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International audience; Curcumin is a natural polyphenol able to bind the amyloid beta peptide, which is related to Alzheimer's disease, and modify its self-assembly pathway. This paper focuses on a multi-disciplinary study that starts from the design of curcumin-like compounds with the key chemical features required for inhibiting amyloid beta aggregation, and reports the effects of these compounds on the in vitro aggregation of amyloid beta peptides. Chemoinformatic screening was performed through the calculation of molecular descriptors that were able to highlight the drug-like profile, followed by docking studies with an amyloid beta peptide fibril. The computational design underlined two different scaffolds that were easily synthesized in good yields. In vitro experiments, ranging from fluorescence spectroscopy and confocal microscopy up to small angle X-ray scattering, provided evidence that the synthesized compounds are able to modify the aggregation pattern of amyloid beta peptides both in the secondary structures, and in terms of the overall structure dimensions. The cytotoxic potential of the synthesized compounds was finally tested in vitro with a model neuronal cell line (LAN5). The overall view of this study suggests new concepts and potential difficulties in the design of novel drugs against diverse amyloidoses, including Alzheimer's disease

Conventional antimicrobial strategies have become increasingly ineffective due to the rapid emergence of antibiotic resistance among pathogenic bacteria. In order to overcome this problem, antimicrobial PhotoDynamic Therapy (PDT) is considered a promising alternative therapy. PDT has a broad spectrum of action and low mutagenic potential. It is particularly effective when microorganisms present endogenous photosensitizing pigments. Helicobacter pylori (Hp), a pathogen notoriously responsible of severe gastric infections (chronic gastritis, peptic ulcer, MALT lymphoma and gastric adenocarcinoma), produces and accumulates the photosensitizers protoporphyrin IX and coproporphyrin, thus it might be a suitable target of antimicrobial PDT. With the aim to design and develop an ingestible LED-based robotic pill for intragastric phototherapy, so that irradiation can be performed in situ without the use of invasive endoscopic light, photophysical studies on the Hp endogenous photosensitizers were carried out. These studies represent an important prerequisite in order to select the most effective irradiation conditions for Hp eradication. The photophysical characterization of Hp porphyrins, including their spectroscopic features in terms of absorption, steady-state and time-resolved fluorescence, was performed on bacterial extracts as well as within planktonic and biofilm growing Hp cells.

Oxyblepharismin is the photo-oxidized form of blepharismin, the chromophore responsible for the photophobic response of heterotrich ciliate Blepharisma japonicum, and represents a nice model for the study of photo-transduction. In this work, we focused on the photophysical characterization of OxyBP, in view of highlighting the main features related to excitation and emission. By a combined experimental and computational approach we identified the main features of absorption and fluorescence emission of the molecule in solvents of different properties, identifying the nature of transitions as well as the possible heterogeneity at ground/excited state. The thorough photophysical characterization of OxyBP is meant to provide the starting point for the elucidation of the photo-transduction pathway in vivo.

Antimicrobial PhotoDynamic Therapy (aPDT) is an emerging strategy aimed at the eradication of bacterial infections, with a special focus on antibiotic-resistant bacteria. This method is easy to apply, not expensive and particularly interesting in case of bacteria that spontaneously produce the required photosensitizers. In the framework of a project aimed at the development of an ingestible pill for the application of aPDT to gastric infections by Helicobacter pylori, a LED-based illuminating prototype (LED-BIP) was purposely designed in order to evaluate the photodamage induced by light of different wavelengths on porphyrin-producing bacteria. This short paper reports about temperature tests performed to assess the maximum exposure time and light dose that can be administered to bacterial cultures inside LED-BIP without reaching temperatures exceeding the physiological range.

