196 results on '"Di Marzio A"'
Search Results
2. Characterization of seed oil from six in situ collected wild Amaranthus species
- Author
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Amara Noor Hussain, Jeroen Geuens, Ann Vermoesen, Mamoona Munir, Duilio Iamonico, Piera Di Marzio, and Paola Fortini
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Amaranthus ,Ecology ,Ecological Modeling ,fatty acids ,Italy ,seeds ,squalene ,statistical analysis ,tocopherols ,sterols ,Agricultural and Biological Sciences (miscellaneous) ,Chemistry ,Biology ,Nature and Landscape Conservation - Abstract
Six Amaranthus species (A. cruentus, A. hybridus, A. hypochondriacus, A. muricatus, A. tuberculatus, and A. viridis) were collected in Italy (wild habitats) from crops and roadsides. Amaranth seed oil was extracted to obtain fractions rich in squalene. Squalene, free fatty acid, tocopherol, and sterol composition and content were investigated in detail. An analysis of variance and principal components was performed. The oil content in the seed ranged from 5.17% (A. muricatus) to 12.20% (A. tuberculatus). The quantity of squalene in the oil varied from 3.43% (A. muricatus) to 6.09% (A. hypochondriacus). The primary sterols were beta-sitosterol, brassicasterol, campesterol, and stigmasterol. The main tocopherols in all the samples were alfa-tocopherol, beta-tocopherol, and delta-tocopherol. Our results exhibited that the smallest seeds (A. tuberculatus) have the highest percentages of oil and squalene, whereas the largest seeds size (A. muricatus) show the lowest percentages. There is also evidence that the samples growing at lower altitudes show the highest concentration of fatty acids. According to our results, the six wild Amaranthus species exhibited similar characteristics to commercial species. This study confirms that the site of the collection has an impact on the oil and squalene content of the Amaranthus species.
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- 2023
3. Oleuropein-Laded Ufasomes Improve the Nutraceutical Efficacy
- Author
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Maria Chiara Cristiano, Francesca Froiio, Antonia Mancuso, Donato Cosco, Luciana Dini, Luisa Di Marzio, Massimo Fresta, and Donatella Paolino
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ufasomes ,linoleic acid ,oleic acid ,oleuropein ,antioxidant activity ,unsaturated fatty acid liposomes ,Chemistry ,QD1-999 - Abstract
Ufasomes are unsaturated fatty acid liposomes made up of oleic and linoleic acids, natural components required in various biological processes. This kind of nanocarrier is characterized by a simple and dynamic structure and is able to improve the bioavailability of unsaturated fatty acids. The aim of this investigation was to evaluate ufasomes as natural compound delivery systems to deliver oleuropein and improve its antioxidant activity. Oleuropein is a phenolic compound mainly present in olives and olive oil, with several biological properties, such as the antioxidant activity. However, to improve their biological activity, antioxidant compounds should be able to cross cell membranes and uniformly incorporate in cells. Because of the great similarity between their constituents and cell membranes, ufasomes could be advantageous carriers for oleuropein delivery. The physico-chemical characteristics of ufasomes were investigated. A regular shape was shown by transmission electron microscopy studies, while the mean sizes were dependent on the ufasomes composition. In vitro studies highlighted that empty ufasomes did not lead to cell mortality at the tested concentrations and a good carrier internalization in CaCo-2 cells, further studies in vitro studies demonstrated that oleuropein-loaded ufasomes were able to enhance the antioxidant activity of the free active substance making this carrier a suitable one for nutraceutical application.
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- 2021
- Full Text
- View/download PDF
4. Flow Cytometry Analysis of Circulating Extracellular Vesicle Subtypes from Fresh Peripheral Blood Samples
- Author
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Marco Marchisio, Pasquale Simeone, Giuseppina Bologna, Eva Ercolino, Laura Pierdomenico, Damiana Pieragostino, Alessia Ventrella, Francesca Antonini, Genny Del Zotto, Daniele Vergara, Christian Celia, Luisa Di Marzio, Piero Del Boccio, Antonella Fontana, Domenico Bosco, Sebastiano Miscia, and Paola Lanuti
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extracellular vesicles ,polychromatic flow cytometry ,biomarkers ,proteomics ,fresh peripheral blood ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Extracellular vesicles (EVs) are released by shedding during different physiological processes and are increasingly thought to be new potential biomarkers. However, the impact of pre-analytical processing phases on the final measurement is not predictable and for this reason, the translation of basic research into clinical practice has been precluded. Here we have optimized a simple procedure in combination with polychromatic flow cytometry (PFC), to identify, classify, enumerate, and separate circulating EVs from different cell origins. This protocol takes advantage of a lipophilic cationic dye (LCD) able to probe EVs. Moreover, the application of the newly optimized PFC protocol here described allowed the obtainment of repeatable EVs counts. The translation of this PFC protocol to fluorescence-activated cell sorting allowed us to separate EVs from fresh peripheral blood samples. Sorted EVs preparations resulted particularly suitable for proteomic analyses, which we applied to study their protein cargo. Here we show that LCD staining allowed PFC detection and sorting of EVs from fresh body fluids, avoiding pre-analytical steps of enrichment that could impact final results. Therefore, LCD staining is an essential step towards the assessment of EVs clinical significance.
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- 2020
- Full Text
- View/download PDF
5. Gold Nanoparticles Radio-Sensitize and Reduce Cell Survival in Lewis Lung Carcinoma
- Author
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Arvind Pandey, Veronica Vighetto, Nicola Di Marzio, Francesca Ferraro, Matteo Hirsch, Nicola Ferrante, Sankar Mitra, Alessandro Grattoni, and Carly S. Filgueira
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gold nanoparticles ,Lewis lung carcinoma ,radio-sensitization ,clonogenic assay ,comet assay ,Chemistry ,QD1-999 - Abstract
It has been suggested that particle size plays an important role in determining the genotoxicity of gold nanoparticles (GNPs). The purpose of this study was to compare the potential radio-sensitization effects of two different sized GNPs (3.9 and 37.4 nm) fabricated and examined in vitro in Lewis lung carcinoma (LLC) as a model of non-small cell lung cancer through use of comet and clonogenic assays. After treatment with 2Gy X-ray irradiation, both particle sizes demonstrated increased DNA damage when compared to treatment with particles only and radiation alone. This radio-sensitization was further translated into a reduction in cell survival demonstrated by clonogenicity. This work indicates that GNPs of both sizes induce DNA damage in LLC cells at the tested concentrations, whereas the 37.4 nm particle size treatment group demonstrated greater significance in vitro. The presented data aids in the evaluation of the radiobiological response of Lewis lung carcinoma cells treated with gold nanoparticles.
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- 2020
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- View/download PDF
6. Current Trends in Simultaneous Determination of Co-Administered Drugs
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Christian Celia, Luisa Di Marzio, Marcello Locatelli, Piera Ramundo, Francesca D’Ambrosio, and Angela Tartaglia
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analytical methods ,chromatographic procedures ,drug associations ,hyphenated techniques ,biologic matrices ,instrument configurations ,Physics ,QC1-999 ,Chemistry ,QD1-999 - Abstract
Recently, high demand of high-throughput analyses with high sensitivity and selectivity to molecules and drugs in different classes with different physical-chemical properties—and a reduction in analysis time—is a principal milestone for novel methodologies that researchers are trying to achieve—especially when analytical procedures are applied to clinical purposes. In addition, to avoid high doses of a single drug that could cause serious side effects, multi-drug therapies are often used to treat numerous diseases. For these reasons, the demand for methods that allow the rapid analysis of mixed compounds has increased in recent years. In order to respond to these needs, new methods and instruments have been developed. However, often the complexity of a matrix can require a long time for the preparation and processing of the samples. Different problems in terms of components, types of matrices, compounds and physical-chemical complexity are encountered when considering drugs association profiles for quantitative analyses. This review addresses not only recently optimized procedures such as chromatographic separation, but also methods that have allowed us to obtain accuracy (precision and trueness), sensitivity and selectivity in quantitative analyses for cases of drug associations.
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- 2020
- Full Text
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7. Detection and Quantification of eDNA-Associated Bacterial Membrane Vesicles by Flow Cytometry
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Valentina Puca, Eva Ercolino, Christian Celia, Giuseppina Bologna, Luisa Di Marzio, Gabriella Mincione, Marco Marchisio, Sebastiano Miscia, Raffaella Muraro, Paola Lanuti, and Rossella Grande
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bacterial membrane vesicles ,flow cytometry ,lactobacillus reuteri ,helicobacter pylori ,biofilm ,extracellular dna (edna) ,extracellular vesicles ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Bacteria generate membrane vesicles, which are structures known as extracellular vesicles (EVs), reported to be involved in different pathogenic mechanisms, as it has been demonstrated that EVs participate in biofilm formation, cell-to-cell communication, bacteria–host interactions, and nutrients supply. EVs deliver nucleic acids, proteins, and polysaccharides. It has been reported that Helicobacter pylori (H. pylori) and Lactobacillus reuteri (L. reuteri), of both planktonic and biofilm phenotypes, produce EVs carrying extracellular DNA (eDNA). Here, we used polychromatic flow cytometry (PFC) to identify, enumerate, and characterize EVs as well as the eDNA-delivering EV compartment in the biofilm and planktonic phenotypes of H.pylori ATCC 43629 and L. reuteri DSM 17938. Biofilm formation was demonstrated and analyzed by fluorescence microscopy, using a classical live/dead staining protocol. The enumeration of EVs and the detection of eDNA-associated EVs were performed by PFC, analyzing both whole samples (cells plus vesicles) and EVs isolated by ultracentrifugation confirm EVs isolated by ultracentrifugation. PFC analysis was performed relying on a known-size beaded system and a mix of three different fluorescent tracers. In detail, the whole EV compartment was stained by a lipophilic cationic dye (LCD), which was combined to PKH26 and PicoGreen that selectively stain lipids and DNA, respectively. Fluorescence microscopy results displayed that both H. pylori and L. reuteri produced well-structured biofilms. PFC data highlighted that, in both detected bacterial species, biofilms produced higher EVs counts when paralleled to the related planktonic phenotypes. Furthermore, the staining with PicoGreen showed that most of the generated vesicles were associated with eDNA. These data suggest that the use of PFC, set according to the parameters here described, allows for the study of the production of eDNA-associated EVs in different microbial species in the same or several phases of growth, thus opening new perspectives in the study of microbial derived EVs in clinical samples.
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- 2019
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8. The impact of vinyl chloride exposure on the health of Italian workers: an evaluation from SIREP compliance data
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Stefania Massari, Davide Di Marzio, Marisa Corfiati, Sergio Iavicoli, Alessandro Marinaccio, and Alberto Scarselli
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Male ,Proportionate mortality ,Health, Toxicology and Mutagenesis ,Vinyl Chloride ,010501 environmental sciences ,Toxicology ,Disease cluster ,01 natural sciences ,Vinyl chloride ,Occupational safety and health ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Occupational Exposure ,Environmental health ,Humans ,Industry ,Medicine ,Carcinogen ,0105 earth and related environmental sciences ,General Environmental Science ,Exposure assessment ,business.industry ,Low dose ,Public Health, Environmental and Occupational Health ,030210 environmental & occupational health ,chemistry ,Carcinogens ,Female ,Occupational exposure ,business ,Plastics - Abstract
The aim of the study is to investigate exposures to vinyl chloride monomer (VCM) at workplace in Italy and the related burden of diseases. Measurements data was collected from a nation-wide occupational exposure registry (SIREP, 1996-2016). Potentially exposed workers were estimated for some industrial sectors. Concurrent exposures were investigated using cluster analysis. Proportionate mortality ratios were calculated linking data to national mortality statistics (2005-2015). Overall 8,460 measurements were analyzed. Most exposures occurred in the manufacture of chemicals and plastic products. A total of 12,148 workers potentially exposed was estimated (64% male). Concurrent exposures were detected frequently (83%). An elevated proportion of deaths for liver cancer was found in male exposed workers. Although VCM is a known carcinogen for humans, there are still many exposure situations, albeit at low doses, in the chemical and plastic industries.
- Published
- 2021
9. Doxorubicin Hydrochloride-Loaded Nonionic Surfactant Vesicles to Treat Metastatic and Non-Metastatic Breast Cancer
- Author
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Donato Cosco, Constantin Mircioiu, Maria Chiara Cristiano, Nicola d'Avanzo, Barbara Ruozi, Massimo Fresta, Christian Celia, Martina Di Francesco, and Luisa Di Marzio
- Subjects
Drug ,MDA-MB-468 ,Chemistry ,General Chemical Engineering ,media_common.quotation_subject ,Cancer ,General Chemistry ,Pharmacology ,medicine.disease ,Metastatic breast cancer ,Article ,medicine ,Doxorubicin Hydrochloride ,Distribution (pharmacology) ,Nanocarriers ,Cytotoxicity ,QD1-999 ,media_common - Abstract
Doxorubicin hydrochloride (DOX) is currently used to treat orthotropic and metastatic breast cancer. Because of its side effects, the use of DOX in cancer patients is sometimes limited; for this reason, several scientists tried designing drug delivery systems which can improve drug therapeutic efficacy and decrease its side effects. In this study, we designed, prepared, and physiochemically characterized nonionic surfactant vesicles (NSVs) which are obtained by self-assembling different combinations of hydrophilic (Tween 20) and hydrophobic (Span 20) surfactants, with cholesterol. DOX was loaded in NSVs using a passive and pH gradient remote loading procedure, which increased drug loading from ∼1 to ∼45%. NSVs were analyzed in terms of size, shape, size distribution, zeta potential, long-term stability, entrapment efficiency, and release kinetics, and nanocarriers having the best physiochemical parameters were selected for further in vitro tests. NSVs with and without DOX were stable and showed a sustained drug release up to 72 h. In vitro studies, with MCF-7 and MDA MB 468 cells, demonstrated that NSVs, containing Span 20, were better internalized in MCF-7 and MDA MB 468 cells than NSVs with Tween 20. NSVs increased the anticancer effect of DOX in MCF-7 and MDA MB 468 cells, and this effect is time and dose dependent. In vitro studies using metastatic and nonmetastatic breast cancer cells also demonstrated that NSVs, containing Span 20, had higher cytotoxicity than NSVs with Tween 20. The resulting data suggested that DOX-loaded NSVs could be a promising nanocarrier for the potential treatment of metastatic breast cancer.
