Effects of copper and zinc on growth, feeding and oxygen consumption of Farfantepenaeus paulensis postlarvae (Decapoda: Penaeidae)

The effect of chronic exposure (35 days) to sub-lethal concentrations of copper (17-212 ppb) and zinc (41-525 ppb) on growth of Farfantepenaeus paulensis postlarvae 17 days old (PL(17)) was analysed. The effects of acute exposure of PL(17) to the same metal on food ingestion and oxygen consumption were also evaluated. Studies were performed using copper and zinc singly, and in a mixture of equipotent concentrations (1:2.5). Chronic exposure to copper (85 and 212 ppb) and zinc (106, 212 and 525 ppb) reduced PL(17) growth. Acute exposure to copper (212 ppb) and zinc (525 ppb) reduced the number of Artemia sp. predated during 30 min and the positive feeding response induced by L-isoleucine. Despite of the lower positive feeding response when PL(17) were exposed to zinc, a significant difference from control condition was not seen. Oxygen consumption was reduced by all copper and zinc concentrations tested. The mean reduction was approximately 32%. The copper zinc-mixture did not modify food consumption and feeding response, or the oxygen consumption of the PL(17). The inhibition of food and oxygen consumption induced by copper and zinc could explain, at least in part, the long-term reduction of growth observed in chronically exposed PL(17). Our results also suggest that the inhibition of food consumption induced by copper is possibly due to an effect on chemosensory mechanisms. Finally, an antagonism between copper and zinc was observed, when were employed to analyse feeding behaviour and aerobic metabolism after acute exposure.     

Characterization of di- and monosulfated, unsaturated heparin disaccharides with terminal N-sulfated 1,6-anhydro-beta-D-glucosamine or N-sulfated 1,6-anhydro-beta-D-mannosamine residues

Modified heparin disaccharides were obtained by the alkaline treatment of a solution containing the disulfated heparin disaccharide DeltaHexA-alpha-(1-->4)-D-GlcNSO(3),6SO(3). Their structures were characterized by one- and two-dimensional NMR spectroscopy: DeltaHexA-alpha-(1-->4)-1,6-anhydro-GlcNSO(3), DeltaHexA-alpha-(1-->4)-1,6-anhydro-ManNSO(3) and DeltaHexA-alpha-(1-->4)-ManNSO(3),6OSO(3). NMR spectroscopy, in combination with HPLC, provided the composition of the mixture. Characteristic NMR signals of the disaccharides were identified, even at low levels, in a high field of (1)H-(13)C correlation NMR spectra (HSQC) of a low molecular weight heparin (LMWH) obtained by beta-elimination (alkaline hydrolysis) of heparin benzyl ester, providing a more complete structural profile of this class of compounds.     

Phytoremediation of Cd,Ni, Pb and Zn by Salvinia minima

Most metals disperse easily in environments and can be bioconcentrated in tissues of many organisms causing risks to the health and stability of aquatic ecosystems even at low concentrations. The use of plants to phytoremediation has been evaluated to mitigate the environmental contamination by metals since they have large capacity to adsorb or accumulate these elements. In this study we evaluate Salvinia minima growth and its ability to accumulate metals. The plants were cultivated for about 60 days in different concentrations of Cd, Ni, Pb and Zn (tested alone) in controlled environmental conditions and availability of nutrients. The results indicated that S. minima was able to grow in low concentrations of selected metals (0.03 mg L^?1 Cd, 0.40 mg L^?1 Ni, 1.00 mg L^?1 Pb and 1.00 mg L^?1 Zn) and still able to adsorb or accumulate metals in their tissues when cultivated in higher concentrations of selected metals without necessarily grow. The maximum values of removal metal rates (mg m² day^?1) for each metal (Cd = 0.0045, Ni = 0.0595, Pb = 0.1423 e Zn = 0.4046) are listed. We concluded that S. minima may be used as an additional tool for metals removal from effluent.     

Cytotoxicity and site-specific DNA damage induced by nitroxyl anion (NO(-)) in the presence of hydrogen peroxide. Implications for various pathophysiological conditions.

Nitroxyl anion (NO(-)), the one-electron reduction product of nitric oxide (NO(.)), is formed under various physiological conditions. We have used four different assays (DNA strand breakage, 8-oxo-deoxyguanosine formation in calf thymus DNA, malondialdehyde generation from 2'-deoxyribose, and analysis of site-specific DNA damage using (32)P-5'-end-labeled DNA fragments of the human p53 tumor suppressor gene and the c-Ha-ras-1 protooncogene) to study the effects of NO(-) generated from Angeli's salt on DNA damage. It was found that strong oxidants are generated from NO(-), especially in the presence of H(2)O(2) plus Fe(III)-EDTA or Cu(II). NO(.) released from diethylamine-NONOate had no such effect. Distinct effects of hydroxyl radical (HO(.)) scavengers and patterns of site-specific DNA cleavage caused by Angeli's salt alone or by Angeli's salt, H(2)O(2) plus metal ion suggest that NO(-) acts as a reductant to catalyze the formation of the HO(.) from H(2)O(2) plus Fe(III) and formation of Cu(I)-peroxide complexes with a reactivity similar to HO(.) from H(2)O(2) and Cu(II). Angeli's salt and H(2)O(2) exerted synergistically cytotoxic effects to MCF-7 cells, determined by lactate dehydrogenase release assay. Thus NO(-) may play an important role in the etiology of various pathophysiological conditions such as inflammation and neurodegenerative diseases, especially when H(2)O(2) and transition metallic ions are present.     

