Spray Congealing of Pharmaceuticals: Study on Production of Solid Dispersions Using Box-Behnken Design

This work aimed at evaluating the spray congealing method for the production of microparticles of carbamazepine combined with a polyoxylglyceride carrier. In addition, the influence of the spray congealing conditions on the improvement of drug solubility was investigated using a three-factor, three-level Box-Behnken design. The factors studied were the cooling air flow rate, atomizing pressure, and molten dispersion feed rate. Dependent variables were the yield, solubility, encapsulation efficiency, particle size, water activity, and flow properties. Statistical analysis showed that only the yield was affected by the factors studied. The characteristics of the microparticles were evaluated using X-ray powder diffraction, scanning electron microscopy, differential scanning calorimetry, and hot-stage microscopy. The results showed a spherical morphology and changes in the crystalline state of the drug. The microparticles were obtained with good yields and encapsulation efficiencies, which ranged from 50 to 80% and 99.5 to 112%, respectively. The average size of the microparticles ranged from 17.7 to 39.4 mu m, the water activities were always below 0.5, and flowability was good to moderate. Both the solubility and dissolution rate of carbamazepine from the spray congealed microparticles were remarkably improved. The carbamazepine solubility showed a threefold increase and dissolution profile showed a twofold increase after 60 min compared to the raw drug. The Box-Behnken fractional factorial design proved to be a powerful tool to identify the best conditions for the manufacture of solid dispersion microparticles by spray congealing.

A utilização da tomografia computadorizada de feixe cônico na reabilitação das fissuras palatinas

A Tomografia Computadorizada de Feixe Cônico vem revolucionando o diagnóstico por imagem na Odontologia. As características permitem um acesso de visualização das imagens que facilita muitos procedimentos. Sua aplicabilidade na Odontologia é diversa, podendo ser útil para as diversas especialidades, porem há controversas na literatura a cerca da correta prescrição desse exame com a justificativa pertinente. Os pacientes com anomalias craniofaciais apresentam algumas particularidades que tornam o seu processo de reabilitação mais complexo e extenso. A utilização da TCFC na reabilitação desses pacientes é importante para diversas especialidades dentre elas a ortodontia, implantodontia, cirurgia ortognática e também otorrinolaringologia. Entretanto devemos considerar que desde sempre os protocolos imaginológicos envolviam e eram estabelecidos com as radiografias convencionais (panorâmica, telerradiografia, periapical e oclusal), os benefícios das imagens na terceira dimensão são bem conhecidos, porem há sempre um aumento da dose de radiação, visto que são exames que abrangem muitas vezes áreas maiores e trabalham com parâmetros de exposição mais elevados. O principio do ALARA deve sempre ser utilizado quando se trata de prescrição de exames que utilizam radiação ionizante, portanto a TCFC deve ser indicada quando houverem benefícios com riscos minimizados. Nessa palestra serão abordadas as vantagens e desvantagens do uso da tomografia computadorizada de feixe cônico frente aos exames radiográficos convencionais no processo reabilitador e os protocolos de indicação baseados na literatura pautados na responsabilidade para o uso de tal tecnologia.

An experimental and theoretical approach to correct for the scattered radiation in an X-ray computer tomography system for industrial applications

The main problem connected to cone beam computed tomography (CT) systems for industrial applications employing 450 kV X-ray tubes is the high amount of scattered radiation which is added to the primary radiation (signal). This stray radiation leads to a significant degradation of the image quality. A better understanding of the scattering and methods to reduce its effects are therefore necessary to improve the image quality. Several studies have been carried out in the medical field at lower energies, whereas studies in industrial CT, especially for energies up to 450 kV, are lacking. Moreover, the studies reported in literature do not consider the scattered radiation generated by the CT system structure and the walls of the X-ray room (environmental scatter). In order to investigate the scattering on CT projections a GEANT4-based Monte Carlo (MC) model was developed. The model, which has been validated against experimental data, has enabled the calculation of the scattering including the environmental scatter, the optimization of an anti-scatter grid suitable for the CT system, and the optimization of the hardware components of the CT system. The investigation of multiple scattering in the CT projections showed that its contribution is 2.3 times the one of primary radiation for certain objects. The results of the environmental scatter showed that it is the major component of the scattering for aluminum box objects of front size 70 x 70 mm2 and that it strongly depends on the thickness of the object and therefore on the projection. For that reason, its correction is one of the key factors for achieving high quality images. The anti-scatter grid optimized by means of the developed MC model was found to reduce the scatter-toprimary ratio in the reconstructed images by 20 %. The object and environmental scatter calculated by means of the simulation were used to improve the scatter correction algorithm which could be patented by Empa. The results showed that the cupping effect in the corrected image is strongly reduced. The developed CT simulation is a powerful tool to optimize the design of the CT system and to evaluate the contribution of the scattered radiation to the image. Besides, it has offered a basis for a new scatter correction approach by which it has been possible to achieve images with the same spatial resolution as state-of-the-art well collimated fan-beam CT with a gain in the reconstruction time of a factor 10. This result has a high economic impact in non-destructive testing and evaluation, and reverse engineering.