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- 2021
10. Overcoming Nanoparticle-Mediated Complement Activation by Surface PEG Pairing
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Lin-Ping Wu, Christian Celia, Santosh Aryal, Jaehong Key, Sara Esposito, Martina Pannuzzo, Seyed Moein Moghimi, Paolo Decuzzi, and Luisa Di Marzio
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Complement lectin pathway ,Chemistry ,Mechanical Engineering ,Nanoparticle ,Bioengineering ,02 engineering and technology ,General Chemistry ,Surface engineering ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,Polyethylene Glycols ,Complement system ,Pairing ,PEG ratio ,PEGylation ,Biophysics ,Nanoparticles ,General Materials Science ,0210 nano-technology ,Complement Activation ,Deposition (chemistry) - Abstract
Many PEGylated nanoparticles activate the complement system, which is an integral component of innate immunity. This is of concern as uncontrolled complement activation is potentially detrimental and contributes to disease pathogenesis. Here, it is demonstrated that, in contrast to carboxyPEG
- Published
- 2020
11. Sarcopenia and Vitamin D Deficiency in Patients with Crohn's Disease: Pathological Conditions That Should Be Linked Together
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Francesco Palmese, Pierluigi Cataleta, Maria Giulia Sama, Rossella Del Toro, Marco Domenicali, Giulia Di Marzio, Palmese F., Del Toro R., Di Marzio G., Cataleta P., Sama M.G., and Domenicali M.
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Vitamin ,Crohn’s disease ,medicine.medical_specialty ,Sarcopenia ,Nutritional assess-ment ,skeletal muscle function ,Review ,Disease ,malnutrition ,Gastroenterology ,Inflammatory bowel disease ,vitamin D deficiency ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Crohn Disease ,inflammatory bowel disease ,Internal medicine ,Vitamin D and neurology ,Prevalence ,Medicine ,Humans ,TX341-641 ,030212 general & internal medicine ,Vitamin D ,Dietary Supplement ,Crohn's disease ,Nutrition and Dietetics ,business.industry ,Nutrition. Foods and food supply ,medicine.disease ,Vitamin D Deficiency ,Malnutrition ,chemistry ,nutritional assessment ,Dietary Supplements ,030211 gastroenterology & hepatology ,business ,Human ,Food Science - Abstract
Sarcopenia is a prevalent condition in patients with Crohn’s disease (CD), representing an independent predictor factor for the development of major postoperative complications. Thus, a proper assessment of the muscle strength, by using different validated tools, should be deemed an important step of the clinical management of these patients. Patients with CD are frequently malnourished, presenting a high prevalence of different macro- and micro-nutrient deficiencies, including that of vitamin D. The available published studies indicate that vitamin D is involved in the regulation of proliferation, differentiation, and regeneration of muscle cells. The relationship between vitamin D deficiency and sarcopenia has been extensively studied in other populations, with interesting evidence in regards to a potential role of vitamin D supplementation as a means to prevent and treat sarcopenia. The aim of this review was to find studies that linked together these pathological conditions.
- Published
- 2021
12. Nano-bio interface between human plasma and niosomes with different formulations indicates protein corona patterns for nanoparticle cell targeting and uptake
- Author
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Massimo Fresta, Francesco Salvatore, Stefania Orrù, Christian Celia, Esther Imperlini, Armando Cevenini, Annalisa Mandola, Luisa Di Marzio, Maddalena Raia, Imperlini, E., Celia, C., Cevenini, A., Mandola, A., Raia, M., Fresta, M., Orru, S., Di Marzio, L., and Salvatore, F.
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Biodistribution ,animal structures ,Protein Corona ,02 engineering and technology ,Proteomics ,03 medical and health sciences ,Drug Delivery Systems ,Nanoparticle ,In vivo ,Humans ,General Materials Science ,Tissue Distribution ,Niosome ,Liposomes ,Nanoparticles ,030304 developmental biology ,0303 health sciences ,Chemistry ,Vesicle ,021001 nanoscience & nanotechnology ,Liposome ,Polysorbates ,Drug delivery ,Biophysics ,0210 nano-technology ,Drug Delivery System ,Human - Abstract
Unraveling the proteins interacting with nanoparticles (NPs) in biological fluids, such as blood, is pivotal to rationally design NPs for drug delivery. The protein corona (PrC), formed on the NP surface, represents an interface between biological components and NPs, dictating their pharmacokinetics and biodistribution. PrC composition depends on biological environments around NPs and on their intrinsic physicochemical properties. We generated different formulations of non-ionic surfactant/non-phospholipid vesicles, called niosomes (NIOs), using polysorbates which are biologically safe, cheap, non-toxic and scarcely immunogenic. PrC composition and relative protein abundance for all designed NIOs were evaluated ex vivo in human plasma (HP) by quantitative label-free proteomics. We studied the correlation of the relative protein abundance in the corona with cellular uptake of the PrC-NIOs in healthy and cancer human cell lines. Our results highlight the effects of polysorbates on nano-bio interactions to identify a protein pattern most properly aimed to drive the NIO targeting in vivo, and assess the best conditions of PrC-NIO NP uptake into the cells. This study dissected the biological identity in HP of polysorbate-NIOs, thus contributing to shorten their passage from preclinical to clinical studies and to lay the foundations for a personalized PrC. This journal is
- Published
- 2021
13. Influence of drug/lipid interaction on the entrapment efficiency of isoniazid in liposomes for antitubercular therapy: a multi-faced investigation
- Author
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Domenico Truzzolillo, Federico Bordi, Maria Carafa, Angelo Sarra, Francesca Sciolla, Silvia Trabalzini, Carlotta Marianecci, Luisa Di Marzio, Edouard Chauveau, Simona Sennato, CNR-ISC Sede Sapienza, Piazzale A. Moro 2, I-00185 Rome, Italy, Laboratoire Charles Coulomb (L2C), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Dipartimento di Chimica e Tecnologie farmaceutiche, Universita di Roma, Piazzale A. Moro 5, I-00185 Rome, Italy, Dipartimento di Farmacia, Universita G. d Annunzio, Via dei Vestini, 66100 Chieti, Italy, Dipartimento di Fisica, La Sapienza Universita di Roma, and Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]
- Subjects
Drug ,Laser transmission spectroscopy ,media_common.quotation_subject ,Drug Compounding ,Antitubercular Agents ,FOS: Physical sciences ,02 engineering and technology ,Calorimetry ,Condensed Matter - Soft Condensed Matter ,010402 general chemistry ,Unilamellar liposomes ,01 natural sciences ,Entrapment ,Colloid and Surface Chemistry ,Therapeutic index ,Mesoscale and Nanoscale Physics (cond-mat.mes-hall) ,medicine ,Drug-lipid interaction ,Isoniazid ,Scattering techniques ,Static light scattering ,Physics - Biological Physics ,Physical and Theoretical Chemistry ,media_common ,Liposome ,Condensed Matter - Mesoscale and Nanoscale Physics ,Chemistry ,Vesicle ,Phosphatidylglycerols ,Surfaces and Interfaces ,General Medicine ,021001 nanoscience & nanotechnology ,bacterial infections and mycoses ,0104 chemical sciences ,3. Good health ,Biological Physics (physics.bio-ph) ,Liposomes ,Biophysics ,Soft Condensed Matter (cond-mat.soft) ,[PHYS.PHYS.PHYS-CHEM-PH]Physics [physics]/Physics [physics]/Chemical Physics [physics.chem-ph] ,Nanocarriers ,0210 nano-technology ,[PHYS.COND.CM-SCM]Physics [physics]/Condensed Matter [cond-mat]/Soft Condensed Matter [cond-mat.soft] ,Biotechnology ,medicine.drug - Abstract
Hypothesis. Isoniazid is one of the primary drugs used in tuberculosis treatment. Isoniazid encapsulation in liposomal vesicles can improve drug therapeutic index and minimize toxic and side effects. In this work, we consider mixtures of hydrogenated soy phosphatidylcholine/phosphatidylglycerol (HSPC/DPPG) to get novel biocompatible liposomes for isoniazid pulmonary delivery. Our goal is to understand if the entrapped drug affects bilayer structure. Experiments. HSPC-DPPG unilamellar liposomes are prepared and characterized by dynamic light scattering, $\zeta$-potential, fluorescence anisotropy and Transmission Electron Microscopy. Isoniazid encapsulation is determined by UV and Laser Transmission Spectroscopy. Calorimetry, light scattering and Surface Pressure measurements are used to get insight on adsorption and thermodynamic properties of lipid bilayers in the presence of the drug. Findings. We find that INH-lipid interaction can increase the entrapment capability of the carrier due to isoniazid adsorption. The preferential INH-HSPC dipole-dipole interaction promotes modification of lipid packing and ordering and favors the condensation of a HSPC-richer phase in molar excess of DPPG. Our findings highlight the importance of fundamental investigations of drug-lipid interactions for the optimal design of liposomal nanocarriers., Comment: 28 pages (main manuscript + supplementary information)
- Published
- 2021
14. Flow Cytometry Analysis of Circulating Extracellular Vesicle Subtypes from Fresh Peripheral Blood Samples
- Author
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Sebastiano Miscia, Damiana Pieragostino, Pasquale Simeone, Domenico Bosco, Eva Ercolino, Piero Del Boccio, Marco Marchisio, Francesca Antonini, Giuseppina Bologna, Christian Celia, Laura Pierdomenico, Luisa Di Marzio, Daniele Vergara, Genny Del Zotto, Antonella Fontana, Paola Lanuti, Alessia Ventrella, Marchisio, M., Simeone, P., Bologna, G., Ercolino, E., Pierdomenico, L., Pieragostino, D., Ventrella, A., Antonini, F., Del Zotto, G., Vergara, D., Celia, C., Di Marzio, L., Del Boccio, P., Fontana, A., Bosco, D., Miscia, S., and Lanuti, P.
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0301 basic medicine ,Proteomics ,Polychromatic flow cytometry ,polychromatic flow cytometry ,030204 cardiovascular system & hematology ,Extracellular vesicles ,Sensitivity and Specificity ,Catalysis ,Article ,Flow cytometry ,Inorganic Chemistry ,lcsh:Chemistry ,03 medical and health sciences ,Plasma ,0302 clinical medicine ,proteomics ,Basic research ,medicine ,Animals ,Humans ,Physical and Theoretical Chemistry ,Particle Size ,Molecular Biology ,lcsh:QH301-705.5 ,Spectroscopy ,medicine.diagnostic_test ,Chemistry ,Organic Chemistry ,Liquid Biopsy ,Reproducibility of Results ,biomarkers ,General Medicine ,Extracellular vesicle ,Biomarker ,fresh peripheral blood ,Cell sorting ,Flow Cytometry ,Peripheral blood ,Computer Science Applications ,Cell biology ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Potential biomarkers ,Fresh peripheral blood ,extracellular vesicles - Abstract
Extracellular vesicles (EVs) are released by shedding during different physiological processes and are increasingly thought to be new potential biomarkers. However, the impact of pre-analytical processing phases on the final measurement is not predictable and for this reason, the translation of basic research into clinical practice has been precluded. Here we have optimized a simple procedure in combination with polychromatic flow cytometry (PFC), to identify, classify, enumerate, and separate circulating EVs from different cell origins. This protocol takes advantage of a lipophilic cationic dye (LCD) able to probe EVs. Moreover, the application of the newly optimized PFC protocol here described allowed the obtainment of repeatable EVs counts. The translation of this PFC protocol to fluorescence-activated cell sorting allowed us to separate EVs from fresh peripheral blood samples. Sorted EVs preparations resulted particularly suitable for proteomic analyses, which we applied to study their protein cargo. Here we show that LCD staining allowed PFC detection and sorting of EVs from fresh body fluids, avoiding pre-analytical steps of enrichment that could impact final results. Therefore, LCD staining is an essential step towards the assessment of EVs clinical significance.
- Published
- 2020
- Full Text
- View/download PDF
15. A Simple and Quick Method for Loading Proteins in Extracellular Vesicles
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Sierra A. Walker, Joy Wolfram, Sara Busatto, Dalila Iannotta, and Luisa Di Marzio
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protein delivery ,Pharmaceutical Science ,lcsh:Medicine ,lcsh:RS1-441 ,02 engineering and technology ,Exosome ,Article ,lcsh:Pharmacy and materia medica ,03 medical and health sciences ,Drug Discovery ,CRISPR ,exosome ,Cas9 ,030304 developmental biology ,0303 health sciences ,Chemistry ,Intercellular transport ,Electroporation ,Drug delivery ,Extracellular vesicles ,Protein delivery ,lcsh:R ,Transfection ,021001 nanoscience & nanotechnology ,Cell biology ,drug delivery ,Molecular Medicine ,0210 nano-technology ,extracellular vesicles ,Intracellular - Abstract
Extracellular vesicles (EVs) mediate intercellular transport of biomolecular cargo in the body, making them promising delivery vehicles for bioactive compounds. Genetic engineering of producer cells has enabled encapsulation of therapeutic proteins in EVs. However, genetic engineering approaches can be expensive, time-consuming, and incompatible with certain EV sources, such as human plasma and bovine milk. The goal of this study was to develop a quick, versatile, and simple method for loading proteins in EVs post-isolation. Proteins, including CRISPR associated protein 9 (Cas9), were bound to cationic lipids that were further complexed with MDA-MB-231 cell-derived EVs through passive incubation. Size-exclusion chromatography was used to remove components that were not complexed with EVs. The ability of EVs to mediate intracellular delivery of proteins was compared to conventional methods, such as electroporation and commercial protein transfection reagents. The results indicate that EVs retain native features following protein-loading and obtain similar levels of intracellular protein delivery as conventional methods, but display less toxicity. This method opens up opportunities for rapid exploration of EVs for protein delivery.
