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In this work, we grouped phytoplankton species according to their cell geometric shape for the first time in lake ecosystems. Field multiannual data were analyzed in order to understand the phytoplankton cell geometric shape distribution along the seasonal environmental variability in two Mediterranean lakes, a shallow natural lake and a deeper artificial lake. We characterized seasonally each lake on the basis of dominant phytoplankton cell geometric shapes, detecting the following common highlights comparing the two ecosystems: a complex and globular geometric shape (Cone + Half Sphere) with high mean volume and low mean surface-to-volume ratio in winter (at the deepest mixing layer); a variety of geometric shapes in spring (at the lowest nutrient concentrations), with a relevant contribution of simple and compact geometric shapes (Sphere and Cube) with low mean volume and high mean surface-to-volume ratio; a simple and elongated geometric shape (Cylinder) in summer (at thermal stratification conditions); a variety of geometric shapes in autumn (at intermediate environmental conditions), when the geometric shapes detected were at similar size. We provide new insights on how phytoplankton cell geometric shapes with similar and different sizes relate to the seasonal environmental variations in lake ecosystems, a topic relatively neglected in literature.

Porous platinum is a frequently used catalyst material in electrosynthesis and a robust broadband absorber in thermoelectrics. Pore size distribution and localization determine its properties by a large extent. However, the pore formation mechanism during the growth of the material remains unclear. In this work we elucidate the mechanism underlying electrochemical growth of nanoporous platinum layers and its control by ionic concentration and current density during electrolysis. The electrode kinetics and reduction steps of PtCl4 on platinum electrodes are investigated by cyclic voltammetry and impedance measurements. Cyclic voltammograms show three reduction steps: two steps relate to the platinum cation reduction, and one step relates to the hydrogen reduction. Hydrogen is not involved in the reduction of PtCl4, however it enables the formation of nanopores in the layers. These findings contribute to the understanding of electrochemical growth of nanoporous platinum layers in isopropanol with thickness of 100 nm to 500 nm. Porous platinum is a frequently used electrocatalyst and thermoelectric material, but the growth mechanism of nanopores in platinum layers is still not fully understood. Here, the authors show that hydrogen is not involved in the reduction process of PtCl4, however it enables the formation of nanopores.

Objective A handheld device was developed and qualified for in vivo human skin evaluation using laser speckle imaging technology. Methods Each laser speckle device prototype allows the choice of up to three different laser wavelengths in the range of 400 nm to 800 nm in total. Speckle pattern analysis gives various speckle parameters, for example, speckle contrast, speckle size, speckle modulation or fractal dimension. The developed laser speckle device prototypes were evaluated investigating three skin issues. Results We receive reproducible results from the speckle imaging device. For skin ageing, we found significant changes within three age groups. The effect of a methyl nicotinate treatment was clearly visible and quantifiable using a moorFLPI device as well as our speckle imaging device. In terms of basal cell carcinoma diagnosis, we found significant differences between normal and diseased skin, even though the number of samples was limited. Conclusion As shown with first application examples, it was possible to demonstrate the potential of the method for skin evaluation in vivo.

According to the methodological standards established by Marine Strategy Framework Directive, the assessment for the pelagic habitat under the Biodiversity Descriptor should be carried out at the regional or sub-regional level. In the case of Mediterranean Sea, the sub-regional assessment seems optimal to take into account biogeographic differences in species composition and functional characteristics. Previous research has shown that phytoplankton diversity indicators are efficient for reliable environmental assessments, although more effort has been recommended to test these indicators on a wide spatial scale to cover wider gradients of natural and anthropogenic pressures. In this work, a set of eight diversity indices was tested against the pressure levels within a common data set of the structure and abundance of phytoplankton communities from the Adriatic, Ionian and Aegean Seas. Expert knowledge was used to define four categories of impacts that take into account partial pressures, such as point and non-point pollution, industry, ports and fisheries. At the level of the common data set, most of the diversity, evenness and dominance indices could only distinguish between the highest level of impact and the rest of impact categories. These indices maintained the distinction between two levels of subsequently dichotomised impacts (no to low impact vs. high impact) across latitudinal and longitudinal gradients. On average, the indices were less sensitive to impacts in the northernmost and westernmost areas than in the southernmost and easternmost areas, although they still showed a significant response. The results also suggest that phytoplankton communities become more uniform and less dominated by a single taxon as sampling depth increases at sites with low impact, while evenness and dominance at impacted sites remain similar at all depths. In order to establish meaningful definitions of good environmental status and targets for pelagic habitats in the Mediterranean Sea, it is necessary to establish spatially specific thresholds by additional examination of indices of good performance.

