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In this study, dissolved organic carbon (DOC) data and optical properties (absorbance and fluorescence) of DOM, weekly collected in the Arno River for 2 years, are used to investigate the main processes determining DOM temporal dynamics in a small Mediterranean river, with torrential hydrology and medium-high human impact, and to quantify the contribution of this river to Med Sea carbon budget. A clear seasonal cycle of DOM, with DOC values ranging between 170 and 490 μM, was observed. Optical properties indicates that DOM quality in the river is different depending on the season; terrestrial humic-like substances prevail in winter, when discharge and floods are the main drivers of DOM concentration and quality, whereas autochthonous protein-like substances prevail in spring and summer, when biological processes dominate. Our results provide a robust estimate of the DOC flux to the Med Sea (9.6 · 10 9  g DOC yr -1 ) and of its range of variability (12.95 · 10 9 -5.12 · 10 9  g DOC yr -1 ). The 80% of this flux was generally delivered during autumn/winter with significant amounts ascribed to single flood events (up to 26% in 2014). This study, by providing a rich dataset on water quantity and quality and by quantifying the importance of the hydrological regime on DOC transport, represents an important step toward a quantitative modeling of the Arno River., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study reports the first information on extracellular enzymatic activity (EEA) combined with a study of DOM dynamics at the Arno River mouth. DOM dynamics was investigated from both a quantitative (dissolved organic carbon, DOC) and a qualitative (absorption and fluorescence of chromophoric DOM, CDOM) perspective. The data here reported highlight that the Arno River was an important source of both DOC and CDOM for this coastal area. CDOM optical properties suggested that terrestrial DOM did not undergo simple dilution at the river mouth but, other physical-chemical and biological processes were probably at work to change its molecular characteristics. This observation was further supported by the "potential" enzymatic activity of β-glucosidase (BG) and leucine aminopeptidase (LAP). Their Vmax values were markedly higher in the river water than in the seawater and their ratio suggested that most of the DOM used by microbes in the Arno River was polysaccharide-like, while in the seawater it was mainly protein-like., (© 2013. Published by Elsevier B.V. All rights reserved.)

Human transferrin (Tf) and saporin-6 (Sap), a ribosome inactivating protein from Saponaria officinalis, were chemically conjugated: the reaction generated two chimeras (called Tf-Sap) that proved to be cytotoxic to HepG2 cells. Electrophoretic and chromatographic analysis revealed that the two conjugates contained saporin and Tf in a 2:1 or 1:1 molar ratio (140 and 110 KDa, respectively). Free saporin is essentially nontoxic, whereas Tf-Sap efficiently kills HepG2 cells, although its ID50 (= 6 nM) is 1000-fold greater than that of ricin. Intracellular transport of these toxins was followed by in vivo fluorescence video microscopy, preparing the conjugates starting from rhodamine isothiocyanate-labeled saporin. Image analysis of living HepG2 cells exposed to fluorescent Tf-Sap revealed that the endocytotic pathway involving passage through secondary endosomes is dictated by Tf and is different from that of ricin (the dimeric toxin from Ricinus communis), which is delivered to the Golgi apparatus, the probable site of activation. We discuss whether differences in toxicity between ricin and Tf-Sap can be attributed to the different mechanisms of transport and activation.

The mechanism of proximal tubule potassium reabsorption was studied by stopped-flow microperfusion and determination of potassium activities by ion-sensitive resin microelectrodes. The proximal tubule was unable to establish transepithelial potassium gradients. Perfusion with 20 mM K+ turned the lumen 3 mV more negative, an effect abolished by Ba2+. The half-time for K+ activities to reach their stationary level after perfusion with 1 mM K+ was significantly increased by Ba2+ from 4.25 +/- 0.14 s to 11.0 +/- 1.71 s, and after perfusion with 20 mM K+, from 5.43 +/- 0.20 to 12.53 +/- 0.90 s. These data indicate that a significant fraction of potassium is transferred across proximal tubule epithelium by a transcellular, K(+)-channel-dependent route.