- Published
- 2021
16. Oleuropein-Laded Ufasomes Improve the Nutraceutical Efficacy
- Author
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Francesca Froiio, Donato Cosco, Luciana Dini, Luisa Di Marzio, Maria Chiara Cristiano, Donatella Paolino, Massimo Fresta, and Antonia Mancuso
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linoleic acid ,Antioxidant ,General Chemical Engineering ,medicine.medical_treatment ,Linoleic acid ,antioxidant activity ,02 engineering and technology ,030226 pharmacology & pharmacy ,Article ,lcsh:Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Nutraceutical ,Oleuropein ,medicine ,General Materials Science ,Food science ,Unsaturated fatty acid ,Liposome ,021001 nanoscience & nanotechnology ,Oleic acid ,lcsh:QD1-999 ,chemistry ,oleic acid ,oleuropein ,unsaturated fatty acid liposomes ,ufasomes ,Nanocarriers ,0210 nano-technology - Abstract
Ufasomes are unsaturated fatty acid liposomes made up of oleic and linoleic acids, natural components required in various biological processes. This kind of nanocarrier is characterized by a simple and dynamic structure and is able to improve the bioavailability of unsaturated fatty acids. The aim of this investigation was to evaluate ufasomes as natural compound delivery systems to deliver oleuropein and improve its antioxidant activity. Oleuropein is a phenolic compound mainly present in olives and olive oil, with several biological properties, such as the antioxidant activity. However, to improve their biological activity, antioxidant compounds should be able to cross cell membranes and uniformly incorporate in cells. Because of the great similarity between their constituents and cell membranes, ufasomes could be advantageous carriers for oleuropein delivery. The physico-chemical characteristics of ufasomes were investigated. A regular shape was shown by transmission electron microscopy studies, while the mean sizes were dependent on the ufasomes composition. In vitro studies highlighted that empty ufasomes did not lead to cell mortality at the tested concentrations and a good carrier internalization in CaCo-2 cells, further studies in vitro studies demonstrated that oleuropein-loaded ufasomes were able to enhance the antioxidant activity of the free active substance making this carrier a suitable one for nutraceutical application.
- Published
- 2021
- Full Text
- View/download PDF
17. Extracellular vesicle therapeutics from plasma and adipose tissue
- Author
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Dalila Iannotta, Luisa Di Marzio, Joy Wolfram, Man Yang, and Christian Celia
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Stromal cell ,Cell ,Biomedical Engineering ,Pharmaceutical Science ,Adipose tissue ,Bioengineering ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Extracellular vesicles ,Article ,extracellular vesicles ,isolation methods ,plasma ,therapeutic ,medicine ,General Materials Science ,Chemistry ,Mesenchymal stem cell ,Extracellular vesicle ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Cell biology ,medicine.anatomical_structure ,Cell culture ,0210 nano-technology ,Intracellular ,Biotechnology - Abstract
Extracellular vesicles (EVs) are cell-released lipid-bilayer nanoparticles that contain biologically active cargo involved in physiological and pathological intercellular communication. In recent years, the therapeutic potential of EVs has been explored in various disease models. In particular, mesenchymal stromal cell-derived EVs have been shown to exert anti-inflammatory, anti-oxidant, anti-apoptotic, and pro-angiogenic properties in cardiovascular, metabolic and orthopedic conditions. However, a major drawback of EV-based therapeutics is scale-up issues due to extensive cell culture requirements and inefficient isolation protocols. An emerging alternative approach to time-consuming and costly cell culture expansion is to obtain therapeutic EVs directly from the body, for example, from plasma and adipose tissue. This review discusses isolation methods and therapeutic applications of plasma and adipose tissue-derived EVs, highlighting advantages and disadvantages compared to cell culture-derived ones.
- Published
- 2021
18. Gold Nanoparticles Radio-sensitize and Reduce Cell Survival in Lewis Lung Carcinoma
- Author
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Sankar Mitra, Nicola Di Marzio, Nicola Ferrante, Veronica Vighetto, Alessandro Grattoni, Arvind Pandey, Matteo Hirsch, and Carly S. Filgueira
- Subjects
Comet assay ,Chemistry ,Colloidal gold ,Cancer research ,Lewis lung carcinoma ,biology_other ,Clonogenic assay ,Cell survival - Abstract
It has been suggested that particle size plays an important role in determining the genotoxicity of gold nanoparticles (GNPs). The purpose of this study was to compare the potential radio-sensitization effects of two different sized GNPs (3.9 and 37.4 nm) fabricated and examined in vitro in Lewis Lung carcinoma (LLC) as a model of non-small cell lung cancer through use of comet and clonogenic assays. After the treatment of 2Gy X-ray irradiation, both particle sizes demonstrated increased DNA damage when compared to treatment with particles only and radiation alone. This radio-sensitization was further translated into a reduction in cell survival demonstrated by clonogenicity. This work indicates that GNPs of both sizes induce DNA damage in LLC cells at the tested concentrations, whereas the 37.4 nm particle size treatment group demonstrated greater significance in vitro. The presented data aids in the evaluation of the radiobiological response of Lewis Lung carcinoma cells treated with gold nanoparticles.
- Published
- 2020
19. Validation via challenge test of a dynamic growth-death model for the prediction of Listeria monocytogenes kinetics in Pecorino di Farindola cheese
- Author
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Romina Romantini, Luca Candeloro, Diana Neri, Luigi Iannetti, Gino Angelo Santarelli, Patrizia Tucci, Violeta Di Marzio, Annamaria Conte, Gabriella Centorotola, Francesco Pomilio, Giacomo Migliorati, Romolo Salini, and Anna Franca Sperandii
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Microorganism ,Population ,medicine.disease_cause ,Microbiology ,Models, Biological ,03 medical and health sciences ,chemistry.chemical_compound ,Listeria monocytogenes ,Cheese ,Lactobacillales ,Raw Foods ,medicine ,Animals ,Food science ,education ,030304 developmental biology ,0303 health sciences ,education.field_of_study ,biology ,030306 microbiology ,food and beverages ,Ripening ,General Medicine ,Raw milk ,biology.organism_classification ,Lactic acid ,Kinetics ,Milk ,chemistry ,Italy ,Food Microbiology ,Rennet ,Bacteria ,Food Science - Abstract
Pecorino di Farindola is a typical cheese produced in the area surrounding the village of Farindola, located in the Abruzzo Region (central Italy), unique among Italian cheese because only raw ewe milk and pig rennet are used for its production. In the literature it is well documented that raw milk is able to support the growth of pathogenic microorganisms such as Listeria monocytogenes. Predictive microbiology can be useful in order to predict growth-death kinetics of pathogenic bacteria, on the basis of known environmental conditions. Aim of this study was to compare predictions obtained from a model, originally designed to predict the kinetics of L. monocytogenes in the dynamic growth-death environment of drying fresh sausage, with the results of challenge tests performed during the ripening of Pecorino di Farindola produced from artificially contaminated raw ewe milk. A challenge test was carried out using ewe raw milk inoculated with L. monocytogenes, in order to produce Pecorino di Farindola cheese stored at 18 °C for 149 days of ripening. During the ripening period, pH and aw values decreased in all samples analysed; lactic acid bacteria become the prevailing microbial population, while for L. monocytogenes a period of stability (neither growth nor death) followed the initial situation. The growth inhibition and the following inactivation may mostly be due to competition with the autochthonous microbiota and to the reduction of water activity. Mathematical modelling was used in order to predict microbial kinetics in the dynamic ripening environment, joining growth and death patterns in a continuous way, and including the highly uncertain growth/no growth range separating the two regions. The effect of lactic acid bacteria on the growth of pathogens was also included. Predicted microbial kinetics were satisfactory, as confirmed by the absence of statistically significant difference between observed and predicted values (p > 0.05). The present study proved, via challenge tests, that a dynamic growth/death model, previously used for a meat product, can be fruitfully used in cheese characterized by active competitive microbiota and progressive drying during ripening.
- Published
- 2020
20. Immunogenicity of polyethylene glycol based nanomedicines: mechanisms, clinical implications and systematic approach
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Christian Celia, Maria Carafa, Hélder A. Santos, Massimo Fresta, Antonella Barone, Luisa Di Marzio, and Nicola d'Avanzo
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Pharmacology ,0303 health sciences ,Chemistry ,Immunogenicity ,Biochemistry (medical) ,anti-PEG antibodies ,complement activation ,hypersensitivity reactions ,immunogenicity ,Pharmaceutical Science ,Medicine (miscellaneous) ,02 engineering and technology ,Polyethylene glycol ,021001 nanoscience & nanotechnology ,Pegylated Liposomal Doxorubicin ,Complement system ,03 medical and health sciences ,chemistry.chemical_compound ,Splenic Marginal Zone ,Pharmacology (medical) ,0210 nano-technology ,Stealth liposomes ,Genetics (clinical) ,030304 developmental biology - Published
- 2020
21. Magnetoelectrochemistry and asymmetric electrochemical reactions
- Author
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Suryakant Mishra, Roberto Giovanardi, Marzia di Marzio, and Francesco Tassinari
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Materials science ,Working electrode ,Chiral imprinting ,chiral imprinting ,chemistry.chemical_element ,Electrochemistry ,Redox ,Asymmetric reaction ,Electronic, Optical and Magnetic Materials ,chemistry.chemical_compound ,Nickel ,Enantiopure drug ,chemistry ,Chemical engineering ,Electrodeposition ,Chemistry (miscellaneous) ,Electrode ,Chiral nickel ,Materials Chemistry ,Tartaric acid ,electrodeposition ,Magnetoelectrochemistry ,asymmetric reaction ,chiral nickel - Abstract
Magnetoelectrochemistry is a branch of electrochemistry where magnetic fields play a vital role in the oxidation and reduction process of the molecules. When it comes to spin-dependent electrochemistry (SDE), becomes a new paradigm. This work presents electrochemical response during the “chiral imprinting” on working electrodes and the effects of potentiostatic and galvanostatic methods. We explore the use of the SDE concept, which is implemented for chiral-ferromagnetic (CFM) hybrid working electrodes, and we compare various electrochemical parameters affecting the quality of deposition. We electrochemically co-deposited nickel (Ni) with a chiral compound (tartaric acid) in its enantiopure forms (L and D), which allows us to obtain a chiral co-deposited nickel-tartaric acid (Ni-LTA or Ni-DTA) working electrode.
- Published
- 2020
22. Early retinal flow changes after vitreoretinal surgery in idiopathic epiretinal membrane using swept source optical coherence tomography angiography
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Agbeanda Aharrh-Gnama, Rossella D'Aloisio, Marta Di Nicola, Pasquale Viggiano, Rodolfo Mastropasqua, Cesare Mariotti, Paolo Carpineto, Lisa Toto, Enrico Borrelli, Carla Iafigliola, and Guido Di Marzio
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0301 basic medicine ,medicine.medical_specialty ,genetic structures ,medicine.medical_treatment ,Vitrectomy ,Ilm peeling ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Ophthalmology ,medicine ,retinal vessel density ,business.industry ,Length density ,Retinal ,General Medicine ,Optical coherence tomography angiography ,Vitreoretinal surgery ,medicine.disease ,vitreoretinal surgery ,swept source optical coherence tomography angiography ,030104 developmental biology ,chemistry ,idiopathic epiretinal membrane ,030221 ophthalmology & optometry ,sense organs ,Epiretinal membrane ,business ,Perfusion - Abstract
(1) Background: The aim of this observational cross-sectional work was to investigate early retinal vascular changes in patients undergoing idiopathic epiretinal membrane (iERM) surgery using swept source optical coherence tomography angiography (SS-OCTA), (2) Methods: 24 eyes of 24 patients who underwent vitrectomy with internal limiting membrane (ILM) peeling were evaluated pre- and postoperatively using SS-OCTA system (PLEX Elite 9000, Carl Zeiss Meditec Inc., Dublin, CA, USA). For each eye, five 6x6-mm OCTA volume scans were acquired by two observers independently. The en face images of superficial capillary plexus (SCP) were then exported to imageJ and a semi-automated algorithm was used for subsequent quantitative analysis. Perfusion density (PD), vessel length density (VLD), vessel diameter index (VDI) and vessel tortuosity (VT) of SCP were evaluated in both the parafoveal (2.5 mm diameter) and perifoveal areas (5.5 mm diameter), (3) Results: At OCTA analysis statistically significant differences were found between controls and diseased eyes for all parameters in parafoveal and perifoveal regions (p<, 0.001, p<, 0.05) except for perifoveal VLD. During 6-month follow up, both anatomical/perfusion and functional parameters showed a statistically significant improvement if compared to preoperative values. In detail, at one month post vitrectomy, VLD and VT significantly changed in parafoveal region (p=0.043, p=0.045), while PD and VDI showed a trend of increase in both parafoveal and perifoveal region. At 6 months after surgery, PD, VLD and VT of parafoveal region significantly improved (p=0.021, p=0.018, p=0.047 respectively). (4) Conclusions: SS-OCTA provides a quantitative and qualitative analysis of the superficial capillary plexus allowing for early vascular changes assessment after vitrectomy with iERM and ILM peeling.
- Published
- 2019
23. pH-sensitive niosomes: Effects on cytotoxicity and on inflammation and pain in murine models
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Maria Carafa, Donatella Paolino, Jessica Ponti, Stefano Pieretti, Elena Del Favero, Alessia Bogni, Federica Rinaldi, François Rossi, Valeria Rondelli, Carlotta Marianecci, and Luisa Di Marzio
- Subjects
BALB 3T3 Cells ,02 engineering and technology ,Pharmacology ,01 natural sciences ,Mice ,Pulmonary surfactant ,Drug Discovery ,animal ,anti-nociceptive/anti-inflammatory activity ,Cells, Cultured ,Pain Measurement ,ibuprofen ,Liposome ,Chemistry ,Vesicle ,Anti-Inflammatory Agents, Non-Steroidal ,Biological activity ,General Medicine ,Hydrogen-Ion Concentration ,021001 nanoscience & nanotechnology ,lidocaine ,cytotoxicity ,0210 nano-technology ,Research Article ,antinociceptive/anti-inflammatory activity ,Pain ,010402 general chemistry ,anti-inflammatory agents ,non-steroidal ,PH-sensitive niosomes ,animals ,anti-inflammatory agents, non-steroidal ,BALB 3T3 cells ,cells, cultured ,fibroblasts ,humans ,hydrogen-ion concentration ,inflammation ,liposomes ,mice ,pain ,pain measurement ,surface-active agents ,disease models, animal ,pharmacology ,drug discovery3003 pharmaceutical science ,Surface-Active Agents ,In vivo ,Animals ,Humans ,Niosome ,cultured ,Inflammation ,disease models ,lcsh:RM1-950 ,Fibroblasts ,0104 chemical sciences ,Disease Models, Animal ,HaCaT ,lcsh:Therapeutics. Pharmacology ,Liposomes ,cells ,pH-sensitive niosomes ,Fetal bovine serum - Abstract
pH-sensitive nonionic surfactant vesicles (niosomes) by polysorbate-20 (Tween-20) or polysorbate-20 derivatized by glycine (added as pH sensitive agent), were developed to deliver Ibuprofen (IBU) and Lidocaine (LID). For the physical-chemical characterization of vesicles (mean size, size distribution, zeta potential, vesicle morphology, bilayer properties and stability) dynamic light scattering (DLS), small angle X-ray scattering and fluorescence studies were performed. Potential cytotoxicity was evaluated on immortalized human keratinocyte cells (HaCaT) and on immortalized mouse fibroblasts Balb/3T3. In vivo antinociceptive activity (formalin test) and anti-inflammatory activity tests (paw edema induced by zymosan) in murine models were performed on drug-loaded niosomes. pH-sensitive niosomes were stable in the presence of 0 and 10% fetal bovine serum, non-cytotoxic and able to modify IBU or LID pharmacological activity in vivo. The synthesis of stimuli responsive surfactant, as an alternative to add pH-sensitive molecules to niosomes, could represent a promising delivery strategy for anesthetic and anti-inflammatory drugs.
- Published
- 2017
24. Detection and Quantification of eDNA-Associated Bacterial Membrane Vesicles by Flow Cytometry
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Christian Celia, Raffaella Muraro, Sebastiano Miscia, Eva Ercolino, Giuseppina Bologna, Paola Lanuti, Gabriella Mincione, Marco Marchisio, Di Marzio L, Puca, and Rossella Grande
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0301 basic medicine ,DNA, Bacterial ,Limosilactobacillus reuteri ,030106 microbiology ,Lactobacillus reuteri ,Catalysis ,Article ,biofilm ,Flow cytometry ,lcsh:Chemistry ,Inorganic Chemistry ,03 medical and health sciences ,Extracellular Vesicles ,Fluorescence microscope ,medicine ,Physical and Theoretical Chemistry ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,medicine.diagnostic_test ,biology ,Helicobacter pylori ,Chemistry ,Vesicle ,flow cytometry ,Organic Chemistry ,Cell Membrane ,Biofilm ,General Medicine ,biology.organism_classification ,Computer Science Applications ,Staining ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Biochemistry ,Nucleic acid ,extracellular DNA (eDNA) ,bacterial membrane vesicles ,Bacteria - Abstract
Bacteria generate membrane vesicles, which are structures known as extracellular vesicles (EVs), reported to be involved in different pathogenic mechanisms, as it has been demonstrated that EVs participate in biofilm formation, cell-to-cell communication, bacteria–host interactions, and nutrients supply. EVs deliver nucleic acids, proteins, and polysaccharides. It has been reported that Helicobacter pylori (H. pylori) and Lactobacillus reuteri (L. reuteri), of both planktonic and biofilm phenotypes, produce EVs carrying extracellular DNA (eDNA). Here, we used polychromatic flow cytometry (PFC) to identify, enumerate, and characterize EVs as well as the eDNA-delivering EV compartment in the biofilm and planktonic phenotypes of H.pylori ATCC 43629 and L. reuteri DSM 17938. Biofilm formation was demonstrated and analyzed by fluorescence microscopy, using a classical live/dead staining protocol. The enumeration of EVs and the detection of eDNA-associated EVs were performed by PFC, analyzing both whole samples (cells plus vesicles) and EVs isolated by ultracentrifugation confirm EVs isolated by ultracentrifugation. PFC analysis was performed relying on a known-size beaded system and a mix of three different fluorescent tracers. In detail, the whole EV compartment was stained by a lipophilic cationic dye (LCD), which was combined to PKH26 and PicoGreen that selectively stain lipids and DNA, respectively. Fluorescence microscopy results displayed that both H. pylori and L. reuteri produced well-structured biofilms. PFC data highlighted that, in both detected bacterial species, biofilms produced higher EVs counts when paralleled to the related planktonic phenotypes. Furthermore, the staining with PicoGreen showed that most of the generated vesicles were associated with eDNA. These data suggest that the use of PFC, set according to the parameters here described, allows for the study of the production of eDNA-associated EVs in different microbial species in the same or several phases of growth, thus opening new perspectives in the study of microbial derived EVs in clinical samples.
- Published
- 2019
25. Comparative ecotoxicity of single and binary mixtures exposures of nickel and zinc on growth and biomarkers of Lemna gibba
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M. E. Sáenz, S Martinez, J. L. Alberdi, and W. D. Di Marzio
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Frond ,Antioxidant ,CA MODEL ,Health, Toxicology and Mutagenesis ,Lemna gibba ,medicine.medical_treatment ,chemistry.chemical_element ,Management, Monitoring, Policy and Law ,Toxicology ,BINARY MIXTURE ,LEMNA GIBBA ,Nickel ,Toxicity Tests ,medicine ,ANTIOXIDANT ENZYMES ,Ecotoxicology ,Araceae ,HEAVY METAL ,Food science ,Cadmium ,biology ,purl.org/becyt/ford/2.8 [https] ,Chemistry ,General Medicine ,biology.organism_classification ,Oxidative Stress ,Zinc ,purl.org/becyt/ford/2 [https] ,Catalase ,Toxicity ,biology.protein ,Ecotoxicity ,Biomarkers ,Water Pollutants, Chemical - Abstract
The aim of the present study was to investigate the toxicity effects of cadmium–nickel (Cd–Ni) after single and mixtures exposures over the macrophyte Lemna gibba. Effects were assessed on growth, as frond number and fresh weight and biochemical parameters, such as total protein content and activity of antioxidant enzymes. Plants were exposed to single Cd and Ni in concentrations that ranged between 0.13–33 mg/L and 0.18 and 11.82 mg/L, respectively. For binary mixtures, individual metal IC50 values were used for selection of the evaluated concentrations. The experimental design consisted in three different ratios based on the concept of toxic units (TU), each ratio was evaluated by five different concentrations. Both single and mixture treatments were performed for 7 days following the conditions according to OECD (2006). Single and mixture exposures affected plant growth and the biomarkers of the antioxidant response. Growth parameters showed a differential sensitivity after individual metal exposures. Cd was more toxic for L. gibba plants when fresh weight was considered, but on the contrary, considering frond number, Ni was the most toxic metal. IC50-7d, based on growth rate calculated on frond number were 17.8 and 2.47 mg/L, and on fresh weight were 1.08 and 3.89 mg/L, for Cd and Ni respectively. LOEC values for Cd were obtained at 2.06 and 1.03 mg/L, for frond number and fresh weight, respectively; while for Ni, these values were 0.92 and 11.82 mg/L. The three evaluated ratios for binary mixtures resulted in a high toxicity considering the same response variables in single metal exposures. Ratio 1 (2/3 TU Cd–1/3 TU Ni) was the most toxic considering both frond number and fresh weight, showing percentage inhibition of growth rates of 96 and 90%, respectively for the highest concentration. A modification of the protein content was observed in single, but especially in the mixture treatments. The activity of catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APOX; EC 1.11.1.11) and guaiacol peroxidase (GPOX; EC 1.11.1.7) was also affected in single and mixtures assays. APOX and GPOX showed a higher increase of its activities respect the controls after mixture treatments than for single metal treatments. Such optimization of the antioxidant system could be one of the causes of the antagonistic toxicity observed in mixture exposures. Concentration addition (CA) reference model, based on frond number, in Cd–Ni mixtures was not a good predictor to evaluate toxicity from dissolved metal concentration since the results showed that toxicity was less than additive, with an average of ΣTU = 2.17. The observed antagonisms resulted to be stronger in mixtures with higher metal concentrations.
- Published
- 2019
26. pH-responsive chitosan based hydrogels affect the release of dapsone: Design, set-up, and physicochemical characterization
- Author
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Salette Reis, Sofia A. Costa Lima, Maria Chiara Cristiano, Alexandre C.C. Vieira, Bruno Sarmento, Domingos Ferreira, Christian Celia, Luisa Di Marzio, Alessandro Silveri, Donatella Paolino, and Luíse L. Chaves
- Subjects
Chemical Phenomena ,02 engineering and technology ,Polyethylene glycol ,Biochemistry ,Chitosan ,03 medical and health sciences ,chemistry.chemical_compound ,Structural Biology ,Leprosy ,medicine ,Interpenetrating polymer network ,Solubility ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Drug Carriers ,technology, industry, and agriculture ,Hydrogels ,General Medicine ,Hydrogen-Ion Concentration ,021001 nanoscience & nanotechnology ,Controlled release ,Drug Liberation ,chemistry ,Self-healing hydrogels ,Glutaraldehyde ,Swelling ,medicine.symptom ,0210 nano-technology ,Rheology ,Dapsone ,Nuclear chemistry - Abstract
Dapsone (DAP) is a bactericidal agent used in the treatment of leprosy, caused by Mycobacterium leprae. Despite its therapeutic potential, DAP has low solubility, which results in allow therapeutic index and a high microbial resistance. Recently, new approaches were used to increase the DAP solubility. In particular, the use of interpenetrating polymer network (IPN)-hydrogels based chitosan (CS) for the controlled release of DAP provides some advantages because they can modify their swelling properties and network structures as a response to environmental stimuli. The aim of this study was to synthesize and physicochemically characterize pH-responsive chitosan/polymer hydrogels to control the release of DAP. For this reason, different combination of polymers, such as polyvinyl pyrrolidone, polyethylene glycol and hydroxypropyl methylcellulose, and concentrations of the cross-linking agents (glutaraldehyde) were used and then blended to the CS. The resulting hydrogels were evaluated in terms of physicochemical and swelling properties, rheological analysis and in vitro release of DAP at different pHs (1.2-6.8). Hydrogels were further characterized by Fourier transformed infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) analysis. pH-responsive DAP-loaded hydrogels may represent the set-up for developing potential oral formulations for the treatment of leprosy caused by Mycobacterium leprae.
- Published
- 2019
27. The role of freshwater copepods in the environmental risk assessment of caffeine and propranolol mixtures in the surface water bodies of Spain
- Author
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Leonor Nozal Martínez, Walter Dario Di Marzio, Tiziana Di Lorenzo, Andrea Castaño-Sánchez, Patricia Garcia-Doncel, Diana M. P. Galassi, and Sanda Iepure
- Subjects
Health, Toxicology and Mutagenesis ,0208 environmental biotechnology ,Fresh Water ,02 engineering and technology ,010501 environmental sciences ,01 natural sciences ,Ciencias de la Tierra y relacionadas con el Medio Ambiente ,chemistry.chemical_compound ,CONCENTRATION ADDITION ,PHARMACEUTICAL AND PERSONAL CARE PRODUCTS ,Oceanografía, Hidrología, Recursos Hídricos ,Concentration addition ,Chemistry (all) ,General Medicine ,Propranolol ,Pollution ,Crustaceans ,ERA ,Health ,Environmental chemistry ,Species sensitivity distribution ,SPECIES SENSITIVITY DISTRIBUTION ,Caffeine ,CIENCIAS NATURALES Y EXACTAS ,medicine.drug ,Environmental Engineering ,Risk Assessment ,Copepoda ,medicine ,TD Environmental technology. Sanitary engineering ,Animals ,Environmental Chemistry ,CRUSTACEANS ,Toxicology and Mutagenesis ,Pharmaceutical and personal care products ,0105 earth and related environmental sciences ,Environmental risk assessment ,Public Health, Environmental and Occupational Health ,General Chemistry ,QL Zoology ,020801 environmental engineering ,chemistry ,Spain ,Environmental science ,Surface water ,Water Pollutants, Chemical - Abstract
In this study we aimed at assessing: (i) the environmental risk posed by mixtures of caffeine and propranolol to the freshwater ecosystems of Spain; (ii) the sensitivity of freshwater copepod species to the two compounds; (iii) if the toxicity of caffeine and propranolol to freshwater copepods contributes to the environmental risk posed by the two compounds in the freshwater bodies of Spain. The environmental risk was computed as the ratio of MECs (i.e. the measured environmental concentrations) to PNECs (i.e. the respective predicted no-effect concentrations). The effects of caffeine and propranolol on the freshwater cyclopoid Diacyclops crassicaudis crassicaudis were tested both individually and in binary mixtures. Propranolol posed an environmental risk in some but not in all the surface water ecosystems of Spain investigated in this study, while caffeine posed an environmental risk to all the investigated freshwater bodies, both as single compound and in the mixture with propranolol. Propranolol was the most toxic compound to D. crassicaudis crassicaudis, while caffeine was non-toxic to this species. The CA model predicted the toxicity of the propranolol and caffeine mixture for this species. D. crassicaudis crassicaudis was much less sensitive than several other aquatic species to both compounds. The sensitivity of D. crassicaudis crassicaudis does not increase the environmental risk posed by the two compounds in the freshwater bodies of Spain, however, further testing is recommended since the effect of toxicants on freshwater copepods can be more pronounced under multiple stressors and temperature increasing due to climate change. Fil: Di Lorenzo, Tiziana. Consiglio Nazionale delle Ricerche; Italia. Istituto di Ricerca sugli Ecosistemi Terrestri; Italia Fil: Castaño Sánchez, Andrea. Universidad de Alcalá; España Fil: Di Marzio, Walter Dario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján. Departamento de Ciencias Básicas; Argentina Fil: García Doncel, Patricia. Universidad de Alcalá; España Fil: Nozal Martínez, Leonor. Universidad de Alcalá; España Fil: Galassi, Diana Maria Paola. Universita degli Studi dell'Aquila; Italia Fil: Iepure, Sanda. Universidad de Alcalá; España
- Published
- 2019
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28. Influence of the Supramolecular Micro-Assembly of Multiple Emulsions on their Biopharmaceutical Features and In vivo Therapeutic Response
- Author
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Cinzia Anna Ventura, Luisa Di Marzio, Donato Cosco, Maria Carafa, Maria Chiara Cristiano, Massimo Fresta, Donatella Paolino, and Felisa Cilurzo
- Subjects
Pharmacology ,Chromatography ,medicine.diagnostic_test ,Chemistry ,Clinical Biochemistry ,Permeation ,Membrane ,medicine.anatomical_structure ,In vivo ,Oil droplet ,Spectrophotometry ,Drug Discovery ,Drug delivery ,Emulsion ,Stratum corneum ,medicine ,Molecular Medicine - Abstract
The ability of some surfactants to self-assemble in a water/oil bi-phase environment thus forming supramolecular structure leading to the formation of w/o/w multiple emulsions was investigated. The w/o/w multiple emulsions obtained by self-assembling (one-step preparation method) were compared with those prepared following the traditional two-step procedure. Methyl-nicotinate was used as a hydrophilic model drug. The formation of the multiple emulsion structure was evidenced by optical microscopy, which showed a mean size of the inner oil droplets of 6 μm and 10 μm for one-step and two-step multiple emulsions, respectively. The in vitrobiopharmaceutical features of the various w/o/w multiple emulsion formulations were evaluated by means of viscosimetry studies, drug release and in vitro percutaneous permeation experiments through human stratum corneum and viable epidermis membranes. The self-assembled multiple emulsions allowed a more gradual percutaneous permeation (a zero-order permeation rate) than the two-step ones. The in vivotopical carrier properties of the two different multiple emulsions were evaluated on healthy human volunteers by using the spectrophotometry of reflectance, an in vivonon invasive method. These multiple emulsion systems were also compared with conventional emulsion formulations. Our findings demonstrated that the multiple emulsions obtained by self-assembling were able to provide a more sustained drug delivery into the skin and hence a longer therapeutic action than two-step multiple emulsions and conventional emulsion formulations. Finally, our findings showed that the supramolecular micro-assembly of multiple emulsions was able to influence not only the biopharmaceutical characteristics but also the potential in vivotherapeutic response.