LifeWatch Italy, the Italian node of LifeWatch ERIC, has promoted and stimulated the debate on the use of semantics in biodiversity data management. Actually, biodiversity and ecosystems data are very heterogeneous and need to be better managed to improve the actual scientific knowledge extracted, as well as to address the urgent societal challenges concerning environmental issues. LifeWatch Italy has realized the Phytoplankton Virtual Research Environment (hereafter Phytoplankton VRE), a collaborative working environment supporting researchers to address basic and applied studies on phytoplankton ecology. The Phytoplankton VRE provides the IT infrastructure to enable researchers to obtain, share and analyse phytoplankton data at a level of resolution from individual cells to whole assemblages. A semantic approach has been used to address data harmonisation, integration and discovery: an interdisciplinary team has developed a thesaurus on phytoplankton functional traits and linked its concepts to other existing conceptual schemas related to the specific domain.

Some authors define a giant ovarian cyst as having more than a 10 cm diameter . Huge cysts are usually benign or have a low-degree of malignancy . Laparoscopy is considered the gold standard approach to manage benign ovarian cysts but a major factor that will make the surgeon decide to perform or not perform a laparotomy is the size of the ovarian mass. We describe the case of an 86-year-old female patient diagnosed and treated for a giant ovarian cyst. The patient presented with complaints of recently aggravated pain in the left hypochondrium. The main symptom was a giant pelvic and abdominal mass that occupied all the quadrants. The CT scan identified the giant tumor as an ovarian cyst and also revealed a Bochdaleck hernia. Other investigations revealed: arterial hypertension, right bundle branch block, and chronic cardiac failure. The surgical treatment of choice was minimal laparotomy followed by suction of the content of the cyst, excision of the cyst’s wall and left adnexectomy. Despite the advanced anesthesiological risks, the patient developed no postoperative complications. Patients can be operated on successfully even in old age and in complicated cases with good perioperative anaesthetic evaluation and careful election of the surgical technique.

The throughput of spontaneous Raman spectroscopy for cell identification applications is limited to the range of one cell per second because of the relatively low sensitivity. Surface-enhanced Raman scattering (SERS) is a widespread way to amplify the intensity of Raman signals by several orders of magnitude and, consequently, to improve the sensitivity and throughput. SERS protocols using immuno-functionalized nanoparticles turned out to be challenging for cell identification because they require complex preparation procedures. Here, a new SERS strategy is presented for cell classification using non-functionalized silver nanoparticles and potassium chloride to induce aggregation. To demonstrate the principle, cell lysates were prepared by ultrasonication that disrupts the cell membrane and enables interaction of released cellular biomolecules to nanoparticles. This approach was applied to distinguish four cell lines – Capan-1, HepG2, Sk-Hep1 and MCF-7 – using SERS at 785 nm excitation. Six independent batches were prepared per cell line to check the reproducibility. Principal component analysis was applied for data reduction and assessment of spectral variations that were assigned to proteins, nucleotides and carbohydrates. Four principal components were selected as input for classification models based on support vector machines. Leave-three-batches-out cross validation recognized four cell lines with sensitivities, specificities and accuracies above 96%. We conclude that this reproducible and specific SERS approach offers prospects for cell identification using easily preparable silver nanoparticles.

High environmental variability is a feature of transitional water ecosystems, which also affects temporal and spatial patterns of phytoplankton. The Lagoon of Venice is characterised by a wide range of environmental conditions, high phytoplankton species diversity and abundance variability. Here we focus on morphological traits and specifically, for the first time in transitional waters, on phytoplankton geometric shape. Our hypothesis is that despite high taxonomic diversity, the prevalent features of the lagoon (shallow, permanently nutrient-enriched, turbulent and light-attenuated) should select a small number of dominant morphological types and adaptive strategies. We tested the hypothesis on a 13-year (1998-2010) phytoplankton data series and related abiotic factors. Seven shapes were found to be dominant and widespread in space and time in the lagoon, with two adaptive strategies: high surface area-to-volume ratio (S/V) together with low greatest axial linear dimension (GALD), and low S/V together with high GALD. Certain morphological traits, which allow a high S/V (from 0.8 to >1), appear quite constant: (i) small size (GALD < 15 ?m), in particular in round-shaped organisms, and (ii) high GALD together with shape attenuation, within the same species or across a range of species. We observed that the lagoon regime selects for the coexistence of competing species with similar morphological traits.

Laser speckle patterns were used for characterization of skin topography (DermaSpec, 13N13841). A set of parameters were obtained for skin surface characterization.Preliminary data has already shown differences between skin tumor and healthy skin areas.

The factors that influence the composition of marine epiphytic microalgal assemblages are poorly-understood. To address this short-coming, 93 samples were collected from four distinct regions in the Florida Keys National Marine Sanctuary (FKNMS) during winter and summer months to test the model that epiphytic microalgal communities are influenced by environmental gradients related to different sites, seasons, and host macrophyte species. One hundred and eighty-three morphotypes from 13 classes (7 phyla) were identified, dominated by 106 Bacillariophyta (77 identified to species equivalent or below), 37 Cyanophyta (13 identified to species equivalent or below), and 30 Dinophyta (21 identified to species equivalent or below). The largest proportion of variability in epiphytic communities was related to physico-chemical parameters (37%), followed by site location (ocean- versus bayside; 15%), seasonal differences (11%), and host macrophyte species (10%). Four physico-chemical variables were found to be most influential: wave height, temperature, ammonium concentration, and salinity . Only six out of 616 epiphyte – host comparisons exhibited significant differences in individual epiphyte taxon abundance between different host species (within site and season), further demonstrating that host-specificity was not strongly evident in this study. Overall, the results of this (sub)tropical study indicate that changing environmental characteristics between sites and seasons were the primary drivers influencing epiphyte community composition. Similar findings were found in an accompanying study of phytoplankton and other studies from temperate and (sub)polar regions, suggesting that common, underlying processes exist among these disparate environments.

