Magnetic quantum dots in biotechnology – synthesis and applications

Quantum dots (QDs) have great promise in biological imaging, and as this promise is realized, there has been increasing interest in combining the benefits of QDs with those of other materials to yield composites with multifunctional properties. One of the most common materials combined with QDs is magnetic materials, either as ions (e.g. gadolinium) or as nanoparticles (e.g. superparamagnetic iron oxide nanoparticles, SPIONs). The fluorescent property of the QDs permits visualization, whereas the magnetic property of the composite enables imaging, magnetic separation, and may even have therapeutic benefit. In this review, the synthesis of fluorescent–magnetic nanoparticles, including magnetic QDs is explored; and the applications of these materials in imaging, separations, and theranostics are discussed. As the properties of these materials continue to improve, QDs have the potential to greatly impact biological imaging, diagnostics, and treatment.     

“Use of acidophilic bacteria of the genus Acidithiobacillus to biosynthesize CdS fluorescent nanoparticles (quantum dots) with high tolerance to acidic pH”

The use of bacterial cells to produce fluorescent semiconductor nanoparticles (quantum dots, QDs) represents a green alternative with promising economic potential. In the present work, we report for the first time the biosynthesis of CdS QDs by acidophilic bacteria of the Acidithiobacillus genus. CdS QDs were obtained by exposing A. ferrooxidans, A. thiooxidans and A. caldus cells to sublethal Cd²⁺ concentrations in the presence of cysteine and glutathione. The fluorescence of cadmium-exposed cells moves from green to red with incubation time, a characteristic property of QDs associated with nanocrystals growth. Biosynthesized nanoparticles (NPs) display an absorption peak at 360 nm and a broad emission spectra between 450 and 650 nm when excited at 370 nm, both characteristic of CdS QDs. Average sizes of 6 and 10 nm were determined for green and red NPs, respectively. The importance of cysteine and glutathione on QDs biosynthesis in Acidithiobacillus was related with the generation of H₂S. Interestingly, QDs produced by acidophilic bacteria display high tolerance to acidic pH. Absorbance and fluorescence properties of QDs was not affected at pH 2.0, a condition that totally inhibits the fluorescence of QDs produced chemically or biosynthesized by mesophilic bacteria (stable until pH 4.5–5.0). Results presented here constitute the first report of the generation of QDs with improved properties by using extremophile microorganisms.     

Autometallographic tracing of Hg–S quantum dots in frogs exposed to inorganic mercury

The histochemical distribution of mercury in the kidneys and gut of frogs (Rana ridibunda) exposed to inorganic mercury was analyzed with autometallography (AMG). It was found that most mercury in the kidneys accumulated in the proximal convoluted tubules as Hg–S nanocrystal, while control animals were totally void of AMG grains. In the gut the highest concentration of mercury was observed in the large intestine. The AMG grains were primarily located in the apical part of the absorptive cells, although rather high concentrations of silver enhanced mercury quantum dots were also detected in a special cell type of gut epithelium and the glycocalyx. A certain amount of AMG grains were detected in the lumen of the gut. We hypothesize that this pool of quantum dots results from sloughed off epithelial cells and macrophages. Such still intact cells and red blood cells containing AMG grains were also found in the lumen of the gut.     

Determination of 17β-oestradiol by fluorescence immunoassay with streptavidin-conjugated quantum dots as label

A sensitive and selective method for the determination of 17β-oestradiol by fluorescence immunoassay (FIA) was established on the basis of quantum dots (QDs) as label. The complex of biotin-labelled anti-rabbit IgG and strepavidin conjugated by quantum dots (QD-SA) was regarded as a probe in this system and the strepavidin-biotin system as signal amplification system. After optimising the conditions of the immunoreaction, such as the concentration of the reagent and the pH of the buffer solution, the linear range and the limit of detection of 17β-oestradiol were 0.01-10,000 ng ml⁻¹ and 0.00542 ng ml⁻¹, respectively. This method was applied to determine oestradiol in water samples, with the percent recoveries in the range of 86-113%.     

