Copper at the Fungal Pathogen-Host Axis

Fungal infections are responsible for millions of human deaths annually. Copper, an essential but toxic trace element, plays an important role at the host-pathogen axis during infection. In this review, we describe how the host uses either Cu compartmentalization within innate immune cells or Cu sequestration in other infected host niches such as in the brain to combat fungal infections. We explore Cu toxicity mechanisms and the Cu homeostasis machinery that fungal pathogens bring into play to succeed in establishing an infection. Finally, we address recent approaches that manipulate Cu-dependent processes at the host-pathogen axis for antifungal drug development.     

Involvement of Microorganisms in the Formation of Carbonate Speleothems in the Cervo Cave (L'Aquila-Italy)

Much is known about the bacterial precipitation of carbonate rocks, but comparatively little is known about the involvement of microbes in the formation of secondary mineral structures in caves. We hypothesized that bacteria isolated from calcareous stalactites, which are able to mediate CaCO₃ precipitation in vitro, play a role in the formation of carbonate speleothems. We collected numerous cultivable calcifying bacteria from calcareous speleothems from Cervo cave, implying that their presence was not occasional. The relative abundance of calcifying bacteria among total cultivable microflora was found to be related to the calcifying activity in the stalactites. We also determined the δ ¹³C and δ ¹⁸ O values of the Cervo cave speleothems from which bacteria were isolated and of the carbonates obtained in vitro to determine whether bacteria were indeed involved in the formation of secondary mineral structures. We identified three groups of biological carbonates produced in vitro at 11°C on the basis of their carbon isotopic composition: carbonates with δ ¹³C values (a) slightly more positive, (b) more negative, and (c) much more negative than those of the stalactite carbonates. The carbonates belonging to the first group, characterized by the most similar δ ¹³C values to stalactites, were produced by the most abundant strains. Most of calcifying isolates belonged to the genus Kocuria. Scanning electron microscopy showed that dominant morphologies of the bioliths were sherulithic with fibrous radiated interiors. We suggest a mechanism of carbonate crystal formation by bacteria.     

A Model Sheet Mineral System to Study Fungal Bioweathering of Mica

The general objective of this research was to examine fungal interactions with silicate minerals within the context of their roles in bioweathering. To achieve this, we used muscovite, a phyllosilicate mineral (KAl₂[(OH)₂|AlSi₃O₁₀]), in the form of a mineral sheet model system for ease of experimental manipulation and microscopic examination. It was found that test fungal species successfully colonized and degraded the surface of muscovite sheets in both laboratory and field experiments. After colonization by the common soil fungus Aspergillus niger, a network of hyphae covered the surface of the muscovite, and mineral dissolution or degradation was clearly evidenced by a network of fungal “footprints” that reflected coverage by the mycelium. For natural soil incubations, microorganisms associated with muscovite sheet material included biofilms of fungi and bacteria on the surface, while mineral encrustation or adhesion to microbial structures was also observed. Our results show that muscovite sheet is a good model mineral system for examination of microbial colonization and degradation, and this was demonstrated using laboratory and field systems, providing more evidence for the bioweathering significance of fungal activities in the context of silicate degradation and soil formation and development. The approach is also clearly applicable to other rock and mineral-based substrates and a variety of free-living and symbiotic microbial systems.     

The Role of Copper in Protein Foams

Chefs have known that whipping egg white proteins (EWP) in a copper bowl will improve foam stability. The improved stability is attributed to a copper–conalbumin complex or alteration of sulfhydryl reactivity. Whey proteins bind copper and show copper-induced changes in disulfide bonds; therefore, they may also be responsive to whipping in a copper bowl. EWP and whey protein isolate (WPI) solutions were whipped in the presence of 1 mM CuSO₄ or in a copper bowl with and without sugar followed by overrun and yield stress measurements and angel food cake formation. Dilational elasticity and surface tension were also measured for WPI solutions. Whipping in a copper bowl or adding 1 mM CuSO₄ significantly improved stability of EWP foams while having no effect on WPI foams. Copper caused disulfide-linked dimer formation of β-lactoglobulin and decreased dilational elasticity and surface tension, but these modifications were insufficient to change the bulk properties of foams. The addition of 10 mM CuSO₄ to WPI solutions was sufficient to increase foam stability to levels similar to EWP; however, the more stable foams formed less stable cakes. It was concluded that the effect of whipping in a copper bowl on foam properties is mainly dependent on the specific proteins forming the foam. Paper no. FSR-07-29 of the Journal Series of the Department of Food Science, North Carolina State University, Raleigh, NC 27695-7624. Presented at the 2nd International Symposium: Delivery of Functionality in Complex Food Systems: Physically-inspired Approaches From Nanoscale to Microscale, October 8–10, 2007.     

用铁蛋白合成纳米粒子的研究进展

铁蛋白是一种生物储铁蛋白 ,其储存铁的特性和储存铁在生物体内的特殊形态 ,通过相对温和、简便的生物、化学、物理过程 ,可合成多种具有特异的力、热、光、电、磁等特性的纳米粒子 ,并可用来构建纳米级的分子器件     

真菌介导纳米银生物合成的研究进展

纳米银(Silver nanoparticles,AgNPs)是指三维结构中至少有一维在1-100 nm范围内的金属银材料,在光学、电学、催化和医药等领域得到了广泛应用.利用真菌生物合成AgNPs相比于传统的化学、物理合成方法,具有反应条件温和、绿色低毒、环境友好等优势备受关注.综述了近年来合成AgNPs真菌种类的研究...     

Saposin Lipid Nanoparticles: A Highly Versatile and Modular Tool for Membrane Protein Research

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Transformation of Copper Oxychloride Fungicide into Copper Oxalate by Tolerant Fungi and the Effect of Nitrogen Source on Tolerance

Aspergillus niger and Penicillium chrysogenum were able to grow on Czapek Dox medium amended with elevated concentrations [up to 500 ppm active ingredient (ai)] of the fungicide copper oxychloride. Solubilization of the fungicide in solid medium was evident by the appearance of a clear (halo) zone underneath and around the growing colonies. The halo formed with A. niger, grown on fungicide-containing nitrate nitrogen medium, was found subsequently to enclose concentric rings of newly crystalline precipitate. These crystals were extracted, examined by scanning electron microscopy and IR, and identified as copper oxalate. The supplemented nitrogen source to the medium greatly affected both fungicide solubilization and fungal tolerance. Ratios of fungicide solubilization rate (R(S)) in relation to the colony growth rate (R(G)) were significantly higher on ammonium than on nitrate nitrogen medium for both fungal strains. Growth ratios (the colony extension rate in the presence of a given concentration of the fungicide in relation to the control colony growth rate) of A. niger were markedly lower on ammonium than on nitrate nitrogen medium. The cellular copper contents, taken up from the fungicide, and the medium titratable acidity were higher in ammonium than in nitrate medium for both fungi. These results suggested fungal possession of variable tolerance mechanisms to this fungicide by complexation and/or precipitation of copper in the medium. Additionally, this work emphasizes the activity of fungi in transformation of insoluble inorganic metal-containing fungicides into insoluble organic metal compounds, which has a potentiality in metal cycling in biogeochemical and environmental context.     

Biomineralization of Fungal Hyphae with Calcite (CaCO3) and Calcium Oxalate Mono- and Dihydrate in Carboniferous Limestone Microcosms

The formation of biogenic fabrics in limestone by two fungi, Serpula himantioides and a polymorphic fungal isolate from limestone identified as a Cephalotrichum (syn. Doratomyces) sp., was investigated. The fungal cultures were grown in laboratory microcosms consisting of Carboniferous limestone and after 21 d incubation at 25°C, biomineralization of fungal filaments was observed. Environmental electron scanning microscopy (ESEM) and X-ray micro-analysis (EDXA) of crystalline precipitates on the hyphae of S. himantioides demonstrated that the secondary crystals exhibited different crystalline forms but were similar in elemental composition to the original limestone. Powder X-ray diffraction (XRD) of crystalline precipitates showed they were composed of a mixture of calcite (CaCO ₃ ) and calcium oxalate monohydrate (CaC ₂ O ₄ · H ₂ O). Analysis of crystals precipitated on the hyphae of the limestone isolate, using ESEM and EDXA, showed that the crystals exhibited similar morphological characteristics and elemental composition to the original limestone. XRD showed that they were composed solely of calcite (CaCO ₃ ) or of calcite with some calcium oxalate dihydrate (CaC ₂ O ₄ · 2H ₂ O). These results provide direct experimental evidence for the precipitation of calcite (CaCO ₃ ) and also secondary mycogenic minerals, on fungal hyphae in low nutrient calcareous environments, and suggest that fungi may play a wider role in the biogeochemical carbon cycle than has previously been appreciated.     

微生物铁蛋白的研究进展

铁蛋白作为一种重要的铁储存蛋白,在不同的微生物体中普遍存在.通过对典型的微生物铁蛋白分子(FTN)的结构及其功能的归纳分析发现,铁蛋白依赖其独特的结构特点,在铁的补充、转运、氧化、成核和储存中扮演着重要作用,也对生物体内的多种生物化学反应影响显著.同时借助基因工程技术对铁蛋白进行相应的分子改造,增加了其作为纳米载体的应...     

Discrete Nanoparticles of Ruta Graveolens Induces the Bacterial and Fungal Biofilm Inhibition

Abstract

Ruta graveolens silver nanoparticles (AgNPs) showed the color change within 30 min and characterized using UV–visible spectra, Fourier Transform Infrared (FTIR), X-ray Diffraction (XRD) and Transmission Electron Microscopy (TEM). UV–visible spectrum of R. graveolens AgNPs showed the sharp peak at the wavelength of 440–560 nm. XRD patterns confirmed that crystalline nature of R. graveolens AgNPs and FTIR results revealed that phytochemical reaction of these R. graveolens is responsible for the synthesis of AgNPs. TEM results showed the size of the R. graveolens AgNPs around 30–50 nm with spherical and triangular nature. Further, the antibacterial and antibiofilm activity of R. graveolens AgNPs showed the effective inhibitory activity against clinically important Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. Our findings suggest that R. graveolens AgNPs can be exploited toward the development of potential antibacterial agents for various biomedical and environmental applications.     

Protein chips for high-throughput doping screening in athletes

The biological significance of protein interactions, their method of generation and reliability is briefly reviewed. Protein interaction networks adopt a scale-free topology that explains their error tolerance or vulnerability, depending on whether hubs or peripheral proteins are attacked. Networks also allow the prediction of protein function from their interaction partners and therefore, the formulation of analytical hypotheses. Comparative network analysis predicts interactions for distantly related species based on conserved interactions, even if sequences are only weakly conserved. Finally, the medical relevance of protein interaction analysis is discussed and the necessity for data integration is emphasized.     

Nonreductive Biomineralization of Uranium(VI) Phosphate Via Microbial Phosphatase Activity in Anaerobic Conditions

The remediation of uranium from soils and groundwater at Department of Energy (DOE) sites across the United States represents a major environmental issue, and bioremediation has exhibited great potential as a strategy to immobilize U in the subsurface. The bioreduction of U(VI) to insoluble U(IV) uraninite has been proposed to be an effective bioremediation process in anaerobic conditions. However, high concentrations of nitrate and low pH found in some contaminated areas have been shown to limit the efficiency of microbial reduction of uranium. In the present study, nonreductive uranium biomineralization promoted by microbial phosphatase activity was investigated in anaerobic conditions in the presence of high nitrate and low pH as an alternative approach to the bioreduction of U(VI). A facultative anaerobe, Rahnella sp. Y9602, isolated from soils at DOE's Oak Ridge Field Research Center (ORFRC), was able to respire anaerobically on nitrate as a terminal electron acceptor in the presence of glycerol-3-phosphate (G3P) as the sole carbon and phosphorus source and hydrolyzed sufficient phosphate to precipitate 95% total uranium after 120 hours in synthetic groundwater at pH 5.5. Synchrotron X-ray diffraction and X-ray absorption spectroscopy identified the mineral formed as chernikovite, a U(VI) autunite-type mineral. The results of this study suggest that in contaminated subsurfaces, such as at the ORFRC, where high concentrations of nitrate and low pH may limit uranium bioreduction, the biomineralization of U(VI) phosphate minerals may be a more attractive approach for in situ remediation providing that a source of organophosphate is supplied for bioremediation.     

Protein microarray: A key approach of proteomics

Proteins are vital parts of living organisms, for they consist of the main components of physiological metabolic pathways. The expression and modification status of proteins in cells are time- and space-dependent based upon functions as well as responses to stress. A subject aiming at understanding of protein status under certain physiological condition in a large scale, termed proteomics, is fundamentally important in life science. Although the proteomic investigation mainly relies on electrophoresis, liquid chromatography, and mass spectrometry, protein microarray is emerging as one of new techniques, and its power is being recognized in the field recently. Herein, we highlight progress in protein microarray, both its generation and application.     

The Role of Histidine Residues in the Non-Enzyme Covalent Attachment of Glucose and Ascorbic Acid to Protein

Copper ions have been suggested to play a role in the non-covalent glycosylation (glycation) of proteins via transition metal-catalysed oxidations. We have further investigated “autoxidative glycosylation” by comparison of the behaviour of dog and bovine serum albumin with respect to the oxidative reactions of glucose and ascorbate. The proteins possess similar numbers of total amino residues available for glucose attachment but dog serum albumin contains fewer histidine groups and also lacks a high affinity copper-binding site. We find that the higher copper-binding capacity of bovine serum albumin is reflected in a lower rate of ascorbate oxidation as well as less protein oxidative damage than is the case for dog serum albumin. We also observe that modification of bovine serum albumin histidine groups by diethylpyrocarbonate enhances ascorbate-mediated protein fluorophore formation.     

The Role of the Yolk Sac in Copper Metabolism during Rat Embryogenesis

Using the immunoblotting method, the synthesis of two copper-transporting P1-type ATPases, ATP7A (a candidate for the product of the Menkes disease gene) and ATP7B (a presumed product of the Wilson disease gene), in the yolk sac cells of rat embryos at days 11 and 20 of embryogenesis was demonstrated. Concomitantly, yolk sac cells produce ceruloplasmin, a soluble copper-transporting glycoprotein, a proportion of which in secreted proteins progressively diminishes, attaining 5.2% at day 11 and 3.1% at day 20 of development. At different stages of embryogenesis, yolk sac cells synthesize two molecular forms of [¹⁴C]-ceruloplasmin, one of which is secreted towards the embryo, whereas the other, towards the decidual membrane. Two forms of ceruloplasmin secreted in polar directions differ in the rate of secretion. The role of the yolk sac as a key organ controlling the delivery and secretion of copper in the embryo during the postimplantation period is discussed.