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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

A comparison of extraction techniques for the determination of metals in soil

Burcham, Shannon Tomoe 14 August 2009 (has links)
A growing issue in groundwater studies concerns the distinction between anthropological and geological metal contaminants in soil. Current extraction methods, such as hot acid extraction, result in the digestion of the sample matrix which may obscure the origin of the contaminant. Since metals from anthropological sources are most often adsorbed to the surface of the soil sediment, the dissolution of the geological matrix is not necessary to release the contaminat metals. The resolution to this problem may lie in the use of chelating agents, where the adsorbed metals can be removed from the sediments without disrupting the geological matrix. This would allow the surface bound metal contaminants to be differentiated from the geological metals which are a natural part of the soil's composition. The basis of this study is a comparison of a hot acid extraction method vs. a chelation extraction method for the determination of metals in soils. Statistical comparisons on the precision and accuracy of the these methods on a reference soil, salt amended soil, and sewage sludge amended soil is discussed. / Master of Science
2

Kartläggning och nedbrytning av komplexbildare i ett integrerat kartongbruk / Mapping and degradation of chelating agents in an integrated board mill

Löfgren, Lina, Olsson, Robert January 2006 (has links)
<p>Skoghalls Bruk fick i juni 2005 tillstånd av miljödomstolen att öka kartongproduktionen från 665 000 till 800 000 ton kartong per år. Villkoren för tillståndet var att under en prövotidsperiod bl.a. utreda möjligheten att minska utsläpp till omkringliggande vatten.</p><p>Syftet med examensarbetet var att kartlägga Skoghalls Bruks avloppssystem med avseende på komplexbildarna EDTA (etylendiamintetraättiksyra) och DTPA (dietylentriaminpentaättiksyra), kväve samt fosfor.</p><p>Dessutom genomfördes nedbrytningsförsök i luftade dammar i laboratorieskala.</p><p>För att få en klarare bild av nedbrytningen i Skoghalls Bruks luftade damm gjordes även en molekylviktsfraktionering på inkommande och utgående avloppsvatten.</p><p>Komplexbildare används inom pappers- och massaindustrin för sin förmåga att binda till sig metalljoner som kan vara skadliga i processen. Metalljoner katalyserar nedbrytningen av väteperoxid i blekningen och påskyndar bildandet av aldehyder i kartongen vilket kan leda till lukt- och smakpåverkan.</p><p>EDTA och DTPA anses vara stabila ämnen, vilket innebär att de är biologiskt svårnedbrytbara. Under den litteraturstudie som genomfördes fanns motsägande uppgifter angående nedbrytningen av komplexbildare, där vissa menade att de genomgår biologisk nedbrytning, men att det sker mycket långsamt. Andra påstod att komplexbildare inte bryts ner överhuvudtaget.</p><p>Komplexbildare kan elimineras genom fotokemisk oxidativ nedbrytning. Då EDTA och DTPA bildar komplex med Fe3+ blir de fotokemiskt labila och oxideras till mindre föreningar vid belysning med ultraviolett ljus. Sammanfattningsvis gav litteraturstudien att det mest effektiva sättet att minska komplexbildare i avloppsvatten innan det släpps till recipienten är att UV-bestråla vid optimala förhållanden, dvs. rätt pH och eventuellt tillsats av Fe3+ om det behövs.</p><p>Provtagning av avloppsvatten i Skoghalls Bruks reningsanläggning gjordes under 6 veckor för att kartlägga utsläppen och de olika delströmmarnas bidrag.</p><p>Nedbrytningsförsök genomfördes i två luftade dammar i laboratorieskala. För att studera temperaturens inverkan konstruerades en sommar- och en vinterdamm med temperaturerna 27 respektive 16°C. Efter referensförsöket, som gjordes för att se eventuella skillnader mellan försöken, undersöktes inverkan av ultraviolett ljus på nedbrytningen av komplexbildare. Sedan tillsattes Fe3+ i form av järnklorid i ett försök att öka den fotokemiska nedbrytningen. Alla försök pågick under två veckor vardera.</p><p>I syfte att undersöka partikelstorleksfördelningen före och efter nedbrytning i Skoghalls Bruks luftade damm gjordes en molekylviktsfraktionering. Inkommande och utgående avloppsvatten från dammen ultrafiltrerades och analyserades med avseende på COD (Chemical Oxygen Demand), komplexbildare, kväve och fosfor.</p><p>Kartläggningen visade att EDTA bröts ner effektivt i reningsanläggningen medan nedbrytningen av DTPA var obetydlig. EDTA reducerades med 88 % över den luftade dammen.</p><p>Avlopp från kartongmaskinerna samt CTMP-fabriken var de avlopp som innehöll mest EDTA. Största mängden DTPA återfanns i blekerifiltratet och i KM 8:s avlopp.</p><p>Skoghalls Bruk släppte ut komplexbildare motsvarande 0,188 kg/ton kartong, i form av 100 % syra i obunden form, under karteringsperioden vilket var klart lägre än gränsvärdet som är 1,6 kg 100 % syra/ton kartong.</p><p>Kväve reducerades med 67 % över den luftade dammen. Det nuvarande gränsvärdet för kväveutsläpp till recipient är 500 kg/dygn, men ett av målen med prövotidsutredningen är att minska utsläppen till 300 kg/dygn. Enligt resultaten från karteringsperioden var kväveutsläppet 204 kg/dygn vilket uppfyller målet för prövotidsutredningen. Av det totala kväveutsläppet kom 39 kg/dygn från komplexbildare. Utsläppet av kväve från mixeriavloppet var 56 kg/dygn till recipient vilket var klart högre än förväntat.</p><p>Den totala fosforreduceringen över reningsanläggningen var 78 %. Utsläppet av fosfor till recipient var 19 kg/dygn vilket är klart lägre än gränsvärdet som är 50 kg/dygn.</p><p>På grund av komplexbildaranalysens mätosäkerhet var det svårt att få entydiga resultat från laboratorieförsöken. Det fastställdes dock att DTPA var svårare att bryta ner än EDTA och att järntillsats i kombination med ultraviolett ljus gav en positiv effekt på nedbrytningen av båda komplexbildarna. Även nedbrytningen av COD, kväve och fosfor ökade vid järntillsats.</p><p>Att DTPA är svårare att bryta ner än EDTA framkom även av resultaten från molekylviktsfraktioneringen. Det visade sig också att EDTA och DTPA inte är helt vattenlösliga utan kan binda till fast material. Största delen av komplexbildarna fanns dock i fraktionen med partiklar med molekylvikt mindre än 1 kDa. Kväve och fosfor fanns i de låg- och högmolekylära fraktionerna.</p> / <p>In June of 2005 Skoghall Mill got permission from the Environmental Court to increase their production of board from 665 000 tons to 800 000 tons per year. The condition for the permission was to, during a trial period, investigate the possibility to decrease the emissions to the surrounding water. The purpose of this master thesis is to do a mapping of Skoghall Mill’s waste water treatment system regarding the chelating agents EDTA (ethylenediaminetetraacetic acid) and DTPA (diethylenetriaminepentaacetic acid), nitrogen and phosphor.</p><p>Also degradation-trials were made with aerated lagoons in laboratory-scale.</p><p>To get a more complete view of the degradation in Skoghalls Mills aerated lagoon a molecular weight fractionation of the incoming and the outgoing water was made.</p><p>Chelating agents are used in the pulp- and paper industry due to its ability to bind metal-ions which are hazardous for the process. The metal-ions catalyze the decomposition of hydrogen peroxide in the bleaching process and the forming of aldehydes in the board which can lead to problems with taste and odour.</p><p>EDTA and DTPA are regarded to be very stable, which means that they are hard to degrade biologically. There was contradictory information about the degradation of chelating agents during the literature survey. Some say that chelating agents are degraded biologically but very slowly. Others say that they don’t degrade at all.</p><p>Chelating agents can be eliminated through photochemical oxidative degradation. When EDTA and DTPA form complexes with Fe3+ they become photolabile and oxidize to smaller compounds at illumination with ultraviolet light. To sum up the literature survey, the most efficient way to reduce chelating agents in sewage water before discharge to receiving waters, is UV treatment at optimal conditions i.e., at the right pH and with addition of Fe3+ if necessary.</p><p>The sampling of the waste water in Skoghalls waste water treatment plant was made during 6 weeks to map the emissions and the contribution of the different sub-streams.</p><p>The degradation-trials were made in two aerated lagoons in laboratory-scale. To study the influence of temperature a summer- and a winter-lagoon were made and the temperatures were sat to, respectively, 27 and 16°C. After the reference-trial, which was made to determine possible differences between the trials, the influence of ultraviolet light on degradation of chelating agents were investigated. In an attempt to enhance the photochemical degradation, Fe3+ in the form of ferric chloride was then added. All trials went on for two weeks each.</p><p>In order to investigate the particle size distribution, before and after degradation in Skoghall Mill’s aerated lagoon, a molecule weight fractionation was made. Incoming and outgoing sewage water from the lagoon was ultra-filtrated and analyzed regarding COD (Chemical Oxygen Demand), chelating agents, nitrogen and phosphor.</p><p>The mapping showed that EDTA was efficiently degraded in the waste water treatment plant, while degradation of DTPA was insignificant. EDTA was reduced by 88 % trough the aerated lagoon.</p><p>Waste water from the board machines and the CTMP-plant contained the largest amount EDTA. DTPA was recovered mostly in the bleaching filtrate and KM 8’s waste water.</p><p>The emission of chelating agents, counted as acid, from Skoghall Mill during the period of the mapping was 0,188 kg/ton board which is well below the emission limit value of 1,6 kg/ton board.</p><p>Nitrogen was reduced by 67 % through the aerated lagoon. The present emission limit value for nitrogen is 500 kg/day. According to the results from the period of the mapping the emission of nitrogen was 204 kg/day which is well below the emission limit value. The contribution of chelating agents to the nitrogen emission was 39 kg/day. 56 kg nitrogen /day were recovered in the waste water from the causticizing area, which is remarkably higher than expected.</p><p>The total reduction of phosphor through the waste water treatment plant was 78 %. The emission of phosphor to receiving waters was 19 kg/day which is well below the emission limit value of 50 kg/day.</p><p>Due to the insecurity in measurement of chelating agents, it was difficult to retain unambiguous results from the laboratory-trials. However it was determined that DTPA was more difficult to degrade than EDTA and that a combination of ultraviolet light and addition of ferric iron had a favourable effect on the degradation of both chelating agents. Also the degradation of COD, nitrogen and phosphor increased when ferric iron was added.</p><p>The fact that DTPA are more difficult to degrade than EDTA was also stated in molecular weight fractionation. It was also shown that EDTA and DTPA weren’t totally water-soluble but can bind to solid material. The largest amount of chelating agents was recovered in the fraction with particles whose molecular weight is less than 1 kDa. Nitrogen and phosphor were recovered in the low- and high-molecular fractions.</p>
3

