The causes of spatial patterning of mounds of a fungus-cultivating termite: results from nearest-neighbour analysis and ecological studies

Little is known about processes regulating population dynamics in termites. We investigated the distribution of mound-colonies of the fungus-cultivating termite Macrotermes bellicosus (Smeathman) in two habitats in the Comoé National Park (Côte d'Ivoire) with nearest-neighbour analysis differentiating between different age classes. These results were compared with ecological data on processes influencing population dynamics. High mound densities were recorded in shrub savannah while only a few mounds were found in gallery forest. Mounds were distributed randomly in both habitats when all mounds were considered together, and when inhabited and uninhabited mounds were treated separately. However, distinctive non-random patterns were revealed in the savannah when we distinguished between different age classes. Small, young colonies were aggregated when they coexisted with larger, older colonies, which were more regularly distributed. This indicates that the distribution of older colonies is influenced by intraspecific competition whereas that of younger colonies is influenced by opposing factors that lead to aggregation. This is in accordance with ecological data. Food is a limiting resource for large colonies, while patchily distributed appropriate microclimatic conditions seem to be more important for young colonies. Colonies that had formerly coexisted (i.e. living colonies and recently dead colonies) showed aggregated, random and regular distribution patterns, suggesting several causes of mortality. Colonies that had never had contact with each other were randomly distributed and no specific regulation mechanism was implicated. These results show that different age classes seem to be regulated by different processes and that separation between age classes is necessary to reveal indicative spatial patterns in nearest-neighbour analysis.     

Cytotoxic activity of nucleoside diphosphate kinase secreted from Mycobacterium tuberculosis.

Pathogenicity of Mycobacterium tuberculosis is closely related to its ability to survive and replicate in the hostile environment of macrophages. For some pathogenic bacteria, secretion of ATP-utilizing enzymes into the extracellular environment aids in pathogen survival via P2Z receptor-mediated, ATP-induced death of infected macrophages. A component of these enzymes is nucleoside diphosphate kinase (Ndk). The ndk gene was cloned from M. tuberculosis H37Rv and expressed in Escherichia coli. Ndk was secreted into the culture medium by M. tuberculosis, as determined by enzymatic activity and Western blotting. Purified Ndk enhanced ATP-induced macrophage cell death, as assayed by the release of [14C]adenine. A catalytic mutant of Ndk failed to enhance ATP-induced macrophage cell death, and periodate-oxidized ATP (oATP), an irreversible inhibitor of P2Z receptor, blocked ATP/Ndk-induced cell death. Purified Ndk was also found to be autophosphorylated with broad specificity for all nucleotides. Conversion of His117-->Gln, which is part of the nucleotide-binding site, abolished autophosphorylation. Purified Ndk also showed GTPase activity. Collectively, these results indicate that secreted Ndk of M. tuberculosis acts as a cytotoxic factor for macrophages, which may help in dissemination of the bacilli and evasion of the immune system.     

Antioxidant protein 2 prevents methemoglobin formation in erythrocyte hemolysates.

Antioxidant protein 2 (AOP2) is a member of a family of thiol-specific antioxidants, recently renamed peroxiredoxins, that evolved as part of an elaborate system to counteract and control detrimental effects of oxygen radicals. AOP2 is found in endothelial cells, erythrocytes, monocytes, T and B cells, but not in granulocytes. AOP2 was found solely in the cytoplasm and was not associated with the nuclear or membrane fractions; neither was it detectable in plasma. Further experiments focused on the function of AOP2 in erythrocytes where it is closely associated with the hemoglobin complex, particularly with the heme. An investigation of the mechanism of this interaction demonstrated that the conserved cysteine-47 in AOP2 seems to play a role in AOP2-heme interactions. Recombinant AOP2 prevented induced as well as noninduced methemoglobin formation in erythrocyte hemolysates, indicating its antioxidant properties. We conclude that AOP2 is part of a sophisticated system developed to protect and support erythrocytes in their many physiological functions.     

