Comparative folate metabolism in humans and malaria parasites (part II): activities as yet untargeted or specific to Plasmodium

The folate pathway represents a powerful target for combating rapidly dividing systems such as cancer cells, bacteria and malaria parasites. Whereas folate metabolism in mammalian cells and bacteria has been studied extensively, it is understood less well in malaria parasites. In two articles, we attempt to reconstitute the malaria folate pathway based on available information from mammalian and microbial systems, in addition to Plasmodium-genome-sequencing projects. In part I, we focused on folate enzymes that are already used clinically as anticancer drug targets or that are under development in drug-discovery programs. In this article, we discuss mammalian folate enzymes that have not yet been exploited as potential drug targets, and enzymes that function in the de novo folate-synthesis pathway of the parasite--a particularly attractive area of attack because of its absence from the mammalian host.     

Verification of a European corn borer (Lepidoptera: Crambidae) loss equation in the major corn production region of the northeastern United States

Field studies in Pennsylvania and Maryland were conducted during 2000, 2001, and 2002 to test the applicability of published yield loss relationships developed in central Pennsylvania for European corn borer, Ostrinia nubilalis (Hübner), management in warmer, longer season corn, Zea mays L., grain production regions of the northeastern United States. Both isoline hybrids and non-Bt lead hybrids were compared against Bacillus thuringiensis (Bt) hybrids to measure effects of the pest on yield. The European corn borer economic analysis model was used to make site-specific predictions of loss per European corn borer larva for comparison with measured yield loss per larva. Although the model did not predict loss per larva at a field level, it did predict loss at a regional level. The model predicted an overall percentage of yield loss per larva of 2.69+/-0.12% over the region, which was similar to the measured yield loss per larva of 2.66+/-0.59% for isoline hybrids and 3.08+/-0.51% for lead hybrids. The model, on average, provided a good prediction of percentage of yield loss per larva within the climatic zones of 1100-1700 degree-days (DD) (base threshold 12.5 degrees C). Our results suggest that the yield loss relationship developed in Central Pennsylvania, when matched to the timing of third instar second generation European corn borer stalk tunneling is adequate for major corn grain production zones of the northeast United States.     

Shortcomings of protein removal prior to high performance liquid chromatographic analysis-A case study using method development for BAY 11-7082

During the analytical method development for BAY 11-7082 ((E)-3-[4-methylphenylsulfonyl]-2-propenenitrile), using HPLC-MS-MS and HPLC-UV, we observed that the protein removal process (both ultrafiltration and precipitation method using organic solvents) prior to HPLC brought about a significant reduction in the concentration of this compound. The use of a structurally similar internal standard, BAY 11-7085 ((E)-3-[4-t-butylphenylsulfonyl]-2-propenenitrile), was not effective in compensating for the loss of analyte as the extent of reduction was different to that of the analyte. We present here a systematic investigation of this problem and a new validated method for the determination of BAY 11-7082.     

Spatial and bathymetric trends in Harpacticoida (Copepoda) community structure in the Northern Gulf of Mexico deep-sea

Harpacticoid copepod community structure was analyzed at 43 stations in the northern Gulf of Mexico deep-sea to test regional and bathymetric patterns of diversity in relation to environmental variables and topographic complexity of the continental slope. Depth, longitude, and proximity to the Florida Escarpment significantly affect average phylogenetic diversity, but basins and canyons do not. Multivariate analysis reveals a significant inverse relationship between diversity and POM flux, which is confirmed by significant region-scale depth and longitude differences. Although species richness declines linearly with increasing depth, the expected number of species (rarefraction) is maximized at approximately 1200 m, and average taxonomic and phylogenetic diversity continue to increase with depth, suggesting greater morphological or functional harpacticoid diversity with increasing depth. Most stations have unique species compositions, suggesting high regional (2200 species) and global (10⁵-10⁶ species) diversity by extrapolation. Therefore, processes maintaining harpacticoid diversity in the northern Gulf of Mexico deep-sea seem to rely on both small-scale dispersal and large-scale food supply mechanisms.     

Variation between freshwater and terrestrial fungal communities on decaying bamboo culms

Fungal communities on decaying culms of a bamboo host (Phyllostachys bambusoides) from freshwater and adjacent terrestrial habitats were identified. Collections were made at Xiao Bai Long Mountain, Yiliang, Yunnan, China in the winter and summer. In each collection, 100 similar-sized bamboo culms were collected, comprising 50 submerged samples from a stream and 50 terrestrial samples from adjacent riparian vegetation. A total of 82 fungal taxa were recorded from the samples, including 30 ascomycetes and 52 anamorphic fungi. The frequency of occurrence of these fungi were recorded and the Shannon–Weiner indices (H′) were applied to evaluate fungal diversity. The results showed that variation of the fungal diversity between the summer and winter collections was insignificant (0.2<p<0.5). Fungal diversity on submerged bamboo however, was significantly higher than that on terrestrial bamboo (p<0.001). Further findings were that: (1) some commonly recorded freshwater and terrestrial taxa were found in both habitats, but overall there were only 15 overlapping species between the two habitats; (2) the dominant species in each habitat were considerably different, and (3) only a few fungi were dominant, while most species were rare, being recorded only once or twice. Factors responsible for the distribution patterns and variations in composition of the fungal communities are discussed.     