Optical detection of choline and acetylcholine based on H₂O₂-sensitive quantum dots

In this paper, we have constructed a simple, rapid and sensitive biosensor for detection of choline and acetylcholine (ACh) based on the hydrogen peroxide (H(2)O(2))-sensitive quantum dots (QDs). The detection limit for choline was 0.1 μM and the linear range was 0.1-0.9 μM and 5-150 μM, respectively. The detection limit for ACh was found to be 10 μM and the linear range was 10-5000 μM. The wide linear ranges were shown to be suitable for routine analyses of choline and ACh. Possible mechanism of the fluorescence of QDs quenched by H(2)O(2) was an electron transfer (ET) process. The experimental conditions of biosensors were optimized, and anti-interference ability was also presented. We also detected the choline in milk samples and the linear range was 5-150 μM. The detection linear range of ACh in serum was 10-140 μM. Most importantly, the recovery of choline in milk and ACh in serum samples were both close to 99%. The excellent performance of this biosensor showed that the method can be used in practice detection of choline and ACh.     

Do endogeic earthworms change plant competition? A microcosm study

Plants compete for limited resources. Although nutrient availability for plants is affected by resource distribution and soil organisms, surprisingly few studies investigate their combined effects on plant growth and competition. Effects of endogeic earthworms (Aporrectodea jassyensis), root-knot nematodes (Meloidogyne incognita) and the spatial distribution of ¹⁵N labelled grass litter on the competition between a grass (Lolium perenne), a forb (Plantago lanceolata) and a legume (Trifolium repens) were investigated in the greenhouse. Earthworms promoted N uptake and growth of L. perenne. Contrastingly, shoot biomass and N uptake of T. repens decreased in the presence of earthworms. P. lanceolata was not affected by the earthworms. We suggest that earthworms enhanced the competitive ability of L. perenne against T. repens. Nematodes increased the proportion of litter N in each of the plant species. Litter distribution (homogeneous vs. patch) did not affect the biomass of any plant species. However, P. lanceolata took up more ¹⁵N, when the litter was homogeneously mixed into the soil. The results suggest that endogeic earthworms may affect plant competition by promoting individual plant species. More studies including decomposers are necessary to understand their role in determining plant community structure.     

How do earthworms affect microfloral and faunal community diversity?

Much of the work regarding earthworm effects on other organisms has focused on the functional significance of microbial-earthworm interactions, and little is known on the effects of earthworms on microfloral and faunal diversity. Earthworms can affect soil microflora and fauna populations directly and indirectly by three main mechanisms: (1) comminution, burrowing and casting; (2) grazing; (3) dispersal. These activities change the soil's physico-chemical and biological status and may cause drastic shifts in the density, diversity, structure and activity of microbial and faunal communities within the drilosphere. Certain organisms and species may be enhanced, reduced or not be affected at all depending on their ability to adapt to the particular conditions of different earthworm drilospheres. A large host of factors (including CaCO₃, enzymes, mucus and antimicrobial substances) influence the ability of preferentially or randomly ingested organisms to survive (or not) passage through the earthworm gut, and their resultant capacity to recover and proliferate (or not) in earthworm casts. Small organisms, particularly microflora and microfauna, with limited ability to move within the soil, may benefit from the (comparatively) long ranging movements of earthworms. Microflora and smaller fauna appear to be particularly sensitive to earthworm activities, and priming effects enhancing nutrient release, particularly in casts, are common. Larger fauna such as microarthropods, enchytraeids and Isopods may be enhanced under some conditions (e.g., in earthworm middens), but in other cases earthworm activity may lead to a decrease in their populations due to competition for food (microbes and organic materials), and spatial and temporal changes in food abundance. Nevertheless, considering the presently available data, the beneficial interactions of earthworms and microflora and fauna appear to far outweigh the potential negative effects. However, much is still unknown regarding the interactions of earthworms of different ecological categories on the diversity and function of microfloral and faunal communities, and much more interdisciplinary research is needed to assess the potential role of earthworms in regulating the diversity of microflora and fauna in soil systems and the potentially beneficial or harmful effects this regulation may have on ecosystem function and plant growth in different ecosystems.     

