Studio di fase I sulla reinfusione intraepatica di cellule staminali CD 133+ altamente purificate nei pazienti con malattia epatica terminale

Numerose evidenze sperimentali hanno dimostrato il contributo delle cellule staminali di derivazione midollare nei processi di rigenerazione epatica dopo danno tissutale. E’ cresciuto pertanto l’interesse sul loro potenziale impiego in pazienti con cirrosi. Questo studio si propone di valutare la fattibilità e la sicurezza della reinfusione intraepatica di cellule staminali midollari autologhe CD133+ in 12 pazienti con insufficienza epatica terminale definita da un punteggio di Model for End Stage of Liver Disease (MELD) compreso tra 17 e 25. L’efficacia in termini di funzionalità epatica rappresenta un obiettivo secondario. Previa mobilizzazione nel sangue periferico mediante somministrazione di granulocyte-colony stimulating factor (G-CSF) alla dose di 7,5 mcg/Kg/b.i.d. e raccolta per leucoaferesi, le cellule CD133+ altamente purificate vengono reinfuse in arteria epatica a partire da 5x104/Kg fino a 1x106/kg. Nei tre giorni successivi si somministra G-CSF per favorire l’espansione e l’attecchimento delle cellule. Durante la mobilizzazione, la reinfusione e nei 12 mesi successivi i pazienti sono sottoposti a periodici controlli clinici, laboratoristici e strumentali e ad attenta valutazione di effetti collaterali. Lo studio è tuttora in corso e ad oggi, 11 pazienti sono stati sottoposti a reinfusione e 4 hanno completato i 12 mesi di follow-up. Il G-CSF è stato ben tollerato e ha consentito di ottenere una buona espansione cellulare. Dopo la reinfusione sono stati documentati un ematoma inguinale e due episodi transitori di encefalopatia portosistemica. Durante il follow-up 4 pazienti sono stati trapiantati e 2 sono morti. Non è stata osservata alcuna modificazione significativa degli indici di funzione epatica. Questi risultati preliminari confermano la possibilità di mobilizzare e reinfondere un numero adeguato di cellule staminali di derivazione midollare in pazienti con malattia epatica in stadio terminale.

Valutazione del rischio ambientale associato alla presenza di metalli pesanti (Cd, Cu, Ni, Pb, Zn) nei sedimenti della Pialassa dei Piomboni.

I metalli pesanti presenti nei sedimenti marini possono essere rilasciati alla colonna d’acqua, in seguito a variazioni delle condizioni ambientali, ed entrare nelle catene trofiche degli organismi marini. A partire da una serie di campioni di sedimenti superficiali provenienti dalla Pialassa dei Piomboni (sito SIC-ZPS, a nord della Provincia di Ravenna), prelevati tra novembre e dicembre del 2012, è stato valutato il rischio potenziale associato alla presenza di metalli pesanti (Cd, Cu, Ni, Pb e Zn) per il biota residente nei sedimenti. È stato valutato il comportamento del rapporto SEM/AVS (contenuto di metalli simultaneamente estratti (SEM) e solfuri acidi volatili (AVS)) per ciascuno dei metalli oggetto dello studio. La metodica analitica ha previsto il trattamento dei campioni con HCl 6M, sotto flusso di azoto, e successivamente la lettura del contenuto di solfuro (raccolto in soluzioni di NaOH) e dei metalli pesanti allo spettrometro. Dal valore dei parametri chimico-fisici ottenuti nel momento del campionamento, è evidente che la zona interna della laguna risulta meno influenzata dalle maree rispetto a quella lungo il canale di navigazione. Le concentrazioni di metalli potenzialmente biodisponibili hanno evidenziato una distribuzione eterogenea. Del set di campioni analizzati, soltanto tre presentano un contenuto totale di metalli potenzialmente biodisponibili superiore al contenuto di solfuri labili (∑SEM/AVS>1), per cui la presenza di metalli bivalenti potrebbe rappresentare un rischio per il biota. Come suggerito da diversi autori, si è proceduto con l’ulteriore normalizzazione del rapporto (∑SEM/AVS) con il contenuto di carbonio organico relativo ad ogni campione di sedimento, escludendo possibili rischi associati alla presenza di metalli pesanti.