Low-diversity fungal assemblage in an Antarctic Dry Valleys soil

The McMurdo Dry Valleys of Antarctica present extreme environmental challenges. Life is restricted to patchy occurrence of lichens, mosses and invertebrates, plus soil microbial communities. Fungi have been described in lichen symbioses but relatively little is known about the occurrence of free-living soil fungi in the Dry Valleys. A challenge in estimating fungal species richness has been the extent to which estimates based on either cultivation or environmental DNA reflect the active assemblage in cold-arid soils. Here, we describe analysis for inland Dry Valleys soil of environmental DNA and RNA (cDNA) to infer total and putative metabolically active assemblages, respectively, plus cultivation approaches using a variety of laboratory growth conditions. Environmental sequences indicated a highly restricted assemblage of just seven phylotypes that affiliated phylogenetically within two known genera, Helicodendron and Zalerion, plus previously unidentified fungal phylotypes. None of the commonly encountered molds and mitosporic genera recorded from maritime Antarctic locations were encountered. A striking difference was observed in the frequency of recovery for phylotypes between libraries. This suggests that both species richness and beta diversity estimates based on DNA libraries have the potential to misinform putatively active assemblages. Cultivation yielded a cold-tolerant Zalerion strain that affiliated with DNA and RNA library clones, and a psychrotrophic yeast (Debaryomyces hansenii), which was not detected using either culture-independent approach.     

Thermal acclimation,heat shock and photoperiod: Do these factors interplay in the adaptive responses of crab neuromuscular systems to temperature?

Evidence is reviewed demonstrating that the adaptive responses to temperature made by the walking leg neuromuscular system of crabs (Carcinus maenas) are in response to local temperature and not in response to hierarchical influences by the CNS and hormonal systems.     

Comparative population genetic analysis of bocaccio rockfish Sebastes paucispinis using anonymous and gene-associated simple sequence repeat loci

Comparative population genetic analyses of traditional and emergent molecular markers aid in determining appropriate use of new technologies. The bocaccio rockfish Sebastes paucispinis is a high gene-flow marine species off the west coast of North America that experienced strong population decline over the past 3 decades. We used 18 anonymous and 13 gene-associated simple sequence repeat (SSR) loci (expressed sequence tag [EST]-SSRs) to characterize range-wide population structure with temporal replicates. No F(ST)-outliers were detected using the LOSITAN program, suggesting that neither balancing nor divergent selection affected the loci surveyed. Consistent hierarchical structuring of populations by geography or year class was not detected regardless of marker class. The EST-SSRs were less variable than the anonymous SSRs, but no correlation between F(ST) and variation or marker class was observed. General linear model analysis showed that low EST-SSR variation was attributable to low mean repeat number. Comparative genomic analysis with Gasterosteus aculeatus, Takifugu rubripes, and Oryzias latipes showed consistently lower repeat number in EST-SSRs than SSR loci that were not in ESTs. Purifying selection likely imposed functional constraints on EST-SSRs resulting in low repeat numbers that affected diversity estimates but did not affect the observed pattern of population structure.     

Pineal photoreceptor cells are required for maintaining the circadian rhythms of behavioral visual sensitivity in zebrafish

In non-mammalian vertebrates, the pineal gland functions as the central pacemaker that regulates the circadian rhythms of animal behavior and physiology. We generated a transgenic zebrafish line [Tg(Gnat2:gal4-VP16/UAS:nfsB-mCherry)] in which the E. coli nitroreductase is expressed in pineal photoreceptor cells. In developing embryos and young adults, the transgene is expressed in both retinal and pineal photoreceptor cells. During aging, the expression of the transgene in retinal photoreceptor cells gradually diminishes. By 8 months of age, the Gnat2 promoter-driven nitroreductase is no longer expressed in retinal photoreceptor cells, but its expression in pineal photoreceptor cells persists. This provides a tool for selective ablation of pineal photoreceptor cells, i.e., by treatments with metronidazole. In the absence of pineal photoreceptor cells, the behavioral visual sensitivity of the fish remains unchanged; however, the circadian rhythms of rod and cone sensitivity are diminished. Brief light exposures restore the circadian rhythms of behavioral visual sensitivity. Together, the data suggest that retinal photoreceptor cells respond to environmental cues and are capable of entraining the circadian rhythms of visual sensitivity; however, they are insufficient for maintaining the rhythms. Cellular signals from the pineal photoreceptor cells may be required for maintaining the circadian rhythms of visual sensitivity.