Tailoring the physicochemical properties of the metallic multijunction nanolayers is a prerequisite for the development of microelectronics. From this perspective, a desired lower reflectance of infrared radiation was achieved by an electrochemical deposition of porous platinum in nonaqueous media on silver mirror supported nickel-chrome and nickel-titanium metallic films with incremental decreasing thicknesses from 80-10 nm. The electro-assembled architectures were examined by means of scanning electron microscopy and Fourier transform infrared spectroscopy and it was observed that the layer and sublayer thicknesses and resistivities have a substantial effect upon the porous platinum morphology and its optical properties. It is here reported that the augmentation of the metallic layer electrical conductivity determines the electroformation of more compact platinum nanolayers. Moreover, the platinum black coating of metallic nanolayers causes a considerable decrease of the reflectance in the region from 1000-8000 cm-1.

Size spectra exhibit common patterns of variation and predictable responses to pressures across ecosystem types, functional guilds and taxonomic groups. Here, we extend the size spectra approach to phytoplankton ecological status assessment in transitional waters by developing, testing and validating a multi-metric index of size spectra sensitivity (ISS-phyto), which integrates size structure metrics with others such as phytoplankton diversity, biomass and sensitivity of size classes to anthropogenic disturbance. The ability of various theoretical models of size spectra sensitivity to discriminate between disturbed and undisturbed ecosystems and levels of anthropogenic stress was evaluated. We used data on phytoplankton samples collected in 14 Mediterranean and Black sea transitional water ecosystems (coastal lagoons) from Italy, Albania, Greece, Bulgaria and Romania, and compared the models’ efficiency by looking at their pressure–impact response along salinity and enrichment gradients, the latter quantified as variations in dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP). Data from a fifteenth Mediterranean lagoon was used for external validation purposes. Right asymmetric models of size class sensitivity, implying higher sensitivity of smaller cell size classes, were found to contribute to the ISS-phyto multimetric tool more effectively than symmetric and left asymmetric models, distinguishing disturbed from undisturbed lagoons and disturbed from undisturbed stations within the same lagoon. When based on right asymmetric sensitivity models, i.e., those that were most efficient in identifying anthropogenic impacts, ISS-phyto also showed the best fit of pressure–response relationships along the salinity and enrichment gradients; at low to high levels of impact ISS response was driven by size class sensitivity and at very high impacts by phytoplankton biomass. A scheme for the classification of Ecological Quality Status based on ISS-phyto is proposed and validated. The validation procedure found that ISS-phyto is an effective and sensitive monitoring tool, robust, easy to apply and to inter-calibrate among laboratories.

Summary Confocal laser scanner microscopy coupled with an image analysis system was used to directly determine the shape and calculate the biovolume of phytoplankton organisms by constructing 3D models of cells. The study was performed on Biceratium furca (Ehrenberg) Vanhoeffen, which is one of the most complex-shaped phytoplankton. Traditionally, biovolume is obtained from a standardized set of geometric models based on linear dimensions measured by light microscopy. However, especially in the case of complex-shaped cells, biovolume is affected by very large errors associated with the numerous manual measurements that this entails. We evaluate the accuracy of these traditional methods by comparing the results obtained using geometric models with direct biovolume measurement by image analysis. Our results show cell biovolume measurement based on decomposition into simple geometrical shapes can be highly inaccurate. Although we assume that the most accurate cell shape is obtained by 3D direct biovolume measurement, which is based on voxel counting, the intrinsic uncertainty of this method is explored and assessed. Finally, we implement a data-driven formula-based approach to the calculation of biovolume of this complex-shaped organism. On one hand, the model is obtained from 3D direct calculation. On the other hand, it is based on just two linear dimensions which can easily be measured by hand. This approach has already been used for investigating the complexities of morphology and for determining the 3D structure of cells. It could also represent a novel way to generalize scaling laws for biovolume calculation. Lay description MICROALGAE IN 3D - Phytoplankton are an extremely diverse, polyphyletic group of microscopic photosynthetic microalgae. They account for approximately half of Earth's primary productivity and differ in their edibility and nutritional value for higher trophic levels. Finally, many phytoplankton species can produce toxins that negatively affect water quality and higher levels of trophic food web. Phytoplankton cell shape and biovolume are commonly used as size descriptor in the study of phytoplankton ecology because of their potentiality to explain the organization of ecological communities and predict their reorganizations under global change. Traditionally, phytoplankton cell shape and biovolume is not measured directly but is obtained from a standardized set of geometric models based on linear dimensions measured by light microscopy. This commonly used method allows visualization and measurement in two dimensions (2D) yielding no information at all on the third dimension of phytoplankton cells. Inaccurate biovolume assessment resulting from geometric approximation leads to erroneous interpretation of eco-physiological processes and morpho-functional traits. Here, we propose a new method based on confocal microscopy coupled with an image analysis system (NIS Elements AR software, Nikon) to determine directly shape and biovolume by means 3D confocal images. We evaluate the accuracy of current methods by comparing the results obtained using geometric models with direct biovolume and shape. We also propose a simplified approach for calculating the biovolume of cells that might with time replace those based on geometrically composed shapes.

A first synoptic and trans-domain overview of plankton dynamics was conducted across the aquatic sites belonging to the Italian Long-Term Ecological Research Network (LTER-Italy). Based on published studies, checked and complemented with unpublished information, we investigated phytoplankton and zooplankton annual dynamics and long-term changes across domains: from the large subalpine lakes to mountain lakes and artificial lakes, from lagoons to marine coastal ecosystems. This study permitted identifying common and unique environmental drivers and ecological functional processes controlling seasonal and long-term temporal course. The most relevant patterns of plankton seasonal succession were revealed, showing that the driving factors were nutrient availability, stratification regime, and freshwater inflow. Phytoplankton and mesozooplankton displayed a wide interannual variability at most sites. Unidirectional or linear long-term trends were rarely detected but all sites were impacted across the years by at least one, but in many case several major stressor(s): nutrient inputs, meteo-climatic variability at the local and regional scale, and direct human activities at specific sites. Different climatic and anthropic forcings frequently co-occurred, whereby the responses of plankton communities were the result of this environmental complexity. Overall, the LTER investigations are providing an unparalleled framework of knowledge to evaluate changes in the aquatic pelagic systems and management options.

In the last few decades, functional trait-based approaches have undergone an extraordinary expansion in phytoplankton ecology, due to the relative simplicity and the well-defined traits that determine the ecological niche of these organisms. A large quantity of heterogeneous and distributed data has been produced on phytoplankton traits and their use could be made more effective and efficient if data harmonization and interoperability would be improved. The use of controlled vocabularies and thesauri is an acknowledged good practice to establish the foundation for semantic interoperability, a critical requirement for reuse and sharing of data. In fact, thesauri, collectively constructed, bypass ambiguity issues in natural language, facilitating the identification and integration of the information available in multiple data sources and allowing both scientists and computer applications to interpret more effectively the meaning of data. Here we present a semantic resource on phytoplankton functional traits: the PhytoTraits thesaurus ( http://thesauri.lifewatchitaly.eu/PhytoTraits/index.php ). PhytoTraits is the result of the interdisciplinary collaboration of experts both from the phytoplankton functional domain and from information and communication technologies, working together within LifeWatch Italy, the Italian node of the e-science European infrastructure for biodiversity and ecosystem research. PhytoTraits is the first initiative to deal with the semantics of phytoplankton functional traits, focusing on morpho-functional traits towards standardized bio-volume assessment. It reflects the agreement of a scientific expert community to fix semantic properties (e.g. label, definition) of approximately 120 traits. Following semantic web standard technologies, the thesaurus was implemented in Simple Knowledge Organization System (SKOS), a common data model based on the Resource Description Framework (RDF). PhytoTraits is freely available online, it can be queried through a SPARQL endpoint ( http://thesauri.lifewatchitaly.eu/PhytoTraits/sparql.php ) and is also accessible via API ( http://thesauri.lifewatchitaly.eu/PhytoTraits/services.php ) for integration with other systems. If adopted as a standard and rigorously applied and enriched by the scientific community, PhytoTraits, providing harmonized concepts with associated unique and resolvable URIs, has the potential to significantly reduce the barriers to data discovery, integration, and exchange.

Porous platinum nanolayers on atomic force microscopy (AFM) probes were prepared by means of micro-electrochemistry of platinum chloride in non-aqueous solution. The silicon nitride AFM probes were previously sputtered with conductive layers of Ti, Ag, AlSiCu alloy and NiCr 5050. Homogeneous and continuous nanolayers of porous platinum (200 nm thick) were constructed on the NiCr/SixNy AFM probes. The porous platinum electrochemically layered AFM probes exhibited a low reflectance of 5% due to their absorption of infrared radiation in the wavelength region of 2–20 μm. When integrated into a microelectric circuit, this nanostructured crystalline system has the potential to convert absorbed thermal energy into electricity.