Kartläggning och nedbrytning av komplexbildare i ett integrerat kartongbruk / Mapping and degradation of chelating agents in an integrated board mill

Löfgren, Lina, Olsson, Robert January 2006 (has links)
Skoghalls Bruk fick i juni 2005 tillstånd av miljödomstolen att öka kartongproduktionen från 665 000 till 800 000 ton kartong per år. Villkoren för tillståndet var att under en prövotidsperiod bl.a. utreda möjligheten att minska utsläpp till omkringliggande vatten. Syftet med examensarbetet var att kartlägga Skoghalls Bruks avloppssystem med avseende på komplexbildarna EDTA (etylendiamintetraättiksyra) och DTPA (dietylentriaminpentaättiksyra), kväve samt fosfor. Dessutom genomfördes nedbrytningsförsök i luftade dammar i laboratorieskala. För att få en klarare bild av nedbrytningen i Skoghalls Bruks luftade damm gjordes även en molekylviktsfraktionering på inkommande och utgående avloppsvatten. Komplexbildare används inom pappers- och massaindustrin för sin förmåga att binda till sig metalljoner som kan vara skadliga i processen. Metalljoner katalyserar nedbrytningen av väteperoxid i blekningen och påskyndar bildandet av aldehyder i kartongen vilket kan leda till lukt- och smakpåverkan. EDTA och DTPA anses vara stabila ämnen, vilket innebär att de är biologiskt svårnedbrytbara. Under den litteraturstudie som genomfördes fanns motsägande uppgifter angående nedbrytningen av komplexbildare, där vissa menade att de genomgår biologisk nedbrytning, men att det sker mycket långsamt. Andra påstod att komplexbildare inte bryts ner överhuvudtaget. Komplexbildare kan elimineras genom fotokemisk oxidativ nedbrytning. Då EDTA och DTPA bildar komplex med Fe3+ blir de fotokemiskt labila och oxideras till mindre föreningar vid belysning med ultraviolett ljus. Sammanfattningsvis gav litteraturstudien att det mest effektiva sättet att minska komplexbildare i avloppsvatten innan det släpps till recipienten är att UV-bestråla vid optimala förhållanden, dvs. rätt pH och eventuellt tillsats av Fe3+ om det behövs. Provtagning av avloppsvatten i Skoghalls Bruks reningsanläggning gjordes under 6 veckor för att kartlägga utsläppen och de olika delströmmarnas bidrag. Nedbrytningsförsök genomfördes i två luftade dammar i laboratorieskala. För att studera temperaturens inverkan konstruerades en sommar- och en vinterdamm med temperaturerna 27 respektive 16°C. Efter referensförsöket, som gjordes för att se eventuella skillnader mellan försöken, undersöktes inverkan av ultraviolett ljus på nedbrytningen av komplexbildare. Sedan tillsattes Fe3+ i form av järnklorid i ett försök att öka den fotokemiska nedbrytningen. Alla försök pågick under två veckor vardera. I syfte att undersöka partikelstorleksfördelningen före och efter nedbrytning i Skoghalls Bruks luftade damm gjordes en molekylviktsfraktionering. Inkommande och utgående avloppsvatten från dammen ultrafiltrerades och analyserades med avseende på COD (Chemical Oxygen Demand), komplexbildare, kväve och fosfor. Kartläggningen visade att EDTA bröts ner effektivt i reningsanläggningen medan nedbrytningen av DTPA var obetydlig. EDTA reducerades med 88 % över den luftade dammen. Avlopp från kartongmaskinerna samt CTMP-fabriken var de avlopp som innehöll mest EDTA. Största mängden DTPA återfanns i blekerifiltratet och i KM 8:s avlopp. Skoghalls Bruk släppte ut komplexbildare motsvarande 0,188 kg/ton kartong, i form av 100 % syra i obunden form, under karteringsperioden vilket var klart lägre än gränsvärdet som är 1,6 kg 100 % syra/ton kartong. Kväve reducerades med 67 % över den luftade dammen. Det nuvarande gränsvärdet för kväveutsläpp till recipient är 500 kg/dygn, men ett av målen med prövotidsutredningen är att minska utsläppen till 300 kg/dygn. Enligt resultaten från karteringsperioden var kväveutsläppet 204 kg/dygn vilket uppfyller målet för prövotidsutredningen. Av det totala kväveutsläppet kom 39 kg/dygn från komplexbildare. Utsläppet av kväve från mixeriavloppet var 56 kg/dygn till recipient vilket var klart högre än förväntat. Den totala fosforreduceringen över reningsanläggningen var 78 %. Utsläppet av fosfor till recipient var 19 kg/dygn vilket är klart lägre än gränsvärdet som är 50 kg/dygn. På grund av komplexbildaranalysens mätosäkerhet var det svårt att få entydiga resultat från laboratorieförsöken. Det fastställdes dock att DTPA var svårare att bryta ner än EDTA och att järntillsats i kombination med ultraviolett ljus gav en positiv effekt på nedbrytningen av båda komplexbildarna. Även nedbrytningen av COD, kväve och fosfor ökade vid järntillsats. Att DTPA är svårare att bryta ner än EDTA framkom även av resultaten från molekylviktsfraktioneringen. Det visade sig också att EDTA och DTPA inte är helt vattenlösliga utan kan binda till fast material. Största delen av komplexbildarna fanns dock i fraktionen med partiklar med molekylvikt mindre än 1 kDa. Kväve och fosfor fanns i de låg- och högmolekylära fraktionerna. / In June of 2005 Skoghall Mill got permission from the Environmental Court to increase their production of board from 665 000 tons to 800 000 tons per year. The condition for the permission was to, during a trial period, investigate the possibility to decrease the emissions to the surrounding water. The purpose of this master thesis is to do a mapping of Skoghall Mill’s waste water treatment system regarding the chelating agents EDTA (ethylenediaminetetraacetic acid) and DTPA (diethylenetriaminepentaacetic acid), nitrogen and phosphor. Also degradation-trials were made with aerated lagoons in laboratory-scale. To get a more complete view of the degradation in Skoghalls Mills aerated lagoon a molecular weight fractionation of the incoming and the outgoing water was made. Chelating agents are used in the pulp- and paper industry due to its ability to bind metal-ions which are hazardous for the process. The metal-ions catalyze the decomposition of hydrogen peroxide in the bleaching process and the forming of aldehydes in the board which can lead to problems with taste and odour. EDTA and DTPA are regarded to be very stable, which means that they are hard to degrade biologically. There was contradictory information about the degradation of chelating agents during the literature survey. Some say that chelating agents are degraded biologically but very slowly. Others say that they don’t degrade at all. Chelating agents can be eliminated through photochemical oxidative degradation. When EDTA and DTPA form complexes with Fe3+ they become photolabile and oxidize to smaller compounds at illumination with ultraviolet light. To sum up the literature survey, the most efficient way to reduce chelating agents in sewage water before discharge to receiving waters, is UV treatment at optimal conditions i.e., at the right pH and with addition of Fe3+ if necessary. The sampling of the waste water in Skoghalls waste water treatment plant was made during 6 weeks to map the emissions and the contribution of the different sub-streams. The degradation-trials were made in two aerated lagoons in laboratory-scale. To study the influence of temperature a summer- and a winter-lagoon were made and the temperatures were sat to, respectively, 27 and 16°C. After the reference-trial, which was made to determine possible differences between the trials, the influence of ultraviolet light on degradation of chelating agents were investigated. In an attempt to enhance the photochemical degradation, Fe3+ in the form of ferric chloride was then added. All trials went on for two weeks each. In order to investigate the particle size distribution, before and after degradation in Skoghall Mill’s aerated lagoon, a molecule weight fractionation was made. Incoming and outgoing sewage water from the lagoon was ultra-filtrated and analyzed regarding COD (Chemical Oxygen Demand), chelating agents, nitrogen and phosphor. The mapping showed that EDTA was efficiently degraded in the waste water treatment plant, while degradation of DTPA was insignificant. EDTA was reduced by 88 % trough the aerated lagoon. Waste water from the board machines and the CTMP-plant contained the largest amount EDTA. DTPA was recovered mostly in the bleaching filtrate and KM 8’s waste water. The emission of chelating agents, counted as acid, from Skoghall Mill during the period of the mapping was 0,188 kg/ton board which is well below the emission limit value of 1,6 kg/ton board. Nitrogen was reduced by 67 % through the aerated lagoon. The present emission limit value for nitrogen is 500 kg/day. According to the results from the period of the mapping the emission of nitrogen was 204 kg/day which is well below the emission limit value. The contribution of chelating agents to the nitrogen emission was 39 kg/day. 56 kg nitrogen /day were recovered in the waste water from the causticizing area, which is remarkably higher than expected. The total reduction of phosphor through the waste water treatment plant was 78 %. The emission of phosphor to receiving waters was 19 kg/day which is well below the emission limit value of 50 kg/day. Due to the insecurity in measurement of chelating agents, it was difficult to retain unambiguous results from the laboratory-trials. However it was determined that DTPA was more difficult to degrade than EDTA and that a combination of ultraviolet light and addition of ferric iron had a favourable effect on the degradation of both chelating agents. Also the degradation of COD, nitrogen and phosphor increased when ferric iron was added. The fact that DTPA are more difficult to degrade than EDTA was also stated in molecular weight fractionation. It was also shown that EDTA and DTPA weren’t totally water-soluble but can bind to solid material. The largest amount of chelating agents was recovered in the fraction with particles whose molecular weight is less than 1 kDa. Nitrogen and phosphor were recovered in the low- and high-molecular fractions.
4

Valor diagnóstico del radiorrenograma con DTPA-TC99m para la estimación de tasa de filtración glomerular en relación a la ecuación de cockroft gault en pacientes mayores de 25 años con enfermedad renal crónica hospital Central PNP Luis N. Sáenz en el periodo marzo – mayo 2015