Studio delle interazioni dei neutroni nell'esperimento xenon1t

La presenza di materia oscura nell'universo venne ipotizzata negli anni '30 a seguito di alcune anomalie nei risultati sperimentali ottenuti in astrofisica e cosmologia. La distribuzione di materia ottenuta non concordava infatti con i dati provenienti dalle osservazioni astronomiche e gli scienziati ipotizzarono l'esistenza di un tipo di materia, denominata appunto materia oscura, che interagisse debolmente con la radiazione elettromagnetica e fosse quindi interamente invisibile ai telescopi, sia che fossero a radiofrequenze che operanti nel campo del visibile o con raggi gamma e X, ma che producesse effetti gravitazionali. Nel corso degli anni si sono aggiunte ulteriori evidenze a sostegno dell'esistenza di materia oscura, provenienti anche dallo studio della cosmologia, e numerosi esperimenti (tra cui XENON, IGEX, DAMA/LIBRA) sono stati condotti per cercare di determinare tipo e massa delle particelle o la loro abbondanza (PLANCK). Il lavoro di questa tesi consiste in uno studio delle interazioni dei neutroni con lo xenon per l'esperimento XENON1T. I neutroni costituiscono un fondo particolarmente pericoloso per l'esperimento, in quanto producono uno scattering direttamente sul nucleo allo stesso modo delle particelle di materia oscura. Nel lavoro svolto sono state dapprima analizzate le caratteristiche delle singole interazioni con lo xenon contenuto nella camera, per poi passare ad uno studio più specifico sul comportamento dei neutroni prodotti dai fotomoltiplicatori del rivelatore. In seguito alle analisi svolte è stato possibile caratterizzare il fondo di neutroni in modo più preciso, permettendo di determinare alcuni criteri di selezione per il loro riconoscimento.

Analisi dello spettro di massa invariante j/psi pi+ pi- in collisioni p-p a sqrt(s) = 8 tev con il rivelatore cms

In questo lavoro di tesi è stato studiato lo spettro di massa invariante del sistema J/psi pi+ pi-, m(J/psi pi+ pi-), in collisioni protone-protone a LHC, con energia nel centro di massa sqrt(s)) pari a 8 TeV, alla ricerca di nuovi stati adronici. Lo studio è stato effettuato su un campione di dati raccolti da CMS in tutto il 2012, corrispondente ad una luminosità integrata di 18.6 fb-1. Lo spettro di massa invariante m(J/psi pi+ pi-), è stato ricostruito selezionando gli eventi J/psi->mu+ mu- associati a due tracce cariche di segno opposto, assunte essere pioni, provenienti da uno stesso vertice di interazione. Nonostante l'alta statistica a disposizione e l'ampia regione di massa invariante tra 3.6 e 6.0 GeV/c^2 osservata, sono state individuate solo risonanze già note: la risonanza psi(2S) del charmonio, lo stato X(3872) ed una struttura più complessa nella regione attorno a 5 GeV/c^2, che è caratteristica della massa dei mesoni contenenti il quark beauty (mesoni B). Al fine di identificare la natura di tale struttura, è stato necessario ottenere un campione di eventi arricchito in adroni B. È stata effettuata una selezione basata sull'elevata lunghezza di decadimento, che riflette la caratteristica degli adroni B di avere una vita media relativamente lunga (ordine dei picosecondi) rispetto ad altri adroni. Dal campione così ripulito, è stato possibile distinguere tre sottostrutture nello spettro di massa invariante in esame: una a 5.36 GeV/c^2, identificata come i decadimenti B^0_s-> J/psi pi+ pi-, un'altra a 5.28 GeV/c^2 come i candidati B^0-> J/psi pi+ pi- e un'ultima allargata tra 5.1 e 5.2 GeV/c^2 data da effetti di riflessione degli scambi tra pioni e kaoni. Quest'ultima struttura è stata identificata come totalmente costituita di una combinazione di eventi B^0-> J/psi K+ pi- e B^0_s-> J/psi K+ K-.