Spins, moments and radii of cd isotopes

The complex nature of the nucleon-nucleon interaction and the wide range of systems covered by the roughly 3000 known nuclides leads to a multitude of effects observed in nuclear structure. Among the most prominent ones is the occurence of shell closures at so-called ”magic numbers”, which are explained by the nuclear shell model. Although the shell model already is on duty for several decades, it is still constantly extended and improved. For this process of extension, fine adjustment and verification, it is important to have experimental data of nuclear properties, especially at crucial points like in the vicinity of shell closures. This is the motivation for the work performed in this thesis: the measurement and analysis of nuclear ground state properties of the isotopic chain of 100−130Cd by collinear laser spectroscopy.rnrnThe experiment was conducted at ISOLDE/CERN using the collinear laser spectroscopy apparatus COLLAPS. This experiment is the continuation of a run on neutral atomic cadmium from A = 106 to A = 126 and extends the measured isotopes to even more exotic species. The required gain in sensitivity is mainly achieved by using a radiofrequency cooler and buncher for background reduction and by using the strong 5s 2S1/2 → 5p 2P3/2 transition in singly ionized Cd. The latter requires a continuous wave laser system with a wavelength of 214.6 nm, which has been developed during this thesis. Fourth harmonic generation of an infrared titanium sapphire laser is achieved by two subsequent cavity-enhanced second harmonic generations, leading to the production of deep-UV laser light up to about 100 mW.rnrnThe acquired data of the Z = 48 Cd isotopes, having one proton pair less than the Z = 50 shell closure at tin, covers the isotopes from N = 52 up to N = 82 and therefore almost the complete region between the neutron shell closures N = 50 and N = 82. The isotope shifts and the hyperfine structures of these isotopes have been recorded and the magnetic dipole moments, the electric quadrupole moments, spins and changes in mean square charge radii are extracted. The obtained data reveal among other features an extremely linear behaviour of the quadrupole moments of the I = 11/2− isomeric states and a parabolic development in differences in mean square nuclear charge radii between ground and isomeric state. The development of charge radii between the shell closures is smooth, exposes a regular odd-even staggering and can be described and interpreted in the model of Zamick and Thalmi.

Fecal calprotectin correlates more closely with the Simple Endoscopic Score for Crohn's disease (SES-CD) than CRP, blood leukocytes, and the CDAI

Studies evaluating the correlation between the widely used Simple Endoscopic Score for Crohn's disease (SES-CD) and noninvasive markers are scarce. The aim of this study was to evaluate the correlation between the SES-CD and fecal calprotectin, C-reactive protein (CRP), blood leukocytes, and the Crohn's disease activity index (CDAI).

Variation in stress reactivity affects cage-induced stereotypies in female CD-1 (ICR) mice

Stereotypies in captive animals typically occur under conditions that are stressful for the animals, and there is some anecdotal evidence that stress levels during early stereotypy development predict later stereotypy levels. Based on this and on the involvement of stress in the behavioural sensitization to psychostimulant drugs, it has been hypothesized that stereotypy development might be causally related to stress. To address this question further, we used mice of the commercial outbred stock CD-1 (ICR) and mice of two lines derived from the outbred CD-1 (ICR) strain by selective breeding for high (HR) and low (LR) stress reactivity, respectively, and examined whether genetically driven variation in stress reactivity is associated with variation in the expression of cage-induced stereotypies. From 21 days of age, 10 females of each line were housed in pairs under standard laboratory conditions until they were video recorded for stereotypic behaviour and tested for corticosterone responses in a stress reactivity test (SRT) at 12 weeks of age. As expected, HR females showed a significantly stronger corticosterone response in the SRT than LR females, while ICR females were intermediate. Unexpectedly, however, both HR and LR females showed very low levels of stereotypic behaviour, while ICR females developed the high levels of stereotypies typical for this strain of mouse. Consequently, there was no significant relationship between measures of acute corticosterone reactivity and stereotypy performance, but a trend for reduced recovery of the corticosterone response in the ICR line suggests that variation in recovery rather than the acute response might predict stereotypy levels in these mice. (C) 2011 Elsevier B.V. All rights reserved.

