitats containing variable levels of leaf litter thickness – with reduced leaf litter simulating disturbance – to investigate whether reduced seed limitation overcomes the establishment limitation posed by litter cover. We do this across gradients in understory light, moisture and temperature, and quantify germination, survival, and then per capita adult biomass and reproduction in order to understand the implications for invasion across the landscape. We find that the combined effects of seed and establishment limitation influence recruitment; however, propagule pressure overwhelms the inhibitory effects of leaf litter thickness. Leaf litter reduces germination by 22–57% and seedling survival by 13–15% from that observed on bare soil. However, density‐dependent reproduction compensates as 1–3 plants can produce far more seeds (approx. 525) than are required for persistence. As such, just a few plants may establish in understory forest habitat and subsequently overwhelm establishment barriers with copious propagule production. These results, for a widespread, invasive plant, are consistent with the emerging perspective for native plants that seed and establishment limitation jointly influence recruitment. The ability for an exotic plant species to compensate for low population densities with high per capita seed production, that then overrides establishment limitations, makes its invasive potential daunting. Further work is required to test if this is a common mechanism underlying plant invasions.     

Protein-based identification of quantitative trait loci associated with malignant transformation in two HER2+ cellular models of breast cancer

Background

A contemporary view of the cancer genome reveals extensive rearrangement compared to normal cells. Yet how these genetic alterations translate into specific proteomic changes that underpin acquiring the hallmarks of cancer remains unresolved. The objectives of this study were to quantify alterations in protein expression in two HER2+ cellular models of breast cancer and to infer differentially regulated signaling pathways in these models associated with the hallmarks of cancer.

Results

A proteomic workflow was used to identify proteins in two HER2 positive tumorigenic cell lines (BT474 and SKBR3) that were differentially expressed relative to a normal human mammary epithelial cell line (184A1). A total of 64 (BT474-184A1) and 69 (SKBR3-184A1) proteins were uniquely identified that were differentially expressed by at least 1.5-fold. Pathway inference tools were used to interpret these proteins in terms of functionally enriched pathways in the tumor cell lines. We observed "protein ubiquitination" and "apoptosis signaling" pathways were both enriched in the two breast cancer models while "IGF signaling" and "cell motility" pathways were enriched in BT474 and "amino acid metabolism" were enriched in the SKBR3 cell line.

Conclusion

While "protein ubiquitination" and "apoptosis signaling" pathways were common to both the cell lines, the observed patterns of protein expression suggest that the evasion of apoptosis in each tumorigenic cell line occurs via different mechanisms. Evidently, apoptosis is regulated in BT474 via down regulation of Bid and in SKBR3 via up regulation of Calpain-11 as compared to 184A1.     

Relationships of hepatic and pancreatic biomarkers with the cholestatic syndrome and tumor stage in pancreatic cancer

We analyzed relationships of hepatic and pancreatic biomarkers with the cholestatic syndrome and tumor stage in exocrine pancreatic cancer (N = 183). Information on laboratory tests and on signs and symptoms was obtained from medical records and patient interviews. Bilirubin, aspartate aminotransferase (AST), γ-glutamyltransferase (GGT) and alkaline phosphatase were lower in tumor stage IV. The association was due to the relationship between cholestatic syndrome and earlier presentation of patients. There was no association between hepatic biomarkers and stage when adjusting by cholestatic syndrome. Relationships of hepatic and pancreatic biomarkers with pancreatic symptoms and tumor stage must be controlled in "-omics" and other studies using biomarkers.     

Grass Invasions Across a Regional Gradient are Associated with Declines in Belowground Carbon Pools

