POLITICHE DI AMMISSIONE E GESTIONE DEI FLUSSI MIGRATORI DA LAVORO IN ITALIA DALLA TURCO-NAPOLITANO AD OGGI.

L'obiettivo di questo elaborato è quello di analizzare gli strumenti di cui si è dotata l'Italia al fine di regolare i flussi immigratori da lavoro. Dall'analisi svolta emerge la difficoltà dei governi italiani ad abbandonare una gestione emergenziale e contingente del fenomeno in favore di una maggiore consapevolezza di fronte all'immigrazione, in generale, e quella economica in particolare, in quanto elemento strutturale del tessuto socio-economico italiano. Le difficoltà oggettive, incontrate nella stesura dell'elaborato, sono frutto della complessità del fenomeno di per sé di natura transnazionale e che coinvolge una moltitudine di soggetti, oltre a presentare molteplici aspetti, da quelli socio-economici a quelli prettamente politici, fortemente intrecciati tra loro. Data la complessità di questo versante della politica migratoria, ho scelto una chiave di lettura storica, in quanto permette di evidenziare il profondo legame tra la discussione politica, caratterizzata da una forte polarizzazione ideologica, e gli interventi attuati in questa materia. All'interno di questi equilibri s'inserisce, inoltre, il crescente ruolo dell'Unione Europea che chiede, nonostante le resistenze dei singoli Stati, di rinunciare a una porzione di sovranità col fine di creare una politica migratoria comune. Nel primo capitolo vengono analizzati l'origine del fenomeno e la nascita della politica migratoria in Italia. Con l'approvazione della legge Turco-Napolitano nel 1998, e il conseguente raggruppamento delle leggi all'interno del Testo Unico, l'Italia cerca di dotarsi di una legge organica in materia migratoria. Il sistema è incentrato sulla chiamata nominativa del lavoratore ancora all'estero, che avviene secondo i limiti numerici definiti dalla programmazione dei flussi ed attuati attraverso il cosiddetto decreto-flussi. Questo meccanismo dimostra, fin dalla nascita, notevoli inadeguatezze e carenze, per cui si deve ricorrere sistematicamente a strumenti di regolarizzazione ex post di una presenza irregolare che tende a riprodursi, anche a causa del lavoro sommerso che funge da potente fattore d'attrazione dei flussi, e tutto questo perché, sia i datori di lavoro che i lavoratori immigrati, non trovano conveniente utilizzare i canali d'ingresso regolari. Nel secondo capitolo il punto di partenza è rappresentato dalla riforma attuata dalla legge Bossi-Fini nel 2001. Tale legge è frutto della nuova maggioranza di centro-destra che fa dell'immigrazione un tema cruciale per raccogliere consensi, associandola a situazioni problematiche di sicurezza e ordine pubblico. La legge interviene in maniera sostanziale sui meccanismi di ammissione, inasprendo le condizioni di ingresso con l'introduzione del “contratto di soggiorno” (che subordina la presenza alla disponibilità di un'occupazione lavorativa) e precarizzando il soggiorno dei regolari, dimezzando la durata dei permessi. Ai fattori interni si aggiunge l'influenza dell'Unione Europea. L'allargamento verso i paesi dell'Est rappresenta l'apice delle carenze del meccanismo di ammissione: una larga parte degli ingressi si sottrae al potere regolatorio del sistema delle quote, dal momento che buona parte dei flussi si compone di soggetti che non necessitano più di alcun permesso di soggiorno. La medesima incapacità emerge anche con il presentarsi della crisi economica alla fine del 2008. Infatti, nonostante la drastica riduzione delle quote, i flussi, prevalentemente diretti al servizio alle famiglie, si mantengono elevati e subiscono una battuta d'arresto solo a partire dal 2011. Nel terzo capitolo vengono presi in considerazione i provvedimenti più recenti: l'Accordo d'Integrazione all'interno del Piano per l'Integrazione e la Carta Blu sono frutto del crescente ruolo dell'Unione Europea e dell'affermarsi del nuovo (e più impegnativo) concetto d'integrazione e, pertanto, rappresentano il tentativo dell'azione politica di adeguarsi ai principi della politica migratoria dell'Unione Europea. Tali provvedimenti, tuttavia, hanno suscitato perplessità e critiche, sia in termini ideologici che in termini di attuabilità concreta, data la mancata predisposizione di risorse aggiuntive necessarie. Il quadro che emerge è che l'Italia ha attratto ingenti flussi senza che a questi corrispondesse una crescita economica, e questo è frutto del potere d'attrazione costituito dai fattori socio-demografici, come la bassa natalità, l'invecchiamento della popolazione, l'evoluzione delle aspettative lavorative, soprattutto dei giovani. In conclusione i problemi rimangono aperti, anche se oggi meno visibili principalmente per due fattori. Il primo consiste nella mancanza di un dibattito politico e mediatico, nonché scientifico, causato dalla poca notiziabilità dell'immigrazione regolare, in favore degli aspetti più eclatanti e conflittuali. Il secondo invece è rappresentato dalla battuta d'arresto dei flussi di arrivo, che fa apparire il problema lontano. E così l'azione politica aspetta, finché non si presenterà la prossima emergenza