After double-strand break induction, formation of ?-H2AX foci due to phosphorylation at Ser-139 of histone H2AX represents an early event of the DNA damage response (DDR). ?-H2AX foci are then rapidly dephosphorylated as signal for the subsequent recruitment of effector proteins. The induction and disappearance of the foci can be, therefore, used to monitor the functioning of the DDR machinery in a cell population exposed to genotoxic stress. Here, we investigated the time-course of ?-H2AX in unstimulated or cultured peripheral lymphocytes in vitro treated with UVB, bleomycin and mitomycin C (MMC). Once the mutagen exposure was performed, cells were harvested at different interval times from 0.5 to 5h. The results show that (i) in 20-h stimulated peripheral lymphocytes, UVB irradiation caused extensive and dose-dependent increases in nuclear phosphorylation, and disappearance of ?-H2AX foci progressed, proportionally to the UV fluence, with increasing the harvesting time; (ii) UVB-exposed G0 cells cultured for 20-h post-irradiation displayed low amounts of DNA phosphorylation, depicting a time-course in which the maximum effect was reached at 0.5h and dephosphorylation started after 1h; (iii) treatment of unstimulated lymphocytes with bleomycin sulphate induced an increase in nuclear phosphorylation of several folds higher than that of untreated cells, depicting kinetics comparable to those observed for UVB-exposed G1 cells; (iv) in stimulated cells, MMC caused a severe and dose-dependent high degree of H2AX phosphorylation together with a very slower kinetic of dephosphorylation with respect to the other experimental treatments. This study confirms the feasibility of the ?-H2AX focus assay as a genotoxic end-point and supports the view that the proposed type of analysis should be introduced in biomonitoring studies of human populations. This could also represent a feasible and useful tool in the screening and diagnosis of precancerous states or very early stages of other diseases.

The step-up photophobic response of the heterotrich ciliate Blepharisma japonicum is mediated by a hypericinic pigment, blepharismin, which is not present in any of the known six families of photoreceptors, namely rhodopsins, phytochromes, xanthopsins, cryptochromes, phototropins, and BLUF proteins. Upon irradiation, native cells become light-adapted (blue) by converting blepharismin into the photochemically stable oxyblepharismin (OxyBP). So far, OxyBP has been investigated mainly from a photophysical point of view in vitro, either alone or complexed with proteins. In this work, we exploit the vivid fluorescence of OxyBP to characterize its lifetime emission in blue B. Japonicum cells, on account of the recognized role of the fluorescence lifetime to provide physicochemical insights into the fluorophore environment at the nanoscale. In a biological context, OxyBP modifies its emission lifetime as compared to isotropic media. The phasor approach to fluorescence lifetime microscopy in confocal mode highlights that fluorescence originates from two excited states, whose relative balance changes throughout the cell body. Additionally, Cilia and kinetids, i.e., the organelles involved in photomovement, display lifetime asymmetry between the anterior and posterior part of the cell. From these data, some hypotheses on the phototransduction mechanism are proposed.

Marine and coastal quality assessment, based on test batteries involving a wide array of endpoints, organisms and test matrices, needs for setting up toxicity indices that integrate multiple toxicological measures for decision-making processes and that classify the continuous toxicity response into discrete categories according to the European Water Framework Directive. Two toxicity indices were developed for the lagoon environment such as the Venice Lagoon. Stepwise procedure included: the construction of a database that identified test-matrix pairs (indicators); the selection of a minimum number of ecotoxicological indicators, called toxicological core metrics (CMs-tox) on the basis of specific criteria; the development of toxicity scores for each CM-tox; the integration of the CMs-tox into two indices, the Toxicity Effect Index (TEI), based on the transformation of Toxic Unit (TU) data that were integrated as logarithmic sum, and the Weighted Average Toxicity Index (WATI), starting from toxicity classes integrated as weighted mean. Results from the indices are compared; advantages and drawbacks of both approaches are discussed.

