Successful Dendrimer and Liposome-Based Strategies to Solubilize an Antiproliferative Pyrazole Otherwise Not Clinically Applicable

Water-soluble formulations of the pyrazole derivative 3-(4-chlorophenyl)-5-(4-nitrophenylamino)-1H-pyrazole-4-carbonitrile (CR232), which were proven to have in vitro antiproliferative effects on different cancer cell lines, were prepared by two diverse nanotechnological approaches. Importantly, without using harmful organic solvents or additives potentially toxic to humans, CR232 was firstly entrapped in a biodegradable fifth-generation dendrimer containing lysine (G5K). CR232-G5K nanoparticles (CR232-G5K NPs) were obtained with high loading (DL%) and encapsulation efficiency (EE%), which showed a complex but quantitative release profile governed by Weibull kinetics. Secondly, starting from hydrogenated soy phosphatidylcholine and cholesterol, we prepared biocompatible CR232-loaded liposomes (CR232-SUVs), which displayed DL% and EE% values increasing with the increase in the lipids/CR232 ratio initially adopted and showed a constant prolonged release profile ruled by zero-order kinetics. When relevant, attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR) and nuclear magnetic resonance (NMR) spectroscopy, scanning electron microscopy (SEM) and dynamic light scattering (DLS) experiments, as well as potentiometric titrations completed the characterization of the prepared NPs. CR232-G5K NPs were 2311-fold more water-soluble than the pristine CR232, and the CR232-SUVs with the highest DL% were 1764-fold more soluble than the untreated CR232, thus establishing the success of both our strategies

Introduction of seismic source directivity on hazard map

The seismic hazard maps are mainly influenced by the uncertainty associated to the ground motion predictive equation (GMPE). This uncertainty represents the unexplained part of the ground motion and it is mostly related to the choice of the model’s variables. In fact the representation of the ground motion through the GMPEs is simple compared to the complexity of the physical process involved: if only the magnitude and distance are taken into account, GMPEs predicts isoseismals curves that are expected to be isotropic around the hypocenter or along the fault. Instead, the presence of a fault plane across which a process of failure in shear develops makes this general formulation reliable only on average. In fact this failure is responsible of an asymmetry in the seismic radiation known, since Ben-Menhaem (PhD1961), as directivity effect. While the general knowledge of the earthquakes is treated explicitly in the empirical prediction, specific trends like the directivity effects are hidden in the uncertainty sigma. A way to reduce the sigma is therefore to refine the seismic seismic source description inside the GMPEs (e.g. NGA project, Power et al, Earthquake Spectra, 2008). In this framework we propose a strategy to introduce the directivity in the GMPEs and to study its effect on uncertainties and on hazard maps. For this purpose, we have used two different directivity models acting on the GMPE as corrective factors: one proposed by Somerville et al. (Seis.Res.Lett.1997) and the other one proposed by Spudich and Chiou (Earthquake Spectra 2008).The first factor depends on geometrical parameters and comes from theoretical deduction. The second one includes many source parameters and it is a hybrid factor, which functional formulation is deduced from the theory, calibrated on synthetic simulations and scaled on data. The classic hazard equation is then adapted in order to increase the number of source parameters (i.e. adding one integral over the parametric space for each new variable involved) and taking into account the corrective factors for directivity (Spagnuolo, PhD2010). We present the comparisons of hazard maps depending on the directivity factor and on the probability density functions of the fault strike and of the rupture “laterality”

Detecting long-lasting transients of earthquake activity on a fault system by monitoring apparent stress, ground motion and clustering

Damaging earthquakes result from the evolution of stress in the brittle upper-crust, but the understanding of the mechanics of faulting cannot be achieved by only studying the large ones, which are rare. Considering a fault as a complex system, microearthquakes allow to set a benchmark in the system evolution. Here, we investigate the possibility to detect when a fault system starts deviating from a predefined benchmark behavior by monitoring the temporal and spatial variability of different micro-and-small magnitude earthquakes properties. We follow the temporal evolution of the apparent stress and of the event-specific residuals of ground shaking. Temporal and spatial clustering properties of microearthquakes are monitored as well. We focus on a fault system located in Southern Italy, where the Mw 6.9 Irpinia earthquake occurred in 1980. Following the temporal evolution of earthquakes parameters and their time-space distribution, we can identify two long-lasting phases in the seismicity patterns that are likely related to high pressure fluids in the shallow crust, which were otherwise impossible to decipher. Monitoring temporal and spatial variability of micro-to-small earthquakes source parameters at near fault observatories can have high potential as tool for providing us with new understanding of how the machine generating large earthquakes works

A microseismic study in a low seismicityarea: the 2001 site-response experimentin the Città di Castello Basin (Italy)

A site response experiment was performed in the basin of Città di Castello (a small town in Central Italy) in May 2001. This study is part of a project on the evaluation of seismic hazard in seismogenic areas funded by the Gruppo Nazionale Difesa dai Terremoti (GNDT). The experiment consisted of a dense fixed transect configuration with most of the stations recording in continuous mode, and several ambient noise measurements both in single station and in array configuration spread over the investigated area. The dense transect was composed of 26 seismic stations in a crosswise configuration with a maximum inter-station distance of 250 m. The stations were deployed in the southern part of the basin, from the eastern bedrock outcrop to the western edge, across the town. About 70 earthquakes were recorded during 10 days of deployment, generally low magnitude or regional events. We located 23 earthquakes and 17 of them were located using the waveform similarity approach at 4 stations outside the target area. These 4 stations were part of a dense temporary seismic network involved in a previous experiment of the same project, aimed at performing a high-resolution picture of the local seismicity. Delay analysis on the recorded waveforms allowed us to infer the basin geometry at depth and estimate the S-wave velocity of sediments. Moreover, we evaluated relative site response along the E-W transect by performing a standard spectral ratio. Amplification factors up to 9 are found inside the basin; at frequencies above 5 Hz stations closer to the edges show higher amplification, whereas stations located in the middle of the basin, where the alluvial sediments are thicker (CD11-CD14), show higher amplification below 5 Hz. We considered the average amplification in two frequency bands (1-5 Hz and 5-10 Hz), representative of the resonance frequency for 2-3 storey buildings and 1 storey houses,respectively. Our results suggest that the potential hazard for 2-3 storey buildings is higher in the center of the basin (amplification factor up to 6), and for 1 storey houses is higher at the edges (amplification factor up to 5)

Solvent content of protein crystals from diffraction intensities by Independent Component Analysis

An analysis of the protein content of several crystal forms of proteins has been performed. We apply a new numerical technique, the Independent Component Analysis (ICA), to determine the volume fraction of the asymmetric unit occupied by the protein. This technique requires only the crystallographic data of structure factors as input.Comment: 9 pages, 2 figures, 1 tabl

SseA, a 3-mercaptopyruvate sulfurtransferase from escherichia coli : crystallization and preliminary crystallographic data

SseA, the translation product of the Escherichia coli sseA gene, is a 31 kDa protein endowed with 3-mercaptopyruvate:cyanide sulfurtransferase activity in vitro. As such, SseA is the prototype of a sulfurtransferase subfamily distinguished from the better known rhodanese sulfurtransferases, which display thiosulfate:cyanide sulfurtransferase activity. The physiological role of the two homologous enzyme families, whose catalytic activity is centred on a reactive invariant cysteine, is a matter of debate. In this framework, the forthcoming crystal structure analysis of SseA will be based on the tetragonal crystal form (space group P4(1) or P4(3)) reported here, with unit-cell parameters a = b = 150.2, c = 37.9 Angstrom. The in vivo role and substrate specificity of sulfurtransferase enzymes has been greatly debated. SseA, a 3-mecaptopyruvate: cyanide sulfurtransferase from E. coli, has been crystallized in a tetragonal crystal form suitable for X-ray crystallographic investigations

p38 MAPK and JNK Antagonistically Control Senescence and Cytoplasmic p16INK4A Expression in Doxorubicin-Treated Endothelial Progenitor Cells

Patients treated with low-dose anthracyclines often show late onset cardiotoxicity. Recent studies suggest that this form of cardiotoxicity is the result of a progenitor cell disease. In this study we demonstrate that Cord Blood Endothelial Progenitor Cells (EPCs) exposed to low, sub-apoptotic doses of doxorubicin show a senescence phenotype characterized by increased SA-b-gal activity, decreased TRF2 and chromosomal abnormalities, enlarged cell shape, and disarrangement of F-actin stress fibers accompanied by impaired migratory ability. P16 INK4A localizes in the cytoplasm of doxorubicin-induced senescent EPCs and not in the nucleus as is the case in EPCs rendered senescent by different stimuli. This localization together with the presence of an arrest in G2, and not at the G1 phase boundary, which is what usually occurs in response to the cell cycle regulatory activity of p16INK4A, suggests that doxorubicin-induced p16 INK4A does not regulate the cell cycle, even though its increase is closely associated with senescence. The effects of doxorubicin are the result of the activation of MAPKs p38 and JNK which act antagonistically. JNK attenuates the senescence, p16 INK4A expression and cytoskeleton remodeling that are induced by activated p38. We also found that conditioned medium from doxorubicin-induced senescent cardiomyocytes does not attract untreated EPCs, unlike conditioned medium from apoptotic cardiomyocytes which has a strong chemoattractant capacity. In conclusion, this study provides a better understanding of the senescence of doxorubicin-treated EPCs, which may be helpful in preventing and treating late onset cardiotoxicity

Diversity of Cardiologic Issues in a Contemporary Cohort of Women With Breast Cancer

Background: Women with breast cancer (BC) represent a special population particularly exposed to cardiovascular disease (CVD) risk. However, cardiologic assessment in BC is mostly limited to detection of left ventricular dysfunction cardiotoxicity (LVD-CTX) due to anticancer treatments. Our aim was to comprehensively investigate CV profile and events in a contemporary BC cohort. Methods and Results: Records of BC patients referred for a Cardio-Oncologic evaluation before starting anticancer treatments, between 2016 and 2019, were retrospectively reviewed (n = 508). Information regarding prevalence and control of CV risk factors, and novel CVD diagnoses were extracted. Occurrence of LVD-CTX, CV events other than LVD-CTX and mortality was assessed. Mean age of study population was 64 ± 13 years; 287 patients were scheduled to receive anthracycline and 165 anti-HER2 therapy. Overall, 53% of BC women had ≥2 CV risk factors, and 67% had at least one of arterial hypertension, dyslipidaemia or diabetes mellitus not adequately controlled. Eighteen (4%) patients were diagnosed a previously unknown CVD. Over a mean follow-up of 2.5 ± 1 years, 3% of BC patients developed LVD-CTX, 2% suffered from other CV events and 11% died. CV risk factors were not associated with LVD-CTX, except for family history of CAD. On the contrary, patients with other CV events exhibited a worse CV profile. Those who died more commonly experienced CV events other than LVD-CTX (p = 0.02). Conclusions: BC women show a suboptimal CV risk profile and are at risk of CV events not limited to LVD-CTX. A baseline Cardio-Oncologic evaluation was instrumental to implement CV prevention and to optimize CV therapies

A microseismic study in a low seismicity area of Italy: the Città di Castello 2000-2001 experiment