Cage-induced stereotypies in female ICR CD-1 mice do not correlate with recurrent perseveration

Stereotypies are repetitive, unvarying, apparently purposeless behavioural patterns. They develop in animals kept in barren environments and are highly prevalent in laboratory mice (Mus musculus), yet their underlying mechanisms have remained elusive. In humans, stereotypies are associated with several psychiatric disorders and are thought to reflect dysfunction of inhibition of motor programs mediated by the corticostriatal circuitry, resulting in recurrent perseveration (=inappropriate repetition of behavioural responses). Several studies in captive animals of different species have reported a correlation between stereotypy performance and perseverative behaviour, indicating a similar dysfunction. To examine whether stereotypies in mice correlate with recurrent perseveration and whether they are causally related, we raised 40 female ICR CD-1 mice in either barren or enriched cages from three to either six or 16 weeks of age (2 x 2 factorial design) and assessed stereotypic behaviour in the home cage and recurrent perseveration on a two-choice guessing task. Enrichment significantly reduced stereotypic behaviour both at six and 16 weeks of age and recurrent perseveration increased with age. Although enriched housing reduced the number of repetitions in the guessing task significantly, there was no clear evidence for an effect on recurrent perseveration, and recurrent perseveration did not correlate positively with stereotypy level. These findings indicate either that this test did not measure recurrent perseveration or that cage stereotypies in these mice do not reflect behavioural disinhibition as measured by recurrent perseveration.

Enantioselective analysis of ketamine and its metabolites in equine plasma and urine by CE with multiple isomer sulfated beta-CD

CE with multiple isomer sulfated beta-CD as the chiral selector was assessed for the simultaneous analysis of the enantiomers of ketamine and metabolites in extracts of equine plasma and urine. Different lots of the commercial chiral selector provided significant changes in enantiomeric ketamine separability, a fact that can be related to the manufacturing variability. A mixture of two lots was found to provide high-resolution separations and interference-free detection of the enantiomers of ketamine, norketamine, dehydronorketamine, and an incompletely identified hydroxylated metabolite of norketamine in liquid/liquid extracts of the two body fluids. Ketamine, norketamine, and dehydronorketamine could be unambiguously identified via HPLC fractionation of urinary extracts and using LC-MS and LC-MS/MS with 1 mmu mass discrimination. The CE assay was used to characterize the stereoselectivity of the compounds' enantiomers in the samples of five ponies anesthetized with isoflurane in oxygen and treated with intravenous continuous infusion of racemic ketamine. The concentrations of the ketamine enantiomers in plasma are equal, whereas the urinary amount of R-ketamine is larger than that of S-ketamine. Plasma and urine contain higher S- than R-norketamine levels and the mean S-/R-enantiomer ratios of dehydronorketamine in plasma and urine are lower than unity and similar.

Molecular interaction between perthiolated [beta]-cyclodextrin (CD) and the guests molecules adamantaneacetic acid (AD) and ferroceneacetic acid (FC); and the effect of the interaction on the electron transition of CD anchored particles

The molecular interactions between the host molecule, perthiolated beta-cyclodextrin (CD), and the guest molecules, adamantaneacetic acid (AD) and ferroceneacetic acid (FC), have been inestigated theoretically in both the gas and aqueous phases. The major computations have been carried out at the theoretical levels, RHF/6-31G and B3LYP/6- 31G. MP2 electronic energies were also computed based at the geometries optimized by both the RHF and B3LYP methods in the gas phase to establish a better estimate of the correlation effect. The solvent phase computations were completed at the RHF/6-31G and B3LYP/6-31G levels using the PCM model. The most stable structures optimized in gas phase by both the RHF and B3LYP methods were used for the computations in solution. A method to systematically manipulate the relative position and orientation between the interacting molecules is proposed. In the gas phase, six trials with different host-guest relative positions and orientations were completed successfully with the B3LYP method for both the CD-AD and CD-FC complexes. Only four trials were completed with RHF method. In the gas phase, the best results from the RHF method gives for the association Gibbs free energy (ΔG°) values equal to -32.21kj/mol for CD-AD and -25.73kj/mol for CD-FC. And the best results from the B3LYP method have ΔG° equal to -47.57kj/mol for CD-AD and -41.09kj/mol for CD-FC. The MP2 correction significantly lowers ΔG° based on the geometries from both methods. For the RHF structure, the MP2 computations lowered ΔG° to -60.64kj/mol for CD-AD and -54.10 for CD-FC. For the structure from the B3LYP method, it was reduced to -59.87 kj/mol for CD-AD and -54.84 kj/mol for CDFC. The RHF solvent phase calculations yielded following results: ΔG°(aq) equals 107.2kj/mol for CD-AD and 111.4kj/mol for CD-FC. Compared with the results from the RHF method, the B3LYP method provided clearly better solvent phase results with ΔG° (aq) equal to 38.64kj/mol for CD-AD and 39.61kj/mol for CD-FC. These results qualitatively explain the experimental observations. However quantitatively they are in poor agreement with the experimental values available in the literature and those recently published by Liu et al. And the reason is believed to be omission of hydrophobic contribution to the association. Determining the global geometrical minima for these very large systems was very difficult and computationally time consuming, but after a very thorough search, these were identified. A relevant result of this search is that when the complexes, CD-AD and CD-FC, are formed, the AD and FC molecules are only partially embedded inside the CD cavity. The totally embedded complexes were found to have significantly higher energies. The semiempirical method, ZINDO, was employed to investigate the effect of complexation on the first electronic excitation of CD anchored to a metal nano-particle. The computational results revealed that after complexation to FC, the transition intensity declines to about 25% of the original value, and after complexation with AD, the intensity drops almost 50%. The tighter binding and transition intensity of CD-AD qualitatively agrees with the experimental result that the addition of AD to a solution of CD and FC restores the fluorescence of CD that was quenched by the addition of FC. A method to evaluate the “hydrophobic force” effect is proposed for future work.

The abundance and isotopic composition of Cd in iron meteorites

Cadmium is a highly volatile element and its abundance in meteorites may help better understand volatility-controlled processes in the solar nebula and on meteorite parent bodies. The large thermal neutron capture cross section of 113Cd suggests that Cd isotopes might be well suited to quantify neutron fluences in extraterrestrial materials. The aims of this study were (1) to evaluate the range and magnitude of Cd concentrations in magmatic iron meteorites, and (2) to assess the potential of Cd isotopes as a neutron dosimeter for iron meteorites. Our new Cd concentration data determined by isotope dilution demonstrate that Cd concentrations in iron meteorites are significantly lower than in some previous studies. In contrast to large systematic variations in the concentration of moderately volatile elements like Ga and Ge, there is neither systematic variation in Cd concentration amongst troilites, nor amongst metal phases of different iron meteorite groups. Instead, Cd is strongly depleted in all iron meteorite groups, implying that the parent bodies accreted well above the condensation temperature of Cd (i.e., ≈650 K) and thus incorporated only minimal amounts of highly volatile elements. No Cd isotope anomalies were found, whereas Pt and W isotope anomalies for the same iron meteorite samples indicate a significant fluence of epithermal and higher energetic neutrons. This observation demonstrates that owing to the high Fe concentrations in iron meteorites, neutron capture mainly occurs at epithermal and higher energies. The combined Cd-Pt-W isotope results from this study thus demonstrate that the relative magnitude of neutron capture-induced isotope anomalies is strongly affected by the chemical composition of the irradiated material. The resulting low fluence of thermal neutrons in iron meteorites and their very low Cd concentrations make Cd isotopes unsuitable as a neutron dosimeter for iron meteorites.