A method for extracting a high-quality RNA from Symbiodinium sp.

Good quality RNA is essential for a range of analyses including microarray and gene expression studies. A number of methods for RNA extraction from symbiotic dinoflagellates were assessed for their ability to recover a high-quality RNA applicable for evaluation of gene expression profiles. The recovery and quality of the obtained RNA were evaluated with respect to UV light absorbance profiles and automated microcapillary electrophoretic RNA separation. A modified RNA extraction procedure that combines two existing commercial kits, Trizol and Qiagen RNeasy kits, was efficiently employed for the recovery of a high-quality RNA under specific homogenization conditions. Cell homogenization using glass beads at the speed of 4,500 rpm for up to 6 min resulted in a good RNA recovery and preserved RNA integrity. A high-quality RNA obtained following the described procedure was successfully applied in reverse transcriptase-polymerase chain reaction (PCR) and in quantitative PCR studies. Gene expression profiles were changed with RNA extraction procedure, with the highest transcript numbers at precise conditions of cell homogenization. RNA samples with RNA integrity number values from 6 and above were recommended for downstream applications. This sequence of RNA isolation and RNA evaluation represents a methodological improvement required for functional genomic studies in dinoflagellates

Extraction and analysis of Mycosporine-Like Amino Acids in marine algae

Marine organisms use mycosporine-like amino acids (MAAs) as biological sunscreens for the protection from damaging ultraviolet (UV) radiation and the prevention of oxidative stress. MAAs have been discovered in many different marine and freshwater species including cyanobacteria, fungi, and algae, but also in animals like cnidarian and fishes. Here, we describe a general method for the isolation and characterization of MAA compounds from red algae and symbiotic dinoflagellates isolated from coral hosts. This method is also suitable for the extraction and analyses of MAAs from a range of other algal and marine biota

Differential Regulation by Heat Stress of Novel Cytochrome P450 Genes from the Dinoflagellate Symbionts of Reef-Building Corals▿

Exposure to heat stress has been recognized as one of the major factors leading to the breakdown of the coral-alga symbiosis and coral bleaching. Here, we describe the presence of three new cytochrome P450 (CYP) genes from the reef-building coral endosymbiont Symbiodinium (type C3) and changes in their expression during exposure to severe and moderate heat stress conditions. Sequence analysis of the CYP C-terminal region and two conserved domains, the “PERF” and “heme-binding” domains, confirmed the separate identities of the CYP genes analyzed. In order to explore the effects of different heat stress scenarios, samples of the scleractinian coral Acropora millepora were exposed to elevated temperatures incrementally over an 18-h period (rapid thermal stress) and over a 120-h period (gradual thermal stress). After 18 h of gradual heating and incubation at 26°C, the Symbiodinium CYP mRNA pool was approximately 30% larger, while a further 6°C increase to a temperature above the average sea temperature (29°C after 72 h) resulted in a 2- to 4-fold increase in CYP expression. Both rapid heat stress and gradual heat stress at 32°C resulted in 50% to 90% decreases in CYP gene transcript abundance. Consequently, the initial upregulation of expression of CYP genes at moderately elevated temperatures (26°C and 29°C) was followed by a decrease in expression under the greater thermal stress conditions at 32°C. These findings indicate that in the coral-alga symbiosis under heat stress conditions there is production of chemical stressors and/or transcriptional factors that regulate the expression of genes, such as the genes encoding cytochrome P450 monooxygenases, that are involved in the first line of an organism's chemical defense