Tecniche di analisi digitale dei segnali prodotti da rivelatori per neutroni

La disintegrazione dei nuclei atomici si traduce in una emissione di vari tipi di radiazioni e particelle tra cui neutroni e raggi gamma. La rivelazione dei neutroni comporta l’utilizzo di rivelatori a scintillazione e tecniche di analisi per poter identificare e ottenere informazioni sull’energia dei neutroni. Il processo di scintillazione per la rivelazione dei neutroni consiste nell’interazione con i nuclei del materiale e successiva emissione luminosa dovuta a ionizzazione degli atomi del rivelatore. La luce e in seguito convertita in impulsi elettrici, i quali possono essere analizzati con opportune tecniche. L’emissione di neutroni `e accompagnata da emissione di raggi gamma e quindi `e necessario identificare i neutroni. Rivelatori basati su scintillatori organici vengono spesso impiegati nella spettrometria neutronica ad energie superiori di 0.5 MeV ed in una vasta gamma di applicazioni come la medicina, l’industria e la radioprotezione. La rilevazione dei neutroni `e molto importante nello studio delle reazioni nucleari di bassa energia e nello studio della materia nucleare lontano dalla valle di stabilita. In questo lavoro abbiamo studiato tre algoritmi: Zero Crossing, Charge Comparison e Pulse Gradient Analysis. Questi algoritmi sono stati in seguito applicati all’analisi di un insieme di dati provenienti dalla reazione nucleare 7Li(p,n)7Be. E stato utilizzato uno scintillatore organico liquido BC501. Si `e effettuato un confronto tra le varie tecniche utilizzate per determinare il grado di discriminazione ottenuto con ognuna di esse. I risultati ottenuti permettono di decidere in seguito quale algoritmo si presta ad essere utilizzato anche in altri esperimenti futuri. Il metodo Pulse Gradient Analysis `e risultato il piu` prometente, essendo anche possibile l’utilizzo on-line.

Identification of target genes of the T-box proteins in Drosophila melanogaster

In a prior bioinformatic analysis by Hüyseyin Binbas, potential Tbx targets sequences in wing-related genes have been identified. Guided by this information, enhancer trap/reporter lacZ insertions were characterized by X-gal staining first in wildtype and then in l(1)omb imaginal discs.rnIn several lines I observed an increase in reporter expression in a l(1)omb mutant background. Since Omb is assumed to function predominantly as a transcriptional repressor, this may indicate direct regulation. Repression by Omb was observed e.g. for brk and tkv. These genes are negatively regulated by Dpp, while omb is induced by Dpp. Omb which mediates the effects of Dpp on proliferation could, thus, also mediate the Dpp effect on patterning of the wing disc. However, brk and tkv were not completely derepressed in l(1)omb indicating that Dpp represses these genes also by an Omb-independent mechanism.rnMore frequently I observed loss of reporter expression in an l(1)omb mutant background. In these cases, regulation by Omb presumably is indirect. For example, STAT92E-lacZ expression in the wildtype eye was symmetrically expressed at the dorsal and ventral margins. In l(1)omb, ventral expression was selectively lost. Loss of omb is known to cause ventral overproliferation of the eye by activation of the Jak/STAT pathway. STAT92E expression is negatively regulated by Jak/STAT signaling suggesting that loss of omb activates Jak/STAT further upstream in the pathway.rnRegional overproliferation of eye and wing in the l(1)omb mutant background proved a complicating issue in the search for Omb targets. This effect made it difficult to decide whether an expanded reporter expression pattern was due to tissue expansion or reporter gene derepression. For instance hth-lacZ appeared to expand along the ventral eye disc margin in l(1)omb. Without addtional experiments it cannot be concluded whether this is due to de-repression or to activation in association with the proliferative state. Parallel to my experiments, evidence accumulated in our laboratory that loss of omb may attenuate Wg and Hegehog signaling. Since these diffusible proteins are the main patterning molecules in the wing imaginal disc, with dpp being downstream of Hh, many of the observed effects could be secondary to reduced Wg and Hh activity. Examples are ab-lacZ, Dll-lacZ and vgBE-lacZ (reduced expression on the dorso-ventral boundary) and inv-lacZ (late larval expression in the anterior wing disc compartment is lost) or sal-lacZ. Epistasis experiment will be required to clarifiy these issues.rnFurthermore, loss of omb appeared to induce cell fate changes. It was reported previously that in an omb null mutant, the dorsal determinant apterous (ap) is ectopically expressed in the ventral compartment (an effect I did not observe with the strongly hypomorphic l(1)omb15, indicating strong dose dependence). Ventral repression of ap is maintained by epigenetic mechanisms. The patchy and variable nature of ectopic expression of ap or grn-1.1-lacZ points to an effect of omb on epigenetic stability.rnIn the second part of my thesis, an analysis of Omb expression in the Drosophila embryonic ventral nervous system was performed. Omb was found co-expressed with Eve in the medial aCC and RP2 motorneurons as well as the fpCC interneuron and the mediolateral CQ neurons. Additionally, Omb was detected in the Eg positive NB7-3 GW serotonergic motoneuron and the N2-4 neurons. Omb was not found in Repo positive glial cells. During embryonic stage 14, Omb showed some coepression with Dpn or Pros. At the embryonic stage 16, Omb was expressed in minor subset of Mid and Wg positive cells.

Sviluppo di un sistema di rivelazione per l'esperimento FAMU (fisica degli atomi muonici) basato su scintillatore LaBr3 con fotomoltiplicatore UQE e sistema spettrometrico DSP

In questa tesi si sono studiati e sviluppati i rivelatori di fotoni x per l’esperimento FAMU (Fisica degli Atomi Muonici gruppo 3 dell’INFN) che indaga sulla natura e la struttura del protone. I rivelatori sono stati assemblati e testati con elementi che rappresentano lo stato dell’arte nel campo dei rivelatori di radiazione (scintillatori LaBr3(Ce) e fotomoltiplicatori Hamamatsu™ Ultra Bi-Alcali). È stata anche studiata e sviluppata parte della catena di formatura del segnale. Questa è stata implementata su un chip FPGA dell’ALTERA™ con buoni risultati per quanto riguarda la qualità del filtro; le performance dell’FPGA non hanno consentito di raggiungere la velocità di 500MHz richiesta dall’esperimento costringendo l’implementazione ad una velocità massima di 320MHz.

X-ray phase contrast imaging of metallized and non-metallized biological samples

Imaging of biological samples has been performed with a variety of techniques for example electromagnetic waves, electrons, neutrons, ultrasound and X-rays. Also conventional X-ray imaging represents the basis of medical diagnostic imaging, it remains of limited use in this application because it is based solely on the differential absorption of X-rays by tissues. Coherent and bright photon beams, such as those produced by third-generation synchrotron X-ray sources, provide further information on subtle X-ray phase changes at matter interfaces. This complements conventional X-ray absorption by edge enhancement phenomena. Thus, phase contrast imaging has the potential to improve the detection of structures on images by detecting those structures that are invisible with X-ray absorption imaging. Images of a weakly absorbing nylon fibre were recorded in in-line holography geometry using a high resolution low-noise CCD camera at the ESRF in Grenoble. The method was also applied to improve image contrast for images of biological tissues. This paper presents phase contrast microradiographs of vascular tree casts and images of a housefly. These reveal very fine structures, that remain invisible with conventional absorption contrast only.