Throughout the world, infections caused by bacteria such as Staphylococcus aureus are a major cause of morbidity and mortality. In order to gain some understanding of the complicated physiological link between host and pathogen, modern techniques such as confocal microscopy and sophisticated OMICs technologies are suitable. However, labeling of pathogens such as S. aureus with green fluorescent protein, for example, or the generation of a reliable antibody, which are prerequisites for the application of reproducible isolation techniques, does not always succeed. Here, we present a universal approach for monitoring pathogen traffic after internalization into host cells by fluorescence microscopy and for isolation of bacteria from host-pathogen interaction assays using gold or ferric oxide-core, poly(vinyl alcohol) coated, and fluorescence-labeled nanoparticles (NP). The incubation of S. aureus HG001 with those NP had only minor effects on the bacterial growth in vitro. Quantitative proteome analysis after 24 h of NP incubation revealed that presence of NP provoked only marginal changes in the proteome pattern. The method presented enabled us to investigate the behavior of S. aureus HG001 during infection of S9 human epithelial cells by means of fluorescence microscopy and proteomics using magnetic separation or cell sorting.

Oxygen-sensitive polystyrene layers and nanoparticles have been developed for measuring oxygen in biosamples. Polymer layers were prepared via spin-coating on glass plates while the nanoparticles were synthesized by microemulsion polymerization. Platinum(II)meso-tetra(pentafluorophenyl)porphine (PtTFPP) was used as an oxygen sensor whereas N,N′-bis(1-hexylheptyl)perylene-3,4:9,10-bis-(dicarboximide) (S13) was used as a reference dye. For both, layers and nanoparticles, the sensor response was assessed in aqueous solution as well as in yeast culture. Quenching of porphyrin phosphorescence by oxygen was observed whereas fluorescence of the reference dye remained essentially unaffected. By using the signal of the reference dye fluctuations in the intensity of the excitation light or the nanosensor concentration can be easily corrected. Both sensor systems were compared with respect to their spectral response and their sensitivity.

Since ancient times, noble gold has continuously contributed to several aspects of life from medicine to electronics. It perpetually reveals its new features. We report the finding of a unique form of gold, reticular nanostructured gold (RNG), as an aqueous black colloid, for which we present a one-step synthesis. The reticules consist of gold crystals that interconnect to form compact strands. RNG exhibits high conductivity and low reflection, and these features, coupled with the high specific surface area of the material, could prove valuable for applications in electronics and catalysis. Due to high absorption throughout the visible and infrared domain, RNG has the potential to be applied in the construction of sensitive solar cells or as a substrate for Raman spectroscopy.

We report the assembly and structural characterization of a Y-shaped DNA template incorporating a central biotin moiety. We also report that this template may be used to assemble nanoscale architectures, which demonstrate the potential of this and related approaches to the fabrication of next-generation electronic devices. Of particular significance is the finding that it is possible to selectively metallize the above DNA template to obtain a three-electrode configuration. Also of particular significance is the finding that a biotin modified nanoparticle will recognize and bind selectively the central biotin moiety of the same template, once functionalized by the protein streptavidin.

A simple method for the site-directed immobilization of antibodies on gold substrates has been developed for surface plasmon resonance (SPR) applications. A freshly prepared gold surface on a glass slide was modified with Protein A via homobifunctional cross linker dithiobissuccinimide propionate (DSP) to achieve uniform, stable, and sterically accessible antibody coating. The density of the immobilized antibodies bound to Protein A was examined as a function of the protein concentration, buffer pH, buffer type and reaction time. The suggested scheme was also employed for the modification of gold-coated substrates for quartz crystal microbalance (QCM) applications. The modified gold surface was stable for several weeks and the reproducibility was satisfactory. The current fabrication technique could be applied to construct other immunosensors or biochips.

Certain carboxyl groups of the plasma membrane are involved in tumorgenesis processes. A gold core-hydroxyapatite shell (AuHA) nanocomposite is introduced as chemo-spectroscopic sensor to monitor these carboxyl groups of the cell membrane. Hydroxyapatite (HA) plays the role both of a chemical detector and of a biocompatible Raman marker. The principle of detection is based on chemical interaction between the hydroxyl groups of the HA and the carboxyl terminus of the proteins. The AuHA exhibits a surface enhanced Raman scattering (SERS) signal at 954 cm− 1 which can be used for its localization. The bio-sensing capacity of AuHA towards human skin epidermoid carcinoma (A431) and Chinese hamster ovary (CHO) cell lines is investigated using Raman microspectroscopic imaging. The localization of AuHA on cells is correlated with scanning electron microscopy, transmission electron microscopy and structured illumination fluorescence microscopy. This qualitative approach is a step towards a quantitative study of the proteins terminus. From the Clinical Editor This method would enable further studies on the molecular profiling of the plasma membrane, in an attempt to provide accurate cell identification. Using a gold core-hydroxyapatite shell (AuHA) nanocomposite, the authors in this paper showed the feasibility of detecting and differentiating cell surface molecules by surface enhanced Raman scattering.

The assembly and detailed structural characterization of a Y-shaped DNA template incorporating a central biotin moiety is reported. Also reported is the use of this template to assemble a model protein-functionalized three-electrode architecture. Of particular significance is the finding that a biotin-modified nanoparticle will recognize and selectively bind the central biotin moiety, once functionalized by the protein streptavidin. Potential applications of the above and related DNA templates in the emerging field of nanoelectronics are considered.