New-old hemoglobin-like proteins of symbiotic dinoflagellates

Symbiotic dinoflagellates are unicellular photosynthetic algae that live in mutualistic symbioses with many marine organisms. Within the transcriptome of coral endosymbionts Symbiodinium sp. (type C3), we discovered the sequences of two novel and highly polymorphic hemoglobin-like genes and proposed their 3D protein structures. At the protein level, four isoforms shared between 87 and 97% sequence identity for Hb-1 and 7899% for Hb-2, whereas between Hb-1 and Hb-2 proteins, only 1521% sequence homology has been preserved. Phylogenetic analyses of the dinoflagellate encoding Hb sequences have revealed a separate evolutionary origin of the discovered globin genes and indicated the possibility of horizontal gene transfer. Transcriptional regulation of the Hb-like genes was studied in the reef-building coral Acropora aspera exposed to elevated temperatures (67 degrees C above average sea temperature) over a 24-h period and a 72-h period, as well as to nutrient stress. Exposure to elevated temperatures resulted in an increased Hb-1 gene expression of 31% after 72h only, whereas transcript abundance of the Hb-2 gene was enhanced by up to 59% by both 1-day and 3-day thermal stress conditions. Nutrient stress also increased gene expression of Hb-2 gene by 70%. Our findings describe the differential expression patterns of two novel Hb genes from symbiotic dinoflagellates and their polymorphic nature. Furthermore, the inducible nature of Hb-2 gene by both thermal and nutrient stressors indicates a prospective role of this form of hemoglobin in the initial coralalgal responses to changes in environmental conditions. This novel hemoglobin has potential use as a stress biomarker

Gene expression profiles of cytosolic heat shock proteins Hsp70 and Hsp90 from symbiotic dinoflagellates in response to thermal stress: possible implications for coral bleaching

Unicellular photosynthetic dinoflagellates of the genus Symbiodinium are the most common endosymbionts of reef-building scleractinian corals, living in a symbiotic partnership known to be highly susceptible to environmental changes such as hyperthermic stress. In this study, we identified members of two major heat shock proteins (HSPs) families, Hsp70 and Hsp90, in Symbiodinium sp. (clade C) with full-length sequences that showed the highest similarity and evolutionary relationship with other known HSPs from dinoflagellate protists. Regulation of HSPs gene expression was examined in samples of the scleractinian coral Acropora millepora subjected to elevated temperatures progressively over 18 h (fast) and 120 h (gradual thermal stress). Moderate to severe heat stress at 26°C and 29°C (+3°C and +6°C above average sea temperature) resulted in an increase in algal Hsp70 gene expression from 39% to 57%, while extreme heat stress (+9°C) reduced Hsp70 transcript abundance by 60% (after 18 h) and 70% (after 120 h). Elevated temperatures decreased an Hsp90 expression under both rapid and gradual heat stress scenarios. Comparable Hsp70 and Hsp90 gene expression patterns were observed in Symbiodinium cultures and in hospite, indicating their independent regulation from the host. Differential gene expression profiles observed for Hsp70 and Hsp90 suggests diverse roles of these molecular chaperones during heat stress response. Reduced expression of the Hsp90 gene under heat stress can indicate a reduced role in inhibiting the heat shock transcription factor which may lead to activation of heat-inducible genes and heat acclimation

Extending the diversity of cytochrome P450 enzymes by DNA family shuffling

The cytochrome P450 enzymes involved in xenobiotic metabolism are an excellent starting point for the directed evolution of novel biocatalysts due to their wide substrate specificity. A shuffled library of three highly homologous mammalian genes (for P450 2C9, P450 2C11 and P450 2C19) was constructed by applying a modified DNA family shuffling procedure. The modifications made to the traditional DNA shuffling protocols involved non-random digestion via the use of different combinations of restriction enzymes (REs) followed by isolation of fragments under 300 bp by size-selective filtration. Shuffled cytochrome P450 mutants were co-expressed in Escherichia coli with their redox partner, NADPH-cytochrome P450 reductase (NPR). We report here how non-random fragmentation may help in chimeragenesis within the areas of low sequence similarity such as substrate recognition sites (SRSs) that are generally underrepresented in recombination using the random fragmentation process. Size-selective filtration was used to limit recovery of incompletely digested fragments and consequently minimize the chances for contamination of the shuffled library with parental forms. No parental forms could be detected in the shuffled library using restriction fragment length polymorphism (RFLP) analysis, suggesting the library was free of parental contamination. Sequencing of randomly selected mutants demonstrated a high level of chimeragenesis with on average of 8.0 +/- 2.2 crossovers and a low level of mutagenesis with 5.2 +/- 2.8 spontaneous mutations per similar to 1.5 kbp of the full-length P450 sequence. The proportion of properly folded protein as indicated by the observation of characteristic Fe(II).CO vs. Fe(II) difference spectra was 15% (4/27) of analysed mutants. Screening of the shuffled library for indole oxidation revealed four clones with similar or higher levels of indigo pigment production to those of the parental P450s and two clones with elevated P450 expression. In this paper we present a method for the effective family shuffling of cytochrome P450 enzymes, applicable to the creation of mutant libraries with expanded metabolic diversity and with a significant proportion of functional clones. Crown Copyright (C) 2007 Published by Elsevier B.V. All rights reserved