Porewater plays an important role in sediment toxicity assessment using bioassays, but the most reliable extracting method and the potential contribution of confounding factors to the real toxicity need to be studied. The applicability of bioassays with the early life stages of Paracentrotus lividus, Mytilus galloprovincialis and Crassostrea gigas on porewaters extracted by centrifugation from the Venice Lagoon (Italy) is evaluated and demonstrated: toxicity tests can discriminate the toxicity of porewaters from sites with different kinds and levels of pollution and, using toxicity scores, data are classified in five toxicity classes. Sulphides do not represent a confounding factor in porewater toxicity; in contrast ammonia exhibited some concentrations above the toxicity threshold for sea urchin embryos. Keywords: Sperm cell toxicity test, Embryotoxicity test, Venice Lagoon

Curcumin is a natural polyphenol able to bind the amyloid beta peptide, which is related to Alzheimer's disease, and modify its self-assembly pathway. This paper focuses on a multi-disciplinary study that starts from the design of curcumin-like compounds with the key chemical features required for inhibiting amyloid beta aggregation, and reports the effects of these compounds on the in vitro aggregation of amyloid beta peptides. Chemoinformatic screening was performed through the calculation of molecular descriptors that were able to highlight the drug-like profile, followed by docking studies with an amyloid beta peptide fibril. The computational design underlined two different scaffolds that were easily synthesized in good yields. In vitro experiments, ranging from fluorescence spectroscopy and confocal microscopy up to small angle X-ray scattering, provided evidence that the synthesized compounds are able to modify the aggregation pattern of amyloid beta peptides both in the secondary structures, and in terms of the overall structure dimensions. The cytotoxic potential of the synthesized compounds was finally tested in vitro with a model neuronal cell line (LAN5). The overall view of this study suggests new concepts and potential difficulties in the design of novel drugs against diverse amyloidoses, including Alzheimer's disease.

A rapid index for the quality status assessment of Mediterranean transitional environments is proposed. It is based on the study of macrophyte associations (macroalgae and seagrasses) and the gross determination of some environmental parameters. The index was set up in 19 sampling sites of the Venice lagoon monitored during one year on a monthly basis taking into account the flora, the main contaminants and some other variables indicating the presence of gradients of stress conditions. Results were compared and intercalibrated with measurements carried out in Sacca di Goro, placed in Po delta, and in Lesina and Orbetello lagoons.

Bioassays are routinely employed for sediment quality assessment. In order to be able to effectively use Bioassays responses in regulatory and management frameworks, toxicity scores, which rank toxicity data in defined classes that are continuous and difficult to interpret, should be reliable and suitable tools to support decisions about the presence or absence of toxicity in tested samples and on how toxic a sample is. A statistical approach is needed to define thresholds for toxicity scores. The Minimum Significance Difference (MSD) criterion allowed the evaluation of toxicity thresholds for each test-matrix and organism pair, based on large sets of experimental data. The MSD values were normalized with respect to the control, ranked in ascending order, and the 90th percentile was identified; the Toxicity Threshold (TT) was calculated by subtracting the 90th percentile from 100 and the Toxicity Limit (TL) was estimated as the percentage of control response multiplied by TT. Taking into account sample responses normalized with respect to control (S), when S > TL, the sample is considered nontoxic; when S ≤ TL, toxicity is statistically present. The MSD criterion was applied on a dataset of results from both published and ongoing projects using the validated embryotoxicity tests with the echinoid Paracentrotus lividus, the oyster Crassostrea gigas, and the mussel Mytilus galloprovincialis, on elutriates from sediments of the Venice Lagoon. Once the threshold defining presence or absence of toxicity had been calculated, four other toxicity classes were developed: low (for toxicity data expressed as percentage of effect), medium, high, and very high toxicity (for toxicity data expressed as TU50).

Sperm cell and embryo toxicity tests using the Mediterranean sea urchin Paracentrotus lividus Lmk were performed to assess the toxicity of As3+, Cd2+, Cr3+, Ni2+, Pb2+, Cu2+, Zn2+, and Hg2+. The aim of this study was to improve information about the comparative sensitivity of sea urchin bioassays to the heavy metals, which are an important cause of contamination in the ecosystem of the Lagoon of Venice. Considering the data in mM/L, the order of toxicity is Hg2+ > Cu2+ > Zn2+ > As3+ ≥ Cr3+ ≥ Cd2+ ≥ Pb2+ ≥ Ni2+ for the sperm cell test and Hg2+ ≥ Pb2+ > Cu2+ > Zn2+ > Cd2+ > Ni2+ > As3+ ≥ Cr3+ for the embryo test. New toxicity data for metals expressed as median effective concentration (EC50) and no-observed-effect concentration (NOEC) are reported for the Mediterranean species. Accurate observations of embryotoxic effects at increasing metal concentrations were done, detecting some different behaviors in metal toxicity. Toxicity data compared with water column and pore-water concentrations recorded in the Lagoon of Venice (Italy) demonstrate the potential ability of bioassays using sea urchin to detect important contaminants in this ecosystem.