Maturrano Ascencio, Christian Joseph January 2015 (has links)
I Objetivos.- Determinar el valor diagnostico del estudio de Radiorrenograma con Dtpa – Tc99m para estimar la tasa de filtración glomerular en pacientes con enfermedad renal crónica en el Hospital Central de la Policía Nacional del Perú “Luis N. Sáenz. Diseño: Estudio observacional, descriptivo, prospectivo y de corte transversal. Institución:”; Lima, Perú. Materiales: Historias Clínicas. II Intervenciones.- estudio de historias clínicas de pacientes con una muestra poblacional, no probabilístico. De los cuales por no cumplir con los criterios de inclusión quedaron un total de 34 historias clínicas, siendo estos 20 de sexo masculino y 14 de sexo femenino los cuales se encuentran dentro de un rango de edad, de 25 años a más. La muestra se obtuvo revisando cada historia clínica, estos presentaban un informe de laboratorio con la filtración glomerular y la orden de estudio de Radiorrenograma con DTPA-Tc99m entre el periodo marzo-mayo del 2015. III Principales medidas de resultados.- Sensibilidad, especificidad, valor predictivo positivo y negativo del Radiorrenograma con DTPA-Tc99m para la estimación de filtración glomerular. Resultados: con los datos recabados, se calculó el valor diagnostico del Radiorrenograma con DTPA-Tc99m para la estimación de filtración glomerular. La sensibilidad del Radiorrenograma con DTPA-Tc99m es de 69.23% (IC 95%,40.30% al 98.17%); una especificidad de 90.48% (IC 95%, 75.54% al 100%); un valor predictivo positivo de 81.82%( IC 95%,54.48% al 100%) y un valor predictivo negativo de 82.61% (IC 95%,64.94% al100%) para la estimación de filtrado glomerular. IV Conclusiones.- El Radiorrenograma con DTPA-Tc99m es el estudio isotópico adecuado para la estimación de filtración glomerular en la enfermedad renal crónica. / I. Purpose -. To determine the diagnostic value of Radiorrenograma with DTPA-Tc99m studying, to estimate glomerular filtration rate in patients with chronic kidney disease at Central Hospital of the National Police of Peru " Luis N. Sáenz. Design: Observational, descritptive, porspective and cross-sectioanl study. Institution: Central Hospital of the National Police of Peru "Luis N. Sáenz, Lima, Peru. Materials: Medical Records II. interventions.- study of medical records of patients with a sample population, which no probabilistic. Of which not meet the inclusion criteria were a total of 34 medical records, and these 20 males and 14 females who are within an age range of 25 years more The sample was obtained bye reviewing medical recods each, they presented a report of the laboratory test with glomerular flitration and the order of study of Radiorrenograma with DTPA-Tc99m between the period March – may 2015 III. Main outcome measures-. Sentitivity, specificity, positive an negative predictive value of Radiorrenograma with DTPA-Tc99m for estimating glomerular filtration. Results: With the data collected; the diagnostic value of Radiorrenograma with DTPA-Tc99m for estimating glomerular filtration was calculated.Radiorrenograma with DTPA-Tc99m sensitivy is 69.23 % (95% CI, 40.30% to 98.17%); a specificity of 90.48 % (95% CI, 75.54% to 100%); a positive predictive value of 81.82 % (95% CI, 54.48% to 100%) and negative predictive value of 82.61 % (95% CI, 64.94 % al 100%) for estimating glomerular filtration. IV. Conclusions -. The Radiorrenograma with DTPA - Tc99m is right isotopic study for estimating GFR in chronic renal disease. Keywords: Radiorrenograma; DTPA – Tc99m; glomerular filtration; Chronic kidney diseas
5

Development of a simplified soft-donor technique for trivalent actinide-lanthanide separations

Langford Paden, Madeleine Hilton January 2015 (has links)
The necessity of reprocessing spent nuclear fuel has arisen from increasing awareness and concern for the environment, in addition to the potential of minimising proliferation. A number of different reprocessing techniques are currently being developed around the world to allow useful spent nuclear fuel (SNF) to be recycled and reused and the remaining waste to be treated. One such technique, currently being developed in the USA is the TALSPEAK process, an advanced reprocessing method for the separation of trivalent lanthanide (Ln3+) and minor actinide (MA3+) components. This process, developed in the 1960s at Oak Ridge National Laboratory, uses DTPA to act as a holdback reagent for MA3+, in a lactate buffered aqueous phase at pH 3.6, allowing Ln3+ to be selectively extracted by organophosphate HDEHP into an organic phase of DIPB or dodecane. TALSPEAK is one of the most promising techniques being researched due to its numerous advantages, particularly its relative resistance to radiolysis and its ability to be carried out without the need for high reagent concentrations. Additionally it gives high separation factors, in the region of ~50-100, comparable to other advanced reprocessing methods under development. The chemistry of the process is very complex and not particularly well understood so it would be advantageous to simplify the process by removing the need for a separate holdback reagent and buffer. In collaboration with colleagues at the Idaho National Lab, the use of amino acids as a potential combined buffer and soft donor was investigated. Although it was found that amino acids do not act as holdback reagents in their own right, optimisation of an L-alanine buffered TALSPEAK system with DTPA was found to allow the process to be carried out effectively at a lower pH of 2, which is more preferable for industrial application. As an extension of this, separation studies were carried out using the tripeptide L-glutathione (GSH) to determine its potential for use as a combined buffer and soft-donor. As with the studies with amino acids, it was found that GSH also does not act as a holdback reagent in its own right, however it does interact with Ln-DTPA complexes at pH 4. When optimised at this pH, separation factors of up to 1200 were achieved for Eu3+/Am3+, whilst still maintaining low MA3+ partitioning. However, further studies by ICP-MS and luminescence spectroscopy showed that a GSH buffered system was not effective for extraction of heavier lanthanides, although the results show the potential for further investigation into other short and longer chain peptide buffered systems and possibly lanthanide-lanthanide separations. Further studies were carried on amino acid appended DTPA ligands which were synthesised in a one step reaction in order to create a combined buffer and soft donor. The ligands were found to self-buffer at around pH 2 and allow successful separation of Eu3+/Am3+ (SF ~ 100). The results from initial investigations by luminescence spectroscopy and solvent extraction are promising and are presented here. Further work is needed on these systems in order to optimise their extraction capability and minimise Am3+ partitioning. In the future this work could promote studies for better understanding of TALSPEAK chemistry that could be used in industrial partitioning processes.
6

Zinco para arroz e soja: doses, aproveitamento pelas culturas, fitodisponibilidade e formas desse elemento no solo / Zinc for rice and soybean: rates, crop recovery, bioavailability and element fractions in the soil