Can a highly invasive species re-invade its native community? The paradox of the red shiner

Red shiners (Cyprinella lutrensis) are among the most widespread, ecologically general, and environmentally tolerant fish species in North America, and are highly invasive where they have been introduced outside their native range. However, long-term data on fish assemblages showed that red shiners gradually (1980s to 2006) disappeared from creeks that are direct tributaries of Lake Texoma (Oklahoma, USA) where they are native and historically had been numerically dominant. Following a major flood in 2007, red shiners were detected anew in some of these creeks, but repeatedly disappeared and re-appeared through November 2009. Given their invasive abilities where they are not native, their failure to become re-established prompted us to examine factors that affect their apparent inability to re-invade their native habitat. We established assemblages of five common fish taxa native to Brier Creek in 12 large, outdoor mesocosm stream units. Subsequently, we introduced red shiners at two densities of 10 or 30 per unit, six replicates each, to examine potential effects of propagule pressure on establishment success. Approximately six months later, we ended the experiment and recovered all fish. Red shiners failed to become established in the experimental units, regardless of initial stocking density. They also exhibited much lower survival than other species in the native community, which not only survived well but exhibited some recruitment. Red shiner survival was significantly negatively related to the number of sunfish (Lepomis spp.) that grew to adult size in experimental units, suggesting that predation can inhibit early stages of invasion by red shiners.     

Can monitoring data contribute to the biodiversity-ecosystem function debate? Evaluating data from a highly dynamic ecosystem

One of the most controversially discussed topics in current biodiversity-ecosystem function research is the transfer of results from experimental and theoretical studies to natural ecosystems. At the same time, monitoring data on biodiversity are requested as key indicators for the state of an ecosystem in most environmental evaluation frameworks. We analyse two monitoring data sets comprising information on abundance and biomass of macrozoobenthos communities in the German Wadden Sea in order to evaluate how much information monitoring data on biodiversity provide concerning ecosystem functioning and what implications this information (or the lack thereof) has for future monitoring programmes. Our results show a positive correlation between number of species of macrozoobenthos and its standing stock. Despite differences in overall biomass and individual size in different functional groups, this correlation remained consistent for different feeding guilds and therefore is likely to be independent of certain species traits. Moreover, functional turnover analyses indicate that increasing species richness is needed to maintain biomass levels over increasing periods of time. Whereas our data thus corroborate predictions from theory, we could not determine any causal relationships, because monitoring data commonly include only vague proxies for very few functional parameters, in our case standing biomass as a proxy for production. As to the use of diversity as an indicator for ecosystem functioning, we advise that management decisions are to be based on verified causal relationships and therefore strongly suggest the general incorporation of unambiguous proxies for functional parameters in the measuring campaigns of monitoring programmes.     

Can we determine if the linear nature of quantum mechanics is violated by the perceptual process?

It now appears feasible to be able to subject to experimental test Ghirardi's proposal concerning utilizing superposed photon states to check reduction mechanisms in perceptual processes which may be governed by nonlinear evolution laws. This can be accomplished with existing techniques involving superposed single photon states and living retinal tissue mounted on microelectrode arrays.     

Can geriatrics survive?

Geriatrics has consistently failed to attract enough staff, and hence geriatric units often cannot provide a full service for the elderly. Meanwhile beds in general medical units may be blocked by elderly patients. This division between geriatric and medical units is arbitrary and wasteful. There are no clinical processes or techniques unique to geriatrics, which is probably why the specialty is unattractive; few doctors want to confine their attention only to the elderly. Geriatrics as a separate specialty should therefore be largely abandoned and the care of the elderly reintegrated into general medicine.     