Blackfly occurrence was investigated on thirty plain streams of the low plain of the Friuli Venezia Giulia Region (NE Italy). Four species were found: Simulium erythrocephalum, S. angustitarse, S. ornatum, and S. paraequinum. Blackfly diffusion was larger than that reported in previous studies. Associations were found among Simuliidae species, hydrophyte species and water velocity.

Recent research trends in sediment quality assessment have demonstrated the need for an integrated approach which combines chemical characterisation with biological effects evaluation, both in the laboratory (toxicity bioassays) and in situ (macrobenthic community structure investigations). The Sediment Quality Triad organises these three aspects of sediment quality assessment in a single approach, where each component is complementary to the others. Considering the lack of previous studies, this preliminary work aimed to verify the potential of an integrated approach in a complex ecosystem, the Venetian lagoon. Five sampling stations along a presumed quality gradient were chosen. The chemical analyses of heavy metals and organic pollutants have been compared with the results of the toxicity bioassays (Microtox® solid phase, saline extracts and pore water, and sea urchin Paracentrotus lividus sperm cell bioassay performed with saline extracts) and with the characterisation of the infaunal community structure. The results of this first application pointed out the complementarity of the three approaches and the ability of the integrated approach in discriminating different levels of environmental stress. Nevertheless, deeper investigations both on the response of the macrobenthic community to natural variability and on the environmental toxicity of several sulphur compounds (i.e. elemental sulphur) will be necessary.

— The ciliate Blepharisma japonicum was exposed to artificial polychromatic and monochromatic UV radiation to evaluate the relative roles of UVB (280–315 nm UV radiation) and UVA (315–400 nm UV radiation) in altering its motility and photobehavior and to determine absolute weighting coefficients for these effects in the UVB range. Under polychromatic UV irradiation B. japonicum cells showed a severe reduction of cell speed and of the capability to respond to light stimuli. At low doses, however, UV caused a significant increase in the average velocity of a cell population. The UVB exclusion experiments indicated that UVA does not significantly alter motility and photoresponsiveness. The increase and the subsequent decrease in cell velocity was observed also under monochromatic irradiation at 281, 290 and 300 nm, whereas at 310 nm cells swim faster up to the highest photon flux density used. The cell capability of reacting to photic stimuli, conversely, steadily declined with increasing photon flux density at all the tested UVB wavelengths. The action spectra for the alteration of cell velocity and the impairment of photoresponsiveness show that the lower the irradiation wavelength, the more remarkable are the UVB effects and suggest different targets for the increase and the decrease in cell velocity.

The effect of monochromatic UVB (280–315 nm) irradiation on the photosynthetic activity of the marine green alga Dunaliella salina was investigated by monitoring the rate of oxygen exchange. Samples were irradiated with narrow bands centered at different UV wavelengths (281, 290, 300, 310 and 322 nm). In a first set of experiments the samples were preirradiated keeping constant the irradiance and varying exposure time. By increasing UVB fluence, a wavelength-dependent decrease in the rate of oxygen production was observed. In a second set of experiments oxygen exchange was monitored simultaneously to UVB irradiation. The decline in the rate of oxygen production was sharper at short wavelengths and faster with increasing UVB photon flux density. The photon fluence action spectra derived from the two sets of experiments are in good agreement and indicate a decrease in the UVB-inhibiting effectiveness of two orders of magnitude from about 0.2 (mmol m-2)-1 at 281 nm to about 0.003 (mmol m-2)-1 at 322 nm. The photon flux density action spectrum derived from the second set of experiments indicates a smoother decrease from about 0.2 (pmol s-l m-2)-1 at 281 nm to about 0.05 (pmol s-l m-z)-l at 310 nm.