Multi-photon excitation microscopy

Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. As well, 2PE implementations in noninvasive optical bioscopy or laser-based treatments point out to the relevance in clinical applications. Here we report about some basic aspects related to the phenomenon, implications in three-dimensional imaging microscopy, practical aspects related to design and realization of MPE microscopes, and we only give a list of potential applications and variations on the theme in order to offer a starting point for advancing new applications and developments.     

S.A.M., the Italian Martian Simulation Chamber

The Martian Environment Simulator (SAM “Simulatore di Ambiente Marziano”) is a interdisciplinary project of Astrobiology done at University of Padua. The research is aimed to the study of the survival of the microorganisms exposed to the “extreme” planetary environment. The facility has been designed in order to simulate Mars’ environmental conditions in terms of atmospheric pressure, temperature cycles and UV radiation dose. The bacterial cells, contained into dedicated capsules, will be exposed to thermal cycles simulating diurnal and seasonal Martian cycles. The metabolism of the different biological samples will be analysed at different phases of the experiment, to study their survival and eventual activity of protein synthesis (mortality, mutations and capability of DNA reparing). We describe the experimental facility and provide the perspectives of the biological experiments we will perform in order to provide hints on the possibility of life on Mars either autochthonous or imported from Earth. Presented at: National Workshop on Astrobiology: Search for Life in the Solar System, Capri, Italy, 26 to 28 October, 2005.     

Sexual and seasonal variations in osmoregulation and ionoregulation in the estuarine crab Chasmagnathus granulatus (Crustacea, Decapoda)

The estuarine crab Chasmagnathus granulatus (Crustacea, Decapoda, Brachyura) inhabits salt marshes along the South Atlantic coast from Rio de Janeiro (Brazil) to Patagonia (Argentina). In the present study, salinity tolerance (0-45‰; 16-1325 mOsm/kg H₂O) and hemolymph osmotic and ionic (Na⁺, Cl⁻, and K⁺) regulation in both female and male C. granulatus were analyzed in summer and winter. Results showed that both female and male C. granulatus are euryhaline. Mortality was only observed in extremely low salinity (0‰; 16 mOsm/kg H₂O) for both sexes. For females, the LT₅₀ at 0‰ salinity was similar in summer (20.1 h) and winter (17.4 h). Males were more tolerant to salinity than females in both seasons, and mortality was observed only in summer (LT₅₀ = 50.9 h). Results from freshly collected crabs or long-term (16-day) osmotic and ionic regulation experiments in the laboratory showed that male C. granulatus is a better hyper-osmoregulator than female in summer and winter. However, a hypo-osmoregulatory ability was only observed in females experimentally subjected to salinity 40‰ (1176 ± 11 mOsm/kg H₂O) in both seasons. In both sexes, hyper-osmotic regulation was achieved by hyper-regulating hemolymph Na⁺, Cl⁻, and K⁺ concentration. In females, hypo-osmotic regulation was achieved by hypo-regulating hemolymph Na⁺ and Cl⁻ concentration. Long-term (16-day) osmotic and ionic regulations in different salinities were similar in males or females collected and tested in summer and winter. Despite this lack of a seasonal effect on hemolymph osmoregulatory and ionoregulatory patterns in males or females, a marked seasonal difference in the dynamics of these processes was observed for both sexes. In the first 2 days after hypo-osmotic shock (20‰→5‰; 636→185 mOsm/kg H₂O), variations in female osmolality and ion (Na⁺ and Cl⁻) concentration were larger and faster in winter than in summer, while in males the opposite was observed. Furthermore, a seasonal effect on the crab response to hyper-osmotic shock (20‰→40‰; 636→1176 mOsm/kg H₂O) was only observed in males. A new osmolality and ion (Na⁺ and Cl⁻) concentration steady state was faster achieved in winter than in summer. Regarding sexual differences, females showed a better capacity to hypo-regulate the hemolymph osmolality and Na⁺ concentration than males, even after a sudden increase in salinity (hyper-osmotic shock) in both seasons. On the other hand, males showed a better capacity to hyper-regulate the hemolymph osmolality and Na⁺ concentration than females, even after a sudden decrease in salinity (hypo-osmotic shock), especially in winter. Taken together, results reported in the present study suggest the need to consider both sex and collection season as important factors in future osmotic and ionic regulation studies in estuarine crabs.     

A mesocosm study of aerobic mineralization of seven aquatic macrophytes

Decomposition experiments were performed using the closed bottles method and seven aquatic macrophytes: Cabombafurcata, Cyperusgiganteus, Egerianajas, Eichhorniaazurea, Salviniaauriculata, Oxycaryumcubense and Utriculariabreviscapa. Cultures with lagoon water and macrophytes detritus were incubated at 20 °C under aerobic conditions. Total particulate and dissolved organic carbon and dissolved oxygen concentrations were analyzed. The major findings were: (i) the maximum amount of consumed oxygen (OC_MAX) and deoxigenation rates (k_d) differ among macrophyte decays (OC_MAX: 165–700 mg g⁻¹ DW; k_d: 0.014–0.045 d⁻¹); these differences depends mainly on molecular and elemental composition of detritus; in short period, the cytoplasm fraction of detritus enhanced the catabolic activity; (ii) the kinetic model adopted to describe the oxygen consumption was shown appropriate; the parameter more affected by the chemical differences of detritus was OC_MAX; the constant rates of oxygen consumption (k_d) were relatively smaller in relation to obtained to oxidative decarboxylation processes (ca. 6 times) and presented minimum changes if compared with the detritus variations.