A Sensitive and Inexpensive Yeast Bioassay for the Mycotoxin Zearalenone and Other Compounds with Estrogenic Activity

Zearalenone (ZON) is a nonsteroidal estrogenic mycotoxin produced by plant-pathogenic species of Fusarium. As a consequence of infection with Fusarium culmorum and Fusarium graminearum, ZON can be found in cereals and derived food products. Since ZON is suspected to be a cause of human disease, including premature puberty syndrome, as well as hyperestrogenism in farm animals, several countries have established monitoring programs and guidelines for ZON levels in grain intended for human consumption and animal feed. We developed a low-cost method for monitoring ZON contamination in grain based on a sensitive yeast bioassay. The indicator Saccharomyces cerevisiae strain YZRM7 is unable to grow unless an engineered pyrimidine biosynthetic gene is activated by the expressed human estrogen receptor in the presence of exogenous estrogenic substances. Deletion of the genes encoding ATP-binding cassette (ABC) transporters Pdr5p and Snq2p increases net ZON uptake synergistically. Less than 1 μg of ZON per liter of medium is sufficient to allow growth of the indicator strain. To prevent interference with pyrimidines potentially present in biological samples, we also disrupted the genes FUR1 and URK1, blocking the pyrimidine salvage pathway. The bioassay strain YZRM7 allows qualitative detection and quantification of total estrogenic activity in cereal extracts without requiring further cleanup steps. Its high sensitivity makes this assay suitable for low-cost monitoring of contamination of maize and small grain cereals with estrogenic Fusarium mycotxins.     

Submerged aquatic vegetation correlations with depth and light attenuating materials in a shallow subtropical lake

A 3-year study was done to quantify the biomass of submerged aquatic vegetation (SAV) and its relationship with environmental attributes in Lake Okeechobee, the largest lake in the southeastern United States. Plants were sampled on 21 occasions at sites located along 15 fixed transects around the shoreline, giving rise to 721 observations of SAV species (Chara spp., Vallisneria americana, Hydrilla verticillata, Potamogeton illinoinensis) dry weight biomass. Environmental sampling focused on factors that attenuate light, including phytoplankton chlorophyll a (chl a), total suspended solids (TSS), non-volatile suspended solids (NVSS) and color. Depth and Secchi transparency also were measured. Based on regression analysis, NVSS was considerably more important in attenuating light than chl a or color. Total biomass of SAV varied from 0 to 271 g dw m^–2, with a mean of 4.7 g dw m^–2, and strong dominance by Chara. The SAV biomass was lower than average for Florida lakes, and may reflect the influence of suspended solids on underwater irradiance, as well as high water level in the late 1990s. Dense SAV was found only where depth was < 2 m and TSS < 20–30 mg l^–1. At locations where high biomass of SAV occurred, the plants may have influenced water quality, because concentrations of TSS, NVSS, and chl a were 2–3 fold lower than at sites with no plants. The potential effects of SAV also were apparent at a regional scale. The shoreline region of the lake displayed a pattern of rising and falling chl a and NVSS with water depth. This occurred both at sites with and without plants, suggesting that it may be driven by physical processes, such as water circulation patterns, which are influenced by depth. However, the pattern was dampened at sites with SAV, indicating a potential to influence these attributes of water quality.     

Localization of laminin alpha4-chain in developing and adult human tissues.

Recent studies suggest important functions for laminin-8 (Ln-8; alpha4beta1gamma1) in vascular and blood cell biology, but its distribution in human tissues has remained elusive. We have raised a monoclonal antibody (MAb) FC10, and by enzyme-linked immunoassay (EIA) and Western blotting techniques we show that it recognizes the human Ln alpha4-chain. Immunoreactivity for the Ln alpha4-chain was localized in tissues of mesodermal origin, such as basement membranes (BMs) of endothelia, adipocytes, and skeletal, smooth, and cardiac muscle cells. In addition, the Ln alpha4-chain was found in regions of some epithelial BMs, including epidermis, salivary glands, pancreas, esophageal and gastric glands, intestinal crypts, and some renal medullary tubules. Developmental differences in the distribution of Ln alpha4-chain were detected in skeletal muscle, walls of vessels, and intestinal crypts. Ln alpha4- and Ln alpha2-chains co-localized in BMs of fetal skeletal muscle cells and in some epithelial BMs, e.g., in gastric glands and acini of pancreas. Cultured human pulmonary artery endothelial (HPAE) cells produced Ln alpha4-chain as M(r) 180,000 and 200,000 doublet and rapidly deposited it to the growth substratum. In cell-free extracellular matrices of human kidney and lung, Ln alpha4-chain was found as M(r) 180,000 protein.     

Transmission of Escherichia coli O157:H7 from contaminated manure and irrigation water to lettuce plant tissue and its subsequent internalization.