Escherichia coli (E. coli) photokilling due to the TiO2 under light irradiation in a batch reactor was studied by using of attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and atomic force microscopy (AFM). The ATR-FTIR spectral features and AFM images were analyzed in relation to E. coli viability data. ATR-FTIR is shown to be a suitable technique to follow the structural changes of the E. coli cell membranes during TiO2 photocatalysis. Formation of the peroxidation products due to the photocatalysis of E. coli cell is reported by this technique. Time dependent ATR-FTIR experiments provides the evidence for the changes in the E. coli cell wall membranes as the precursor events leading to bacterial lysis. Under the same experimental conditions used by ATR-FTIR spectroscopy, AFM microscopy was carried out to provide direct evidence for the E. coli lysis taking place under light irradiation after about 1 h in the presence of TiO2. By transmission electron microscopy (TEM), the aggregated TiO2 Degussa P-25 in aqueous solution is shown to interact with the bacteria surface and partly to remain in the aqueous solution at the concentration of 1 mg/ml.

The degradation of 6-chloro- N 2 -ethyl- N 4 -isopropyl-1,3,5-triazine-2,4-diamine (atrazine) by heterogeneous photocatalysis using suspended and supported TiO 2 is reported. The influence of reaction parameters such as catalyst and atrazine concentrations, pH, light intensity and temperature has been investigated and the optimal conditions for the abatement of atrazine have been identified. The primary degradation and the total mineralization of atrazine have been monitored by high performance liquid chromatography (HPLC) and total organic carbon (TOC), respectively. Atrazine got destroyed in both suspended and supported systems, but its complete mineralization is not observed, probably due to the formation of cyanuric acid as a stable product. The activation energy for the degradation reaction of atrazine is determined as 2.6 kcal mol −1 . The efficiencies of photodegradation in both suspended and supported systems (the latter in a 150 ml flow reactor) are compared and the results indicate that the catalytic activity of TiO 2 is not affected when it is immobilized using a functionalized polymer. It is observed that the durability of the supported catalyst activity is reduced after six or seven runs, but it could be fully re-activated by a simple heat treatment at 480 °C for 1 h. The findings clearly suggest that immobilized photocatalysts could indeed be developed for photocatalysis, without compromising on the activity of the primary catalyst particles.

A tyrosinase-modified Pt electrode, based on physical entrapment of the enzyme in agar–agar gel, was constructed and used to investigate the inhibitory effect of six carboxylic acids. At an applied potential of −50 mV versus saturated calomel electrode (SCE), the bioelectrode develops a fast, steady state response, linearly correlated with the phenol concentration up to 10 mg/l, with a sensitivity of 3.7 nA l/mg. A kinetic analysis of the amperometric response to phenol, recorded in the absence and in the presence of carboxylic acids (benzoic, 3-bromobenzoic, 4-ethylbenzoic, acetic, phenylacetic, 2-naphthylacetic acids), revealed that for the first four compounds the inhibition process corresponds to an uncompetitive one. Using the Lineweaver–Burk linearization the inhibition constants as well as the inhibition coefficients were calculated for the strong inhibitors: benzoic, 3-bromobenzoic, 4-ethylbenzoic and acetic acids.

An all-fibre based Raman-on-chip setup is introduced which enables analysis of solutions and trapped particles without microscopes or objectives. Beside the novel quartz microfluidic chip, innovative multi-core single-mode fibres with integrated fibre Bragg gratings are used for detection. The limit of quantitation is 7.5 mM for urea and 2.5 mM for nicotine with linear Raman spectroscopy. This is an improvement of more than two orders of magnitude compared with previous fibre-based microfluidic Raman detection schemes. Furthermore, our device was combined with optical traps to collect Raman-on-chip spectra of spherical polymer beads.

The geometric shape is traditionally used to calculate phytoplankton cell measurements (e.g. biovolume), but it can also play an important role in determining community distributions. Little is known about how geometric shapes relate to other morphological traits or to the environment. We explored whether shapes and related morphological traits are selected by environmental forcing. For this, samples were collected seasonally at 21 stations in coastal-marine waters of the Salento Peninsula (Italy). Phytoplankton taxa were classified in terms of geometric shape, biovolume (organism size) and surface-to-volume ratio (S:V). The relationship between greatest axial linear dimension (GALD) and S:V was assessed for each shape. A Canonical Correspondence Analysis (CCA) was performed to evaluate phytoplankton shape distribution on temporal and spatial scales. Phytoplankton community was characterized by high morphological diversity. GALD and S:V were inversely related in most of the shapes. CCA showed that phytoplankton shape distribution was influenced more by seasonal than by spatial variation: elongated shapes characterized the cold period; rounded and combined shapes the warmer period. Most of the shapes showed conservatism of the S:V and trade-off with the size. Geometric shapes represent an interesting feature to be considered in trait-based approaches to study phytoplankton distributions in aquatic ecosystems.

The investigation of the plasma membrane with intercorrelated multiparameter techniques is a prerequisite for understanding its function. Presented here, is a simultaneous electrochemical and topographic study of the cell membrane using a miniaturized amperometric enzymatic biosensor. The fabrication of this biosensor is also reported. The biosensor combines a scanning force microscopy (AFM) gold-coated cantilever and an enzymatic transducer layer of peroxidases (PODs). When these enzymes are brought in contact with the substrate, the specific redox reaction produces an electric current. The intensity of this current is detected simultaneously with the surface imaging. For sensor characterization, hydroquinone-2-carboxylic acid (HQ) is selected as an intrinsic source of H(2)O(2). HQ has been electrochemically regenerated by the reduction of antraquinone-2-carboxylic acid (AQ). The biosensor reaches the steady state value of the current intensity in 1 ± 0.2s.

Fluorescent ion indicators are widely used to measure ion concentrations in living cells. However, despite considerable efforts in synthesizing new compounds, no ratiometric sodium indicator is available that can be excited at visible wavelengths. Ratiometric indicators have an advantage in that measured fluorescence intensities can be corrected for fluctuations of the indicator concentration and the illumination intensity, which is not possible when non-ratiometric indicators are used. One way to circumvent this problem is to measure fluorescence lifetimes, which are independent of these factors. Another way to overcome the disadvantages of a non-ratiometric indicator dye is to embed it, together with a reference dye, into nanoparticles. By relating the indicator fluorescence to the fluorescence of the reference dye, inhomogeneities in the nanosensor concentration or the illumination intensity can be cancelled out reliably. In this study we compare the benefits and drawbacks of these approaches. © 2010 Copyright SPIE - The International Society for Optical Engineering.

In this study, we describe the preparation and evaluation of new fluorescent sensor nanoparticles for the ratiometric measurement of chloride concentrations. Both a chloride-sensitive dye (lucigenin) and a reference dye (sulforhodamine derivative) were incorporated into polyacrylamide nanoparticles via inverse microemulsion polymerization and investigated for their response to chloride ions in buffered suspension as well as in living cells. The fluorescence intensity of lucigenin reversibly decreased in the presence of chloride ions due to a collisional quenching process, which can be described with the Stern-Volmer equation. The determined Stern-Volmer constant K SV for the quenching of lucigenin incorporated into particles was found to be 53 M (-1) and is considerably smaller than the Stern-Volmer constant for quenching of free lucigenin ( K SV = 250 M (-1)) under the same conditions. To test the nanosensors in living cells, we incorporated them into Chinese hamster ovary cells and mouse fibroblasts by using the conventional lipofectamin technique and monitored the response to changing chloride concentrations in the cell.

1. Individual size has important implications for the physiology and ecology of phytoplankton. The analysis of size structure is a common approach for studying structure and function of planktonic ecosystems, with applications for the monitoring and conservation of aquatic ecosystem health. Several ways to describe the size structure of phytoplankton communities are used, including size-abundance relationships (SAR), non-normalized and normalized biomass size spectra (BSS), non-normalized and normalized number size spectra (NSS). 2. The main goal of this study was to test the effectiveness of the different tools proposed for describing and monitoring phytoplankton guilds in transitional waters. Comparative analyses were undertaken to evaluate (a) the consistency of each tool in describing the phytoplankton guilds, in terms of statistical significance and agreement with theoretical expectations; and (b) the effectiveness of each tool to discriminate conditions/cases. 3. Eleven transitional water bodies occurring along the eastern and western coasts of the Adriatic Basin were sampled in autumn 2004 and spring 2005. 4. Results obtained highlight that: (1) NSS globally showed higher statistical goodness of fit than BSS; (2) slopes of BSS and NSS, both normalized and non-normalized, were less negative than expected following the linear biomass hypothesis; and (3) the non-normalized NSS shape descriptors were best able to discriminate environmental heterogeneity both between and within transitional water ecosystems. 5. Parameters of NSS are suggested to be consistent descriptors of phytoplankton size structure and potentially important tools for monitoring and conservation of transitional water ecosystems.

Several linear and branched DNA structures from 80-200 nm with a biotine molecule in the middle have been prepared. These structures have been decorated by addition of positively charged gold nanoparticles carrying 4-(dimethylamino)pyridine ligands. Streptavidin binds to the central biotine molecule introducing a 20 nm gap in the structure in which a biotinylated nanoparticle can be introduced. The simplest structure (80 nm, linear) is formed by 4 oligonucleotides. By changing some of these components changes on length, shape, and recognition system easily can be introduced., Project financed by E.E.C.C., IST-1999-11974, BIOAND and STRP 014006 Nano3D and Spanish Ministry of Education BFU2005-23719-E, NAN2004-09415-C05-03 and BFU2004-02048.

