Il giudizio di ottemperanza: attuale rilevanza, applicazione e ruolo sistematico nel processo amministrativo

Il presente lavoro ha ad oggetto la funzione svolta, nel sistema di giustizia amministrativa, dal giudizio di ottemperanza ossia l’istituto garante dell’attuazione dei provvedimenti giurisdizionali ineseguiti dall'amministrazione. A tal fine, si ripercorre l’evoluzione storica e l’attuale disciplina contenuta nel C.p.a. In particolare, ci si sofferma sui provvedimenti ottemperabili, tra cui il decreto del Presidente della Repubblica in sede di ricorso straordinario, sulle azioni di condanna esperibili nel giudizio di esecuzione, sull'ottemperanza di chiarimenti e sulle disposizioni dedicate al giudice ed al Commissario. In seguito, il giudizio di ottemperanza viene inquadrato all'interno della giurisdizione di merito di cui si menzionano le origini, l’attuale disciplina ed il dibattuto significato da attribuire alla categoria. Il giudizio di ottemperanza, in quanto una delle più importanti ipotesi di giurisdizione estesa al merito, viene delineato per macro categorie, anche attraverso una dettagliata analisi della giurisprudenza più recente. Vengono esaminate le pronunce amministrative relative alla natura, oggetto, riedizione del potere amministrativo, giudice e Commissario. Infine, nell'ottica di un inquadramento sistematico ed al fine di comprendere le future prospettive del giudizio di ottemperanza, quest’ultimo viene messo a confronto con le maggiori novità introdotte nel giudizio di legittimità, tra cui i mezzi che consentono un’anticipazione dell’esecuzione alla fase di cognizione

High-efficiency novel extraction process of target polyphenols using enzymes in hydroalcoholic media

Agro-industrial by-products are a sustainable source of natural additives that can replace the synthetic ones in the food industry. Grape pomace is an abundant by-product that contains about 70% of the grape’s polyphenols. Polyphenols are natural antioxidants with multiple health-promoting properties. They are secondary plant metabolites with a wide range of solubilities. Here, a novel extraction process of these compounds was developed using enzymes that specifically liberates target polyphenols in the appropriate hydroalcoholic mixture. Tannase, cellulase, and pectinase retained 22, 60, and 52% of their activity, respectively, in ethanol 30% v/v. Therefore, extractions were tested in ethanol concentrations between 0 and 30% v/v. Some of these enzymes presented synergistic effects in the extraction of specific polyphenols. Maximum yield of gallic acid was obtained using tannase and pectinase enzymes in ethanol 10% v/v (49.56 ± 0.01 mg L−1 h−1); in the case of p-coumaric acid, by cellulase and pectinase treatment in ethanol 30% v/v (7.72 ± 0.26 mg L−1 h−1), and in the case of trans-resveratrol, by pectinase treatment in ethanol 30% v/v (0.98 ± 0.04 mg L−1 h−1). Also, the effect of enzymes and solvent polarity was analysed for the extraction of malvidin-3-O-glucoside, syringic acid, and quercetin. Previous studies were mainly focused on the maximization of total polyphenols extraction yields, being the polyphenolic profile the consequence but not the driving force of the optimization. In the present study, the basis of a platform for a precise extraction of the desire polyphenols is provided.Fil: Piazza, Dana Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Meini, María Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Recovery of phenolic antioxidants from Syrah grape pomace through the optimization of an enzymatic extraction process