Leite, Camila Marcon de Carvalho 14 January 2015 (has links)
A deficiência de Zn no solo causa efeitos indesejáveis na produção agrícola, pois a baixa disponibilidade deste micronutriente para as plantas promove a diminuição da atividade enzimática, além da deficiência deste elemento na alimentação, que pode levar ao estado de subnutrição. Tendo em vista a problemática do Zn no sistema solo-planta e suas variações nos compartimentos do solo, é importante a avaliação de sua fitodisponibilidade e as frações do solo que este elemento está associado. O objetivo deste trabalho foi avaliar a fitodisponibilidade e a compartimentalização de Zn no solo, para as culturas de arroz (Oryza sativa L.) e soja (Glycine max L. Merrill) e avaliar o efeito das doses de Zn sobre a nutrição e exportação deste nutriente pela cultura. Utilizou-se como plantas teste as culturas de arroz e soja para avaliar o efeito das doses de Zn sobre a nutrição e translocação deste nutriente até os grãos. Para tanto, uma amostra de um Latossolo Vermelho, textura argilosa da região de Piracicaba (SP) foi utilizada e ZnCl2 (marcado com 65Zn) como fonte. O experimento foi conduzido em casa de vegetação em DIC, com cinco doses de Zn (0, 1, 2, 4 e 8 mg kg-1 de solo), com quatro repetições. O experimento foi conduzido até a formação de grãos e foi realizada determinação de Zn por Espectrômetria de Absorção Atômica após digestão nitroperclórica e contagem do 65Zn nas partes da planta: parte aérea (PA) e panícula (P), para arroz e PA, vagem (V) e grão (G), para soja. Calculou-se a quantidade de Zn proveniente da fonte (Znpf) nas partes das plantas e o aproveitamento do Zn da fonte pelas culturas (Ap). Nas amostras de solo foram realizadas extrações por DTPA (ZnDTPA) e Mehlich-1 (ZnM1) em duas subamostragens (t1 e t2), antes da semeadura e florescimento, respectivamente. O fracionamento de Zn foi realizado em amostras de t2 nas frações: trocável (ZnTroc); ligado a carbonatos (ZnCarb); a matéria orgânica (ZnMO); a óxidos (ZnOxi) e residual (Znres). Adicionalmente, foi realizada análise do teor pseudo-total de Zn (ZnPST). Os dados obtidos foram submetidos à análise de variância pelo teste-F a 95 % de probabilidade, ajuste das variáveis em função das doses por regressões e teste de média e análises de correlações entre as principais variáveis respostas. O Zn acumulado total na planta se ajustou à regressão linear em função do aumento das doses, entretanto ao analisar as partes separadamente, só houve diferença entre as doses para a variável PA em ambas as culturas. O Znpf total nas plantas apresentou incremento com a adição das doses crescentes de Zn ao solo, entretanto, eu aproveitamento foi baixo, 12 e 8,75 % para arroz e soja, respectivamente. As doses de ZnCl2 adicionadas ao solo, aumentaram a concentração de Zn presente nas frações ZnTroc > ZnMO > ZnCarb, em ordem decrescente. O Zn total acumulado nas plantas de arroz e soja apresentam correlações crescentes para os extratores DTPA e M1 nas duas subamostragens (t1 e t2), em função das doses avaliadas. O Zn extraído pelo DTPA ou M1, apresentaram correlação significativa com o Zn extraído nas frações, na ordem decrescente, ZnTroc > ZnCarb > ZnMO / Zn deficiency in soil causes undesirable effects on agricultural production because the low availability of this micronutrient for plants promotes the decrease in enzymatic activity besides the deficiency of this element in food, which can lead to a state of malnutrition. Considering the problem of Zn in soil-plant system and its variations in soil compartments, it is important to evaluate its bioavailability and forms in soil which this element is associated. The objective of this study was to evaluate Zn bioavailability and compartmentalization in the soil, to the rice (Oryza sativa L.) and soybean (Glycine max L. Merrill) crops and evaluate the effect of Zn levels on nutrition and exportation of this nutrient by the crops. As test plant rice and soybeans were used to evaluate the effect of Zn rates on nutrition and the nutrient translocation to the grains. For this purpose, a sample of an Oxisol (FAO), an clay texture from Piracicaba (SP) region, was used and ZnCl2 (labelled with 65Zn) as a source. The experiment was conducted in a greenhouse in CRD, with five rates of Zn (0, 1, 2, 4 and 8 mg kg-1 soil), with four repetition. The experiment was conducted until the formation of grains and Zn analyzis were performed by Atomic Absorption Spectrophotometery after nitricperchloric digestion and 65Zn counting of parts: shoot (PA) and panicle (P) for rice and PA, beans pods (Pd) and grains (G) for soybeans. The amount of Zn derived from the source (Znpf) was calculated in the plant parts and the recovery of the Zn by the crops (Ap). In the soil samples, extractions were performed by DTPA (ZnDTPA) and Mehlich-1 (ZnM1) methods in two sub samples (t1 and t2) prior to seeding and flowering, respectively. The Zn fractionation was performed on samples of t2 in the fractions: exchangeable (ZnTroc); linked to carbonates (ZnCarb); to organic matter (ZnMO); to oxides (ZnOxi) and residual (Znres). Additionally, analysis of the pseudo-total content of Zn (ZnPST) was performed. Data obtained was subjected to analysis of variance by the F-test at 95% probability, adjusting the variables depending on the rates for regression and average test and analysis of correlations between the main variables. The total cumulative Zn in the plant set to the linear regression in terms of increasing rates, however when analyzing the parts separately, just the PA showed differences between doses, for both crops. The total Znpf increased with the increasing rates, however its recoveries were low, approximately 12% for rice and 8.75 % for soybean. The ZnCl2 rates in soil increased the Zn concentration in the fractions: ZnTroc > ZnMO > ZnCarb, in descending order. The total cumulative Zn in the rice and soybean plants are growing correlations for extractors DTPA and M1 for two sub samples (t1 and t2), depending on the rates evaluated. The Zn extracted by DTPA or M1 were significantly correlated with Zn extracted in fractions in descending order: ZnTroc > ZnCarb > ZnMO
7

Estudo de conjugação do anticorpo anti-CD20 para marcação com radionuclídeos metálicos ou lantanídeos / The study of conjugation of anti-CD20 monoclonal antibody for labeling with metalic or lanthanides radionuclides