Regulation of XMyoDa transcription by the binding of XMEF2A to the TATA box

MEF2 is a $\underline{\rm m}$yocyte-specific $\underline{\rm e}$nhancer-binding $\underline{\rm f}$actor that binds a conserved DNA sequence, CTA(A/T)$\sb4$TAG. A MEF2 binding site in the XMyoDa promoter overlaps with the TATA box and is required for muscle specific expression. To examine the potential role of MEF2 in the regulation of MyoD transcription during early development, the appearance of MEF2 binding activity in developing Xenopus embryos was analyzed with the electrophoretic mobility shift assay. Two genes were isolated from a X. Laevis stage 24 cDNA library that encode factors that bind the XMyoDa TFIID/MEF2 site. Both genes are highly homologous to each other, belong to the MADS ($\underline{\rm M}$CM1-$\underline{\rm A}$rg80-agamous-$\underline{\rm d}$eficiens-$\underline{\rm S}$RF) protein family, and most highly related to the mammalian MEF2A gene, hence they are designated as XMEF2A1 and XMEF2A2. Proteins encoded by both cDNAs form specific complexes with the MEF2 binding site and show the same binding specificity as the endogenous MEF2 binding activity. XMEF2A transcripts accumulate preferentially in developing somites after the appearance of XMyoD transcripts. XMEF2 protein begins to accumulate in somites at tailbud stages. Transcriptional activation of XMyoD promoter by XMEF2A required only the MADS box and MEF2-specific domain when XMEF2A is bound at the TATA box. However, a different downstream transactivation domain was required when XMEF2A activates transcription through binding to multiple upstream sites. These results suggest that different activation mechanisms are involved, depending on where the factor is bound. Mutations in several basic amino acid clusters in the MADS box inhibit DNA binding suggesting these amino acids are essential for DNA binding. Mutation of Thr-20 and Ser-36 to the negatively charged amino acid residue, aspartic acid, abolish DNA binding. XMEF2A activity may be regulated by phosphorylation of these amino acids. A dominant negative mutant was made by mutating one of the basic amino acid clusters and deleting the downstream transactivation domain. In vivo roles of MEF2 in the regulation of MyoD transcription were investigated by overexpression of wild type MEF2 and dominant negative mutant of XMEF2A in animal caps and assaying for the effects on the level of expression of MyoD genes. Overexpression of MEF2 activates the transcription of endogenous MyoD gene family while expression of a dominant negative mutant reduces the level of transcription of XMRF4 and myogenin genes. These results suggest that MEF2 is downstream of MyoD and Myf5 and that MEF2 is involved in maintaining and amplifying expression of MyoD and Myf5. MEF2 is upstream of MRF4 and myogenin and plays a role in activating their expression. ^

Geochemistry and physics on sediment cores from the Peru Basin

We studied preservation/dissolution cycles and paleoproductivity in eight sediment cores from the Peru Basin south of the highly productive surface waters of the eastern equatorial Pacific. Stratigraphy is based on stable oxygen isotopes and on combined magnetostratigraphy and biostratigraphy. Sediment cores which span the last 8 m.y., were retrieved during cruise 79 with RV SONNE close to the carbonate compensation depth (CCD). In general, sediments show Pacific-type carbonate cycles. We interpret a pronounced carbonate peak between 6 and 7 Ma as the result of a western and northern extension of the highly productive Peru Current. Decreased carbonate contents from the late Miocene to the late Pliocene might be associated with a slow contraction of the latitudinal extent of the high-productivity belt north of the study areas. During the Pliocene, carbonate variations showed 400 kyr cycles indicating the growth and decay of ice sheets, which should have been associated with pulsations of the Antarctic ice cap. An abrupt collapse of the carbonate system occurred at 2.4 Ma. Higher frequency variations of the carbonate record indicate the major increase of the northern hemisphere glaciation. During the Quaternary, carbonate fluxes are high during glacials and low during interglacials. Large amplitude variations with long broad minima and maxima, associated with small migrations of the lysocline and the CCD (< 200 m), are indicative of the preservation/dissolution history in the Peru Basin. During the early Pleistocene, climatic forcing by the 41 kyr obliquity cycle is not observed in the carbonate record. During the last 800 kyr, variability in the carbonate record was dominated by the 100 kyr eccentricity cycle. Fluxes of biogenic material (calcium carbonate, organic carbon, opal, and barium) were greatest during glacials, which imply higher productivity and export production of the Peru Current during cold climatic periods. Dissolution was greatest during interglacials as inferred from the relatively poor preservation of planktonic foraminifera and from the low accumulation rate of carbonate. After the Mid-Brunhes Event (400 ka), we observe a plateaulike shift to enhanced dissolution and to intensified productivity.

X-ray fotos of 10 sediment cores from the Baltic Sea

Various types of trace fossils have been studied on radiographs of sediment cores from the Western Baltic since the early nineteen sixties. lnvestigations on the endo- und epifauna including their habitats and population densities carried out independently by biologists helped to identify the processes of their formation and classify the structures. Biogenic traces are ubiquitous in both sandy and muddy sediments of the Great Belt, where the bottom waters are weil oxygenated through inflows from the North Sea (through Kattegat-Skagerrak). Almost all types of bioturbation structures encountered in the Kiel Bay are also observed in a variety of shapes and forms, and can be considered as representative for the Western Baltic area. Polychaetes, bivalves and echinoderms were recorded at the sediment surface of the cores, and also in living positions in the sediment. The different types of burrows having distint outlines ('trace fossis'), and those having indistinct outlines ('biodeformational strctures'), and their varying abundance encountered in the area have been linked, wherever possible, to the sediment type, and to the macro-benthos.