Definition of hydrologic regimes in regulated rivers for aquatic biodiversity conservation.

Negative ecological effects of water withdrawal activities from lotic systems have led to the need for the restoration of naturally shaped flow schemes. As a first step in this direction, individuation of sustainable minimum flow volumes can limit extreme alteration of ecosystem structure and functionality. The definition of such minimum flow values, to be released downstream each water diversion structure, must follow the understanding and the quantification of ecological alteration related to flow reduction. This thesis focuses on the study of instream ecological effects of water withdrawal from lowland rivers (Ticino and Adda rivers), with the purpose to define adequate indicators for the identification of ecological effects of hydrological alteration. The application of such indicators should be useful for the definition of environmentally sustainable minimum flow schemes. For this purpose, national protocols for the study of biological communities were applied and critically analyzed. These monitoring criteria and tools appeared inadequate for addressing the hydrological assessment as postulated in this work and for the studied geographical context. Alternative sampling and analysing methods are proposed, following two different directions: (1) a different approach in the use of a common and well known structural indicator (macroinvertebrate fauna) and (2) the integration of such structural descriptors with a functional approach that considers the river quality through ecosystem methabolism measures. Although their widespread use, connected to simple field and laboratory application, macroinvertebrates appeared to be an inadequate tool for hydrological alteration monitoring. However, interesting considerations about relationship between community density and richness and environmental disturbance could be made. Presented data about physico-chemical parameters, collected through the open-channel method, show an influence of flow on ecosystem functional processes, mainly related to influence on aquatic vegetation. Collected data were useful to define an easy method to assess general response of the ecosystem to hydrological modifications, which future application could lead to interesting results and to an alternative approach for the application of WFD (Dir. 2000/60/EC) requirements

Thermal regime of a highly regulated Italian river (Ticino River) and implications for aquatic communities

Thermal alteration is one of the adverse effects of flow regulation sharpened in recent years due to climate changes. In this work, we characterize the thermal regime of a highly regulated river located in Northern Italy, which is the emissary of Lake Maggiore. The thermal characteristics of the study reach are influenced by the presence of two dams and by climate warming. In the 15-year monitored period (1999-2013) a significant increase of the mean annual river temperature (i.e., +1.8°C) was indeed recorded. Minimum and maximum water temperatures were detected in correspondence of low-flow periods in February and August, respectively. During August the temperature reached a maximum of 27°C, that is a value below the upper thermal limit of tolerance for most of the aquatic taxa collected in the study area. Moreover, the local presence of seeps and the interaction with groundwater play an important ecological role by guaranteeing patches of cool water to the aquatic communities. Nevertheless, the sensitive early life stages of fish species that spawn in spring/summer may be affected by the high summer temperatures and the expected future climate warming. The wider range of temperatures preferred by alien fish species could result into a competitive disadvantage for autochthonous species. The current minimum flows established by local laws as environmental flows appeared to be able to guarantee an acceptable thermal alteration in morphologically heterogeneous reaches where river/groundwater interaction occurred, at least within the current climatic conditions.</p

Pullulan Based Bioconjugates for Ocular Dexamethasone Delivery

Posterior segment eye diseases are mostly related to retinal pathologies that require pharmacological treatments by invasive intravitreal injections. Reduction of frequent intravitreal administrations may be accomplished with delivery systems that provide sustained drug release. Pullulan-dexamethasone conjugates were developed to achieve prolonged intravitreal drug release. Accordingly, dexamethasone was conjugated to similar to 67 kDa pullulan through hydrazone bond, which was previously found to be slowly cleavable in the vitreous. Dynamic light scattering and transmission electron microscopy showed that the pullulan-dexamethasone containing 1:20 drug/glucose unit molar ratio (10% w/w dexamethasone) self-assembled into nanoparticles of 461 +/- 30 nm and 402 +/- 66 nm, respectively. The particles were fairly stable over 6 weeks in physiological buffer at 4, 25 and 37 degrees C, while in homogenized vitreous at 37 degrees C, the colloidal assemblies underwent size increase over time. The drug was released slowly in the vitreous and rapidly at pH 5.0 mimicking lysosomal conditions: 50% of the drug was released in about 2 weeks in the vitreous, and in 2 days at pH 5.0. In vitro studies with retinal pigment epithelial cell line (ARPE-19) showed no toxicity of the conjugates in the cells. Flow cytometry and confocal microscopy showed cellular association of the nanoparticles and intracellular endosomal localization. Overall, pullulan conjugates showed interesting features that may enable their successful use in intravitreal drug delivery.Peer reviewe

First record of quagga mussel, Dreissena bugensis Andrusov, 1897, in Italy: morphological and genetic evidence in Lake Garda

Here, we report the first record of quagga mussel, Dreissena bugensis Andrusov, 1897, in Italy. This mollusc is native of the Dnieper River drainage of Ukraine and is one of the most aggressive invaders in freshwater ecosystems. The spread of quagga mussels throughout Europe and Northern America begun during the 1940s and 1989, respectively. Individuals of quagga mussel were identified in February and March 2022 in two stations located in the shallow (Bardolino) and deeper basins (Castelletto di Brenzone) of Lake Garda. Samples, collected with an Eckman grab, allowed identifying individuals of quagga mussel attached on macrophytes, stones and other mollusc shells. The identification of the individuals was carried out both through the analysis of morphological characters and genetic and phylogenetic analyses using the mitochondrial COI gene. The discovery of quagga mussel is the last of a long series of reports of non-indigenous species introduced into Lake Garda. Lacking direct connection by rivers or canals with other waterbodies colonized by D. bugensis at the northern side of the Alps, a more probable cause of introduction of this new species into Lake Garda can be due to unintentional overland transport through recreational boats and fishing gear. With this new discovery, Lake Garda has confirmed its pivotal role as a southern Alpine corridor for the introduction of non-indigenous species previously established at the northern border of the Alps. Considering its high colonization rate and compared with other invasion patterns observed in Europe and North America, it can be assumed that D. bugensis will soon establish itself with dominant populations throughout Lake Garda. At the same time, it can also be assumed that it will soon spread to other Italian water bodie

Physical PEGylation to Prevent Insulin Fibrillation

Insulin is one of the most marketed therapeutic proteins worldwide. However, its formulation suffers from fibrillation, which affects the long-term storage limiting the development of novel devices for sustained delivery including portable infusion devices. We have investigated the effect of physical PEGylation on structural and colloidal stability of insulin by using 2 PEGylating agents terminating with polycyclic hydrophobic moieties, cholane and cholesterol: mPEG5kDa-cholane and mPEG5kDa-cholesterol, respectively. Microcalorimetric analyses showed that mPEG5kDa-cholane and mPEG5kDa-cholesterol efficiently bind insulin with binding constants (Ka) of 3.98 104 and 1.14 105 M-1, respectively. At room temperature, the 2 PEGylating agents yielded comparable structural stabilization of \u3b1-helix conformation and decreased dimerization of insulin. However, melting studies showed that mPEG5kDa-cholesterol has superior stabilizing effect of the protein conformation than mPEG5kDa-cholane. Furthermore, the fibrillation study showed that at a 1:1 and 1:5 insulin/polymer molar ratios, mPEG5kDa-cholesterol delays insulin fibrillation 40% and 26% more efficiently, respectively, as compared to mPEG5kDa-cholane which was confirmed by transmission electron microscopy imaging. Insulin was released from the mPEG5kDa-cholane and mPEG5kDa-cholesterol assemblies with comparable kinetic profiles. The physical PEGylation has a beneficial effect on the stabilization and shielding of the insulin structure into the monomeric form, which is not prone to fibrillation and aggregation