The composition of plant communities everywhere now likely comprises alien as well as native species, and those aliens that become invasive have wide-ranging impacts on the structure and function of recipient ecosystems. These impacts include perturbations to soil carbon (C) cycling, but the direction and magnitude of impacts are species and climate dependent, making it difficult to generalize whether a specific invader will promote losses or gains in soil C stocks. Generalizations of a specific invader??s impacts are necessary; however, because the range of an invader can encompass thousands of square kilometers, meaning their effects can have broad, regional consequences. To quantify broad-scale and context-dependent impacts of a specific invader, multi-site investigations that capture and measure local and regional environmental heterogeneity are necessary. Using this approach, we show that a widespread grass invader of forest understories is associated with declines in soil C during infilling (spreading within the invaded range). Across the 36 study sites, total soil C stocks declined (P?=?0.113) by approximately 12% (estimated mean?±?SD, uninvaded: 2,429?±?512.9 vs. invaded: 2,140?±?520.7?g?C?m^?2). The decline in total soil C is driven by a significant (P?=?0.047) reduction in the native-derived, mineral-associated soil C fraction. This fraction, whose mass and slow turnover makes it an important C store, is approximately 15% lower in invaded (estimated mean?±?SD: 1,560?±?400.4?g?C?m^?2) than uninvaded plots (1,826?±?398.1?g?C?m^?2). Notably, declines in this C fraction are only apparent at 21 of the sites, reflecting how environmental heterogeneity in other variables (specifically pH, soil moisture, and clay content) are important to quantify to determine invader impacts across a region. The 26% decline in microbial biomass with invasion (P?=?0.011; estimated mean?±?SD, uninvaded: 10.05?±?1.79 vs. invaded: 7.40?±?1.80?g?C?m^?2) is also dependent on site characteristics (pH), and reductions are greater where the invader occurs at higher densities. Reductions in microbial biomass and soil C with invasion suggest that grass invasion will alter soil C cycling and decrease forest-C stores across the study region, although invader effects at a specific-site will be dependent on environmental context.     

Applicability of indicator monitoring to ecological risk assessment

Although ecological risk assessment (ERA) and environmental monitoring would seem to be potentially complimentary activities, they have been disjunct in practice. This is because of differences in goals and products. Environmental monitoring determines status and trends in indicators to determine whether the environment is improving. ERA estimates effects of stressors on endpoint attributes to support decision making. Indicators are, by definition, indicative of some unmeasured condition. Assessment endpoints are valued properties of the environment that are susceptible to stressors of concern. Indicators are justified by the logic of the monitoring program, which may be self-referential. Assessment endpoints are justified by their potential susceptibility and by environmental policies and public values. Indicators are often expressed in terms of indices or scores that obscure the actual condition of the environment. Because assessment endpoints must be clear to decision makers and the public, they require real units of actual environmental properties. Monitoring programs are peripherally concerned about causal relationships, while risk assessment is devoted to elucidating causal relationships. As a result, risk assessments may use the results of monitoring studies, but only after disaggregating the indicators to their components and choosing those that are appropriate. Monitoring programs could be more useful if they used a risk-based approach to address important problems rather than simply tracking indicators.     

90 Initial characterization of the terminal domains of linear plasmid-like dna's from ernodesmis verticillata (chlorophyta)

Phylogenetic analyses inferred from rbcL sequences indicates that specimens of Caloglossa (Harvey) G. Martens collected from Vermilion Bay, Louisiana and Galveston, Texas, reported as C. leprieurii (Montagne) G. Martens (1869) constitutes a separate taxon. Studies are presently being conducted to compare this northwestern Gulf of Mexico taxon with material of C. intermedia Kamiya et West in Kamiya et al. (2000, J. Phycol. 36:411-420) from the Western Atlantic coast and with C. apomeiotica West et Zuccarello in West et al. (1994, Bot. Mar. 37:381–390) from Baja California, Mexico.     

Identification of Listeria monocytogenes Genes Expressed in Response to Growth at Low Temperature

Listeria monocytogenes is a food-borne bacterial pathogen that is able to grow at refrigeration temperatures. To investigate microbial gene expression associated with cold acclimation, we used a differential cDNA cloning procedure known as selective capture of transcribed sequences (SCOTS) to identify bacterial RNAs that were expressed at elevated levels in bacteria grown at 10°C compared to those grown at 37°C. A total of 24 different cDNA clones corresponding to open reading frames in the L. monocytogenes strain EGD-e genome were obtained by SCOTS. These included cDNAs for L. monocytogenes genes involved in previously described cold-adaptive responses (flaA and flp), regulatory adaptive responses (rpoN, lhkA, yycJ, bglG, adaB, and psr), general microbial stress responses (groEL, clpP, clpB, flp, and trxB), amino acid metabolism (hisJ, trpG, cysS, and aroA), cell surface alterations (fbp, psr, and flaA), and degradative metabolism (eutB, celD, and mleA). Four additional cDNAs were obtained corresponding to genes potentially unique to L. monocytogenes and showing no significant similarity to any other previously described genes. Northern blot analyses confirmed increased steady-state levels of RNA for all members of a subset of genes examined during growth at a low temperature. These results indicated that L. monocytogenes acclimation to growth at 10°C likely involves amino acid starvation, oxidative stress, aberrant protein synthesis, cell surface remodeling, alterations in degradative metabolism, and induction of global regulatory responses.