Chemical analyses and bioassays were used in conjunction to assess the quality of sediments of the Venice lagoon. Organic micropollutants (polycyclic aromatic hydrocarbons [PAHs] polychlorinated biphenyls [PCBs], and chlorinated pesticides) were extracted from sediment samples and analyzed by gas chromatography after fractionation into classes of compounds. The Vibrio fischeri test was used to assess the acute toxicity of sediment extracts. The test was applied to organic extracts before cleanup and to extracts purified from sulfur and fractionated into single classes of compounds. Extracts before purification were much more toxic than single fractions. In particular, sulfur was toxic to V. fischeri. For PAHs and PCBs the 50% effective concentration (EC50) and EC20 values were determined using natural and spiked extracts (EC50 = 0.5 μg/ml and 1.4 μg/ml, respectively; EC20 = 0.13 μg/ml and 0.17 μg/ml). Sensitivity limits of the method for these compounds were also estimated as was an EC50 value of elemental sulfur dissolved in ethanol (0.022 μg/ml). A mathematical model was used to fit the concentration–response data to a sigmoid curve.

A key question to answer studying the biological effects of ultraviolet radiation on planktonic micro-organisms is whether they can perceive UV-B radiation as a sensory signal, likewise they do with visible light. We have faced this problem performing an individual-cell analysis of Blepharisma japonicum photomotile responses to UV-B stimuli. Our results on spectral responsiveness and on the effects of a photoresponse inhibitor indicate that B. japonicum is capable to perceive and transduce UV-B radiation as an environmental sensory stimulus, which it escapes from gathering in shadowed areas. Similar UV-B avoidance motile reactions could serve as a behavioural defence mechanism contributing to avoid harmful overexposure to UV-B.

Vibrio fisheri test has been used to evaluate the toxicity of contaminated sediment of the Venice lagoon. The following classes of organic microcontaminants were considered in this study: aliphatic hydrocarbons (AHs), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs) and chlorinated pesticides. Contaminants were extracted from samples with n‐hexane/dichloromethane in a sonication bath. Extracts were then purified and fractionated with column chromatography. A solvent exchange into ethanol was made before testing, to reduce toxicity. Preliminary tests were performed to set up the procedure to apply Vibrio fisheri to organic extracts. The application of the test with organic extracts was made at different steps of clean‐up and fractionation into classes of compounds. The purpose of the study was to set up a procedure to comparatively assess sediment contamination with chemical analyses and toxicity bioassays.

Laser-driven electron accelerators based on the Laser Wakefield Acceleration process has entered a mature phase to be considered as alternative devices to conventional radiofrequency linear accelerators used in medical applications. Before entering the medical practice, however, deep studies of the radiobiological effects of such short bunches as the ones produced by laser-driven accelerators have to be performed. Here we report on the setup, characterization and first test of a small-scale laser accelerator for radiobiology experiments. A brief description of the experimental setup will be given at first, followed by an overview of the electron bunch characterization, in particular in terms of dose delivered to the samples. Finally, the first results from the irradiation of biological samples will be briefly discussed.

In the coloured ciliate Blepharisma japonicum , step-up photophobic responses are triggered by the endogenous pigment blepharismin. Blepharismin, red in dark-grown cells, is intracellularly photooxidized into a blue form (oxyblepharismin), still acting as photosensing pigment. With the aim of correlating the spectroscopic properties of blepharismin and oxyblepharismin in vivo with their photophysiological features, optical absorption, steady-state and time-resolved fluorescence spectra have been measured on cell suspensions. Both in blepharismin and oxyblepharismin in their physiological molecular environment, three fluorescent species have been observed, with virtually the same lifetimes ( ∼ 0.2 ns, ∼ 1.0 ns, ∼ 3.5 ns), but significantly different relative amplitudes. In red cells the long-living component has a very low relative amplitude (∼ 4%) and the short-living one is largely predominant (> 78%), whereas in blue cells the slowly decaying species has a slightly higher relative amplitude (∼ 40%) than the intermediately (∼ 31%) and the fast decaying species (∼ 29%). Together with the spectral width of time-gated spectra, these data are discussed in connection with current hypotheses on the structures of the chromophores. No meaningful difference in the above-mentioned spectroscopic parameters was observed after 30 min of UV-B irradiation, showing that no significant difference exists between red and blue blepharismin as far as UV-B lability is concerned.

We present a laser-driven source of electron bunches with average energy 260 keV and picosecond duration, which has been setup for radiobiological tests covering the previously untested sub-MeV energy range. Each bunch combines high charge with short duration and sub-millimeter range into a record instantaneous dose rate, as high as 10(9) Gy s(-1). The source can be operated at 10 Hz and its average dose rate is 35 mGy s(-1). Both the high instantaneous dose rate and high level of relative biological effectiveness, attached to sub-MeV electrons, make this source very attractive for studies of ultrafast radiobiology on thin cell samples. The source reliability, in terms of shot-to-shot stability of features such as mean energy, bunch charge and transverse beam profile, is discussed, along with a dosimetric characterization. Finally, a few preliminary biological tests performed with this source are presented. .

the effects of artificial UV-B irradiation on motility, viability, photomotile responses and photoreceptor spectroscopic properties of the red ciliate Blepharisma japonicum were investigated. Both the mean velocity of the cells and the percentage of motile cells exhibiting step-up photophobic responses significantly decreased, increasing UV-B irradiation time. No marked difference with dark-kept control samples was observed in cells irradiated with visible light. Absorption and fluorescence measurements on cell suspensions indicated that no UV-B induced bleaching or spectroscopically detectable damage of the photoreceptor chromophore took place. The experimental results suggest that the UV-B target is a component of the photosensory transduction chain different from the photopigment itself and that cell vitality is damaged by a direct effect of UV-B, without any significant contribution from a pigment-sensitized photodynamic reaction.

Photoinhibition of photosynthesis was investigate in the marine macroalga Ulva laetevirens by measuring chlorophyll flourescence (PAM) and oxygen evolution. After exposure to unfiltered solar radiation the ratio of the variable flourescence to the maxiaml flourescence F v /F m (F v = F m -F o ) as well as the oxygen production declined rapidly with increasing duration of the exposure. Fluence rate-response curves for the oxygen production showed a decrease in the slope with exposure time and a shift of the compensation point to higher fluence rates as well as a decrease in the photosunthetic oxygen production. Solar radiation deprived of most of the UV-B (long pass above 320nm) induced a similar degree of photoinhibition of the ratio F v /F m as unfiltered solar radiation, whereas the oxygen production was less affected. The photoinhibitory effect of solar radiation was diminished when the solar radiation was filtered by a filter which passes wavelengths above 400nm. Even though UV-B accounts for a very small fraction of solar radiation, it has a considerable effect on photosynthesis. In addition, UV-A and visible radiation contribute to photoinhibitionin Ulva .

Laser flash photolysis spectra of chromophore extracts from Blepharisma japonicum cells have been measured. The variation of the triplet state spectra with pH is reported and the related pK a , values of 2–3 for deprotonation of the excited triplet state was determined. The anionic Blepharismin triplet state does not appear to readily undergo one-electron reduction by mild reducing agents, but undergoes oxidation by benzoquinone. Pulse radiolysis has been used to independently determine the spectra of the one-electron reduced and one-electron-oxidised forms of Blepharismin.

Rivers have been considered for a long time simply as vehicles transporting water and not as ecosystems. In Italy there has been a massive transformation of the lotic habitats, particularly with regard to the riparian zones and to the quality of waters. The structure and function of the populations and communities of organisms represent indicators of the changes which take place in the environment. Numerous indices, based on the study of the macroinvertebrate communities, have been proposed for evaluating the quality of the river environments in Europe. In Italy, a concrete result of the river monitoring programme, based on analysis of the macroinvertebrate communities, is the biological quality maps of hydrographic networks. The macroinvertebrate communities play an important role in the process of transforming the organic material input and as indicators of the quality and structural and functional integrity of the lotic environment.

