Introduction The 80kD glycoprotein transferrin (TF) and its related receptor (TFR1) play a major role in the recruitment by cancer cells of factors for their multiplication, adhesion, invasion and metastatic potential. Though primarily designed to bind iron and then be internalised into cells with its receptor, TF can also bind most transition metals such as Co, Cr, Mn, Zr, Ni, Cu, V, In & Ga. Under certain conditions TF binds Ti (IV) even more tightly than it does Fe and that this occurs at the N-lobe (as distinct from C) of apoTF. Further, under physiological conditions the species Fe(C)Ti(N)-TF may provide the route for Ti entry into cells via TFR1 (1). Thus, the radiometal PET reporter isotope 45Ti with an ‘intermediate’ (~hrs) half-life suited to tracking cell-focused biological mechanisms is an attractive option for elucidating cellular mechanisms involving TF binding and internalisation, at least in (preclinical) animal models. 45Ti (T½ = 3.08 hr; + branching ratio = 85 %; mean β+ energy = 439keV, no significant dose-conferring non-511keV γ-emissions) was produced using the reaction 45Sc(p,n)45Ti by irradiating (monoisotopic) scandium discs with an energy-degraded proton beam produced by an 18MeV isochronous medical cyclotron. Separation and purification was achieved with an hydroxylamine hydrochloride functionalised resin. Comparative microPET imaging was performed in an immunodeficient mouse model, measuring biodistributions of the radiolabels 45Ti-oxalate and 45Ti-human-TF (45Ti-h-TF), out to 6hr post-injection. Materials and Methods High purity 15mm diameter scandium disc foils (99.5%, Goodfellow, UK) each thickness 0.100 ± 0.005 mm (55 mg) were loaded into an in-house constructed solid-targetry system mounted on a 300mm external beam line utilising helium-gas and chilled water to cool the target body (2). The proton beam was degraded to 11.7 MeV using a graphite disc integrated into the graphite collimator. This energy abolishes the competing ‘contaminant’ reactions 45Sc(p,n+p)44Sc and 45Sc(p,2n)44Ti. Beam current was measured using the well documented 65Cu(p,n)65Zn reaction. Calculations showed that the chosen energy is close to the optimal primary energy (~12 MeV) for maximising the (thin-target) yield from a 0.100 mm thick target. For separation of Ti from the Sc target two methods were examined; (i) ion exchange column separation using 2000 mg AG 50W-X8 resin conditioned with 10mL 9M HCl. Disc is dissolved in 1 mL of 9M HCl, which at completion of reaction is pipetted into column. Successive 1 mL volumes of 9M HCl are added, and subsequent elutions collected. (ii) Following Gagnon et al., (3) a method employing hydroxylamine hydro-chloride functionalised resin (’hydroxamate method’) was applied, similar to its use in our hands for purification and separation of 89Zr (2) following its original description for 89Zr by Holland et al., (4). Disc dissolved in 2mL 6M HCl, then diluted to 2M. Elute through column to waste fraction 1 (w1 – see FIG. 1). Then elute 6 mL of 2M HCl through column to w2, followed by 6 mL of traceSELECT H2O to w3. Finally, elute Ti into successive 1 mL product fractions (p1, 2 etc.) using 5 mL of 1M oxalic acid. This procedure takes approximate 1 hr. 45Ti in elution vials was measured using γ-spectroscopy. Sc in the same vials was determined later using ICP-MS. Results A typical production run using a beam current of 40 μA for 60min on a 0.100mm-thick disc produced an activity of 1.83 GBq. Radionuclidic analysis of an irradiated disc using calibrated cryo-HPGe γ-spectroscopy revealed T½ = 2.97–3.19 hr (95% CI) for 45Ti, and with contaminant 44Sc < 0.19 %, with no other isotopes detected. Despite systematic adjustments to column conditions satisfactory chemical separation was not achieved using the ion exchange column method (i), despite previous reports of its success (5). Typical results of separation using the successful hydroxamate method (ii) are shown on the FIGURE 1. It is seen that significant portion of 45Ti is lost in the initial washing steps leading to waste collection. N = 4 replicate experiments showed a variation (SD) of 10 % of the mean in each elu-tion fraction. Subsequent ICP-MS of the same elutions for (cold) Sc showed approximately 80 % by mass appeared in w1 and 20 % in w2, with negligible total mass (total fraction ~1/6000) of Sc in product (p1–4) vials. However, the FIG. 1 shows that a total of only 30% of the original activity of 45Ti (corrected to EOB) is available in the product vials, with the vial of highest specific activity (p1) containing 14 %. However, using a stack of 2×0.100mm thick Sc discs as a target yields isotope of adequate specific activity with-out need for concentration, for subsequent labelling and small-animal imaging purposes. In a ‘proof-of-principle’ experiment, two groups of healthy Balb/c-nu/nu female adult mice were administered with 45Ti radiotracers. The first group (N = 3) received approximately 20 MBq IP of 45Ti-oxalate buffered to pH = 7.0, and under-went microPET/CT imaging (Super Argus PET, Sedecal, Spain) out to 6hr post-injection, plus biodistribution analysis of radioactivity by dis-section at sacrifice (6hr). The second group (N = 3) received approximately 20 MBq IP of 45Ti-h-TF and were also studied to 6hr post-injection, followed by radioactive analysis after dissection at sacrifice. Organ and tissue biodistributions of the two groups at 6hr were similar but with 45Ti-oxalate showing slightly greater affinity for bone. Biodistribution by dissection results broadly confirmed the findings from PET images. However, TLC results suggested that similarity of radiolabel biodistributions of the two groups may be due to contamination of the TF radiolabel with non-conjugated Ti at time of injection. An alternative explanation is dechelation in vivo of 45Ti from 45Ti-h-TF. Conclusion Despite significant loss of 45Ti to the waste fractions of the separation process (total 53 %, corrected to EOB), 45Ti of acceptable specific activity and high radionuclidic purity has been produced from the reaction 45Sc(p,n)45Ti, with separation and purification of the product by hydroxamate column chemistry, confirming an earlier report. Though microPET in vivo imaging using 45Ti-based radiolabels was shown to be feasible, the similarity in the results for the label 45Ti-h-TF compared with ‘raw’ 45Ti-oxalate suggests further investigations. These may include a direct comparison of in vivo 45Ti-h-TF small-animal imaging plus post-dissection biodistribution with the same procedures using 89Zr labelled h-apotransferrin (6).

Thesis (Ph. D.)--Ohio State University, 2006. / Full text release at OhioLINK's ETD Center delayed at author's request

Orientador: Celso Ulysses Davanzo / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-05T16:24:43Z (GMT). No. of bitstreams: 1 Lucato_MelissaUmata_D.pdf: 2256384 bytes, checksum: fd4564569486d21c7ee858263e6a0d83 (MD5) Previous issue date: 2005 / Resumo: O presente trabalho teve como principal objetivo a elaboração de metodologias analíticas que fornecessem informações diretas referentes aos problemas analisados, utilizando simultaneamente métodos não destrutivos de obtenção de espectros (FTIR) e calibração multivariada (PLS) com seleção de variáveis (AIgoritmo Genético e AIgoritmo das Projeções Sucessivas). Para efetivar tal objetivo foram estudados dois diferentes sistemas analíticos: I-Análise dos combustíveis comerciais (diesel, gasolina e álcool etílico automotivo): Foram analisadas propriedades de amostras comerciais dos combustíveis como: densidade, tipos de hidrocarbonetos, curva de destilação, propriedades de octanagem, entre vários outros, a partir de espectros de FTIR-ATR. Os valores de referência foram obtidos a partir de normas ASTM, que foram correlacionados com resultados da calibração multivariada através do método PLS e MLR, com e sem seleção de variáveis. Os erros obtidos apresentaram valores inferiores aos da reprodutibilidade do método padrão para a maioria das propriedades. Além disso, os resultados entre os modelos propostos foram comparados estatisticamente pelo testeF. II -Estudo das propriedades de degradação do PET, Poli(tereftalato de etileno): Foram determinadas a viscosidade intrínseca [h] e a concentração de grupos carboxílicos terminais (CEG) através da técnica de refletância difusa e técnica fotoacústica no infravermelho com calibração multivariada (PLS). Os modelos foram comparados estatisticamente pelo teste-F. Os resultados apontaram um método alternativo às laboriosas análises convencionais que empregam reagentes tóxicos, gerando resíduos / Abstract: This research project was mainly intended to elaborate analytical methodologies that supplied referring direct information to the analyzed problems, using simultaneously non destructive methods of attainment of spectroscopy (FTIR) and multivaried calibration (PLS) with variable selection (Genetic Algorithm and Successive Projections Algorithm). To accomplish this objective, two different analytical systems had been studied: 1- Analysis of commercial fuels (diesel, gasoline and automotive etílic alcohol): - Properties of commercial samples of fuels had been analyzed as: density, types of hydrocarbons, curve of destillation, octane number properties, among several others, from spectra of FTIR. The foundation values had been gotten from ASTM norms, that had been correlated with results of the multivariate calibration through PLS and MLR methods, using or not variable selection. The obtained errors had presented inferior values to the ones for reprodutibilidade of the standard method for the majority of the properties. Moreover, the results among the considered models were statistically compared by the F test . II- Study of the Properties of PET's degradation, Poly(ethylene therphtalate) : - Intrinsic viscosity [h] and the concentration of terminal carboxylic groups had been determined (CEG) by the technique of diffuse reflectance and photoacoustic technique in the infrared region with multivaried calibration (PLS). The models were statistically compared by the F test. The results point to an alternative method for the laborious conventional analyses that use toxic reagents and generates residues / Doutorado / Quimica Analitica / Doutor em Ciências

Orientador: Carlos Alberto Rodrigues Anjos / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-25T04:26:54Z (GMT). No. of bitstreams: 1 Botti_LauraCostaMoreira_M.pdf: 1490815 bytes, checksum: dc0cfefc2c575ceab1d1f3b15b4e0ec6 (MD5) Previous issue date: 2014 / Resumo: Considerada a principal causa de perda da qualidade em óleos e gorduras, a oxidação leva à formação de sabores e odores indesejáveis, perda de funcionalidade e valor nutricional em especial do azeite de oliva (AO). Assim, o objetivo deste trabalho foi avaliar a eficiência de embalagens de vidro (transparente e âmbar) e polietileno tereftalato (PET) aliado à tecnologia de barreira ao oxigênio e à radiação UV, como opção à manutenção da estabilidade do AO. Inicialmente foi feita a caracterização dos materiais, seguida do envase das amostras e estocagem de metade delas em caixas de papelão mantidas na ausência de luz. Ao mesmo tempo, a outra metade foi exposta à iluminação intermitente por períodos de 12h e intensidade luminosa de 1000 lux durante seis meses. Ao longo do tempo foram feitas as seguintes análises: acidez, índice de peróxido, cor L*a*b*, cor Lovibond, extinção específica a 232 (K232) e 270nm (K270), clorofila, carotenoides, fenóis totais e ?-tocoferol. Durante o período analisado, houve aumento da acidez, índice de peróxido, K232 e K270 para todas as amostras, sendo as embalagens convencionais transparentes as mais susceptíveis e a condição sob iluminação a mais crítica. Em contrapartida, a cor, o teor de clorofila, carotenoides e fenóis totais foram reduzidos com o tempo e também sofreram influência da luz e do material de embalagem. A degradação do ?-tocoferol foi influenciada até o segundo mês pelas condições de estocagem e tipo de material da embalagem. A partir de então, os índices de tocoferol foram reduzidos de forma drástica em todos os sistemas de embalagem e provavelmente não foram detectados pela metodologia. De forma geral, sob o abrigo da luz, tanto a embalagem de PET com barreira ao oxigênio como o vidro foram efetivas nos primeiros meses, mostrando-se como opção para produtos que não necessitem de elevada proteção. Entretanto, a embalagem de vidro âmbar mostrou-se a mais eficiente devido a suas características intrínsecas e ao amplo espectro de proteção para amostras quando expostas à luz. A partir dos resultados, conclui-se que aditivos do tipo estabilizantes de radiação UV não foram efetivos nas condições de estudo, porém o uso de absorvedores de oxigênio aliado a tecnologia de blendas pode ser uma alternativa para a área de embalagens de azeite e óleos. Assim, é essencial que se pense no uso complementar de diferentes tecnologias em prol da manutenção da qualidade e estabilidade dos alimentos / Abstract: he oxidation is considered the main cause of quality loss in oils and, consequently, leads to the formation of undesirable flavors and odors, loss of nutritional value and functionality in particular olive oil (OO). This work aimed to evaluate the efficiency of glass containers (clear and amber) and polyethylene terephthalate (PET) combined with the improvement of oxygen barrier technology and UV radiation, as an option to maintain the stability of OO. At first, the materials were characterized, the samples were bottled and then half of them stored in cardboard boxes kept in the dark. At the same time, the other half was exposed to intermittent periods of 12 hours light and light intensity of 1000 lux for six months. Acidity, peroxide value, color L * a * b *, color Lovibond, specifc extinction at 232 (K232) and 270nm (K270), chlorophyll, carotenoids, total phenols and ? ¿ tocopherol were analyzed over time. During the reporting period, there was an increase in acidity, peroxide value, K232 and K270 for all samples, with the conventional transparent packaging being the most susceptible and the illumination condition, the most critical. In contrast, the color, chlorophyll content, carotenoids and total phenolics were reduced with time and were also affected by light and the packaging material. The degradation of ?-tocopherol was influenced by storage conditions and type of packaging material until the second month. Thereafter, the tocopherol levels were reduced drastically in all packaging systems and probably were not detected by the methodology. In general, in the dark, both the PET packaging with oxygen barrier such as glass were effective in the first months, showing as an option for products that do not require high protection. However, the amber glass packaging due to their inherent and broad-spectrum protection from light features proved to be the most efficient when exposed to light. The results evaluation of this research showed that the stabilizing additive type of UV radiation were not effective under the conditions of study, but the use of oxygen absorbers combined with blends technology can be an alternative to the oil packaging. Thus, it is essential to think about the complementary use of different technologies for the maintenance of the quality and stability of food / Mestrado / Tecnologia de Alimentos / Mestra em Tecnologia de Alimentos

Poly(ADP-ribose)polymerase-1 and -2 (PARP1/2) are nuclear proteins involved in DNA repair. Tumors with defects in homologous recombination, including BRCA1- and BRCA2-deficient cancers, have been shown to be sensitive to PARP inhibition. The Weissleder group has synthesized fluorescent and radioactive derivatives of the PARP1/2 inhibitor AZD2281. We hypothesized that fluorescent and radioactive AZD2281-based imaging agents would quantify PARP1/2 expression in vitro and in vivo. To test this hypothesis, a panel of pancreatic ductal adenocarcinoma and ovarian carcinoma cell lines were characterized by immunocytochemistry for PARP1/2 expression. AZD2281-derived fluorescence signal correlated with anti-PARP antibody fluorescence signal strength in vitro. Four cell lines representing a range of PARP1/2 expression levels were then xenografted into Nu/Nu mice. Mice bearing four tumor types each were imaged with AZD2281-derived imaging agents, sacrificed, and their tumors excised for stand-alone imaging and Western blot. AZD2281-derived signal correlated with tumor PARP1/2 expression determined by Western blot, indicating that PARP1/2 expression level is a determinant of fluorescent signal strength and SUVs of AZD2281-derived agents in vivo. These data indicate that AZD2281-derived agents are useful tools for quantifying intracellular PARP1/2 both in vitro and in vivo, which could one day enable prospective identification of tumors likely to respond to PARP inhibitors.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21294号 / 医博第4383号 / 新制||医||1030(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 佐藤 俊哉, 教授 今中 雄一, 教授 増永 慎一郎 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

No description available.

El incremento y la consecuente acumulación de desechos plásticos hacen necesaria la implementación de alternativas para su manejo y disposición. El poli(tereftalato de etileno), PET, es uno de los plásticos más utilizados, especialmente en la elaboración de botellas descartables que, debido a su uso masivo y a su alta resistencia a la degradación, hacen conveniente el desarrollo de métodos para su reciclaje. Este trabajo presenta una manera de afrontar la acumulación de residuos de PET utilizando el producto de la despolimerización en la obtención de resinas de poliéster insaturado que, entre sus diversas aplicaciones, pueden utilizarse para el encapsulamiento de residuos que hacen necesario evitar el riesgo de su liberación al ambiente debido a su peligrosidad. En primer lugar, se estudió la despolimerización de desechos de PET con etilenglicol variando las cantidades de catalizador y el tiempo de reacción. Se logró hasta 82% de rendimiento del monómero tereftalato de bis(2-hidroxietileno), BHET, con 0,7% de acetato de cinc y 3 horas de reacción. Además, se determinó que también se produce el dímero de BHET entre 5 - 6%. La caracterización de ambos productos se realizó por espectroscopía infrarroja, de resonancia magnética nuclear (RMN) y de masas. Posteriormente, se sintetizaron diversas resinas de poliéster insaturado utilizando el BHET obtenido de la despolimerización del PET, anhidrido maleico (AM), ácido adípico (AA) y un glicol. Los glicoles utilizados fueron etilenglicol (EG), dietilenglicol (DG), propilenglicol (PG) y 2-etil-1,3-hexanodiol (EHD). Las resinas estudiadas estuvieron compuestas por las combinaciones de glicol-AM, glicol-AM-BHET, glicol-AM-AA y glicol-AM-BHET-AA a modo de compararlas y analizar cada una de ellas en el encapsulamiento de sales. La caracterización de los poliésteres insaturados se realizó mediante análisis de RMN y se logró elucidar las estructuras químicas de las resinas sintetizadas con etilenglicol y dietilenglicol. Adicionalmente, se calcularon los pesos moleculares promedio de los poliésteres a partir del análisis de grupos terminales -OH y -COOH que varían entre 488 a 1023 g/mol. Las resinas sintetizadas con EHD poseen los pesos moleculares más altos. Finalmente, se evaluó la capacidad de las resinas sintetizadas como matriz de encapsulamiento. Para ello se utilizó cloruro de sodio como modelo del compuesto a encapsular. Las probetas se prepararon mezclando la resina de poliéster, la sal y estireno. El entrecruzamiento se llevó a cabo con un iniciador (peróxido de metil etil cetona) y un catalizador (octoato de cobalto). La capacidad de encapsulamiento se determinó sumergiendo las probetas por ocho semanas en agua destilada. Se encontró que la presencia de BHET en la resina brinda mayor resistencia e impermeabilidad a las matrices poliméricas. Además, las probetas formadas con las resinas PG-AM-BHET y EHD-AM-BHET retienen el 80% de sal luego de estar sumergidas en agua y las de DG-AM-AA retienen la menor cantidad de sal (13%). / Tesis

Progression of Alzheimer's disease has been associated with the deposition of aggregated amyloid beta (Aβ) protein in the brain. Though first described in post-mortal tissue, the development of Aβ specific tracers for positron emission tomography (PET) permits in-vivo mapping of its distribution in the brain. One of the well-known and early-developed tracers is the Pittsburgh Compound B (PiB) (Klunk et al., 2004). However, the challenge with PiB lies in the stability of the radioisotope 11C. 11C's short half-life of only 20 minutes hinders its transportation and usage at imaging facilities that are not in close proximity with the radioisotopes manufacturer. Recently, an alternative Aβ tracer has been developed, Florbetapir (Wong et al, 2010.), with a half-life of 110 minutes that should allow wider accessibility to imaging sites while improve the detection of Aβ. To define better the specificity and utility of Florbetapir, we propose to utilize existing PET data acquired with the radioactive tracer Florbetapir from the Alzheimer's disease Neuroimaging Initiative (ADNI). Our goal is to characterize the symmetry of Aβ protein deposition in the brain of patients with Alzheimer's disease. While a previous study has investigated this issue using PiB, Florbetapir has not been used. Our project will involve data post-processing by segmenting out non-brain tissues. Segmented data is then normalized by the pixel intensity and a distribution curve is created using MathCad program. In addition, we will calculate the asymmetry score for Regions of Interest. This will permit comparison of the uptakes of tracer between brain hemispheres to be made. Results from our project can provide insight into Florbetapir's binding affinity for Aβ. In addition, Florbetapir's potential as a better alternative to PiB can also be evaluated.