The transmission of Escherichia coli O157:H7 from manure-contaminated soil and irrigation water to lettuce plants was demonstrated using laser scanning confocal microscopy, epifluorescence microscopy, and recovery of viable cells from the inner tissues of plants. E. coli O157:H7 migrated to internal locations in plant tissue and was thus protected from the action of sanitizing agents by virtue of its inaccessibility. Experiments demonstrate that E. coli O157:H7 can enter the lettuce plant through the root system and migrate throughout the edible portion of the plant.     

Late Serpukhovian (Namurian A) microfacies and carbonate microfossils from the Carboniferous of Nötsch (Austria)

Summary The Carboniferous of N?tsch (Austria), divided into Erlachgraben, Badstub and N?tsch Formations, is composed of a thick sequence of dominantly siliciclastic deepsea sediments. Intercalated marly and silty limestones in the upper Erlachgraben Formation consist of bioclastic wackestones and algal wackestones/packstones which contain a diverse fossil assemblage of formainifers, algae and pseudo-algae. These microfossils are accurately described and documented, and three species of algae are established:Principia fluegeli n. sp.,Paraepimastopora noetschensis n. sp., andNanopora pseudofragilissima n. sp. Based on the occurrence of both important species of the foraminifers Lasiodiscoidea (Howchinia gibba andEolasiodiscus dilatatus), and also on the presence ofEndothyranopsis plana, of the lastEarlandia ex. gr.vulgaris and of the firstEostaffella exp gr.postmosquensis, the upper Erlachgraben Formation is dated as late Serpukhovian (goniatite biozone E 2 of the Namurian A; Arnsbergian stage, corresponding to the Zapaltyubinsky of the standard Russian sequence; foraminiferal biozones 18 or Cf 7 of Belgium, or Cf 16 of the Donbass). Compared to the Pyrenees and the Donbass region, the algal flora of the Carboniferous of N?tsch seems to be relatively endemic. Algae and foraminifers originally inhabited a shallow carbonate ramp and were transported and redeposited in a deep-water environment by gravity flows. The formainifers most probably migrated from the Donbass region along the shelf of a narrow seaway to N?tsch.     

A critical epitope for substrate recognition by the nucleosome remodeling ATPase ISWI

The ATPase ISWI is the catalytic core of several nucleosome remodeling complexes, which are able to alter histone–DNA interactions within nucleosomes such that the sliding of histone octamers on DNA is facilitated. Dynamic nucleosome repositioning may be involved in the assembly of chromatin with regularly spaced nucleosomes and accessible regulatory sequence elements. The mechanism that underlies nucleosome sliding is largely unresolved. We recently discovered that the N-terminal ‘tail’ of histone H4 is critical for nucleosome remodeling by ISWI. If deleted, nucleosomes are no longer recognized as substrates and do not stimulate the ATPase activity of ISWI. We show here that the H4 tail is part of a more complex recognition epitope which is destroyed by grafting the H4 N-terminus onto other histones. We mapped the H4 tail requirement to a hydrophilic patch consisting of the amino acids R₁₇H₁₈R₁₉ localized at the base of the tail. These residues have been shown earlier to contact nucleosomal DNA, suggesting that ISWI recognizes an ‘epitope’ consisting of the DNA-bound H4 tail. Consistent with this hypothesis, the ISWI ATPase is stimulated by isolated H4 tail peptides ISWI only in the presence of DNA. Acetylation of the adjacent K₁₂ and K₁₆ residues impairs substrate recognition by ISWI.     

Methylation at arginine 17 of histone H3 is linked to gene activation

The nuclear hormone receptor co-activator CARM1 has the potential to methylate histone H3 at arginine residues in vitro. The methyltransferase activity of CARM1 is necessary for its co-activator functions in transient transfection assays. However, the role of this methyltransferase in vivo is unclear, given that methylation of arginines is not easily detectable on histones. We have raised an antibody that specifically recognizes methylated arginine 17 (R17) of histone H3, the major site of methylation by CARM1. Using this antibody we show that methylated R17 exists in vivo. Chromatin immunoprecipitation analysis shows that R17 methylation on histone H3 is dramatically upregulated when the estrogen receptor-regulated pS2 gene is activated. Coincident with the appearance of methylated R17, CARM1 is found associated with the histones on the pS2 gene. Together these results demonstrate that CARM1 is recruited to an active promoter and that CARM1-mediated R17 methylation on histone H3 takes place in vivo during this active state.