The iron oxide-hydroxyapatite (FeOxHA) nanoparticles reported here differ from those reported before by their advantage of homogeneity and simple preparation; moreover, the presence of carboxymethyldextran (CMD), together with hydroxyapatite (HA), allows access to the cellular membrane, which makes our magnetic apatite unique. These nanoparticles combine magnetic behavior, Raman label ability and the property of interaction with the cellular membrane; they therefore represent an interesting material for structural differentiation of the cell membrane. It was observed by Raman spectroscopy, scanning electron microscopy (SEM) and fluorescence microscopy that FeOxHA adheres to the plasma membrane and does not penetrate the membrane. These insights make the nanoparticles a promising material for magnetic cell sorting, e.g. in microfluidic device applications.

Reported is the use of DNA to template the assembly of nanowires and protein-functionalized nanogap electrodes. Specifically, the use of DNA to template the assembly of gold nanowires between conventionally patterned gold contacts on a silicon wafer substrate. Also the use of DNA to template the assembly of protein-functionalized gold nanogap electrodes on a silicon wafer substrate. Of particular significance is the finding that suitably modified gold nanoparticles recognize and bind selectively the protein-functionalized nanogap between the above electrodes and are localized there. This and related work forms part of a broader effort directed toward the development of the alternative bottom-up fabrication technologies that will be needed to extend Moore's Law beyond 2012.

Sensitive amperometric biosensors for phenols compounds, based on tyrosinase (polyphenoloxidase, PPO) immobilized on a Pt electrode in an electropolymerized poly-amphiphilic pyrrole matrix or cross-linked with glutaraldehyde, were constructed and compared. Steady-state amperometric measurements, performed at -50 mV vs. SCE in aqueous phosphate buffer containing LiClO(4) 0.1 M (pH 7) as well as in a chloroform solution containing 0.1 M C(6)H(5)CH(2)N(CH(3))(3)Cl, were used in order to compare the electroanalytical and kinetic parameters of the investigated amperometric biosensors in aqueous and nonaqueous media. It was established that the polypyrrole matrix has a higher efficiency for enzyme retention resulting in higher bioelectrode sensitivity, both in aqueous buffer (690 microA M(-1)) and in chloroform (149 microA M(-1)).

Two different approaches, both exploiting two enzymes cooperative functioning, to enhance the sensitivity of tyrosinase (PPO) based biosensor for amperometric detection of phenols have been compared. For this purpose, one monoenzyme electrode (PPO) and two bienzyme electrodes (PPO and d -glucose dehydrogenase, GDH; PPO and horseradish peroxidase, HRP) were constructed using agar–agar gel as enzyme immobilization matrix. The biosensors responses for l -tyrosine detection were recorded at −50 mV versus saturated calomel electrode (SCE). The highest sensitivity (74 mA M −1 ) was observed for the PPO–GDH couple, while that recorded for PPO–HRP couple system was only 32 times higher than that measured for monoenzyme electrode (0.01 mA M −1 ). The ability of the PPO-, PPO–GDH-, PPO–HRP-based biosensors to assay phenols was demonstrated by quantitative determination of phenol, 1,2-dihydroxybenzene, 1,3-dihydroxybenzene, 1,4-dihydroxybenzene, 2-amino-3 (4-hydroxyphenyl) propanoic acid, 2-hydroxytoluene, 3-hydroxytoluene, 4-hydroxytoluene, 4-clorophenol, 3-clorophenol, 2-clorophenol, 4-hydroxybenzoic acid.

We report a nanoparticulate composite TiO2–BaTiO3 film which exhibits an increased antibacterial photocatalytic activity under visible light. The pure BaTiO3, TiO2 or their mixture do not attain a significant photocatalytic capacity under visible light. However, when these oxides are simultaneously synthesized under controlled conditions the resulting crystals exhibit a high catalytic effect. The effect of this material on microorganism lysis is studied and the mechanisms for the observed damage are investigated. Attenuated total reflection Fourier transform infrared spectroscopy provides the evidence of chemical changes (formation of carbonyl and carboxylic groups) in the cell membranes under visible light by TiO2–BaTiO3 but not by TiO2. Cyclic voltammetry demonstrates that peroxidation occurs in the absence of UV light and in the presence of TiO2–BaTiO3 but not in the presence of TiO2 alone. Atomic force microscopy reveals the morphological changes of the cells in this process.

In this study, we describe the design of new ratiometric fluorescent nanosensors, whose architecture is based on a gold core surrounded by a poly(vinylalcohol)-polyacetal shell. To the gold core, indicator dyes and reference dyes are attached via a cysteine linker. This nanosensor architecture is flexible with regards to the number and types of fluorophore linkages possible. The robust poly(vinylalcohol)-polyacetal shell protects the fluorophores linked to the core from non-specific interactions with intracellular proteins. The nanosensors developed in this way are biocompatible and can be easily incorporated into mammalian cells without the use of transfection agents. Here, we show the application of these nanosensors for intracellular pH and sodium ion measurements. © 2010 IOP Publishing Ltd.