Phylogenetic analysis of genes involved in mycosporine-like amino acid biosynthesis in symbiotic dinoflagellates

Mycosporine-like amino acids (MAAs) are multifunctional secondary metabolites involved in photoprotection in many marine organisms. As well as having broad ultraviolet (UV) absorption spectra (310–362 nm), these biological sunscreens are also involved in the prevention of oxidative stress. More than 20 different MAAs have been discovered so far, characterized by distinctive chemical structures and a broad ecological distribution. Additionally, UV-screening MAA metabolites have been investigated and used in biotechnology and cosmetics. The biosynthesis of MAAs has been suggested to occur via either the shikimate or pentose phosphate pathways. Despite their wide distribution in marine and freshwater species and also the commercial application in cosmetic products, there are still a number of uncertainties regarding the genetic, biochemical, and evolutionary origin of MAAs. Here, using a transcriptome-mining approach, we identify the gene counterparts from the shikimate or pentose phosphate pathway involved in MAA biosynthesis within the sequences of the reef-building coral symbiotic dinoflagellates (genus Symbiodinium). We also report the highly similar sequences of genes from the proposed MAA biosynthetic pathway involved in the metabolism of 4-deoxygadusol (direct MAA precursor) in various Symbiodinium strains confirming their algal origin and conserved nature. Finally, we reveal the separate identity of two O-methyltransferase genes, possibly involved in MAA biosynthesis, as well as nonribosomal peptide synthetase and adenosine triphosphate grasp homologs in symbiotic dinoflagellates. This study provides a biochemical and phylogenetic overview of the genes from the proposed MAA biosynthetic pathway with a focus on coral endosymbionts

Mycosporine-Like Amino Acids from Coral Dinoflagellates▿

Coral reefs are one of the most important marine ecosystems, providing habitat for approximately a quarter of all marine organisms. Within the foundation of this ecosystem, reef-building corals form mutualistic symbioses with unicellular photosynthetic dinoflagellates of the genus Symbiodinium. Exposure to UV radiation (UVR) (280 to 400 nm) especially when combined with thermal stress has been recognized as an important abiotic factor leading to the loss of algal symbionts from coral tissue and/or a reduction in their pigment concentration and coral bleaching. UVR may damage biological macromolecules, increase the level of mutagenesis in cells, and destabilize the symbiosis between the coral host and their dinoflagellate symbionts. In nature, corals and other marine organisms are protected from harmful UVR through several important photoprotective mechanisms that include the synthesis of UV-absorbing compounds such as mycosporine-like amino acids (MAAs). MAAs are small (<400-Da), colorless, water-soluble compounds made of a cyclohexenone or cyclohexenimine chromophore that is bound to an amino acid residue or its imino alcohol. These secondary metabolites are natural biological sunscreens characterized by a maximum absorbance in the UVA and UVB ranges of 310 to 362 nm. In addition to their photoprotective role, MAAs act as antioxidants scavenging reactive oxygen species (ROS) and suppressing singlet oxygen-induced damage. It has been proposed that MAAs are synthesized during the first part of the shikimate pathway, and recently, it has been suggested that they are synthesized in the pentose phosphate pathway. The shikimate pathway is not found in animals, but in plants and microbes, it connects the metabolism of carbohydrates to the biosynthesis of aromatic compounds. However, both the complete enzymatic pathway of MAA synthesis and the extent of their regulation by environmental conditions are not known. This minireview discusses the current knowledge of MAA synthesis, illustrates the diversity of MAA functions, and opens new perspectives for future applications of MAAs in biotechnology