165 research outputs found
Estrazione di metalli pesanti da terreni contaminati: recupero della soluzione di lavaggio
No full textIn questo lavoro sono riportati i risultati di prove di estrazione, che simulano un processo di “in situ soil flushing”, effettuate su un terreno permeabile, inquinato artificialmente in laboratorio da piombo e rame. Come agente chelante è stato scelto il sale bisodico dell’ EDTA (E-Na2), proprio per la forte tendenza dell’ acido etilendiamminotetracetico (EDTA) a formare complessi stabili con i metalli pesanti.
Il piombo ed il rame, aggiunti separatamente al terreno, sono, invece tra gli elementi più frequenti nei casi di contaminazione di terreni industriali
Ottimazione delle capacità ossidative nel processo Fenton elettrogenerato
No full textIl reattivo di Fenton, costituito da una miscela di perossido di idrogeno e solfato ferroso, appartiene alla numerosa categoria di sistemi ossidanti che basano la loro attività sulla formazione del radicale ossidrile, una specie estremamente instabile ma dotata di uno straordinario potenziale di ossidazione:
H2O2 + Fe2+ → Fe3+ + HO• + HO- k=51 M-1s-1
La “versione” elettrochimica del reattivo di Fenton1 rappresenta una via per contrastare il rapido decadimento della concentrazione dei radicali ossidrile al procedere degli step reattivi che si verifica nel metodo chimico. In particolare la costante generazione in situ del perossido di idrogeno secondo la reazione
O2 + 2H+ + 2e- H2O2
ovvia ai suoi consistenti consumi nel sistema chimico e la rigenerazione catodica degli ioni Fe2+ secondo la reazione
Fe3+ + e- → Fe2+ Eo=+0.771 V vs SHE
permette di rifornire continuamente il sistema di tale reattivo ovviando alla difficoltà di smaltimento degli ioni ferrici presenti in grosse concentrazioni nel Fenton chimico. Nei precedenti lavori sul Fenton elettrogenerato2 le esigue concentrazioni di substrato da degradare non hanno reso necessaria una particolare ottimizzazione della concentrazione dei reagenti, che d’altra parte è risultata essere di fondamentale importanza nel Fenton chimico3.
Il presente lavoro vuole pertanto studiare l’influenza della concentrazione dei reagenti e dei loro rapporti per una ottimizzazione del Fenton elettrogenerato applicato alla degradazione di alti carichi di substrato anche nell’ottica di un’applicazione a reflui reali
Heavy metal leaching from clayey soil: mechanism of exchange and competitive effects
No full textThis study deals with an experimentation performed with the aim to assess the feasibility of a process for the remediation of copper and iron contaminated soil. Experimental tests were carried out on an artificially contaminated soil, the leaching of copper and iron was performed by flushing an aqueous solution of EDTA: the conditions for metals extraction were evaluated. The effectiveness of soil conditioning prior EDTA extraction was also investigated. Results show that a preliminary soil conditioning with the NaNO3 solution was necessary to avoid variations in soil permeability and to allow the percolation of the extractant solution
Modification of some haematological and haematochemical parameters in horse during long distance rides.