- Published
- 2015
29. Nose to Brain Delivery: New Trends in Amphiphile-Based 'Soft' Nanocarriers
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Maria Carafa, Carlotta Marianecci, Federica Rinaldi, Luisa Di Marzio, Donatella Paolino, and Patrizia Nadia Hanieh
- Subjects
Polymers ,Chemistry, Pharmaceutical ,Nanotechnology ,Micelle ,Surface-Active Agents ,Drug Delivery Systems ,Drug Discovery ,Amphiphile ,Animals ,Humans ,Tissue Distribution ,Niosome ,Administration, Intranasal ,Pharmacology ,Drug Carriers ,Liposome ,Chemistry ,Brain ,Pharmaceutical Preparations ,Liposomes ,Drug delivery ,Nanoparticles ,Nasal administration ,Nanocarriers ,Drug carrier ,Hydrophobic and Hydrophilic Interactions - Abstract
The aim of the present paper is to highlight the potential of nasal mucosa as an administration route for targeting the central nervous system, in particular, the brain. Among the formulation strategies for enhance nose to brain drug delivery, the use of colloidal carriers has became a revolutionary approach. These systems should be able to entrap drugs in the desired amount, to penetrate through anatomical barriers, to efficiently release the loaded drugs in the site of action and moreover to show a good physicochemical, biological stability and good biocompatibility. The use of vesicular systems (liposomes and niosomes) together with the use of micelles, in nose to brain delivery are here presented. Vesicle structure is characterized by the presence of a hydrophobic bilayer and an aqueous core that is absent in micelles. Amphiphilic molecules are responsible for soft nanocarriers formation, in particular: liposomes are formed by phospholipids, while niosomes by non-ionic surfactant and micelles by amphiphilic polymers.
- Published
- 2015
30. Nanoliposomes as Multidrug Carrier of Gemcitabine/Paclitaxel for the Effective Treatment of Metastatic Breast Cancer Disease: A Comparison with Gemzar and Taxol
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Maria Chiara Cristiano, Francesca Froiio, Nicola d'Avanzo, Martina Di Francesco, Luisa Di Marzio, Massimo Fresta, and Christian Celia
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Pharmacology ,Gemcitabine/Paclitaxel ,business.industry ,Biochemistry (medical) ,Pharmaceutical Science ,Medicine (miscellaneous) ,Disease ,medicine.disease ,Gemcitabine Hydrochloride ,Metastatic breast cancer ,chemistry.chemical_compound ,Breast cancer ,Paclitaxel ,chemistry ,Cancer research ,medicine ,Effective treatment ,Pharmacology (medical) ,business ,Genetics (clinical) - Published
- 2020
31. Ammonium glycyrrhizate skin delivery from ultradeformable liposomes: A novel use as an anti-inflammatory agent in topical drug delivery
- Author
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Christian Celia, Marcello Locatelli, Maria Chiara Cristiano, Donatella Paolino, Dalila Iannotta, Felisa Cilurzo, Antonella Barone, and Luisa Di Marzio
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Adult ,Surface Properties ,medicine.drug_class ,Administration, Topical ,Anti-Inflammatory Agents ,Ammonium glycyrrhizate ,02 engineering and technology ,Pharmacology ,01 natural sciences ,Anti-inflammatory ,Anti-inflammatory activity ,chemistry.chemical_compound ,Drug Delivery Systems ,Colloid and Surface Chemistry ,In vivo ,0103 physical sciences ,Glycyrrhiza ,Stratum corneum ,medicine ,Humans ,Ammonium ,Colloidal nanocarriers ,Particle Size ,Physical and Theoretical Chemistry ,Sodium Cholate ,Glycyrrhizin ,Topical administration ,Ultradeformable liposomes ,Skin ,Inflammation ,010304 chemical physics ,biology ,Nicotinic Acids ,Surfaces and Interfaces ,General Medicine ,Permeation ,Glycyrrhizic Acid ,021001 nanoscience & nanotechnology ,biology.organism_classification ,Healthy Volunteers ,medicine.anatomical_structure ,chemistry ,Erythema ,Liposomes ,0210 nano-technology ,Biotechnology - Abstract
Glycyrrhiza glabra L. is a native plant of Central and South-Western Asia that is also diffused in the Mediterranean area and contains several bioactive compounds such as: flavonoids, sterols, triterpene and saponins. Glycyrrhizin, containing glycyrrhizic and glycyrrhizinic acids has anti-inflammatory and antiallergic effects that are similar to corticosteroids. Ammonium glycyrrhizinate is a derivative salt of glycyrrhizic acid with similar anti-inflammatory activity that cannot pass through the skin due to its physicochemical properties and molecular weight. Although several nanoformulations, such as ethosomes, are designed to provide a systemic effect through a topical application, there are different limitations to the distribution inside the blood stream. For this reason, ultradeformable liposomes, or transfersomes, are selected to improve the topical delivery of drugs and allow the distribution of payloads in the blood stream because they pass intact through the stratum corneum epidermis barrier, due to the presence of sodium cholate, aqueous cutaneous gradient, and the rapid penetration of transfersomes by cutaneous tight junctions, thus allowing the systemic delivery of different therapeutic cargo in non-occlusive conditions. The aim of this work was the synthesis and physicochemical characterization of the ammonium glycyrrhizinate-loaded ultradeformable liposomes, the evaluation of drug release and permeation through stratum corneum and epidermis barrier. The in vivo anti-inflammatory effect of ammonium glycyrrhizinate-loaded ultradeformable liposomes was tested on human healthy volunteers. The results demonstrated that the ammonium glycyrrhizinate-loaded ultradeformable liposomes decreased the skin inflammation on the human volunteers and the resulting nanoformulations can be used as a potential topical drug delivery system for anti-inflammatory therapy. ☆Parts of these results were presented as a poster communication at the Recent Developments in Pharmaceutical Analysis 2019 (RDPA 2019), Chieti, Italy.
- Published
- 2020
32. Gold Nanoparticles Radio-Sensitize and Reduce Cell Survival in Lewis Lung Carcinoma
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Matteo Hirsch, Alessandro Grattoni, Arvind Pandey, Veronica Vighetto, Carly S. Filgueira, Nicola Di Marzio, Nicola Ferrante, Sankar Mitra, and Francesca Ferraro
- Subjects
DNA damage ,General Chemical Engineering ,medicine.disease_cause ,Article ,lcsh:Chemistry ,03 medical and health sciences ,0302 clinical medicine ,comet assay ,medicine ,Clonogenic assay ,Comet assay ,Gold nanoparticles ,Lewis lung carcinoma ,Radio-sensitization ,General Materials Science ,030304 developmental biology ,0303 health sciences ,Chemistry ,radio-sensitization ,In vitro ,lcsh:QD1-999 ,Colloidal gold ,gold nanoparticles ,030220 oncology & carcinogenesis ,Cancer research ,clonogenic assay ,Particle size ,Genotoxicity - Abstract
It has been suggested that particle size plays an important role in determining the genotoxicity of gold nanoparticles (GNPs). The purpose of this study was to compare the potential radio-sensitization effects of two different sized GNPs (3.9 and 37.4 nm) fabricated and examined in vitro in Lewis lung carcinoma (LLC) as a model of non-small cell lung cancer through use of comet and clonogenic assays. After treatment with 2Gy X-ray irradiation, both particle sizes demonstrated increased DNA damage when compared to treatment with particles only and radiation alone. This radio-sensitization was further translated into a reduction in cell survival demonstrated by clonogenicity. This work indicates that GNPs of both sizes induce DNA damage in LLC cells at the tested concentrations, whereas the 37.4 nm particle size treatment group demonstrated greater significance in vitro. The presented data aids in the evaluation of the radiobiological response of Lewis lung carcinoma cells treated with gold nanoparticles.
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- 2020
33. A Custom-Made Semiautomatic Analysis of Retinal Nonperfusion Areas After Dexamethasone for Diabetic Macular Edema
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Antonio Maria Chiarelli, Rossella D'Aloisio, Mariacristina Parravano, Lisa Toto, Luca Di Antonio, Federica Evangelista, Guido Di Marzio, Arcangelo Merla, Giada D'Onofrio, and Rodolfo Mastropasqua
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0301 basic medicine ,medicine.medical_specialty ,Diabetic macular edema ,Biomedical Engineering ,Ischemia ,widefield OCT angiography ,Article ,Dexamethasone ,Macular Edema ,Widefield OCT angiography ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Ophthalmology ,Diabetes mellitus ,Diabetes Mellitus ,medicine ,Humans ,In patient ,retinal nonperfusion areas ,Diabetic Retinopathy ,business.industry ,Retinal Vessels ,Retinal ,Diabetic retinopathy ,Intravitreal dexamethasone implant ,medicine.disease ,Retinal nonperfusion areas ,030104 developmental biology ,intravitreal dexamethasone implant ,chemistry ,030221 ophthalmology & optometry ,Implant ,diabetic macular edema ,business ,Tomography, Optical Coherence ,medicine.drug - Abstract
Purpose To evaluate the changes of retinal capillary nonperfusion areas and retinal capillary vessel density of the superficial capillary plexus (SCP) and deep capillary plexus in patients with diabetes with diabetic macular edema treated with an intravitreal dexamethasone implant (IDI). Methods We enrolled 28 patients with diabetic retinopathy and diabetic macular edema candidates to IDI. All patients underwent widefield optical coherence tomography angiography with PLEX Elite 9000 device with 15 × 9 mm scans centered on the foveal center at baseline, 1 month, 2 months, and 4 months after IDI. In all the patients, the variation of the retinal capillary nonperfusion areas and of the retinal vessel density of the SCP and deep capillary plexus were calculated using an automatic software written in Matlab (MathWorks, Natick, MA). Results During follow-up, SCP showed a statistically significant reduction of ischemic areas at 1 month after IDI (P = 0.04) and slightly increased not significantly thereafter (P = 0.15). The percentage of nonperfusion areas changed from 11.4% at baseline, to 6.3% at 1 month, 8.1%, at 2 months, and 10.2% at 4 months. The whole vessel density of SCP slightly increased (not significantly) from 35.30% at baseline to 38.00% at 1 month, and then decreased to 37.85% at 2 months and 36.04% at 4 months (P = 0.29). Retinal capillary nonperfusion areas and retinal vessel density at the deep capillary plexus did not change significantly (P = 0.31 and P = 0.73, respectively). Conclusions Widefield optical coherence tomography angiography showed a decrease in retinal capillary nonperfusion areas after dexamethasone implant suggesting a possible drug-related reperfusion of retinal capillaries particularly evident in the early period. Translational relevance A custom-made automatic analysis of retinal nonperfusion areas may allow a better and precise evaluation of ischemic changes after intravitreal therapy.
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- 2020
34. Widefield Swept Source OCTA in Retinitis Pigmentosa
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Guido Di Marzio, Chiara De Nicola, Daniele Libertini, Rodolfo Mastropasqua, Giuseppe Di Martino, Luca Di Antonio, Rossella D'Aloisio, Marta Di Nicola, Giada Ferro, Alfonso Senatore, and Lisa Toto
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medicine.medical_specialty ,Visual acuity ,genetic structures ,Clinical Biochemistry ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,widefield swept source optical coherence tomography angiography ,retinitis pigmentosa ,Ophthalmology ,Retinitis pigmentosa ,medicine ,Retinal perfusion density ,Widefield swept source optical coherence tomography angiography ,030304 developmental biology ,retinal perfusion density ,lcsh:R5-920 ,0303 health sciences ,Retina ,business.industry ,Length density ,Retinal ,medicine.disease ,eye diseases ,Peripheral ,medicine.anatomical_structure ,chemistry ,030221 ophthalmology & optometry ,sense organs ,medicine.symptom ,lcsh:Medicine (General) ,business ,Microperimetry ,Perfusion - Abstract
(1) Background: To evaluate superficial capillary plexus (SCP), deep capillary plexus (DCP), choriocapillaris (CC), perfusion density (PD), and vessel length density (VLD) in macular and near/mid periphery regions in patients with retinitis pigmentosa (RP) using widefield swept source optical coherence tomography angiography (WSS-OCTA). (2) Methods: Twelve RP patients (20 eyes) and 20 age-matched subjects (20 eyes) were imaged with the SS-OCTA system (PLEX Elite 9000, Carl Zeiss Meditec Inc., Dublin, CA, USA). Quantitative analysis was performed in the macular and peripheral regions. The main outcome measures were SCP, DCP, CC, PD, and VLD in central and peripheral areas. (3) Results: Mean visual acuity, central macular thickness, and microperimetry were significantly reduced in RP patients compared to normal subjects (p < 0.05). The perfusion density and VLD of SCP, DCP, and CC were significantly reduced in RP patients compared to normal controls both in the central and peripheral retina (p < 0.05). A significant direct correlation was found in RP patients between PD of the 1.5 mm central retina both in DCP and CC and microperimetry at 4° and 8°. (4) Conclusions: Widefield SS-OCTA shows an impairment of retinal and choroidal perfusion density and vessel length density in central and peripheral retina of RP patients. The reduction of flow features correlates with the macular function.
