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Large scale docking screening and simulations of molecular dynamics for the study of ligand binding in different protein models
No full textMolecular recognition is central to biology, it is the starting point for almost all processes in biological systems; knowledge of the molecular association aids in understanding a variety of pathways taking place in the living cell. Molecular binding is an attractive interaction between two molecules which results in a stable molecular\ud
complex. Among computational applications, docking is the methodology for predicting the complex. There are many docking strategies, fast molecular docking and absolute\ud
binding free energies are applied in this work.\ud
These methodologies have been applied to three biological systems: neuraminidase of H5N1 influenza virus, human Endothelial Protein C receptor (EPCR) and β-tubulin of\ud
the psycrophilic organism, Euplotes focardii. In particular, the first study is a drug design application; two selective ligands for the open conformation of the H5N1\ud
influenza virus neuraminidase has been obtained, performing a comparative binding study with fast molecular docking and specific interaction filters. In the second study structurally stable mutants of EPCR with impaired binding affinity for phosphatidylethanolamine (PTY), will be designed and selected for the analysis of the\ud
ligand role in EPCR function. In the third work the structural adaptation to low temperature of the three β-tubulin isotypes are analysed employing molecular dynamics\ud
simulations.\ud
The applied computational methods allow to obtain accurate predictions in the field of binding analysis. In particular, fast molecular docking allows to obtain the correct binding mode and provide a good approximation of the interaction energy; molecular dynamics supply accurate binding affinities and is a useful tool for the study of the\ud
effect of ligand binding on protein structures
Presence of biogenic amines in the Formaggio di Fossa produced in the Marche region\ud
Get PDFFormaggio di Fossa is an Italian traditional cheese of the Montefeltro area (Emilia Romagna and Marche regions) characterized by a particular step of ripening that is carried out into pits (infossamento) borne in the sandstone. Since the XIV century, the inhabitants were used to set food, especially cereals and cheese, into pits to preserve them during winter and to protect them from invaders. The aim of the present work is to study physical and chemical features of this product with particular reference to the presence of the most important biogenic amines (-Phenylethylamine, Putrescine, Cadaverine, Histamine, Tyramine, Spermine and Spermidine), compared with a control cheese fully ripened in factory. Formaggio di Fossa showed higher values of Aw, pH, humidity, proteins, pH 4,6-soluble nitrogen (NCN) and water soluble nitrogen (NPN) and much lower amounts of fat. Much higher amounts of total biogenic amines were detected in Formaggio di Fossa than in control cheese, where their concentration was very low. Cadaverine, putrescine and tyramine were the most concentrated biogenic amines. Nevertheless, thyramine was present at levels suggested as compatible with GMPs (Good Manufacturing Practices). Histamine was detected at low amounts, far from potentially toxic levels.\ud
Finally, in this work have been studied also the microbiological features and the correlation between BAs and decarboxylase activity of bacteria isolated from the Formaggio di Fossa.\ud
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Pyrethroid insecticides induce oxidative stress, immuno and neuro-toxicity in Wistar rats
No full textPyrethroids (Cypermethrin CY, Permethrin PERM, etc.) are widely used insecticides of low acute toxicity in mammals, however there are serious concerns on the risks related to their exposure. They mainly act on central nervous system and show significant effects on peripheral nerves, muscles and immune system.\ud
In this work, the effects of CY and/or PERM exposure on pup (1/10 LD50, 10 days) and adolescent (1/10 LD50, 60 days) rats were investigated. In particular the implications of pyrethroids exposure on immune and central nervous system were studied.\ud
Pyrethroids impair both monocytes and PMNs respiratory burst. They reduce reactive oxygen species production in activated monocytes while increase their level in PMNs. In vivo antioxidant supplementation (Vitamin E and/or Coenzyme Q10) can protect against the abnormal respiratory burst observed in PMNs from adolescent rats. In vivo and in vitro studies suggest physico-chemical changes at plasma membrane level in leukocytes. Moreover, pyrethroids induce protein and lipid oxidation in white blood cells.\ud
These insecticides increase protein oxidation also in striatum where PERM depletes GSH (both in pups and\ud
adolescent rats). In vivo Vitamin E supplementation restores the GSH level to the control value observed in\ud
adolescent rats. Moreover, antioxidant supplementation protects striata from the PERM induced DNA damage. PERM induces single and double-strand breaks as shown by comet assay. Modified versions of comet assay were performed in order to better characterize PERM induced DNA damage.\ud
With the aim to evaluate the role of mitochondrial superoxide anion on DNA damage, in vitro studies on\ud
striatal sub-mitochondrial particles (SMPs) were performed. In our experimental conditions, PERM inhibits mitochondrial complex I activity. This effect discards the involvement of mitochondria reactive oxygen species in the striatum DNA damage, while focalizes the main role of PERM on other oxidative stress pathways. The pivotal role of oxidative stress in PERM induced damage is underscored by the fact that in vitro GSH supplementation or pre-treatment prevents PERM induced DNA damage
Matrimonio e famiglia non fondata sul matrimonio\ud
No full textFamily not founded on marriage is specifically characterized as respect marriage. The choice to create a family without marriage needs to be grasped as the will of avoiding the specific regulation of marriage.\ud
The social formations which originate need only to be submitted to a solidaristic principle, according to what is prescribed in article 2 of the Italian Constitution. Nevertheless, the topic of family not founded on marriage is often related to atypical couples not admitting to marriage.\ud
We have to avoid that, dealing with that subject, we choose solution in contrast with freedom of person and exigency of guaranteeing the full development of personality into the family. In that way, it’s useful to reflect over the possibility of a «redefinition» of knowledge of marriage, so that it can include also atypical couples as well as it can distinguish itself from other form of cohabitation without marriage.\ud
To correctly configure such a perspective of redefinition, we have to collocate properly the knowledge of marriage in an anthropological and sociological perspective.\ud
In that way, we can recognize the multiplicity of ancestral family forms and marriage forms, and their later evolution. In the same perspective, we have to consider the historical specificity of cohabitation and of family not founded on marriage, as well as their recent evolution.\ud
The necessity of a «redefinition» of the knowledge of marriage is today particularly strong; the debate over that is quite developed in USA.\ud
In such a legal and cultural context, even now – paradoxically - characterized by the importance of religion in lawmaking process, one of the main objections deals with the hypothesis of the former configurability of knowledge of marriage into the religious sphere; in that perspective, the same term «marriage» should be referred to as a determinate cultural ambit, as well as it should be configured as an absolute, immutable, ahistorical element. This objection doesn’t appear binding enough. If we have an excursus over the scriptural assumption who originated the religious knowledge of marriage, in Western society, we can see the evolution of that knowledge, as well as its non-univocity. \ud
Besides, if we consider the enforcement of knowledge of marriage in the early Western society, we realize how important Roman family law has been for the development of Christian tradition.\ud
About the question of a possible «redefinition» of knowledge of marriage, «new natural lawyers», who support tradition, and «liberals», favourable to changing, are deeply in discussion in the United States of America. There is also a minoritarian group, the «progressive critics», who contest the same idea of marriage and the related special rights; they indeed wish an equalization of any kind of family, founded or not founded on marriage.\ud
The necessity of considering the debate dealing with the hypothesis of «redefinition» of knowledge of marriage in contexts different from our legal system is originated by the comprehension of the importance of a comparative perspective in family law. In fact, all phenomena dealing with family are, nowadays inserted in a globalizing dynamic. \ud
A theme, in particular, can be considered as paradigmatic in our research: the possibility of a formal recognition of same-sex couples. It seems to represent – when that kind of unions are admitted in the form of «marriage» - the «point of criticality» respect the traditional knowledge of marriage.\ud
The specific field of research is focused on common law, civil law, and mixing system Countries.\ud
So that, our analysis deals with the United States of America, where decisions over same-sex marriages are getting more and more importance in the last years, as well as scholars started being deeply involved into that subject. It deals also with Spain, a Country quite similar to ours, but where, in a very short period, it has produced a deep, radical change in family law, admitting same-sex marriage and divorce without separation.\ud
The analysis refers also to South Africa, where we can observe a surprising variety of marriage forms. Beyond «common law marriage», a marriage originated from Western legal system, they have the «customary marriage», a traditional, polygynous marriage, and the «civil union marriage», that allow both opposite-sex and same-sex couples to decide if to denominate their union as «marriage» or «civil partnership», with the same legal effects, independently from the denomination.\ud
In the Italian context, before considering a hypothesis of «redefinition» of knowledge of marriage, according with what is happening in the other legal systems we considered, it’s required that we configure marriage, as well as family not founded on marriage, properly, as prescribed by the Italian Constitution. These social formations need to be regarded according with the application of solidaristic value and centrality of the person. Also the European Union’s prescriptions have to be considered. \ud
Only later, having all the elements required, it will be possible to configure the eventuality of the introduction of same-sex marriage, as several lower Courts referred to the Italian Constitutional Court to decide about on the matter.\u
Istruzione,Ricerca ed Open Access. Il Sapere tra liberta', limiti e diritti. Esperienze Scolastiche
No full textWeird facets of ribosome synthesis in stressfully chilled\ud bacteria
No full textLolita Piersimoni, Mara Giangrossi, Claudio O. Gualerzi & Cynthia L. Pon\ud
\ud
Laboratory of Genetics, Department of Biology MCA, University of Camerino, 62032 Camerino (MC), Italy\ud