Characterization of multivalent lactose quantum dots and its application in carbohydrate–protein interactions study and cell imaging

We have previously reported a facile and convenient method for the preparation of a new type of lactose-CdSeS/ZnS quantum dots conjugates (Lac-QDs) that exhibit biocompatibility, noncytotoxicity and specificity to leukocytes. In order to further study the carbohydrate–protein interactions, a series of Lac-QDs with different lactose densities and a PEGylated (n = 3) lactose-QDs conjugate (LacPEG-QDs) with more flexible sugar ligands were prepared. The amount of the sugar molecules on QDs can be determined by NMR, which was in agreement with the results from TGA determination. The formula of the conjugates was determined with ICP-OES. The interactions between the conjugated QDs and the PNA protein were measured using SPR, which revealed that higher lactose density favored binding affinity under the same concentration, and Lac-QDs exhibit higher affinity than LacPEG-QDs. We further used a solid phase assay to assess the anti-adhesion activity of Lac-QDs and LacPEG-QDs on the cell level. The results showed that Lac-QDs had stronger activity in preventing THP1 from adhering to HUVEC than LacPEG-QDs, which was consistent with the SPR results. We reasoned that decrease in the conformational entropy induced by appropriate restriction of sugar flexibility could enhance the binding affinity of glyco-QDs, which implies that entropy change may be the main contributor to the interaction between high valent glyco-QDs and protein. The fabrication of lactose on QDs provides a fluorescent multivalent carbohydrate probe that can be used as mimics of glycoprotein for the study of carbohydrate–protein interactions and cell imaging.     

Detection of human neuronal α7 nicotinic acetylcholine receptors by conjugates of snake α-neurotoxin with quantum dots

Fluorescent derivatives are widely used to study the structure and functions of proteins. Quantum dots (QDs), fluorescent semiconductor nanocrystals, have a high quantum yield and are much more resistant to bleaching compared to organic dyes. Conjugates of α-neurotoxins with QDs were used for visualization of human α7 acetylcholine receptors heterologously expressed in GH4C1 pituitary adenoma cells. Specific staining of cells by the conjugated toxins was observed.     

Quantum dots – a versatile tool in plant science?

An optically stable, novel class of fluorophores (quantum dots) for in situ hybridisation analysis was tested to investigate their signal stability and intensity in plant chromosome analyses. Detection of hybridisation sites in situ was based on fluorescence from streptavidin-linked inorganic crystals of cadmium selenide. Comparison of quantum dots (QDs) with conventional detection systems (Alexa 488) in immunolabeling experiments demonstrated greater sensitivity than the conventional system. In contrast, detection of QDs in in situ hybridisation of several plant chromosomes, using several high-copy sequences, was less sensitve than Alexa 488. Thus, semiconductor nanocrystal fluorophores are more suitable for immunostaining but not for in situ hybridisation of plant chromosomes.     

Is there a biology of quantum information?

This paper briefly considers the notion of a biology of quantum information from a number of complementary points of view. We begin with a very brief look at some of the biomolecular systems that are thought to exploit quantum mechanical effects and then turn to the issue of measurement in these systems and the concomitant generation of information. This leads us to look at the internalist stance and the exchange interaction of quantum particles. We suggest that exchange interaction can also be viewed using ecological ideas related to apparatus-object. This can also help develop the important notion of complementarity in biosystems in relation to the nature and generation of information at the microphysical scale.     

How random is a highly denatured protein?

There has been renewed interest in determining the physicochemical properties of denatured states of proteins. In many denatured states there is evidence for the existence of nonrandom configurational distributions. Here we examine the small-angle neutron scattering profile of yeast phosphoglycerate kinase in the native state and in highly denaturing conditions. We show that the denatured protein scattering profile can be interpreted using a model developed for synthetic polymers in which the chain behaves as a random coil in a good solvent, i.e. with excluded volume interactions. The implications of this result for our appreciation of the protein folding process are discussed.     

Can viruses form biofilms?

The recent finding that the human T-cell leukemia virus type 1 (HTLV-1) encases itself in a carbohydrate-rich adhesive extracellular 'cocoon', which enables its efficient and protected transfer between cells, unveiled a new infectious entity and a novel mechanism of viral transmission. These HTLV-1 structures are observed at the surface of T cells from HTLV-1-infected patients and are reminiscent of bacterial biofilms. The virus controls the synthesis of the matrix, which surrounds the virions and attaches them to the T cell surface. We propose that, similar to bacterial biofilms, viral biofilms could represent 'viral communities' with enhanced infectious capacity and improved spread compared with 'free' viral particles, and might constitute a key reservoir for chronic infections.