Phenolic compounds are highly valuable products that remain trapped in grape pomace, an abundant winery by-product. Therefore, efficient extraction procedures of these compounds represent a route for grape pomace valorisation. Here we performed a screening of the factors affecting the aqueous enzymatic extraction of phenolic compounds from Syrah grape pomace, including the following independent variables: temperature, pH, pectinase, cellulase and tannase; and a subsequent optimization through response surface methodology. At the optimal region, the enzymatic treatment enhanced the extraction yield of phenolics by up to 66% and its antioxidant capacity by up to 80%, reducing the incubation time and enzyme doses in respect to previous studies. We found that tannase raises the antioxidant capacity of the extract by the liberation of gallic acid, while cellulose favours the liberation of p-coumaric acid and malvidin-3-O-glucoside. We also tested the procedure in different grape pomace varieties, verifying its wide applicability.Fil: Meini, María Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química Biológica. Área Biofísica; ArgentinaFil: Cabezudo, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Boschetti, Carlos Eugenio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid

Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer.Fil: Porfiri, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Funcionalidad y Tecnología de Alimentos; ArgentinaFil: Melnichuk, Natasha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Brinatti, César. Universidade Estadual Do Campinas. Instituto de Química.; BrasilFil: Loh, Watson. Universidade Estadual Do Campinas. Instituto de Química.; BrasilFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Canabinoides como uma nova opção terapêutica nas doenças de Parkinson e de Alzheimer: uma revisão de literatura

Os derivados canabinoides podem ser vistos como novos potenciais terapêuticos para o tratamento da doença de Parkinson e Alzheimer. Assim, esta revisão teve como objetivo descrever os efeitos terapêuticos e adversos do uso de canabidiol e de delta-9-tetrahidrocanabinol nas doenças de Parkinson e de Alzheimer. Para tanto, foi realizada uma busca na base de dados Medline no período entre 2007 e 2017. Os descritores utilizados foram (Tetrahydrocannabinol OR Cannabidiol)AND (Parkinson OR Alzheimer) AND (Treatment OR Therapeutics). Os resultados mostraram efeitos terapêuticos promissores do canabidiol e do delta-9-tetrahidrocanabinol nestas doenças, tais como redução de sintomas motores e cognitivos, e ação neuroprotetora. Estes resultados podem ser explicados, em parte, pelos efeitos antioxidante, antiinflamatório, antagonista de receptores CB1, ou pela ativação de receptores PPAR-gama produzido por estas substâncias. Além disso, poucos efeitos adversos foram descritos, como boca seca e sonolência. Nesse contexto, estes resultados evidenciam a necessidadede novas pesquisas a respeito dos efeitos terapêuticos e adversos de canabinoides com maiores doses e períodos de exposição, para quem sabe, em um futuro próximo, ser possível olhar estas substâncias como uma opção terapêutica mais eficaz e segura para estes pacientes

Structural and functional analysis of Aspergillus niger xylanase to be employed in polyethylenglycol/salt aqueous two-phase extraction

The structure and enzymatic activity of Aspergillus niger xylanase were evaluated in different media to establish an appropriate protocol for the extraction of the enzyme in polymer/salt aqueous two-phase systems. Different factors were studied: the concentration and molecular weight (1000, 2000, 4600 and 8000) of polyethyleneglycol, the concentration and type of salt (sodium citrate and potassium phosphate) and pH, time and temperature. Xylanase was stable for 5 h at pH between 2.7 and 9.0 and at temperatures up to 50 °C. Fluorescence spectroscopy and circular dichroism experiments showed that neither the secondary/tertiary structure of the enzyme nor its catalytic activity were significantly altered in the presence of either salt or PEG. Xylanase partitioned into the PEG-rich phase driven by the excluded volume effect. Partitioning was more favorable to the polymer phase in the PEG1000/NaCit system, where Kp was 12 times higher than in the others aqueous two-phase systems. These results demonstrate the potential application of the PEG1000/NaCit system as a first step for the extraction of Aspergillus niger xylanase.Fil: Loureiro, Dana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Tubio, Gisela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Production of grape pomace extracts with enhanced antioxidant and prebiotic activities through solid-state fermentation by Aspergillus niger and Aspergillus oryzae