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We present data indicating that an imbalance of initiation factors/ribosome is generated by a cold-shock-induced decrease of the rate of assembly of ribosomal subunits accompanied by an increased expression of initiation factors. The rRNA synthesized after cold shock (a 37°C→10°C temperature-downshift), unlike that synthesized during exponential growth at 37°C, is transcribed mainly from the P2 promoters, which become activated by stress. Furthermore, our data indicate that both synthesis and maturation of rRNA are substantially retarded after temperature downshift. The amount of rRNA synthesized de novo during and after cold-adaptation represents a fairly large amount of total RNA. The fate of the rRNA synthesized during cold-shock is that of-ending up primarily in the active ribosomes present in the polysomes, while the majority of the rRNA whose synthesis had started immediately before or after the stress ultimately ends up in presumably inactive 70S monomers.\ud
With respect to ribosomal proteins (r-proteins), we show that after cold-shock five r-proteins (S2, S3, S5, S6 and S19) give rise to two distinct spots in 2D-gel electrophoresis. The appearance of new spots for proteins S2, S3, S5 and S6 is due to the lack of some post translational modifications as determined by mass spectrometry analysis. In addition, the data indicate the presence of post translational modifications so far unknown.\ud
We also demonstrate that cold-shock does not affect the stoichiometry of the r-proteins within the 30S ribosomal subunits. The analysis of non-ribosomal proteins associated with the peak of 30S ribosomal subunits (which probably contains precursors of the 50S subunits) becomes specifically enriched in a number of proteins which are or might be involved in assembly and/or maturation of the ribosomal particles and translation. For example, among the initiation factors, IF2 is the only cold-shock induced initiation factor whose level increases in association with the 30S ribosomal subunit and whose function in cold-shock is actually not known. These findings open new perspectives towards the identification of a cold-shock role for this and other proteins. \ud
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Supported by a MIUR PRIN 2007 Grant to CLP\u
Rational design, synthesis and biological evaluation of new molecules directed to neurotransmitter receptors
No full textA broad range of severe pathologies such as pain, anxiety, neurodegeneration, hypertension, asthma, as well as some gastrointestinal diseases, are due to receptor dysfunction. Some of these pathologies have a tremendous social impact as they involve large population segments, in particular aged people. The systems more frequently involved in these pathologies are adenosinic, adrenergic, aminoacidergic, cholinergic (muscarinic and nicotinic), dopaminergic, imidazoline, triptaminergic, and sigma receptors as well as Wnt pathway.\ud
The goals of the research reported in this thesis intended (i) to obtain more information about structure, functions and dysfunctions of neurotransmitter receptors, (ii) to\ud
individuate novel leads to develop useful drugs in the treatment of neuroreceptor dysfunctions. Therefore imidazoline, 1, 4-dioxane and spiropiperidine nuclei were\ud
utilized for designing novel a2-adrenergic (III, IX), imidazoline (IV, VIII, X, XII), and dopaminergic (V), muscarinic (VI), and 5-HT1A-serotoninergic (XI), and sigma ligands (VII), respectively. Finally, new azaindole analogues have been synthesized as RHO kinase inhibitors (I, II)
SOCIAL NETWORKS OF THE ITALIAN MAFIA; THE STRONG AND WEAK PARTS
Get PDFIn this paper, it is argued that well-built, social network system has enabled the Mafia to\ud
reach a certain level of success through three main networks: members, local people, and\ud
politicians. I assert that the role of the executive power of the state has been partially\ud
supportive in this success. Moreover, this paper also concludes that to combat different\ud
Mafia groups, it is essential to know their strong and weak parts. Consequently, it is found\ud
that their well-built network system does not solely comprise of strong parts but that the\ud
weak parts also exist, albeit, that they have not yet played a defective role in the resolution of\ud
the Mafia. Therefore, this paper suggests that the illustration of both the strong and weak\ud
parts of these networks can have prominent and assisting role in the combat against the\ud
Mafia phenomenon in the future, either by strengthening the weak parts or by weakening the\ud
strong parts of its networks
A Semantic Framework for Declarative and Procedural Knowledge
Get PDFIn any scientic domain, the full set of data and programs has reached an-ome status, i.e. it has grown massively. The original article on the Semantic Web describes the evolution of a Web of actionable information, i.e.\ud
information derived from data through a semantic theory for interpreting the symbols. In a Semantic Web, methodologies are studied for describing, managing and analyzing both resources (domain knowledge) and applications (operational knowledge) - without any restriction on what and where they\ud
are respectively suitable and available in the Web - as well as for realizing automatic and semantic-driven work\ud
ows of Web applications elaborating Web resources.\ud
This thesis attempts to provide a synthesis among Semantic Web technologies, Ontology Research, Knowledge and Work\ud
ow Management. Such a synthesis is represented by Resourceome, a Web-based framework consisting of two components which strictly interact with each other: an ontology-based and domain-independent knowledge manager system (Resourceome KMS) - relying on a knowledge model where resource and operational knowledge are contextualized in any domain - and a semantic-driven work ow editor, manager and agent-based execution system (Resourceome WMS).\ud
The Resourceome KMS and the Resourceome WMS are exploited in order to realize semantic-driven formulations of work\ud
ows, where activities are semantically linked to any involved resource. In the whole, combining the use of domain ontologies and work ow techniques, Resourceome provides a exible domain and operational knowledge organization, a powerful engine for semantic-driven work\ud
ow composition, and a distributed, automatic and\ud
transparent environment for work ow execution