A novel zinc bis(thiosemicarbazone) complex for live cell imaging

Fluorescent zinc complexes have recently attracted a lot of interest owing to their vast applications in cellular imaging. We report the synthesis as well as physical, chemical and biological studies of a novel zinc glyoxalbis(4-methyl-4-phenyl-3-thiosemicarbazone), [Zn(GTSC)]₃, complex. As compared with the well-studied zinc biacetylbis(4-methyl-3-thiosemicarbazone), Zn(ATSM), complex, which was used as a reference, [Zn(GTSC)]₃ had 2.5-fold higher fluorescence. When cellular fluorescence was measured using flow cytometry, we observed that [Zn(GTSC)]₃ had 3.4-fold to 12-fold higher fluorescence than Zn(ATSM) in various cell lines (n = 9) of different tissue origin. Confocal fluorescence microscopy results showed that [Zn(GTSC)]₃ appeared to have a nuclear localization within 30 min of addition to MCF7 cells. Moreover, [Zn(GTSC)]₃ showed minimal cytotoxicity compared with Zn(ATSM), suggesting that [Zn(GTSC)]₃ may be less deleterious to cells when used as an imaging agent. Our data suggest that the novel [Zn(GTSC)]₃ complex can potentially serve as a biocompatible fluorescent imaging agent for live cells.     

Mass-spectrometric characterization of cisplatin binding sites on native and denatured ubiquitin

Because interactions between cisplatin and plasma proteins contribute to drug efficacy and side effects, it is important to understand both the binding sites of cisplatin on the proteins and the formation of protein–cisplatin adducts. Previous results suggest that cisplatin preferentially binds to residues on the protein surface. The present work employed electrospray ionization mass spectrometry (MS) to identify such sites on both native and denatured ubiquitin (Ub). Fourier transform (FT) MS and tandem MS (MS/MS and MS³) enable analysis of Ub–cisplatin adduct digests to locate specific cisplatin binding sites. Results indicate that there are three such binding sites, i.e., M1, T12 and T14, and D32, on native Ub. The intensity of the relevant peaks in the FT-MS spectrum of the native Ub adduct digest demonstrates that residues T12 and T14 comprise the primary cisplatin binding site under the native conditions rather than residue M1 as reported in previous research studies. It is found in the present work, however, that M1 is the primary binding site on denatured Ub. Comparison of cisplatin binding sites on native and denatured Ub in this research demonstrates that the conformation of a protein significantly influences the preference of cisplatin for specific binding sites.     

Purification and characterization of a mucin-binding mycelial lectin from <Emphasis Type=Italic>Aspergillus nidulans</Emphasis> with potent mitogenic activity

Mucin-specific lectin from mycelium of Aspergillus nidulans was purified using anion exchange and gel filtration chromatographic techniques with an overall recovery of 32% and 21.97-fold purification. The purified lectin migrated as a single band in SDS–PAGE with an apparent molecular mass of 34 kDa. Carbohydrate analysis revealed that it is a glycoprotein with total sugar content of 2.54%. Optimal agglutination was observed when serially diluted lectin was incubated with human type O erythrocyte suspension at pH 7.0–8.0 and temperature 20–30°C. Lectin was found to be completely stable within pH 5.0–8.0 and temperature at or below 40°C. Demetallization by extensive dialysis against EDTA did not alter its haemagglutination activity. Lectin activity was reduced to half after 24 h incubation with urea and thiourea, with no such effect of guanidine HCl. The lectin showed potent mitogenic response towards mouse splenocytes, attaining a maximum at 200 μg/ml as compared to untreated control cells. Mitogenic lectins are invaluable tools to assess the functioning of immune cells. None of the microfungal lectin has yet been investigated for mitogenic activity. This is the first report on mitogenic activity of lectin from Aspergillus sp.     

Production of phytohormones by root-associated saprophytic actinomycetes isolated from the actinorhizal plant Ochetophila trinervis

The aim of the present study was to evaluate phytohormone production by symbiotic and saprophytic actinomycetes isolated from the actinorhizal plant Ochetophila trinervis which had previously proved to stimulate nodulation by Frankia. Three saprophytic strains out of 122, isolated from the rhizosphere of this plant with multiple enzymatic activities were selected for plant growth experiments in pots: Streptomyces sp. (BCRU-MM40), Actinoplanes sp. (BCRU-ME3) and Micromonospora sp. (BCRU-MM18). For experiments, the symbiotic N₂-fixing strain Frankia (BCU110501), isolated from nodules of the same actinorhizal plant was used. Phytohormone production was evaluated in supernatant of non-inoculated and inoculated culture media in exponential growth phase. Indole 3-acetic acid (IAA) and gibberellic acid (GA₃) were analyzed by gas chromatography-mass spectrometry (GC–MS), while zeatine (Z) production was determined by gas chromatography-flame ionization detector and high performance liquid chromatography (HPLC fluorescent and UV). The levels of the three phytohormones produced by the saprophytic rhizoactinomycetes were higher than that produced by the symbiotic Frankia strain. Zeatine biosynthesis was higher (μg ml⁻¹) than IAA and GA₃ (ng ml⁻¹), and Micromonospora strain produced the highest levels of these phytohormones. Although O. trinervis has been shown to be intercellularly infected by Frankia without mediation of root hair deformation, when plants were co-inoculated with actinomycetes’ culture, some root hair deformation was observed. This is the first report on identification of IAA, GA₃ and Z in saprophytic actinomycetes and their potential role in plant–microbe interaction.     

