Inhibitory effect of 2,3,5,6-tetrafluoro-4-[4-(Aryl)-1H-1,2,3-triazol-1-yl]benzenesulfonamide derivatives on HIV reverse transcriptase associated rnase H activities

The HIV-1 ribonuclease H (RNase H) function of the reverse transcriptase (RT) enzyme catalyzes the selective hydrolysis of the RNA strand of the RNA:DNA heteroduplex replication intermediate, and represents a suitable target for drug development. A particularly attractive approach is constituted by the interference with the RNase H metal-dependent catalytic activity, which resides in the active site located at the C-terminus p66 subunit of RT. Herein, we report results of an in-house screening campaign that allowed us to identify 4-[4-(aryl)-1H-1,2,3-triazol-1-yl]benzenesulfonamides, prepared by the “click chemistry” approach, as novel potential HIV-1 RNase H inhibitors. Three compounds (9d, 10c, and 10d) demonstrated a selective inhibitory activity against the HIV-1 RNase H enzyme at micromolar concentrations. Drug-likeness, predicted by the calculation of a panel of physicochemical and ADME properties, putative binding modes for the active compounds, assessed by computational molecular docking, as well as a mechanistic hypothesis for this novel chemotype are reported

Quantifying the legacy of the Chinese Neolithic on the maternal genetic heritage of Taiwan and Island Southeast Asia

There has been a long-standing debate concerning the extent to which the spread of Neolithic ceramics and Malay-Polynesian languages in Island Southeast Asia (ISEA) were coupled to an agriculturally driven demic dispersal out of Taiwan 4000 years ago (4 ka). We previously addressed this question using founder analysis of mitochondrial DNA (mtDNA) control-region sequences to identify major lineage clusters most likely to have dispersed from Taiwan into ISEA, proposing that the dispersal had a relatively minor impact on the extant genetic structure of ISEA, and that the role of agriculture in the expansion of the Austronesian languages was therefore likely to have been correspondingly minor. Here we test these conclusions by sequencing whole mtDNAs from across Taiwan and ISEA, using their higher chronological precision to resolve the overall proportion that participated in the “out-of-Taiwan” mid-Holocene dispersal as opposed to earlier, postglacial expansions in the Early Holocene. We show that, in total, about 20 % of mtDNA lineages in the modern ISEA pool result from the “out-of-Taiwan” dispersal, with most of the remainder signifying earlier processes, mainly due to sea-level rises after the Last Glacial Maximum. Notably, we show that every one of these founder clusters previously entered Taiwan from China, 6–7 ka, where rice-farming originated, and remained distinct from the indigenous Taiwanese population until after the subsequent dispersal into ISEA

Chelation motifs affecting metal-dependent viral enzymes: N′-acylhydrazone ligands as dual target inhibitors of HIV-1 Integrase and Reverse Transcriptase Ribonuclease H domain

Human immunodeficiency virus type 1 (HIV-1) infection, still represent a serious global health emergency. The chronic toxicity derived from the current anti-retroviral therapy limits the prolonged use of several antiretroviral agents, continuously requiring the discovery of new antiviral agents with innovative strategies of action. In particular, the development of single molecules targeting two proteins (dual inhibitors) is one of the current main goals in drug discovery. In this contest, metal-chelating molecules have been extensively explored as potential inhibitors of viral metal-dependent enzymes, resulting in some important classes of antiviral agents. Inhibition of HIV Integrase (IN) is, in this sense, paradigmatic. HIV-1 IN and Reverse Transcriptase-associated Ribonuclease H (RNase H) active sites show structural homologies, with the presence of two Mg(II) cofactors, hence it seems possible to inhibit both enzymes by means of chelating ligands with analogous structural features. Here we present a series of N′-acylhydrazone ligands with groups able to chelate the Mg(II) hard Lewis acid ions in the active sites of both the enzymes, resulting in dual inhibitors with micromolar and even nanomolar activities. The most interesting identified N′-acylhydrazone analog, compound 18, shows dual RNase H-IN inhibition and it is also able to inhibit viral replication in cell-based antiviral assays in the low micromolar range. Computational modeling studies were also conducted to explore the binding attitudes of some model ligands within the active site of both the enzymes

Modellistica distribuita di bilancio idrologico per la gestione dell'irrigazione : il caso di studio del consorzio di bonifica Muzza Bassa Lodigiana