Akanji, Akinkunmi Ganiyu 25 April 2013 (has links)
Linfomas são cânceres que se iniciam a partir da transformação maligna de um linfócito no sistema linfático. Os linfomas são divididos em duas categorias principais: os linfomas de Hodgkin e todos os outros linfomas, denominados linfomas não-Hodgkin (LNH). Os pacientes com LNH são comumente tratados com radioterapia apenas ou combinada com quimioterapia utilizando-se de anticorpo monoclonal anti-CD20, principalmente o rituximab (MabThera&reg). O uso de anticorpos monoclonais (Acm) conjugados à quelantes bifuncionais radiomarcados com radionuclídeos metálicos ou lantanídeos é uma realidade de tratamento para portadores de LNH pelo princípio de radioimunoterapia (RIT). Este estudo concentrou-se nas condições de conjugação do anticorpo monoclonal rituximab (MabThera&reg) com grupamentos quelantes bifuncionais DOTA e DTPA. Na marcação dos Acm conjugados com lutécio-177, foram estudadas as condições de pré-purificação do Acm, condições de conjugação, determinação de número de quelantes acoplados à molécula do anticorpo, purificação do anticorpo conjugado, radiomarcação do anticorpo conjugado, com lutécio-177, purificação do anticorpo marcado, a ligação específica in vitro dos compostos marcados às células Raji, e distribuição biológica em camundongos BALB/c sadios. As três metodologias empregadas na pré-purificação do anticorpo (diálise, cromatografia de exclusão molecular com coluna Sephadex G-50 e ultrafiltração) demonstram-se eficientes e proporcionaram recuperação da amostra superior a 90%. A metodologia de ultrafiltração foi considerada a mais simples e prática, podendo ser aplicada a procedimentos rotineiros de produção de radiofármacos. Além disso, proporcionou a recuperação final de amostra de 97% em microlitros. Nas conjugações do anticorpo com os quelantes DOTA e DTPA em razões molares diferentes do Acm:quelante, observou-se número de grupamentos quelantes acoplados à molécula do Acm proporcional à razão molar estudada. Quando foi avaliada a influência de condições diferentes de conjugação no número de quelantes acoplados à molécula do Acm, não foram observadas diferenças significativas, com resultados de pureza radioquímica (PR) inferior a 80% em todas as condições estudadas. Na comparação de métodos de purificação do Acm conjugado, a abordagem inédita apresentada neste estudo, na qual a cromatografia de exclusão molecular foi combinada com a ultrafiltração resultou em maior eficiência na purificação e preservação da estrutura do anticorpo. Nos estudos de radiomarcação do anticorpo conjugado com DOTA e DTPA, os imunoconjugados de DTPA apresentaram, de forma geral, maior eficiência de marcação com resultados reprodutíveis quando comparados com os imunoconjugados de DOTA, considerando-se as diferentes razões molares utilizadas. As metodologias cromatográficas empregadas no controle de pureza radioquímica do composto radiomarcado proporcionaram a discriminação das diferentes espécies radioquímicas no meio de marcação. A metodologia de purificação do composto conjugado e radiomarcado utilizada proporcionou a obtenção de compostos com alta pureza radioquímica, 97,4&plusmn;1,3% (DOTA 1:50) e 98,7&plusmn;0,2% (DTPA 1:50). Nos estudos de ligação específica às células tumorais Raji, o anticorpo conjugado com quelante DTPA nas razões molares de 1:50 e 1:20 apresentaram perfil semelhante de ligação, com aumento da porcentagem de ligação específica proporcional à concentração celular, enquanto que o imunoconjugado na razão molar de 1:10 apresentou alta porcentagem de ligação não específica. Os resultados obtidos nos estudos de biodistribuição in vivo do anticorpo conjugado e radiomarcado nem sempre se mostraram compatíveis com a biodistribuição de anticorpos radiomarcados íntegros. No caso do quelante DOTA, o imunoconjugado obtido a partir da razão molar 1:20, apresentou melhores características de biodistribuição. No caso do quelante DTPA, a razão molar utilizada pareceu refletir diretamente no clareamento sanguíneo do anticorpo e todas as razões molares utilizadas apresentaram instabilidade in vivo. / Lymphomas are malignancies or cancers that start from the malign transformation of a lymphocyte in the lymphatic system. Lymphomas are divided in two major categories: Hodgkin lymphoma and non-Hodgkin lymphoma (NHL). Patient with NHL are generally treated with radiotherapy alone or combined with immunotherapy using monoclonal antibody rituximab (MabThera&reg). Currently, monoclonal antibodies (Mab) conjugated with bifunctional chelate agents and radiolabeled with metallic or lanthanides radionuclides are a treatment reality for patients with NHL by the principle of radioimmunotherapy (RIT). This study focused on the conditions of conjugation of Acm rituximab (MabThera&reg) with bifunctional chelating agents DOTA and DTPA and labeling with 177-luthetium. Various parameters were studied: method of Acm purification, conditions of Acm conjugation and the determination of the number of chelate coupled to the Acm, the purification of the conjugated Mab, labeling conditions with lutetium-177, purification of the radiolabeled immunoconjugate, radiochemical purity (RP), in vitro specific binding determination to Raji cells (Human Burkitt) and biological distribution performed in normal BALB/c mouse. The three methodologies employed in pre-purification of Acm (dialysis, size exclusion chromatograph and ultrafiltration) demonstrated to be efficient; they provided sample recovery exceeding 90%. However, the methodology of ultrafiltration resulted in greater sample recovery and in microliters. The number of chelate attached to the Mab molecule was proportional to the molar ratio studied. When the influence of different conditions of conjugation in the number of chelate bounded to the Mab was studied, no notable differences were observed. The RP < 80% was observed in all the methods applied. Purification of the conjugated antibody by different methods showed that the innovative combination of Sephadex and ultrafiltration methods resulted in higher efficiency of purification. The optimized conditions for purification of the conjugated antibody preserved the protein integrity. Radiolabelling studies of DOTA and DTPA immunoconjugated showed that DTPA derivatives presented, in general, radiochemical yield superior than DOTA conjugated Mab, considering the different molar ratios studied. The chromatographic methods employed in the RP determination were efficient to separate the different radiochemical species presented in the reaction medium. The methodology used in the purification of the labeled Mab resulted in labeled compounds with high radiochemical purity, 97.4&plusmn;1.3% (DOTA 1:50) and 98.7&plusmn;0.2% (DTPA 1:50). Considering specific cell binding assays (Raji cells), the Mab conjugated to DTPA at 1:50 and 1:20 molar ratios presented similar results, and the percent of cell binding were proportional to the cell concentration, whereas the cell binding for 1:10 molar ratio showed high percent of nonspecific cell binding. The results of in vivo biodistribution studies of labeled Mab not always were compatible with the biodistribution of intact radiolabelled antibody. The DOTA immunoconjugated produced at 1:20 molar ratio, showed better performance in biodistribution studies. In the case of DTPA immunoconjugated, the blood clearance seems to be influenced by the molar ratio applied and the immunoconjugated produced with DTPA chelate at different molar ratio resulted in high in vivo instability compounds.
8

Estudo com radioaerossol de DTPA Tecnécio-99m em pacientes portadores de pneumopatia por amiodarona / Study with radiolabeled aerosol 99mTc-DTPA in patients with with amiodarone induced pulmonary disease

Terra Filho, Mario 16 June 1989 (has links)
Com o objetivo de se avaliar a importância do \"clearance\" do dietilenotriamino-pentacetato marcado com Tecnécio 99m (DTPA-Tecnécio-99m) em portadores de pneumopatia por amiodarona foram estudados 40 indivíduos, em quatro grupos. Grupo I: 10 voluntários normais, assintomáticos e não fumantes (8 homens e 2 mulheres), com média de idade de 56,80 anos. Grupo II: 10 voluntários normais, assintomáticos e fumantes (6 homens e 4 mulheres ), com média de idade de 27,50 anos. Grupo III: 10 pacientes não fumantes ( 4 homens e 5 mulheres ), com média de idade de 52,90 anos. Todos faziam uso crônico de amiodarona por via oral. Grupo IV: 10 pacientes portadores de pneumopatia por amiodarona, quatro ex-fumantes, dois fumantes e quatro não fumantes ( 8 homens e 2 mulheres) com média de idade de 52,90 anos. Todos faziam uso de amiodarona por via oral e nenhum fumou nas 4 semanas que precederam o estudo. Após espirometria que constou do registro da curva volume-tempo, todos inalaram 4 ml de solução salina contendo 740 MBq de DTPA Tecnécio-99m, durante cinco minutos. Através de uma c~mara de cintilação computadorizada foram obtidas imagens pulmonares, definindo-se 9 áreas de interesse. Para cada região escolhida foi determinada uma curva de \"clearance\" extraindo-se o valor de meia-vida biológica em minu- tos ( T 1/2 ) e a taxa percentual de \" clearance\" alvéolo capilar do radioaerossol por minuto (K%/min). Observamos que, das variáveis espirométricas consideradas, a capacidade vital forçada (CVF) e o volume expiratório forçado no 1 segundo (VEF1) mostraram diferenças significantes entre os grupos I e IV. A contagem total de radioatividade de ambos os pulmões não mostrou relação com a CVF e o VEF1. O \" clearance \" pulmonar do DTPA Tecnécio-99m foi maior nos grupos 11 e IV, porém não permitindo sua diferenciação. Estes resultados permitem concluir: Os pacientes portadores de pneumonite por amiodaro- na apresentam\" clearance \" alvéolo-capilar de DTPA Tecnécio-99m significativamente maior que os indivíduos do grupo de normais não fumantes. Este fato também se verificou em relação aos pacientes em uso crônico de amiodarona mas sem evidências de pneumopatia. Não é possível diferenciar os fumantes dos portadores de pneumonite por amiodarona através da análise da integridade da barreira alvéolo-epitelial com DTPA Tecnécio-99m. Comparativamente, o estudo da integridade alvéolo-epitelial pelo \"clearance\" pulmonar de DTPA Tecnécio-99m é mais sensível que a espirometria na avaliação da pneumonite por amiodarona, permitindo diferenciar estes pacientes dos que fazem uso crônico da droga / In order to evaluate the role of the clearance of 99m Technetium chelated to diethylenetriamine-penta-acetate ( 99mTc-DTPA) in amiodarone induced pulmonary disease, 40 individuaIs were studied in four groups. Group I: 10 normal non smoking volunteers (8 men and 2 women ), whose mean age was 56.80 years. Group lI: 10 normal smoking volunteers ( 6 men and 4 women ), aging 27.50 years in average. Group III: 10 non smoking patients ( 4 men and women ), aging 52.90 years in average, who were chronically taking oral amiodarone. Group IV: 10 patients with amiodarone induced pul- monary disease (8 men and 2 women), four non-smokers, two smokers and four previous smokers. Their mean age was 62.90 years. AIl of them were taking oral amiodarone and none has smoked in the 4 weeks previous to the study. After spirometry, where a volume-time curve was registered, alI individuaIs inhaled 740 MBq of 99mTc-DTPA diluted in 4 ml of saline, for five minutes. Pulmonary images were obtained in a computadorized scintillation camera and 9 regions of interest were selected. A clearance curve of each region was determined, from which the effective half-life in minutes (T 1/2 ) and the alveolar-capilar clearance rate per minute ( k%/min ) of the radiolabeled aerosol were mea- sured. The spirometric analys disclosed a statistically lower value of the forced vital capacity ( FVC ) and forced expiratory volume in the first second ( FEV1 ) in the patients of group IV when compared to group I. The total radioactivity count for both lungs were not influenced by FVC and FEV1. The 99mTc-DTPA clearance rate was higher in groups 11 and IV, but these two groups could not be statistically differentiated. Based on these results it 1s concluded: patients with amiodarone induced pulmonary pneumonitis have higher clearance rates of 99m Tc-DTPA than normal non smoking controls and than patients taking amiodarone but with no lung toxicity. It is not possible to separate patients with amiodarone induced disease from normal smokers by determining 99m Tc-DTPA clearance rates. The determination of the alveolar-epithelial barrier integrity by 99m Tc-DTPA clearance rate is a more sensitive test than spirometry in the evaluation of amiodarone induced pneumonitis making it possible to differentiate these patients from those who take the drug and have no lung toxicity
9