To test the hypothesis that light signal transduction in the unicellular ciliates Stentor coeruleus and Blepharisma japonicum involves a change in intracellular pH as an initial signal following photoexcitation, we studied the dependence of the photophobic responses of the cells to changes in extracellular pH and to reagents that specifically affect intracellular pH. The extracellular pH can modify not only the intracellular pH, but can even reverse the sign of the pH gradient across the cell membrane. Thus, as predicted by the hypothesis, low extracellular pH reversibly inhibited the photophobic response of the ciliates. The intracellular pH-modulating reagents tested included ammonium chloride, a membrane-permeable weak acid that lowers the intracellular pH, and the protonophores carbonylcyanide m-chlorophenyl-hydrazone (CCCP) and carbonylcyanide p-(trifluoromethoxy)-phenyl-hydrazone (FCCP), which collapse the pH gradient across the cell membrane. The low pH and protonophore treatments caused a gradual inhibition of the photophobic responses in both ciliates. The observed reduction of the responsiveness of the cells to visible light can be attributed to the alteration of the intracellular pH, which is suggested to play a specific role in the photosensory transduction in both Stentor coeruleus and Blepharisma japonicum.

— Photosensory responses in the ciliated protozoan Blepharisma japonicum are mediated by a hypericin-like chromophore, blepharismin, localized in granules distributed under the cell membrane. A blepharismin-binding protein, with an apparcnt molecular weight ranging between 35 and 38 kDa, has been isolated by means of column separations and preparative isoelectric focusing and characterized by means of gel electrophoresis, analytic isoelectric focusing as well as absorption and fluorescence spectroscopy.

When exposed, in the presence of molecular oxygen, to light intensities of the order of 3-30 W m -2 , the ciliate Blepharisma japonicum changes its color from red to blue, because of the photooxidation of the photoreceptor pigment, blepharismin, to oxyblepharismin. Both red- and blue-pigmented cells show step-up photophobic responses. The action spectra of the light-dependent behavior of the red and the blue form of Blepharisma have been determined; their structure is very similar to that of the absorption spectra of the red and blue pigment, respectively. These findings suggest that the photosensing and phototransducing properties of blepharismin are maintained in its photooxidized form, oxyblepharismin

The paper couples the results obtained by applying the expert and the rapid Macrophyte Quality Indices set up to assess the ecological status of the Italian transitional environments according to the requirements by the Water Framework Directive (2000/60/CE). The indices were validated by comparing the composition of the macrophyte assemblages and the values of some bio-physico-chemical parameters of the water column of 20 stations of the Venice lagoon sampled monthly for one year between 2003 and 2005. In 5 stations out of the 20, the ones which fall within the 5 classes of ecological status suggested by the Water Framework Directive, sedimentation rates, sediment grain-size, and nutrient and pollutant (metals, Polychloro-Dibenzo-Dioxins/Furans, Polycyclic Aromatic Hydrocarbons, Pesticides and Polychlorinated Biphenils) concentrations in surface sediments were also determined. Results showed strong relationships between the trends of these environmental parameters and the composition and structure of macrophyte associations, as well as with the Macrophyte Quality Index assessment. Chlorophyceae showed a trend opposite to Rhodophyceae whose presence was concentrated in oxygenated and transparent environments. Chlorophyceae and the species characterised by low scores prevailed in turbid areas where nutrient and pollutant concentrations were high. Results allowed the identification of the conditions of the “reference sites” (confinement areas and sites with high water renewal) and the integration of the dichotomic key used for the application of the R-MaQI.

The effect of the singlet oxygen quencher crocetin on the photomotile responses and photodynamic killing of the ciliated protozoan Blepharisma japonicum was investigated. The percentage of cells surviving photodynamic killing is higher in the presence of the water-soluble carotenoid derivative than in control samples. However, no significant difference is observed in the photobehavior of the control and crocetin-containing samples. These results indicate that blepharismin-sensitized photodynamic reactions occur in Blepharisma, but do not play any meaningful role in initiating photosensory processes.