Mannosylated Polycations Target CD206+Antigen-Presenting Cells and Mediate T-Cell-Specific Activation in Cancer Vaccination

Immunotherapy is deemed one of the most powerful therapeutic approaches to treat cancer. However, limited response and tumor specificity are still major challenges to address. Herein, mannosylated polycations targeting mannose receptor are developed as vectors for plasmid DNA (pDNA)-based vaccines to improve selective delivery of genetic material to antigen presenting cells and enhance immune cell activation. Three diblock glycopolycations (M15A12, M29A25, and M58A45) and two triblock copolymers (M29A29B9 and M62A52B32) are generated by using mannose (M), agmatine (A), and butyl (B) derivatives to target CD206, complex nucleic acids, and favor the endosomal escape, respectively. All glycopolycations efficiently complex pDNA at N/P ratiosPeer reviewe

pH-Controlled Liposomes for Enhanced Cell Penetration in Tumor Environment

An innovative pH-switchable colloidal system that can be exploited for site-selective anticancer drug delivery has been generated by liposome decoration with a new novel synthetic non-peptidic oligo-arginine cell-penetration enhancer (CPE) and a quenching PEGylated counterpart that detaches from the vesicle surface under the acidic conditions of tumors. The CPE module (Arg(4)-DAG) is formed by four arginine units conjugated to a first-generation (G1) 2,2-bis(hydroxymethyl)propionic acid (bis-MPA)/2,2-bis(aminomethyl)propionic acid (bis-AMPA) polyester dendron terminating with 1,2-distearoyl-3-azidopropane for liposome bilayer insertion. The zeta potential of the Arg(4)-DAG-decorated liposomes increased up to +32 mV as the Arg(4)-DAG/lipids molar ratio increased. The Arg(4)-DAG liposome shielding at pH 7.4 was provided by methoxy-PEGS(5 kDa)-polymethacryloyl sulfadimethoxine (mPEG(5) (kDa)-SDM8) with 7.1 apparent pK(a). Zeta potential, surface plasmon resonance and synchrotron small-angle X-ray scattering analyses showed that at pH 7.4 mPEG(5) (kDa)-SDM8 associates with polycationic Arg(4)-DAG-decorated liposomes yielding liposomes with neutral zeta potential. At pH 6.5, which mimics the tumor environment, mPEG(5) (kDa)-SDM8 detaches from the liposome surface yielding Arg(4)-DAG exposure. Flow cytometry and confocal microscopy showed a 30-fold higher HeLa cancer cell association of the Arg(4)-DAG-decorated liposomes compared to non-decorated liposomes. At pH 7.4, the mPEG(5) (kDa)-SDM8-coated liposomes undergo low cell association while remarkable cell association occurred at pH 6.5, which allowed for the controlled intracellular delivery of model macromolecules and small molecules loaded in the liposome under tumor conditions.Peer reviewe

Pattern of domestic violence from 2011 to 2015 in Beira, Mozambique

Background: Violence against women represents a violation of a fundamental human right and is a significant cause of death and disability worldwide. In developing countries, this issue is particularly dramatic and in sub-Saharan Africa were reached 65% of women reporting domestic violence.Objective: In this study, we assessed the burden and pattern of domestic violence registered at Beira Central Hospital, Mozam- bique from 2011 to 2015.Methods: We performed a descriptive analysis of data collected at the CHB Legal Medicine Service.Results: In five years, are recorded a total amount of 1,491 admissions for domestic violence of which 1307 were females. About 80% of all female cases are represented by the 11-40 age range and, in almost 90% the aggressor was the current or past partner. More than 75% were cases of repeated violence and in more than 60% there were minors attending the phenomenon.Conclusion: It is crucial to act immediately and with a multi-disciplinary approach in order to fight domestic violence, especially against women due to its dramatic consequences as isolation, inability to work, loss of wages, lack of participation in regular activities and limited ability to care for themselves and their children.Keywords: Domestic violence, violence against women, sexual violence, physical violence