Estudo com radioaerossol de DTPA Tecnécio-99m em pacientes portadores de pneumopatia por amiodarona / Study with radiolabeled aerosol 99mTc-DTPA in patients with with amiodarone induced pulmonary disease

Mario Terra Filho 16 June 1989 (has links)
Com o objetivo de se avaliar a importância do \"clearance\" do dietilenotriamino-pentacetato marcado com Tecnécio 99m (DTPA-Tecnécio-99m) em portadores de pneumopatia por amiodarona foram estudados 40 indivíduos, em quatro grupos. Grupo I: 10 voluntários normais, assintomáticos e não fumantes (8 homens e 2 mulheres), com média de idade de 56,80 anos. Grupo II: 10 voluntários normais, assintomáticos e fumantes (6 homens e 4 mulheres ), com média de idade de 27,50 anos. Grupo III: 10 pacientes não fumantes ( 4 homens e 5 mulheres ), com média de idade de 52,90 anos. Todos faziam uso crônico de amiodarona por via oral. Grupo IV: 10 pacientes portadores de pneumopatia por amiodarona, quatro ex-fumantes, dois fumantes e quatro não fumantes ( 8 homens e 2 mulheres) com média de idade de 52,90 anos. Todos faziam uso de amiodarona por via oral e nenhum fumou nas 4 semanas que precederam o estudo. Após espirometria que constou do registro da curva volume-tempo, todos inalaram 4 ml de solução salina contendo 740 MBq de DTPA Tecnécio-99m, durante cinco minutos. Através de uma c~mara de cintilação computadorizada foram obtidas imagens pulmonares, definindo-se 9 áreas de interesse. Para cada região escolhida foi determinada uma curva de \"clearance\" extraindo-se o valor de meia-vida biológica em minu- tos ( T 1/2 ) e a taxa percentual de \" clearance\" alvéolo capilar do radioaerossol por minuto (K%/min). Observamos que, das variáveis espirométricas consideradas, a capacidade vital forçada (CVF) e o volume expiratório forçado no 1 segundo (VEF1) mostraram diferenças significantes entre os grupos I e IV. A contagem total de radioatividade de ambos os pulmões não mostrou relação com a CVF e o VEF1. O \" clearance \" pulmonar do DTPA Tecnécio-99m foi maior nos grupos 11 e IV, porém não permitindo sua diferenciação. Estes resultados permitem concluir: Os pacientes portadores de pneumonite por amiodaro- na apresentam\" clearance \" alvéolo-capilar de DTPA Tecnécio-99m significativamente maior que os indivíduos do grupo de normais não fumantes. Este fato também se verificou em relação aos pacientes em uso crônico de amiodarona mas sem evidências de pneumopatia. Não é possível diferenciar os fumantes dos portadores de pneumonite por amiodarona através da análise da integridade da barreira alvéolo-epitelial com DTPA Tecnécio-99m. Comparativamente, o estudo da integridade alvéolo-epitelial pelo \"clearance\" pulmonar de DTPA Tecnécio-99m é mais sensível que a espirometria na avaliação da pneumonite por amiodarona, permitindo diferenciar estes pacientes dos que fazem uso crônico da droga / In order to evaluate the role of the clearance of 99m Technetium chelated to diethylenetriamine-penta-acetate ( 99mTc-DTPA) in amiodarone induced pulmonary disease, 40 individuaIs were studied in four groups. Group I: 10 normal non smoking volunteers (8 men and 2 women ), whose mean age was 56.80 years. Group lI: 10 normal smoking volunteers ( 6 men and 4 women ), aging 27.50 years in average. Group III: 10 non smoking patients ( 4 men and women ), aging 52.90 years in average, who were chronically taking oral amiodarone. Group IV: 10 patients with amiodarone induced pul- monary disease (8 men and 2 women), four non-smokers, two smokers and four previous smokers. Their mean age was 62.90 years. AIl of them were taking oral amiodarone and none has smoked in the 4 weeks previous to the study. After spirometry, where a volume-time curve was registered, alI individuaIs inhaled 740 MBq of 99mTc-DTPA diluted in 4 ml of saline, for five minutes. Pulmonary images were obtained in a computadorized scintillation camera and 9 regions of interest were selected. A clearance curve of each region was determined, from which the effective half-life in minutes (T 1/2 ) and the alveolar-capilar clearance rate per minute ( k%/min ) of the radiolabeled aerosol were mea- sured. The spirometric analys disclosed a statistically lower value of the forced vital capacity ( FVC ) and forced expiratory volume in the first second ( FEV1 ) in the patients of group IV when compared to group I. The total radioactivity count for both lungs were not influenced by FVC and FEV1. The 99mTc-DTPA clearance rate was higher in groups 11 and IV, but these two groups could not be statistically differentiated. Based on these results it 1s concluded: patients with amiodarone induced pulmonary pneumonitis have higher clearance rates of 99m Tc-DTPA than normal non smoking controls and than patients taking amiodarone but with no lung toxicity. It is not possible to separate patients with amiodarone induced disease from normal smokers by determining 99m Tc-DTPA clearance rates. The determination of the alveolar-epithelial barrier integrity by 99m Tc-DTPA clearance rate is a more sensitive test than spirometry in the evaluation of amiodarone induced pneumonitis making it possible to differentiate these patients from those who take the drug and have no lung toxicity
10