The study of the effects of carbon dioxide-induced elevation of hydrogen peroxide toxicity on microbes as a novel tool for water purification

The supercritical concentration of CO₂ (SCCO₂) and a high concentration (3.0%) of molecules of hydrogen peroxide (H₂O₂) are currently being used as antiseptic and antibacterial agents. The fact that low concentrations of CO₂ have an activation effect on functional activity of microbes allows us to predict that CO₂ could elevate the toxic effect of H₂O₂ on cells. To check this hypothesis the dependency of the toxic effect of H₂O₂ on wild type of Escherichia coli K-12 on soluble concentration of CO₂ in culture media was studied. The obtained data show that culture media enriched with CO₂ leads to the increase of toxic effect of H₂O₂ on microbes at both cases when pH is constant and when it changes. So CO₂ in non-supercritical concentration could elevate the toxic effect of H₂O₂ on microbes by the activation of the metabolic processes in microbes. During the experiments we used classical microbiological methods (indirect viable cell counts or counting colony forming units (CFUs)), as well as the method of measuring hydrogen peroxide content in aqueous solution by means of enhanced chemiluminescence method in a peroxidase-luminol-p-iodophenol system. This discovery is concerning to use CO₂/H₂O₂ combination system, which could have implication in the inhibition of growth of microbes in water and the microbiological monitoring of water could provide valuable information for managing the health of exhibition of aqua ecosystems.     

Biodiversity and conservation of insects and other invertebrates

The thematic issue of Biodiversity and Conservation devoted to the biodiversity and conservation of insects and other invertebrates is introduced. The issue comprises 23 original research papers covering diverse habitats from forests to grasslands, ponds and rivers to coasts, and the tropics to boreal regions. Amongst the organisms discussed are ants, bees, beetles, butterflies, crabs, microgastropods, millipedes, spiders, and weevils. Some of the difficulties of conserving the most species-rich groups of eukaryotes, in the face of ignorance as to their identities and positions in ecological processes, are noted and the precautionary principle is seen as a pragmatic and responsible approach.     

The influence of temporal variation on relationships between ecosystem services

A growing literature aims to identify areas of congruence in the provision of multiple ecosystem goods and services. However, little attention has been paid to the effect that temporal variation in the provision of such services may have on understanding of these relationships. Due to a lack of temporally and spatially replicated monitoring surveys, such relationships are often assessed using data from disparate time periods. Utilising temporally replicated data for indices of freshwater quality and agricultural production we demonstrate that through time the biophysical values of ecosystem services may vary in a spatially non-uniform way. This can lead to differing conclusions being reached about the strength of relationships between services, which in turn has implications for the prioritisation of areas for management of multiple services. We present this first analysis to illustrate the effect that the use of such temporally disparate datasets may have, and to highlight the need for further research to assess under what circumstances temporal variation of this sort will have the greatest impact.     

Great-granny’s Garden: a living archive and a sensory garden

Since 2003, the Botanical Garden in Oslo has been involved in a project coordinated by the Norwegian Genetic Resource Centre. The wide range of work supervised by this centre includes conservation of ornamental plants. Our garden has been responsible for the registration and collecting of ornamentals in Southeast-Norway and has a special responsibility for the conservation of Paeonia species and cultivars. As a result of the project, Great-granny’s Garden was opened to the public in 2008. It has two objectives. Firstly, it shall be a living archive of Norway’s horticultural heritage. Although proven hardy, easy to grow, and long-lived, old varieties of traditional ornamentals are rapidly disappearing. We aim to keep these old-fashioned varieties for sustainable use in future horticulture and encourage people to use them in present day gardening, both in new gardens and in the restoration of old ones. Secondly, the garden is designed as a sensory garden for people with dementia, in cooperation with Oslo’s Resource Centre for Dementia and Psychiatric Care of the Elderly. It is enclosed by a picked fence and by shrubs, offers rest on several benches, and has a paved and easy to follow round-walk among traditional garden elements and plants with a lush variety of colours, forms, and scents. A sensory garden stimulates many senses, evokes pleasant emotions, brings out long-forgotten memories, and stimulates communication. Sensory gardens are therefore considered an important tool in the therapy of dementia.     

Non-native and translocated fish species in Serbia and their impact on the native ichthyofauna

A total of 22 fish species have been introduced into the inland waters of Serbia, either intentionally or accidentally. This paper provides a summary of data concerning time and reason of introduction, mode of expansion, degree of acclimatization, impact on native fish and estimated area of recent distribution. Four of the non-native fish species currently occupy more than 51% of Serbian territory while 5 of them occupy between 21–50% of territory. This paper reviews impacts of introduced freshwater fish in Serbia based on collected data.