Recent seismological studies contribute to better understand the first order characteristics of earthquake occurrence in Italy, identifying the potential sites for moderate to large size earthquakes. Ad hoc passive seismic experiments performed in these areas provide information to focus on the location and geometry of the active faults more closely. This information is relevant for assessing seismic hazard and for accurately constraining possible ground shaking scenarios. The area around the Città di Castello Basin, in the Northern Apennines (Central Italy), is characterized by the absence of instrumental seismicity (M > 2.5), it is adjacent to faults ruptured by recent and historical earthquakes. To better understand the tectonics of the area, we installed a dense network of seismic stations equipped with broadband and short period seismometers collecting data continuously for 8 months (October 2000-May 2001). The processing of ~ 900 Gbyte of data revealed a consistent background seismicity consisting of very low magnitude earthquakes (ML < 3.2). Preliminary locations of about 2200 local earthquakes show that the area can be divided into two regions with different seismic behaviour: an area to the NW, in between Sansepolcro and Città di Castello, where seismicity is not present. An area toward the SE, in between Città di Castello, Umbertide and Gubbio, where we detected a high microseismicity activity. These findings suggest a probable different mechanical behaviour of the two regions. In the latter area, the seismicity is confined between 0 and 8 km of depth revealing a rather well defined east-dipping, low angle fault 35 km wide that cuts through the entire upper crust down to 12-15 km depth. Beside an apparent structural complexity, fault plane solutions of background seismicity reveal a homogeneous pattern of deformation with a clear NE-SW extension

Il terremoto del 21 giugno 2013 in Lunigiana. Le attività del coordinamento Sismiko

Il 21 giugno 2013 alle ore 10.33 UTC è stato registrato dalla Rete Sismica Nazionale (RSN) [Amato e Mele, 2008; Delladio, 2011] dell’Istituto Nazionale di Geofisica e Vulcanologia (INGV) un terremoto di magnitudo (ML) 5.2 nel distretto sismico1 denominato “Alpi Apuane” tra i comuni di Minucciano in provincia di Lucca e Fivizzano e Casola in provincia di Massa e Carrara, zona conosciuta come “Lunigiana”. L’evento sismico, localizzato dai sismologi in turno presso la sala di sorveglianza sismica di Roma [Basili, 2011] con coordinate 44.153°N e 10.135° E e una profondità di circa 5 km è stato ben risentito in tutta la penisola centro-settentrionale ed è stato seguito in poche ore da numerosi eventi anche di ML ≥ 3.0 (16 nelle prime 72 ore). Storicamente l’area oggetto della sequenza sismica è stata interessata da numerosi terremoti di magnitudo superiore a 5.0 il più grande dei quali quello avvenuto nel 1920 nella zona della Garfagnana (fonte dati: Catalogo Parametrico dei Terremoti Italiani - CPTI11 [Rovida et al., 2011]), ad una distanza di circa 12 km dal mainshock odierno, interessata anch’essa da una piccola sequenza sismica a gennaio del 2013. In considerazione dell’entità dell’evento e seguendo le procedure definite per le situazioni di emergenza internamente all’INGV anche in accordo con l’Allegato A2 della Convenzione vigente 2012- 20203 fra l’ente e il Dipartimento di Protezione Civile (DPC), è stata attivata la Rete Sismica Mobile della sede INGV di Roma (Re.Mo. [Moretti et al., 2010]). Nell’arco di tempo di poco più un’ora dall’accadimento del mainshock è stata disposta l’installazione di una rete sismica temporanea costituita da sei stazioni a integrazione delle reti sismiche permanenti già presenti in area epicentrale (RSN e Regional Seismic network of North-Western Italy – RSNI [Ferretti et al., 2008; 2010; Eva et al., 2010; Pasta et al., 2011]). Nel contempo sono stati consultati tramite e-mail i referenti delle unità di rete sismica mobile delle altre sedi INGV che nell’ambito del coordinamento “Sismiko” [Moretti et al., 2012] negli ultimi due anni hanno dato la propria disponibilità, in termini di personale e strumentazione, ad intervenire in caso di emergenza sismica; sono stati inoltre contattati i colleghi del Dipartimento di Scienze della Terra, dell’Ambiente e della Vita, dell’Università degli Studi di Genova (DISTAV) i più vicini all’area epicentrale e gestori della RSNI che hanno comunicato loro stessi l’intenzione di installare due stazioni temporanee, una in real-time e una in configurazione stand-alone. In questo lavoro viene descritta l’attività compiuta dalla Rete Sismica Mobile INGV, la tempistica dell’intervento effettuato in sinergia con i colleghi dell’Università di Genova, i dettagli circa l'installazione e la gestione delle stazioni sismiche temporanee nel primo mese di attività e una valutazione del dataset acquisito