No full textThe aim of this study was to evaluated the effect of the low-intensity long-lasting trial on some haematological and haematochemical parameters during an international endurance race. Six clinically healthy and regularly trained Sella Italiana mares were used. On blood sample collected at rest, at 30 km, and 30’ after the trial, haematological (Red Blood Cell, Haemoglobin, Haematocrit; Mean Corpuscolar Volume, Mean Corpuscolar Hemoglobin, Mean Corpuscolar Hemoglobin Concentration, White Blood Cell and Platelets), and haematochemical parameters (Glucose, Aspartate aminotransferase, Total protein and protein fraction (albumin, α1-globulins, α2-globulins, β-globulins and γ-globulins), Triglycerydes, Cholesterol, Creatinine, Urea, Potassium, Sodium and Chloride) were assessed. One way repeated ANOVA showed a statistical significant effect of low-intensity long-lasting trials on the following parameters: Red Blood Cell (p≤0.009), Haematocrit (p≤0.031), Mean Corpuscolar Hemoglobin Concentration (p≤0.013), White Blood Cell (p<0.0001), Platelets (p<0.0001), α1-globulins (p≤0.038) and γ-globulins (p≤0.009), Creatinine (p≤0.023) and Potassium (p≤0.012). Our results confirm the effects of endurance trial on horse metabolism, underlining that haematological and haematochemical profiles could be an useful index for the prevention of many typical diseases of the athlete horse other than in the assessment of the fitness of the endurance horse
Antitumor HPV E7-specific CTL activity elicited by in vivo engineered exosomes produced through DNA inoculation
No full textPaola Di Bonito,1 Chiara Chiozzini,2 Claudia Arenaccio,2 Simona Anticoli,2 Francesco Manfredi,2 Eleonora Olivetta,2 Flavia Ferrantelli,2 Emiliana Falcone,3 Anna Ruggieri,3 Maurizio Federico2 1Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy; 2National AIDS Center, Istituto Superiore di Sanità, Rome, Italy; 3Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità, Rome, Italy Abstract: We recently proved that exosomes engineered in vitro to deliver high amounts of HPV E7 upon fusion with the Nefmut exosome-anchoring protein elicit an efficient anti-E7 cytotoxic T lymphocyte immune response. However, in view of a potential clinic application of this finding, our exosome-based immunization strategy was faced with possible technical difficulties including industrial manufacturing, cost of production, and storage. To overcome these hurdles, we designed an as yet unproven exosome-based immunization strategy relying on delivery by intramuscular inoculation of a DNA vector expressing Nefmut fused with HPV E7. In this way, we predicted that the expression of the Nefmut/E7 vector in muscle cells would result in a continuous source of endogenous (ie, produced by the inoculated host) engineered exosomes able to induce an E7-specific immune response. To assess this hypothesis, we first demonstrated that the injection of a Nefmut/green fluorescent protein-expressing vector led to the release of fluorescent exosomes, as detected in plasma of inoculated mice. Then, we observed that mice inoculated intramuscularly with a vector expressing Nefmut/E7 developed a CD8+ T-cell immune response against both Nef and E7. Conversely, no CD8+ T-cell responses were detected upon injection of vectors expressing either the wild-type Nef isoform of E7 alone, most likely a consequence of their inefficient exosome incorporation. The production of immunogenic exosomes in the DNA-injected mice was formally demonstrated by the E7-specific CD8+ T-cell immune response we detected in mice inoculated with exosomes isolated from plasma of mice inoculated with the Nefmut/E7 vector. Finally, we provide evidence that the injection of Nefmut/E7 DNA led to the generation of effective antigen-specific cytotoxic T lymphocytes whose activity was likely part of the potent, therapeutic antitumor effect we observed in mice implanted with TC-1 tumor cells. In summary, we established a novel method to generate immunogenic exosomes in vivo by the intramuscular inoculation of DNA vectors expressing the exosome-anchoring protein Nefmut and its derivatives. Keywords: nanovesicles, cytotoxic T lymphocytes, HIV-1 Nef, DNA vector
Diclofenac effects in sea urchin gametes and fertilization
No full textThe drugs, agents for diseases therapy and prevention, are ubiquitous environmental pollutants. We analyzed a specific anti-inflammatory, diclofenac, present in soil and water. The study focused on Arbacia lixula (Linnaeus, 1758) and Paracentrotus lividus (Lamarck, 1816) reproductive success. In A. lixula we identified the drug qualitative and quantitative fertilization effects, and if they affect the functional male and female gametes capacity; in P. lividus we analyzed the drug effects and the male and female gametes quantitative integrity. Through the High Liquid Chromatografy we quantitatively evaluated the real diclofenac contribution on A. lixula embryos and on A. lixula and P. lividus reproductive cells. The results confirmed that the diclofenac is an agent that influence the A. lixula fertilization. The embryos shown: morphological aberrations and slowdown of embryonic development with delay cell divisions. The A. lixula gametes, treated with five toxic solutions, showed modifications or alterations: the eggs are lysed and the spermatozoa shown a decrease in sperm speed. In P. lividus male gametes we observed a real decrease in sperm speed but the egg cells, apparently, no have morpho-structural change although it is found a real diclofenac absorption. The study finds that the two species have a diclofenac sensitivity and that the male gametes are more vulnerable and sensitive to high diclofenac concentrations compared to female gametes. Much remains to be done to increase the knowledges despite today we known the potential risks due to the drugs presence in the waters and being that many of these substances have additive or synergistic activity and their effect can be greatly enhanced
N-terminal fatty acids of NEFmut are required for the CD8+ t-cell immunogenicity of in vivo engineered extracellular vesicles
No full textWe recently described a cytotoxic CD8+ T lymphocyte (CTL) vaccine platform based on the intramuscular (i.m.) injection of DNA eukaryotic vectors expressing antigens of interest fused at the C-terminus of HIV-1 Nefmut, i.e., a functionally defective mutant that is incorporated at quite high levels into exosomes/extracellular vesicles (EVs). This system has been proven to elicit strong CTL immunity against a plethora of both viral and tumor antigens, as well as inhibit both transplantable and orthotopic tumors in mice. However, a number of open issues remain regarding the underlying mechanism. Here we provide evidence that hindering the uploading into EVs of Nefmut-derived products by removing the Nefmut N-terminal fatty acids leads to a dramatic reduction of the downstream antigen-specific CD8+ T-cell activation after i.m. injection of DNA vectors in mice. This result formally demonstrates that the generation of engineered EVs is part of the mechanism underlying the in vivo induced CD8+ T-cell immunogenicity. Gaining new insights on the EV-based vaccine platform can be relevant in view of its possible translation into the clinic to counteract both chronic and acute infections as well as tumors
Riattivazione naturale di Caprine herpesvirus 1 (CpHV.1) in capre con infezione latente in Sicilia
No full textTumor cells endowed with professional antigen-presenting cell functions prime PBLs to generate antitumor CTLs
No full textAbstract: Intrinsic genetic instability of tumor cells leads to continuous production of mutated proteins referred to as tumor-specific neoantigens. Generally, they are recognized as nonself products by the host immune system. However, an effective adaptive response clearing neoantigen-expressing cells is lost in tumor diseases. Most advanced therapeutic strategies aim at inducing neoantigen-specific immune activation through personalized approaches. They include tumor cell exome sequencing, human leukocyte antigen (HLA) typing, synthesis, and injection of peptides/RNA with adjuvants. Here, we propose an innovative method to induce a CD8+ T cytotoxic lymphocyte (CTL) immune response against tumor neoantigens bypassing the steps needed in current therapeutic strategies of personalized vaccination. We assumed that tumor cells can be the most efficient and precise factory of major histocompatibility complex (MHC) class I-associated, tumor neoantigen-derived peptides. Hence, endowing tumor cells with professional antigen-presenting functions would prime CD8+ T lymphocytes towards a response against nonself tumor antigens. To explore this possibility, both adenocarcinoma and melanoma human cells were engineered to express both CD80 and CD86 costimulatory molecules. HLA-matched lymphocytes were then primed through cocultivation with the engineered tumor cells. The generation of tumor-specific CD8+ T lymphocytes was tested through the combined analysis of cell activation markers, formation of immunologic synapses, generation of tumor antigen-specific CD8+ T lymphocytes, and cytotoxic activity. Our data consistently indicate that tumor cells endowed with professional antigen-presenting functions can generate an effective tumor-specific CTL immune response. This finding may open avenues towards the development of innovative antitumor immunotherapies. Key messages: We established a novel method to induce antitumor CTLs without a need to identify TAAs and/or tumor neoantigens.This strategy relies on transducing tumor cells with a retroviral vector expressing both CD80 and CD86.In this way, tumor cells prime naïve CD8+ T lymphocytes in a way that CTLs killing the same tumor cells are generated.These findings open the way towards preclinical assays in the perspective to introduce this antitumor immunotherapy strategy in clinic
Reversed-phase capillary electrochromatography of aloins and related constituents of aloe
No full textThe separation of some anthrone-C-glucosyl constituents of aloe was carried out by isocratic capillary electrochromatography using a 50 I~m I. D. column packed with 3 I~m ODS silica gel. In comparison to HPLC mode, CEC resolution of aloins was obtained in a very short time with
a high selectivi b, factor. The influence of mobile phase parameters (pH and ionic strength of the buffer, acetonitrile amount in the eluent etc.) on the retention of aloins was examined in order to improve the separation conditions and to study the retention mechanism. Efficiency was found to increase with decreasing the EOF up to a reduced plate height of about 2 at a reduced linear velocif/of 0.3 mm s 1. This effect was ascribed to the fact that, under the buffer
conditions used (pH 8), aloins are partially anionically charged, with a consequent low mass transfer bel',,veen the mobile and stationary phases. However, resolution remains almost the same in a wide pH range permitting baseline separations of aloins in short analysis times. Since aloins are known to undergo fast degradation reaction dependent on pH, a kinetic study was also carried out. The method was extended to the analysis of aloins in food and cosmetic
products
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