Estudo de conjugação do anticorpo anti-CD20 para marcação com radionuclídeos metálicos ou lantanídeos / The study of conjugation of anti-CD20 monoclonal antibody for labeling with metalic or lanthanides radionuclides

Akinkunmi Ganiyu Akanji 25 April 2013 (has links)
Linfomas são cânceres que se iniciam a partir da transformação maligna de um linfócito no sistema linfático. Os linfomas são divididos em duas categorias principais: os linfomas de Hodgkin e todos os outros linfomas, denominados linfomas não-Hodgkin (LNH). Os pacientes com LNH são comumente tratados com radioterapia apenas ou combinada com quimioterapia utilizando-se de anticorpo monoclonal anti-CD20, principalmente o rituximab (MabThera&reg). O uso de anticorpos monoclonais (Acm) conjugados à quelantes bifuncionais radiomarcados com radionuclídeos metálicos ou lantanídeos é uma realidade de tratamento para portadores de LNH pelo princípio de radioimunoterapia (RIT). Este estudo concentrou-se nas condições de conjugação do anticorpo monoclonal rituximab (MabThera&reg) com grupamentos quelantes bifuncionais DOTA e DTPA. Na marcação dos Acm conjugados com lutécio-177, foram estudadas as condições de pré-purificação do Acm, condições de conjugação, determinação de número de quelantes acoplados à molécula do anticorpo, purificação do anticorpo conjugado, radiomarcação do anticorpo conjugado, com lutécio-177, purificação do anticorpo marcado, a ligação específica in vitro dos compostos marcados às células Raji, e distribuição biológica em camundongos BALB/c sadios. As três metodologias empregadas na pré-purificação do anticorpo (diálise, cromatografia de exclusão molecular com coluna Sephadex G-50 e ultrafiltração) demonstram-se eficientes e proporcionaram recuperação da amostra superior a 90%. A metodologia de ultrafiltração foi considerada a mais simples e prática, podendo ser aplicada a procedimentos rotineiros de produção de radiofármacos. Além disso, proporcionou a recuperação final de amostra de 97% em microlitros. Nas conjugações do anticorpo com os quelantes DOTA e DTPA em razões molares diferentes do Acm:quelante, observou-se número de grupamentos quelantes acoplados à molécula do Acm proporcional à razão molar estudada. Quando foi avaliada a influência de condições diferentes de conjugação no número de quelantes acoplados à molécula do Acm, não foram observadas diferenças significativas, com resultados de pureza radioquímica (PR) inferior a 80% em todas as condições estudadas. Na comparação de métodos de purificação do Acm conjugado, a abordagem inédita apresentada neste estudo, na qual a cromatografia de exclusão molecular foi combinada com a ultrafiltração resultou em maior eficiência na purificação e preservação da estrutura do anticorpo. Nos estudos de radiomarcação do anticorpo conjugado com DOTA e DTPA, os imunoconjugados de DTPA apresentaram, de forma geral, maior eficiência de marcação com resultados reprodutíveis quando comparados com os imunoconjugados de DOTA, considerando-se as diferentes razões molares utilizadas. As metodologias cromatográficas empregadas no controle de pureza radioquímica do composto radiomarcado proporcionaram a discriminação das diferentes espécies radioquímicas no meio de marcação. A metodologia de purificação do composto conjugado e radiomarcado utilizada proporcionou a obtenção de compostos com alta pureza radioquímica, 97,4&plusmn;1,3% (DOTA 1:50) e 98,7&plusmn;0,2% (DTPA 1:50). Nos estudos de ligação específica às células tumorais Raji, o anticorpo conjugado com quelante DTPA nas razões molares de 1:50 e 1:20 apresentaram perfil semelhante de ligação, com aumento da porcentagem de ligação específica proporcional à concentração celular, enquanto que o imunoconjugado na razão molar de 1:10 apresentou alta porcentagem de ligação não específica. Os resultados obtidos nos estudos de biodistribuição in vivo do anticorpo conjugado e radiomarcado nem sempre se mostraram compatíveis com a biodistribuição de anticorpos radiomarcados íntegros. No caso do quelante DOTA, o imunoconjugado obtido a partir da razão molar 1:20, apresentou melhores características de biodistribuição. No caso do quelante DTPA, a razão molar utilizada pareceu refletir diretamente no clareamento sanguíneo do anticorpo e todas as razões molares utilizadas apresentaram instabilidade in vivo. / Lymphomas are malignancies or cancers that start from the malign transformation of a lymphocyte in the lymphatic system. Lymphomas are divided in two major categories: Hodgkin lymphoma and non-Hodgkin lymphoma (NHL). Patient with NHL are generally treated with radiotherapy alone or combined with immunotherapy using monoclonal antibody rituximab (MabThera&reg). Currently, monoclonal antibodies (Mab) conjugated with bifunctional chelate agents and radiolabeled with metallic or lanthanides radionuclides are a treatment reality for patients with NHL by the principle of radioimmunotherapy (RIT). This study focused on the conditions of conjugation of Acm rituximab (MabThera&reg) with bifunctional chelating agents DOTA and DTPA and labeling with 177-luthetium. Various parameters were studied: method of Acm purification, conditions of Acm conjugation and the determination of the number of chelate coupled to the Acm, the purification of the conjugated Mab, labeling conditions with lutetium-177, purification of the radiolabeled immunoconjugate, radiochemical purity (RP), in vitro specific binding determination to Raji cells (Human Burkitt) and biological distribution performed in normal BALB/c mouse. The three methodologies employed in pre-purification of Acm (dialysis, size exclusion chromatograph and ultrafiltration) demonstrated to be efficient; they provided sample recovery exceeding 90%. However, the methodology of ultrafiltration resulted in greater sample recovery and in microliters. The number of chelate attached to the Mab molecule was proportional to the molar ratio studied. When the influence of different conditions of conjugation in the number of chelate bounded to the Mab was studied, no notable differences were observed. The RP < 80% was observed in all the methods applied. Purification of the conjugated antibody by different methods showed that the innovative combination of Sephadex and ultrafiltration methods resulted in higher efficiency of purification. The optimized conditions for purification of the conjugated antibody preserved the protein integrity. Radiolabelling studies of DOTA and DTPA immunoconjugated showed that DTPA derivatives presented, in general, radiochemical yield superior than DOTA conjugated Mab, considering the different molar ratios studied. The chromatographic methods employed in the RP determination were efficient to separate the different radiochemical species presented in the reaction medium. The methodology used in the purification of the labeled Mab resulted in labeled compounds with high radiochemical purity, 97.4&plusmn;1.3% (DOTA 1:50) and 98.7&plusmn;0.2% (DTPA 1:50). Considering specific cell binding assays (Raji cells), the Mab conjugated to DTPA at 1:50 and 1:20 molar ratios presented similar results, and the percent of cell binding were proportional to the cell concentration, whereas the cell binding for 1:10 molar ratio showed high percent of nonspecific cell binding. The results of in vivo biodistribution studies of labeled Mab not always were compatible with the biodistribution of intact radiolabelled antibody. The DOTA immunoconjugated produced at 1:20 molar ratio, showed better performance in biodistribution studies. In the case of DTPA immunoconjugated, the blood clearance seems to be influenced by the molar ratio applied and the immunoconjugated produced with DTPA chelate at different molar ratio resulted in high in vivo instability compounds.

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