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- 2020
35. Design, synthesis and characterization of a PEGylated stanozolol for potential therapeutic applications
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Markku Varjosalo, Cristian Vergallo, Christian Celia, Luisa Di Marzio, Karina Moslova, Felisa Cilurzo, Maria Chiara Cristiano, Massimo Fresta, Hélder A. Santos, Sini Miettinen, Giulia Torrieri, Riccardo Provenzani, Division of Pharmaceutical Chemistry and Technology, Nanomedicines and Biomedical Engineering, Pharmaceutical Design and Discovery group, Institute of Biotechnology, Department of Chemistry, Molecular Systems Biology, Helsinki One Health (HOH), Drug Research Program, and Helsinki Institute of Life Science HiLIFE
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endocrine system diseases ,Chemistry, Pharmaceutical ,medicine.medical_treatment ,Pharmaceutical Science ,02 engineering and technology ,Pharmacology ,030226 pharmacology & pharmacy ,Polyethylene Glycols ,Polyethylene glycol (PEG) derivatives ,Biopolymers ,Anabolic Agents ,0302 clinical medicine ,Drug Stability ,Testosterone ,Cytotoxicity ,Stanozolol ,Chemistry ,PEGylation ,021001 nanoscience & nanotechnology ,3. Good health ,317 Pharmacy ,Receptors, Androgen ,Lipophilicity ,Androgens ,GROWTH ,0210 nano-technology ,Hydrophobic and Hydrophilic Interactions ,medicine.drug ,Cell Survival ,Hormone Replacement Therapy ,Drug Compounding ,Biological Availability ,Steroid ,03 medical and health sciences ,Cell Line, Tumor ,Toxicity Tests ,medicine ,Humans ,MTT assay ,Viability assay ,POLY(ETHYLENE ,Water ,nutritional and metabolic diseases ,IN-VITRO ,Conjugated drug delivery systems ,Bioavailability ,Solubility ,Drug Design - Abstract
Stanozolol (STZ) is a drug used to treat serious disorders like aplastic anemia and hereditary angioedema. It is also indicated as an adjunct therapy for the treatment of vascular disorders and growth failures. Encouraging results obtained using animal models demonstrated that STZ increases bone formation and mineralization, thus improving both density and biomechanical properties. Like natural androgens, such as TST and 5α-dihydrotestosterone (5α-DHT), STZ binds androgen receptor (AR) to activate AR-mediated signalling. Despite its therapeutic effects, this synthetic anabolic-androgenic steroid (AAS), or 5α-DHT derivative, due to its high lipophilicity, is poor soluble in water. Thus, to increase the water solubility and stability of STZ, as well as its bioavailability and efficacy, an innovative PEGylated STZ (STZ conjugated with (MeO-PEG-NH2)10kDa, (MeO-PEG-NH)10kDa-STZ) was synthesized. As confirmed by chromatography (RP-HPLC) and spectrometry (ATR-FTIR, 1H-NMR, elemental CHNS(O) analysis, MALDI-TOF/TOF) analyses, a very pure, stable and soluble compound was obtained. Acetylcholinesterase (AChE) competitive ELISA kit demonstrated that the resulting PEGylated STZ competes against biological TST, especially at lower concentrations. Cytotoxicity of increasing concentrations (1, 10, 25 or 50 µM) of STZ and/or (MeO-PEG-NH)10kDa-STZ was also evaluated for up 80 h by performing the MTT assay on human osteosarcoma Saos-2 cells, which express AR and are responsive to STZ. PEGylation mitigated cytotoxicity of STZ, by increasing the cell viability values, especially at higher drug concentrations. Furthermore, these results suggest that (MeO-PEG-NH)10kDa-STZ is a promising and reliable drug to be used in clinical conditions in which TST is required.
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- 2020
36. Survival, metabolic rates and locomotory activities of a groundwater-obligate copepod species under long-term exposures to tetrachloroethylene
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María Elena Sáenz, Tiziana Di Lorenzo, Giuseppe Messana, Leonardo Piccini, Diana M. P. Galassi, and Walter Darío Di Marzio
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chemistry.chemical_compound ,biology ,chemistry ,Obligate ,Ecology ,Tetrachloroethylene ,General Engineering ,Copepods ,tetrachloroethylene ,traits ,biology.organism_classification ,Copepod ,Groundwater ,Term (time) - Abstract
Volatile organic compounds (VOCs) are known to potentially cause a severe change in the respiratory metabolism of freshwater species, however the effect of these contaminants on groundwater-obligate species has not been investigated to date. Tetrachloroethylene (TCE) is a VOC frequently found in the groundwater bodies of industrialized areas, even years after a contamination event because TCE degradation takes several decades to occur. Contamination from TCE is considered persistent and difficult to remediate. Its high density favors a gravity-driven vertical infiltration into groundwater bodies. The TCE threshold value is 1.1 μg/L in groundwater bodies of Italy. TCE concentration in many Italian groundwater bodies is widely over this legal limit. In this study, we investigated the effect of 1.1 μg/L TCE on the survival, oxygen consumption, and locomotory activities of a groundwater-obligate copepod species. The specimens required for the trials were collected in the Antro del Corchia Cave (Tuscany). We measured the individual-based oxygen consumption of this species as a proxy of possible metabolic reactions to long-term (5 days) exposures to TCE at 8.0°C that is the mean annual temperature of groundwater flowing in the cave. To this end, we used a sealed glass microplate equipped with 24-planar oxygen sensor spots with optical isolation glued onto the bottom of 80-μL wells (Loligo Systems, Denmark) integrated with a 24-channel fluorescence-based respirometry system (SDR Sensor Dish Reader, PreSens, Germany). The system allows simultaneous measurement of 20 replicates and 4 controls. Survival and locomotory activity assessments were performed by counting the number of alive individuals and measuring the number of moving animals in 5 mL glass vials each containing 20 individuals. Preliminary results showed a decreasing in oxygen consumption of the organisms exposed to 1.1 μg/L TCE for 5 days at 8°C respect to the control. However, neither survival nor locomotory activities appeared to have been affected by exposure to TCE. See Suppl. material 1.
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- 2018
37. Mathematical Models as Tools to Predict the Release Kinetic of Fluorescein from Monoglyceride Colloidal Liquid Crystals
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Donatella Paolino, Christian Celia, Felisa Cilurzo, Luisa Di Marzio, Constantin Mircioiu, and Andra Tudose
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life_sciences_other ,chemistry.chemical_compound ,Colloid ,Materials science ,Mathematical model ,chemistry ,Chemical engineering ,Liquid crystal ,Fluorescein ,Monoglyceride ,Kinetic energy - Abstract
The manuscript studies the release kinetic of fluorescein from colloidal liquid crystals made up from monoglyceride and different non-ionic surfactants. The release experiments were carried out under sink conditions, and mathematical models were described as extrapolations from solutions of diffusion equation in different initial and boundary conditions imposed by pharmaceutical formulations. The diffusion equation was solved, using Laplace and Fourier transformed functions for the release kinetic from infinite reservoirs in a semi-infinite medium. Solutions represents a general square root law and can be applied for the release kinetic of fluorescein from monoglyceride colloidal liquid crystals. Akaike, Schwartz and Imbimbo criteria were used to establish the appropriate mathematical model and the hierarchy of performances of different models applied to the release experiments. The Fisher statistic test was applied to perform significance of differences among mathematical models. Differences evaluated by mathematical criteria demonstrated that small or no significant statistic differences were carried out between various applied models and colloidal formulations. Phenomenological models were preferred over to empirical and semi-empirical ones. The general square root model shows that the diffusion-controlled release of fluorescein is the mathematical models extrapolated for monoglyceride colloidal liquids, in the first part of the process.
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- 2018
38. Genotoxic Evidences of Glyphosate and Chlorpyriphos on Eisenia fetida Coelomocytes
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Santiago Martinez, Marcelo L. Larramendy, Silvana Patricia Curieses, María Elena Sáenz, Walter Darío Di Marzio, and J. L. Alberdi
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Eisenia fetida ,biology ,business.industry ,COMMERCIAL FORMULATIONS ,Crop yield ,food and beverages ,General Medicine ,Pesticide ,HERBICIDES ,biology.organism_classification ,DNA DAMAGE ,SUBLETHAL EFFECTS ,SINGLE CELL GELL ELECTROFORESIS ASSAY ,purl.org/becyt/ford/1 [https] ,purl.org/becyt/ford/1.5 [https] ,chemistry.chemical_compound ,Agronomy ,chemistry ,Agriculture ,Glyphosate ,business ,INSECTICIDES - Abstract
The organophosphorus herbicide glyphosate (GLY) and the organophosphate insecticide chlorpyriphos (CPF) are key pesticides in modern management cultures worldwide. Sublethal toxicity of the commercial herbicide formulation Roundup® and the insecticide formulation Terfos® were evaluated on Eisenia fetida coelomocytes exposed under in vivo and ex vivo laboratory conditions. Induction of DNA single-strand breaks evaluated by the single cell gel electrophoresis assay and coelomocyte viability as well as alterations in coelomocyte trophic indexes were employed as endpoints for genotoxicity and cytotoxicity, respectively. Specimens were exposed at concentrations corresponding to recommended pesticide field application rate, and endpoints were evaluated after 7 and 14 days of treatment (in vivo exposure). In addition, coelomocytes were exposed to aqueous leachate of pesticide-contaminated soils during 1 h (ex vivo exposure). Earthworms exposed to Roundup® and Terfos® showed an increased frequency of DNA damage. Also, a decrease of coelomocyte viability and decrease of trophic indexes were observed in all treatments. The results demonstrate that either GLY- and CPF-based formulations exerted genotoxic as well as cytotoxic effects in coelomocytes of E. fetida exposed in vivo and ex vivo. Fil: Curieses, Silvana Patricia. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Sáenz, María Elena. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alberdi, José Luis. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina Fil: Martinez, Ricardo Santiago. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Larramendy, Marcelo Luis. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina Fil: Di Marzio, Walter Dario. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
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- 2018
39. Cyto and Genotoxicity of Positive and Negative Coated Silica Nanoparticles on Celomocytes of Earthworms Eisenia fetida (Oligochaeta, Annelida)
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Pablo Scodeller, W. D. Di Marzio, M. E. Sáenz, J. L. Alberdi, and Silvana Patricia Curieses
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inorganic chemicals ,Eisenia fetida ,biology ,Chemistry ,technology, industry, and agriculture ,General Medicine ,respiratory system ,biology.organism_classification ,medicine.disease_cause ,Silica nanoparticles ,Oligochaeta ,Environmental chemistry ,mental disorders ,medicine ,health care economics and organizations ,Genotoxicity - Abstract
To understand and assess the effects of nanoparticles (NPs) on the environment, should be well established quantitatively the concentration-response relationships. Also relate the potential effects on selected variables of response with the exposure to sub-lethal levels of NPs. In this work, we describe the ecotoxicological evaluation of negative and positive coated silica NPs (Si-Nps) on coelomic cells from Eisenia fetida.
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- 2018
40. Simultaneous quantification of Gemcitabine and Irinotecan hydrochloride in rat plasma by using high performance liquid chromatography-diode array detector
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Nicola Costa, L. Di Marzio, Donatella Paolino, Marcello Locatelli, Massimo Fresta, Donato Cosco, A. Di Tullio, Christian Celia, Felisa Cilurzo, and L. Malatesta
- Subjects
Antimetabolites, Antineoplastic ,Clinical Biochemistry ,Pharmaceutical Science ,Irinotecan ,01 natural sciences ,High-performance liquid chromatography ,Deoxycytidine ,Chemistry Techniques, Analytical ,Analytical Chemistry ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Pharmacokinetics ,Chromatography detector ,Drug Discovery ,Irinotecan Hydrochloride ,medicine ,Animals ,Spectroscopy ,Chromatography, High Pressure Liquid ,Chromatography ,Chemistry ,010401 analytical chemistry ,Plasma ,Antineoplastic Agents, Phytogenic ,Gemcitabine ,0104 chemical sciences ,Rats ,Standard curve ,030220 oncology & carcinogenesis ,Injections, Intravenous ,Camptothecin ,medicine.drug - Abstract
In this manuscript we aimed at the simultaneous separation and quantification of Gemcitabine and Irinotecan hydrochloride (injected both as single components and in combination) from Sprague Dawley rat plasma by using a validated method obtained through the use of a High Performance Liquid Chromatography (HPLC)-diode array detector (DAD). Gemcitabine and Irinotecan hydrochloride were detected and quantified using a Zorbax Extend C-18 column (250 mm × 4.6 mm; 5 μm particle size) in gradient elution mode. The chromatographic analyses were carried out in 15 min. The analytical mode was calibrated and validated in the concentration range from 0.1 to 18 μg/mL both for Gemcitabine and Irinotecan hydrochloride. Sprague Dawley rat plasma was used to perform the analysis. 3-methylxanthine was the internal standard. The weighted-matrix matched standard curves of Gemcitabine and Irinotecan hydrochloride showed a good linearity up to 18 μg/mL. Parallelism tests were also performed to evaluate whether the over-range samples could be analyzed after dilution without affecting the analytical performance. The intra- and inter-day precision (RSD%) values of Gemcitabine and Irinotecan hydrochloride were ≤7.14% and ≤11.5%, respectively. The intra- and inter-day trueness (Bias%) values were in the range from -11.5% to 1.70% for both drugs. The analytical mode performance was further tested after collecting Sprague Dawley rat plasma following a single-dose administration of chemotherapeutics or their association. The validated HPLC-DAD method allowed the simultaneous quantification of Gemcitabine and Irinotecan hydrochloride in the rat plasma, besides the evaluation of the pharmacokinetic parameters and drug delivery.
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- 2018
41. Hierarchical Microplates as Drug Depots with Controlled Geometry, Rigidity, and Therapeutic Efficacy
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Massimo Fresta, Paolo Decuzzi, Rui C. Pereira, Davide Romanelli, Rosita Primavera, Roberto Palomba, Christian Celia, Tiziano Catelani, Daniele Di Mascolo, Luisa Di Marzio, and Martina Di Francesco
- Subjects
Drug ,Materials science ,media_common.quotation_subject ,Geometry ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,Dexamethasone ,3. Good health ,0104 chemical sciences ,PLGA ,chemistry.chemical_compound ,microparticles local delivery rigidity multifunctional particles polymeric material ,Rigidity (electromagnetism) ,chemistry ,Drug Design ,Nanoparticles ,General Materials Science ,Lactic Acid ,0210 nano-technology ,Porosity ,Polyglycolic Acid ,media_common - Abstract
A variety of microparticles have been proposed for the sustained and localized delivery of drugs with the objective of increasing therapeutic indexes by circumventing filtering organs and biological barriers. Yet, the geometrical, mechanical, and therapeutic properties of such microparticles cannot be simultaneously and independently tailored during the fabrication process to optimize their performance. In this work, a top-down approach is employed to realize micron-sized polymeric particles, called microplates (μPLs), for the sustained release of therapeutic agents. μPLs are square hydrogel particles, with an edge length of 20 μm and a height of 5 μm, made out of poly(lactic- co-glycolic acid) (PLGA). During the synthesis process, the μPL Young's modulus can be varied from 0.6 to 5 MPa by changing the PLGA amounts from 1 to 7.5 mg, without affecting the μPL geometry while matching the properties of the surrounding tissue. Within the porous μPL matrix, different classes of therapeutic payloads can be incorporated including molecular agents, such as anti-inflammatory dexamethasone (DEX), and nanoparticles containing imaging and therapeutic molecules themselves, thus originating a truly hierarchical platform. As a proof of principle, μPLs are loaded with free DEX and 200 nm spherical polymeric nanoparticles, carrying DEX molecules (DEX-SPNs). Electron and fluorescent confocal microscopy analyses document the uniform distribution and stability of molecular and nanoagents within the μPL matrix. This multiscale, hierarchical microparticle releases DEX for at least 10 days. The inclusion of DEX-SPNs serves to minimize the initial burst release and modulate the diffusion of DEX molecules out of the μPL matrix. The biopharmacological and therapeutic properties together with the fine tuning of geometry and mechanical stiffness make μPLs a unique polymeric depot for the potential treatment of cancer, cardiovascular, and chronic, inflammatory diseases.
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- 2018
42. SUMO polymeric chains are involved in nuclear foci formation and chromatin organization in Trypanosoma brucei procyclic forms
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Vanina E. Alvarez, Javier G. De Gaudenzi, María Agustina Berazategui, Paula Ana Iribarren, and Lucía Ayelén Di Marzio
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Models, Molecular ,0301 basic medicine ,Amino Acid Motifs ,Biotecnología del Medio Ambiente ,Lysine ,SUMO protein ,Gene Expression ,lcsh:Medicine ,Biochemistry ,purl.org/becyt/ford/1 [https] ,Database and Informatics Methods ,Amino Acids ,lcsh:Science ,Protozoans ,Multidisciplinary ,biology ,Organic Compounds ,Chromosome Biology ,Chemistry ,Eukaryota ,Bioquímica y Biología Molecular ,SUMOylation ,Chromatin ,Cell biology ,Synaptonemal complex ,Telomeres ,Physical Sciences ,Small Ubiquitin-Related Modifier Proteins ,Epigenetics ,Post-translational modification ,Basic Amino Acids ,Sequence Analysis ,CIENCIAS NATURALES Y EXACTAS ,Research Article ,Trypanosoma ,Chromosome Structure and Function ,Bioinformatics ,Transgene ,Trypanosoma brucei brucei ,Saccharomyces cerevisiae ,SUMO CHAINS ,Trypanosoma brucei ,Research and Analysis Methods ,Chromosomes ,Ciencias Biológicas ,03 medical and health sciences ,Protein Domains ,Sequence Motif Analysis ,Trypanosoma Brucei ,Genetics ,Amino Acid Sequence ,purl.org/becyt/ford/1.6 [https] ,Protein Structure, Quaternary ,Cell Nucleus ,Biology and life sciences ,Organic Chemistry ,lcsh:R ,Organisms ,Chemical Compounds ,Proteins ,Cell Biology ,TRYPANOSOMA ,Subcellular localization ,biology.organism_classification ,Parasitic Protozoans ,NUCLEAR FOCI ,030104 developmental biology ,Mutation ,CHROMATIN ORGANIZATION ,lcsh:Q ,Protein Multimerization ,Trypanosoma Brucei Gambiense - Abstract
SUMOylation is a post-translational modification conserved in eukaryotic organisms that involves the covalent attachment of the small ubiquitin-like protein SUMO to internal lysine residues in target proteins. This tag usually alters the interaction surface of the modified protein and can be translated into changes in its biological activity, stability or subcellular localization, among other possible outputs. SUMO can be attached as a single moiety or as SUMO polymers in case there are internal acceptor sites in SUMO itself. These chains have been shown to be important for proteasomal degradation as well as for the formation of subnuclear structures such as the synaptonemal complex in Saccharomyces cerevisiae or promyelocytic leukemia nuclear bodies in mammals. In this work, we have examined SUMO chain formation in the protozoan parasite Trypanosoma brucei. Using a recently developed bacterial strain engineered to produce SUMOylated proteins we confirmed the ability of TbSUMO to form polymers and determined the type of linkage using site-directed mutational analysis. By generating transgenic procyclic parasites unable to form chains we demonstrated that although not essential for normal growth, SUMO polymerization determines the localization of the modified proteins in the nucleus. In addition, FISH analysis of telomeres showed a differential positioning depending on the polySUMOylation abilities of the cells. Thus, our observations suggest that TbSUMO chains might play a role in establishing interaction platforms contributing to chromatin organization. Fil: Iribarren, Paula Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Di Marzio, Lucía Ayelén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Berazategui, Maria Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: de Gaudenzi, Javier Gerardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Alvarez, Vanina Eder. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
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- 2018
43. The ecotoxicity of binary mixtures of Imazamox and ionized ammonia on freshwater copepods: Implications for environmental risk assessment in groundwater bodies
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Marco Cifoni, María Elena Sáenz, Diana M. P. Galassi, Walter Dario Di Marzio, and Tiziana Di Lorenzo
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0106 biological sciences ,Imazamox ,GROUNDWATER ,Health, Toxicology and Mutagenesis ,Fresh Water ,010501 environmental sciences ,Ecotoxicology ,01 natural sciences ,purl.org/becyt/ford/1 [https] ,chemistry.chemical_compound ,purl.org/becyt/ford/1.5 [https] ,Mixture ,Drug Interactions ,STYGOFAUNA ,Groundwater ,IMAZAMOX ,biology ,Ecology ,Imidazoles ,General Medicine ,Pollution ,Health ,Environmental chemistry ,Public Health ,Ammonia ,Copepods ,Stygofauna ,Public Health, Environmental and Occupational Health ,Risk Assessment ,Copepoda ,Species Specificity ,Animals ,COPEPODS ,Toxicology and Mutagenesis ,0105 earth and related environmental sciences ,Environmental risk assessment ,Pollutant ,AMMONIA ,Dose-Response Relationship, Drug ,MIXTURE ,010604 marine biology & hydrobiology ,Environmental and Occupational Health ,Pesticide ,Models, Theoretical ,biology.organism_classification ,chemistry ,Ecotoxicity ,Epigeal ,Copepod ,Water Pollutants, Chemical - Abstract
Groundwater bodies are impacted by substances such as pesticides and N-fertilizers, which usually occur in the environment as complex mixtures rather than isolated pollutants. The threat that these mixtures pose to groundwater-dwelling organisms is still poorly understood. The aims of the present study were to test the acute effect of a binary mixture of a herbicide (Imazamox) and NH4+ on epigean (Eucyclops serrulatus) and hypogean (Diacyclops belgicus) freshwater copepod species. In addition, to evaluate if the effect of the mixture can be explained by referencing non-interaction models or by more complex interaction models; and the implications for groundwater risk assessment. Compared with the action of the compounds evaluated separately, the effects of Imazamox and NH4+ in the binary mixture were more than additive or synergistic for both species. MixTox models evidenced a dose ratio and dose level deviations from concentration addition and independent action traditional models. The hypogean species was three times more sensitive to NH4+ that the epigean species when assayed as a single chemical. However, D. belgicus was only 1.13 times more sensitive than E. serrulatus when NH4+ was assayed in the mixture. The use of an integrated approach for substances that are known to interact in groundwater, should include copepods species as test organisms. Fil: Di Marzio, Walter Dario. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cifoni, Marco. Universita degli Studi dell'Aquila; Italia Fil: Sáenz, María Elena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina Fil: Galassi, Diana M.P.. Universita degli Studi dell'Aquila; Italia Fil: Di Lorenzo, Tiziana. Consiglio Nazionale delle Ricerche; Italia
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- 2018
44. Ultradeformable liposomes as multidrug carrier of resveratrol and 5-fluorouracil for their topical delivery
- Author
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Cinzia Anna Ventura, Donato Cosco, Donatella Paolino, Luisa Di Marzio, Maria Carafa, Massimo Fresta, and Jessica Maiuolo
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Drug ,Skin Neoplasms ,Cell Survival ,Skin Absorption ,media_common.quotation_subject ,Pharmaceutical Science ,Antineoplastic Agents ,Apoptosis ,Human skin ,In Vitro Techniques ,Resveratrol ,Pharmacology ,Administration, Cutaneous ,chemistry.chemical_compound ,Cell Line, Tumor ,Stilbenes ,Skin delivery ,Multi-drug colloidal carriers, Skin cancer, Skin delivery, Supramolecular therapeutics, Ultradeformable liposomes ,medicine ,Stratum corneum ,Humans ,multi-drug colloidal carriers ,Ultradeformable liposomes ,Skin ,media_common ,skin cancer ,supramolecular therapeutics ,integumentary system ,Cell growth ,Cell Cycle ,Actinic keratosis ,medicine.disease ,medicine.anatomical_structure ,chemistry ,Liposomes ,Cancer cell ,Fluorouracil ,Skin cancer - Abstract
Ultradeformable liposomes represent useful formulations able to increase the skin permeation of drug compounds. In this study, resveratrol- and 5-fluorouracil-loaded ultradeformable liposomes were investigated for the potential treatment of non-melanoma skin cancer. The in vitro anticancer activity of ultradeformable liposomes was tested on human skin cancer cells through viability-, cell cycle- and apoptosis-analysis. Furthermore, we tested the percutaneous permeation of ultradeformable liposomes using human stratum corneum and viable epidermis. The co-encapsulation of resveratrol and 5-fluorouracil (multi-drug carrier) in ultradeformable liposomes improved their anticancer activity on skin cancer cells as compared to both the free drug form and the single entrapped agents. These multi-drug ultradeformable liposomes arrest cell proliferation in G1/S, thus modifying the action of 5-fluorouracil and increasing the activity of resveratrol. This effect might depend on the ultradeformable liposomes, which may accumulate in deeper skin layers, thus generating a cutaneous depot from which resveratrol and 5-fluorouracil are gradually released. Resveratrol and 5-fluorouracil co-loaded ultradeformable liposomes could be a new nanomedicine for the treatment of squamous cell carcinoma, i.e., actinic keratosis, Bowen's disease, and keratoacanthoma.
- Published
- 2015
45. Simultaneous determination of eperisone hydrochloride and paracetamol in mouse plasma by high performance liquid chromatography-photodiode array detector
- Author
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Renato C. Barbacane, Carlo Capolupo, Marcello Locatelli, Roberta Cifelli, Cristina Di Legge, Luisa Di Marzio, Christian Celia, Nicola Costa, and Massimo Fresta
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Hydrochloride ,Analytical chemistry ,Administration, Oral ,Validation Studies as Topic ,Biochemistry ,High-performance liquid chromatography ,Analytical Chemistry ,Mice ,chemistry.chemical_compound ,medicine ,Animals ,Protein precipitation ,Tissue Distribution ,Sample preparation ,Chromatography, High Pressure Liquid ,Eperisone ,Acetaminophen ,Detection limit ,Propiophenones ,Chromatography ,Organic Chemistry ,General Medicine ,Reference Standards ,Mice, Inbred C57BL ,Standard curve ,chemistry ,Area Under Curve ,Calibration ,Blood sampling ,medicine.drug - Abstract
This paper reports the validation of a quantitative high performance liquid chromatography-photodiode array (HPLC-PDA) method for the simultaneous analysis, in mouse plasma, of eperisone hydrochloride and paracetamol by protein precipitation using zinc sulphate-methanol-acetonitrile. The analytes were resolved on a Gemini C18 column (4.6 mm × 250 mm; 5 μm particle size) using a gradient elution mode with a run time of 15 min, comprising re-equilibration, at 60°C (± 1°C). The method was validated over the concentration range from 0.5 to 25 μg/mL for eperisone hydrochloride and paracetamol, in mouse plasma. Ciprofloxacin was used as Internal Standard. Results from assay validations show that the method is selective, sensitive and robust. The limit of quantification of the method was 0.5 μg/mL for eperisone hydrochloride and paracetamol, and matrix-matched standard curves showed a good linearity, up to 25 μg/mL with correlation coefficients (r(2))≥ 0.9891. In the entire analytical range the intra and inter-day precision (RSD%) values were ≤ 1.15% and ≤ 1.46% for eperisone hydrochloride, and ≤ 0.35% and ≤ 1.65% for paracetamol. For both analytes the intra and inter-day trueness (bias%) values ranged, respectively, from -5.33% to 4.00% and from -11.4% to -4.00%. The method was successfully tested in pharmacokinetic studies after oral administration in mouse. Furthermore, the application of this method results in a significant reduction in terms of animal number, dosage, and improvement in speed, rate of analysis, and quality of pharmacokinetic parameters related to serial blood sampling.
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- 2015
46. Gel-embedded niosomes: Preparation, characterization and release studies of a new system for topical drug delivery
- Author
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Maria Carafa, C. Di Meo, Carlotta Marianecci, Federica Rinaldi, Franco Alhaique, A.M. Trotta, L. Di Marzio, Pietro Matricardi, and Tommasina Coviello
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xanthan gum ,locust bean gum ,monoammonium glycyrrhizinate ,surfactant vesicles ,reology ,Administration, Topical ,Acrylic Resins ,Polysorbates ,Polysaccharide ,Galactans ,Mannans ,Surface-Active Agents ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Pulmonary surfactant ,Plant Gums ,Niosome ,Physical and Theoretical Chemistry ,Hexoses ,chemistry.chemical_classification ,Chromatography ,Vesicle ,Anti-Inflammatory Agents, Non-Steroidal ,Polysaccharides, Bacterial ,Surfaces and Interfaces ,General Medicine ,Permeation ,Glycyrrhizic Acid ,In vitro ,Solutions ,Drug Liberation ,Kinetics ,chemistry ,Permeability (electromagnetism) ,Delayed-Action Preparations ,Liposomes ,Locust bean gum ,Gels ,Biotechnology - Abstract
In the present paper physical gels, prepared with two polysaccharides, Xanthan and Locust Bean Gum, and loaded with non-ionic surfactant vesicles, are described. The vesicles, composed by Tween20 and cholesterol or by Tween85 and Span20, were loaded with Monoammonium glycyrrhizinate for release experiments. Size and zeta (ζ)-potential of the vesicles were evaluated and the new systems were characterized by rheological and dynamo-mechanical measurements. For an appropriate comparison, a Carbopol gel and a commercial gel for topical applications were also tested. The new formulations showed mechanical properties comparable with those of the commercial product indicating their suitability for topical applications. In vitro release experiments showed that the polysaccharide network protects the integrity of the vesicles and leads to their slow release without disruption of the aggregated structures. Furthermore, being the vesicles composed of molecules possessing enhancing properties, the permeation of the loaded drugs topically delivered can be improved. Thus, the new systems combine the advantages of matrices for a modified release (polymeric component) and those of an easier permeability across the skin (vesicle components). Finally, shelf live experiments indicated that the tested gel/vesicle formulations were stable over 1 year with no need of preservatives.
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- 2015
47. Niosomes as Drug Nanovectors: Multiscale pH-Dependent Structural Response
- Author
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Paolo Decuzzi, Luisa Di Marzio, Maria Carafa, Luciana Dini, Paola Brocca, Elena Del Favero, Federica Rinaldi, Christian Celia, Donatella Paolino, Massimo Fresta, Valeria Rondelli, Antonio Serra, Carlotta Marianecci, Laura Cantù, Marianecci, C, Dimarzio, L, Delfavero, E, Cantù, L, Brocca, P, Rondelli, V, Rinaldi, F, Dini, Luciana, Serra, Antonio, Decuzzi, P, Celia, C, Paolino, D, Fresta, M, Carafa, M, Marianecci, Carlotta, Di Marzio, Luisa, Del Favero, Elena, Cantù, Laura, Brocca, Paola, Rondelli, Valeria, Rinaldi, Federica, Decuzzi, Paolo, Celia, Christian, Paolino, Donatella, Fresta, Massimo, and Carafa, Maria
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Lipid Bilayers ,Polysorbates ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,Spectrum Analysis, Raman ,01 natural sciences ,Pulmonary surfactant ,X-Ray Diffraction ,Scattering, Small Angle ,Electrochemistry ,General Materials Science ,Niosome ,Lipid bilayer ,Spectroscopy ,Liposome ,Chemistry ,Chemistry, Physical ,Vesicle ,Bilayer ,Surfaces and Interfaces ,Hydrogen-Ion Concentration ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,0104 chemical sciences ,Polysorbate ,Membrane ,Cholesterol ,Liposomes ,Biophysics ,Lipid Bilayer ,Nanocarriers ,0210 nano-technology - Abstract
The use of nanocarriers, which respond to different stimuli controlling their physicochemical properties and biological responsivness, shows a growing interest in pharmaceutical science. The stimuli are activated by targeting tissues and biological compartments, e.g., pH modification, temperature, redox condition, enzymatic activity, or can be physically applied, e.g., a magnetic field and ultrasound. pH modification represents the easiest method of passive targeting, which is actually used to accumulate nanocarriers in cells and tissues. The aim of this paper was to physicochemically characterize pH-sensitive niosomes using different experimental conditions and demonstrate the effect of surfactant composition on the supramolecular structure of niosomes. In this attempt, niosomes, made from commercial (Tween21) and synthetic surfactants (Tween20 derivatives), were physicochemically characterized by using different techniques, e.g., transmission electron microscopy, Raman spectroscopy, and small-angle X-ray scattering. The changes of niosome structure at different pHs depend on surfactants, which can affect the supramolecular structure of colloidal nanocarriers and their potential use both in vitro and in vivo. At pH 7.4, the shape and structure of niosomes have been maintained; however, niosomes show some differences in terms of bilayer thicknesses, water penetration, membrane coupling, and cholesterol dispersion. The acid pH (5.5) can increase the bilayer fluidity, and affect the cholesterol depletion. In fact, Tween21 niosomes form large vesicles with lower curvature radius at acid pH; while Tween20-derivative niosomes increase the intrachain mobility within a more interchain correlated membrane. These results demonstrate that the use of multiple physicochemical procedures provides more information about supramolecular structures of niosomes and improves the opportunity to deeply investigate the effect of stimuli responsiveness on the niosome structure.
- Published
- 2016
48. Responses to silver nanoparticles and silver nitrate in a battery of biomarkers measured in coelomocytes and in target tissues of Eisenia fetida earthworms
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García-Velasco Nerea, Urionabarrenetxea Erik, Curieses Silvana Patricia, Sáenz María Elena, Bilbao Eider, Soto Manu, and Di Marzio Walter Darío
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Eisenia fetida ,Silver ,Cell Survival ,Health, Toxicology and Mutagenesis ,0211 other engineering and technologies ,Metal Nanoparticles ,02 engineering and technology ,010501 environmental sciences ,01 natural sciences ,Silver nanoparticle ,Ciencias de la Tierra y relacionadas con el Medio Ambiente ,Coelomocyte Cytotoxicity ,purl.org/becyt/ford/1 [https] ,chemistry.chemical_compound ,Soil ,purl.org/becyt/ford/1.5 [https] ,Metallothionein ,Animals ,Soil Pollutants ,Viability assay ,Oligochaeta ,Eisenia Fetida ,Cytotoxicity ,0105 earth and related environmental sciences ,021110 strategic, defence & security studies ,Soil Ecotoxicology ,biology ,Dose-Response Relationship, Drug ,Antioxidant Response ,Earthworm ,Public Health, Environmental and Occupational Health ,General Medicine ,Silver Nanoparticle ,biology.organism_classification ,Catalase ,Pollution ,Silver nitrate ,Metal Detoxification ,Biochemistry ,chemistry ,Solubility ,Environmental chemistry ,biology.protein ,Silver Nitrate ,Meteorología y Ciencias Atmosféricas ,CIENCIAS NATURALES Y EXACTAS ,Biomarkers - Abstract
The current use and development of applications with silver nanoparticles (Ag NPs) could lead to potential inputs of these NPs to soils. Consequently, it is crucial to understand the ecotoxicological risks posed by Ag NPs in the terrestrial compartment. In the present investigation, the effects produced by PVP-PEI coated Ag NPs were assessed in Eisenia fetida earthworms in comparison with the soluble form (AgNO3). Earthworms were exposed for 1, 3 and 14 days to a range of sublethal concentrations of Ag (0, 0.05 and 50 mg/kg) and at each exposure time, apart from mortality and weight loss of individuals, metallothionein (MT) protein concentration and catalase (CAT) activity were quantified in earthworm tissues. In addition, cellular and molecular level endpoints (cell viability, absolute and relative trophic indices and transcription levels of catalase-cat- and metallothionein-mt-) were measured in coelomocytes extruded from exposed earthworms. Despite the lack of effects in traditional endpoints (mortality and weight loss), Ag NPs and AgNO3 posed changes at lower levels of biological complexity (biochemical, cellular and molecular levels). Both Ag forms induced similar changes in the metal detoxification mechanism (MT, mt) and in the antioxidant response system (CAT, cat) of E. fetida. In contrast, Ag form dependant cytotoxicity and subpopulation ratio alterations (eleocytes/amoebocytes) were recorded in extruded coelomocytes. Complementarily, the use of coelomocytes to assess molecular level endpoints represented a relevant alternative for development of non-invasive biomarkers. Fil: Curieses, Silvana Patricia. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: García Velasco, Nerea. Universidad del País Vasco; España Fil: Urionabarrenetxea, Erik. Universidad del País Vasco; España Fil: Sáenz, María Elena. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Bilbao, Eider. Universidad del País Vasco; España Fil: Di Marzio, Walter Dario. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Soto, Manu. Universidad del País Vasco; España
- Published
- 2017
49. Bioactive isoflavones from Pueraria lobata root and starch: Different extraction techniques and carbonic anhydrase inhibition
- Author
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Daniela Secci, Christian Celia, Andrea Angeli, Claudiu T. Supuran, Luisa Di Marzio, Simona Riga, Stefania Cesa, Adriano Mollica, Andrei Mocan, Marcello Locatelli, and Simone Carradori
- Subjects
Pueraria ,pueraria lobata ,isoflavones ,HPLC-PDA ,microwave-assisted extraction ,liquid ionic ,natural carbonic anhydrase inhibitors ,Food industry ,Starch ,Ionic Liquids ,Toxicology ,01 natural sciences ,Plant Roots ,chemistry.chemical_compound ,Ingredient ,Structure-Activity Relationship ,Carbonic anhydrase ,Humans ,Ultrasonics ,Food science ,Carbonic Anhydrase Inhibitors ,Microwaves ,Chromatography, High Pressure Liquid ,Carbonic Anhydrases ,Chromatography ,biology ,010405 organic chemistry ,business.industry ,Plant Extracts ,010401 analytical chemistry ,Extraction (chemistry) ,Reproducibility of Results ,General Medicine ,Isoflavones ,biology.organism_classification ,Kudzu ,0104 chemical sciences ,Isoenzymes ,chemistry ,biology.protein ,business ,Food Science - Abstract
Kudzu, the dried root of an important edible plant (Pueraria lobata), is used in Traditional Chinese Medicine for the important nutritional value strictly related to its isoflavone derivatives. These compounds characterize the quality of kudzu contained in different preparations, as pharmaceutical ingredient as well as dietary/food supplement (e.g. starch). The optimization of the isoflavones recovery, monitored by HPLC-PDA, through different innovative and conventional extraction techniques, e.g. microwave-assisted, ultrasound-assisted and conventional extraction, represented a suitable challenge in food industry and natural products evaluation. The impact on the isoflavone extraction by using an ionic liquid-assisted procedure was also considered. Furthermore, the inhibitory activity of the most representative isoflavones, isolated from kudzu, was evaluated using four isoforms (I, II, IX and XII) of human carbonic anhydrase (hCA) due to their role in several physiopathological processes.
- Published
- 2017
50. Physicochemical characterization of pH-responsive and fusogenic self-assembled non-phospholipid vesicles for a potential multiple targeting therapy
- Author
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Nicola d'Avanzo, Donatella Paolino, Cinzia Anna Ventura, Massimo Fresta, Luisa Di Marzio, Christian Celia, Martina Di Francesco, Rosita Primavera, Felisa Cilurzo, and Marcello Locatelli
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Polysorbates ,Pharmaceutical Science ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Mice ,Surface-Active Agents ,Drug Delivery Systems ,Pulmonary surfactant ,In vivo ,Cell Line, Tumor ,Neoplasms ,Fluorescence spectroscopy, Fusion assay, Multiple targeting therapy, Niosomes, pH-responsiveness, Physicochemical characterization ,medicine ,Animals ,Humans ,Doxorubicin ,Niosome ,Chemistry ,Biological membrane ,Hydrogen-Ion Concentration ,021001 nanoscience & nanotechnology ,In vitro ,0104 chemical sciences ,RAW 264.7 Cells ,Biophysics ,Nanoparticles ,Doxorubicin Hydrochloride ,Nanocarriers ,0210 nano-technology ,medicine.drug - Abstract
In order to obtain nanocarriers suitable for the delivery of drugs in the treatment of cancer, pH-responsive nanovesicles capable of facilitating fusion (fusogenic nanovesicles) were synthesized and then their physicochemical characteristics were modified. These nanovesicles were made by combining polysorbates having different physicochemical features with the aim of realizing multiple-targeting nanoformulations suitable for in vitro treatment of cancer cells. Tween21 and Tween80 were self-assembled at different molar concentrations resulting in pH-responsive fusogenic nanovesicles with an average size of less than 150 nm, and a narrow size distribution (polydispersity index) value of less than 0.2. Hydrophobic and hydrophilic fluorescent probes were loaded inside the nanovesicles in order to study their pH-responsiveness and fusogenic properties and it was noted that this process did not modify their physicochemical features. The pH-responsiveness and fusogenic assay demonstrated that the nanovesicles containing Tween21 at different molar ratios were pH-responsive and interacted with a synthetic model of a biological membrane supplemented with Ca2+ in the incubation medium. Fifty percent (molar ratio) of Tween21 was replaced with Tween80, since Tween80 can promote the adsorption of apolipoproteins (A–E) onto the surfaces of nanovesicles without altering their pH-responsiveness or fusogenic properties. In fact this equivalent molar concentration of Tween21 and Tween80 also maintained their degree of interaction with the apolipoproteins (A–E). Doxorubicin hydrochloride-loaded nanovesicles were synthesized and physicochemically characterized in order to obtain nanoformulations suitable for anticancer treatment. The therapeutic nanovesicles showed physicochemical properties similar to those of empty nanoformulations, and maintained pH-responsiveness, fusogenic properties and targeting versus the apolipoproteins (A–E). The doxorubicin hydrochloride was loaded into the nanovesicles using both passive and pH gradient remote loading procedures. The latter provided the nanovesicles with an entrapment efficiency percentage of over 30%, which was much higher than the 10% that was obtained using the passive loading procedure. The entrapment efficiency improved up to 60% for the nanovesicles made from the same molar concentration of Tween21 and Tween80. The anticancer activity of doxorubicin hydrochloride-loaded nanovesicles was further tested in vitro using human neuroblastoma (SH-SY5Y) cells which respond to treatment with this chemotherapeutic drug, but the nanovesicles carrying it must cross the BBB by means of specific receptors before the drug can provide a therapeutic effect in vivo. The anticancer activity of these doxorubicin hydrochloride-loaded nanovesicles was time- and dosage- dependent, and the surfactant components making up the nanoformulations was also a determining factor in the efficiency of their activity. These nanovesicles could provide innovative nanotherapeutics for potential in vitro multidrug targeting therapy.
- Published
- 2017
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