LAUREA SPECIALISTICANegli ultimi anni le situazioni di crisi idrica sempre più frequenti hanno evidenziato la necessità di utilizzare l’acqua in modo sempre più oculato. Questo problema si riflette nel cercare una migliore e più adeguata gestione e pianificazione dell’irrigazione da parte dei Consorzi. Infatti, in questo settore si sono sviluppati negli ultimi anni sistemi basati su algoritmi in grado di ottimizzare le decisioni, con l’obiettivo di migliorare l’efficienza nell’uso delle risorse idriche limitate. Il presente lavoro di tesi fa parte del progetto, finanziato dalla Regione Lombardia, “Previsione meteo idrologica per la Gestione Irrigua” (“PRE.G.I.”), che propone la realizzazione di un prodotto di previsione dei contenuti idrici dei suoli e dei fabbisogni irrigui utili alla regolazione parsimoniosa dell’acqua per l’irrigazione, unendo la previsione meteorologica di lungo periodo alla simulazione del bilancio idrico del suolo. Lo strumento PREGI è applicato ai territori della Provincia di Lodi serviti dal Consorzio di Bonifica Muzza Bassa Lodigiana (MBL) ed è sviluppato da Centro Epson Meteo – Meteo Operations Italia s.r.l., MMI s.r.l., il D.I.I.A.R. del Politecnico di Milano e dal Consorzio Bonifica Muzza Bassa Lodigiana (MBL). Prendendo spunto dalle recenti pubblicazioni di letteratura, nel Capitolo 1 viene fornito prima un inquadramento generale sulla necessità di migliorare sempre di più la gestione irrigua e, successivamente, gli obiettivi che la tesi propone. Segue poi una descrizione dell’area di studio con particolare attenzione alle caratteristiche idrologiche del bacino irriguo in esame (Capitolo 2). Nel terzo e nel quarto capitolo viene presentata rispettivamente una descrizione del sistema irriguo del Consorzio MBL nel suo stato attuale e nella sua modellazione. Nel Capitolo 5 sono stati discussi i risultati ottenuti dalle simulazioni effettuate con il modello idrologico FEST-EWB per il periodo compreso tra il 2003 e il 2010. Infine nel Capitolo 6 sono state presentate le conclusioni ottenute a seguito delle analisi svolte e i possibili sviluppi futuri in questo settore.In recent years, the water crisis have highlighted the increasingly frequent need to use water in an ever more discerning. This problem is reflected in the search for a better and more appropriate planning and management of irrigation by the Consortium. In fact, in this area has developed in recent years systems based on algorithms that optimize decisions, with the aim of improving the efficient use of limited water resources. The present work is part of the project, funded by the Region of Lombardy, "Hydrologic Forecast for Irrigation Management" ("PRE.GI"), which proposes the creation of a product for estimating water content of soils and irrigation requirements useful for the economical control of water for irrigation, weather forecasting by combining the long-term simulation of soil water balance. The tool is applied to the territories of the Province of Lodi served by Consorzio di Bonifica Muzza Bassa Lodigiana (MBL) and is developed by the Centro Epson Meteo – Meteo Operations Italia s.r.l., MMI s.r.l., D.I.I.A.R. of Politecnico di Milano and by Consorzio Bonifica Muzza Bassa Lodigiana (MBL). Inspired by the recent publications of literature, in Chapter 1 comes before a general overview on the need to get better and better irrigation management and, subsequently, the objectives that the thesis suggests. Then follows a description of the study area with particular attention to the characteristics of the hydrological basin irrigation under review (Chapter 2). In the third and fourth respectively chapter presents a description of the irrigation system of the MBL Consortium in its current state and its modeling. In Chapter 5 were discussed the results obtained from simulations with the hydrological model EWB-FEST for the period between 2003 and 2010. Finally in Chapter 6 were presented the findings obtained as a result of the analysis carried out and possible future developments in this area

EAD3 standard to codify archival data: some news and many confirmations

Alla luce delle accresciute possibilità di offrire una descrizione archivistica più ricca e articolata e dell’ esperienza maturata negli anni, nell’estate 2015 la Society of American Archivists ha adottato come standard una nuova versione dell’Encoded Archival Description: EAD3. Il contributo, dopo aver ricostruito il processo decisionale e le modalità di revisione che hanno interessato la versione EAD 2002, analizza la nuova versione dello standard nei suoi aspetti maggiormente innovativi. Completa il contributo un’appendice relativa all’impiego di EAD3 all’inventario dell’archivio dell' Ex Ospedale psichiatrico Santa Maria della Pietà di Roma (1979-1999).In consideration of the increased possibilities to provide a richer and a more well-structured archival description and the experience matured in years, in Summer 2015 the Society of American Archivists has adopted the new version of Encoded Archival Description as a standard: EAD3. After retracing the decision process and the review related to the EAD 2002 version, the article examines the new version of the standard in its main original aspects. An appendix on the use of EAD3 in the inventory of Ex Ospedale psichiatrico Santa Maria della Pietà di Roma (1979-1999) archive, completes the essay

A genetic chronology for the Indian Subcontinent points to heavily sex-biased dispersals

Background India is a patchwork of tribal and non-tribal populations that speak many different languages from various language families. Indo-European, spoken across northern and central India, and also in Pakistan and Bangladesh, has been frequently connected to the so-called “Indo-Aryan invasions” from Central Asia ~3.5 ka and the establishment of the caste system, but the extent of immigration at this time remains extremely controversial. South India, on the other hand, is dominated by Dravidian languages. India displays a high level of endogamy due to its strict social boundaries, and high genetic drift as a result of long-term isolation which, together with a very complex history, makes the genetic study of Indian populations challenging. Results We have combined a detailed, high-resolution mitogenome analysis with summaries of autosomal data and Y-chromosome lineages to establish a settlement chronology for the Indian Subcontinent. Maternal lineages document the earliest settlement ~55–65 ka (thousand years ago), and major population shifts in the later Pleistocene that explain previous dating discrepancies and neutrality violation. Whilst current genome-wide analyses conflate all dispersals from Southwest and Central Asia, we were able to tease out from the mitogenome data distinct dispersal episodes dating from between the Last Glacial Maximum to the Bronze Age. Moreover, we found an extremely marked sex bias by comparing the different genetic systems. Conclusions Maternal lineages primarily reflect earlier, pre-Holocene processes, and paternal lineages predominantly episodes within the last 10 ka. In particular, genetic influx from Central Asia in the Bronze Age was strongly male-driven, consistent with the patriarchal, patrilocal and patrilineal social structure attributed to the inferred pastoralist early Indo-European society. This was part of a much wider process of Indo-European expansion, with an ultimate source in the Pontic-Caspian region, which carried closely related Y-chromosome lineages, a smaller fraction of autosomal genome-wide variation and an even smaller fraction of mitogenomes across a vast swathe of Eurasia between 5 and 3.5 ka

Structure of the accretion flow of IX Velorum as revealed by high-resolution spectroscopy

Context: Several high-mass transfer cataclysmic variables show evidence for outflow from the system, which could play an important role in their evolution. We investigate the system IX Vel, which was proposed to show similar characteristics. Aims: We study the structure of the IX Vel system, particularly the structure of its accretion flow and accretion disc. Methods: We use high-resolution time-resolved spectroscopy to construct radial velocity curves of the components in IX Vel, we compute Doppler maps of the system which we use to estimate the temperature distribution maps. Results: We improve the spectroscopic ephemeris of the system and its orbital period P_orb = 0.19392793(3) d. We construct Doppler maps of the system based on hydrogen and helium emission lines and the Bowen blend. The maps show features corresponding to the irradiated face of the secondary star, the outer rim of the accretion disc, and low-velocity components located outside the accretion disc and reaching towards L3. We constructed a temperature distribution map of the system using the Doppler maps of Balmer lines. Apart from the features found in the Doppler maps, the temperature distribution map shows a region of high temperature in the accretion disc connecting the expected position of a bright spot and the inner parts of the disc. Conclusions: We interpret the low-velocity emission found in the Doppler map as emission originating in the accretion disc wind and in an outflow region located in the vicinity of the third Lagrangian point L3. This makes IX Vel a member of the RW Sex class of Cataclysmic Variables

Artificial thymic organoids represent a reliable tool to study T-cell differentiation in patients with severe T-cell lymphopenia

The study of early T-cell development in humans is challenging because of limited availability of thymic samples and the limitations of in vitro T-cell differentiation assays. We used an artificial thymic organoid (ATO) platform generated by aggregating a DLL4-expressing stromal cell line (MS5-hDLL4) with CD34+ cells isolated from bone marrow or mobilized peripheral blood to study T-cell development from CD34+ cells of patients carrying hematopoietic intrinsic or thymic defects that cause T-cell lymphopenia. We found that AK2 deficiency is associated with decreased cell viability and an early block in T-cell development. We observed a similar defect in a patient carrying a null IL2RG mutation. In contrast, CD34+ cells from a patient carrying a missense IL2RG mutation reached full T-cell maturation, although cell numbers were significantly lower than in controls. CD34+ cells from patients carrying RAG mutations were able to differentiate to CD4+CD8+ cells, but not to CD3+TCRαβ+ cells. Finally, normal T-cell differentiation was observed in a patient with complete DiGeorge syndrome, consistent with the extra-hematopoietic nature of the defect. The ATO system may help determine whether T-cell deficiency reflects hematopoietic or thymic intrinsic abnormalities and define the exact stage at which T-cell differentiation is blocked

B-cell reconstitution after lentiviral vector-mediated gene therapy in patients with Wiskott-Aldrich syndrome

Background Wiskott-Aldrich syndrome (WAS) is a severe X-linked immunodeficiency characterized by microthrombocytopenia, eczema, recurrent infections, and susceptibility to autoimmunity and lymphomas. Hematopoietic stem cell transplantation is the treatment of choice; however, administration of WAS gene-corrected autologous hematopoietic stem cells has been demonstrated as a feasible alternative therapeutic approach. Objective Because B-cell homeostasis is perturbed in patients with WAS and restoration of immune competence is one of the main therapeutic goals, we have evaluated reconstitution of the B-cell compartment in 4 patients who received autologous hematopoietic stem cells transduced with lentiviral vector after a reduced-intensity conditioning regimen combined with anti-CD20 administration. Methods We evaluated B-cell counts, B-cell subset distribution, B cell-activating factor and immunoglobulin levels, and autoantibody production before and after gene therapy (GT). WAS gene transfer in B cells was assessed by measuring vector copy numbers and expression of Wiskott-Aldrich syndrome protein. Results After lentiviral vector-mediated GT, the number of transduced B cells progressively increased in the peripheral blood of all patients. Lentiviral vector-transduced progenitor cells were able to repopulate the B-cell compartment with a normal distribution of B-cell subsets both in bone marrow and the periphery, showing a WAS protein expression profile similar to that of healthy donors. In addition, after GT, we observed a normalized frequency of autoimmune-associated CD19+CD21-CD35- and CD21low

Ancient Migratory Events in the Middle East: New Clues from the Y-Chromosome Variation of Modern Iranians

Knowledge of high resolution Y-chromosome haplogroup diversification within Iran provides important geographic context regarding the spread and compartmentalization of male lineages in the Middle East and southwestern Asia. At present, the Iranian population is characterized by an extraordinary mix of different ethnic groups speaking a variety of Indo-Iranian, Semitic and Turkic languages. Despite these features, only few studies have investigated the multiethnic components of the Iranian gene pool. In this survey 938 Iranian male DNAs belonging to 15 ethnic groups from 14 Iranian provinces were analyzed for 84 Y-chromosome biallelic markers and 10 STRs. The results show an autochthonous but non-homogeneous ancient background mainly composed by J2a sub-clades with different external contributions. The phylogeography of the main haplogroups allowed identifying post-glacial and Neolithic expansions toward western Eurasia but also recent movements towards the Iranian region from western Eurasia (R1b-L23), Central Asia (Q-M25), Asia Minor (J2a-M92) and southern Mesopotamia (J1-Page08). In spite of the presence of important geographic barriers (Zagros and Alborz mountain ranges, and the Dasht-e Kavir and Dash-e Lut deserts) which may have limited gene flow, AMOVA analysis revealed that language, in addition to geography, has played an important role in shaping the nowadays Iranian gene pool. Overall, this study provides a portrait of the Y-chromosomal variation in Iran, useful for depicting a more comprehensive history of the peoples of this area as well as for reconstructing ancient migration routes. In addition, our results evidence the important role of the Iranian plateau as source and recipient of gene flow between culturally and genetically distinct population