Grape pomace is a winery by-product that accumulates in high amounts in winemaking industry, which usually found difficulties regarding its disposal. Nonetheless, it has a great proportion of polyphenols which can be recovered to valorise this biomass. Filamentous fungi produce hydrolytic enzymes, which can assist in the liberation of polyphenols. Grape pomace was fermented in solid-state by Aspergillus niger and Aspergillus oryzae with the aim of facilitating the aqueous extraction of polyphenols with antioxidant activity by on-site enzyme production. Both fungi increased the antioxidant activity of the extracts, reaching maximum values of 73.7 ± 0.8 (A. niger) and 109.2 ± 0.5 (A. oryzae) mmol of Trolox equivalents/100 g of grape pomace. During fermentations, relevant enzymes were produced at high yields, A. niger produced a balanced profile of enzymes (cellulase, tannase, and pectinase), while A. oryzae switches to cellulase or tannase selective induction according to the fermentation conditions. Positive correlations were found between enzyme production, polyphenols recovery, and antioxidant activity. The extracts obtained after fermentation promoted the growth of Lactobacillus casei cultures. Therefore, the solid-state fermentation was effective for the simultaneous production of relevant industrial enzymes and grape pomace extracts with antioxidant and prebiotic properties, which have potential as functional food additives.Fil: Meini, María Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Cabezudo, Ignacio. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Galetto, Cecilia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Partitioning of xylanase from Thermomyces lanuginosus in PEG/NaCit aqueous two-phase systems: Structural and functional approach

The structure and catalytic activity of xylanase from Thermomyces lanuginosus were studied in different media (containing polyethylene glycol -PEG- or salt) at different temperatures. The aim was to study how the native structure of the enzyme is affected to understand the partitioning behavior of xylanase in PEG/sodium citrate (PEG/NaCit) aqueous two-phase systems. The presence of PEGs of different molar masses slightly altered the native structure of xylanase, although its catalytic activity was not affected. All the polymers assayed protect the native structure (and catalytic activity) of xylanase against temperature, except for PEG1000. Surface hydrophobicity experiments showed that xylanase favorable interacts with PEGs. Partitioning experiments confirmed this result and demonstrated that PEG1000/NaCit is the best system to partition xylanase from Thermomyces lanuginosus, since the Kp was 17.7 ± 0.3.Fil: Loureiro, Dana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Tubio, Gisela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Characterization of the Interaction Between Pancreatic Trypsin and an Enteric Copolymer as a Tool for Several Biotechnological Applications

Protein-polyelectrolyte complexes are very interesting systems since they can be applied in many long-established and emerging areas of biotechnology. From nanotechnology to industrial processing, these complexes are used for many purposes: to build multilayer particles for biosensors; to entrap and deliver proteins for pharmaceutical applications; to isolate and immobilize proteins. The enteric copolymer poly(methacrylic acid-co-methyl methacrylate) 1:2 (MMA) has been designed for drug delivery although its chemical properties allow to use it for other applications. Understanding the interaction between trypsin and this polymer is very important in order to optimize the mechanism of formation of this complex for different biotechnological applications.The formation of the trypsin-MMA complex was studied by spectroscopy and isothermal titration calorimetry. Structural analysis of trypsin was carried out by catalytic activity assays, circular dichroism and differential scanning calorimetry. Isothermal titration calorimetry experiments showed that the insoluble complex contains 12 trypsin molecules per MMA molecule at pH 5 and they interact with high affinity to form insoluble complexes. Both electrostatic and hydrophobic forces are involved in the formation of the complex. The structure of trypsin is not affected by the presence of MMA, although it interacts with some domains of trypsin affecting its thermal denaturation as seen in the differential scanning calorimetry experiments. Its catalytic activity is not altered. Dynamic light scattering demonstrated the presence of a soluble trypsin-copolymer complex at pH 5 and 8. Turbidimetric assays show that the insoluble complex can be dissolved by low ionic strength and/or pH in order to obtain free native trypsin.Fil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Loureiro, Dana Belen. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Tubio, Gisela. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentin