O fluoroacetato de sódio (FAS), também conhecido como \"composto 1080\", foi um raticida amplamente utilizado no combate de roedores a partir da década de 40, mas que teve seu uso proibido em diversos países, incluindo o Brasil, devido a sua alta toxicidade e ausência de tratamento efetivo nas intoxicações. A despeito deste fato, diversos casos de intoxicação ainda são registrados em seres humanos e animais, em virtude de seu uso ilícito como raticida doméstico ou adição em iscas nas intoxicações criminosas. O FAS, na natureza, pode ser encontrado, também, em algumas plantas tóxicas de grande importância no Brasil, como a Palicourea marcgravii, e possivelmente seja o princípio ativo tóxico de outras plantas de relevância, como a Mascagnia rigida. Neste sentido, este trabalho objetivou o estudo dos aspectos clínicos e anatomopatológicos encontrados nas intoxicações provocadas pelo FAS, em ratos, comparando estes com os encontrados nas intoxicações por extrato aquoso da M. rígida. Também foram comparadas metodologias analíticas para identificação deste agente tóxico utilizando cromatografia em camada delgada (CCD) e cromatografia de alta eficiência (CLAE) com dois tipos de detectores o de UV e o de Condutividade Iônica. Para tanto, quatro grupos contendo seis ratos Wistar, machos, receberam em dose única por via oral 7,0 mg/kg de FAS ou água destilada ou ainda 1,6 ou 3,2 g/kg de extrato aquoso de M. rígida, sendo avaliados os sinais clínicos apresentados por estes animais. Após eutanásia foram coletados fragmentos de cérebro, coração, fígado, rins, pulmão, intestino e baço para exame histopatológico. Tanto os animais que receberam FAS como os que receberam extrato aquoso de M. rígida apresentaram sintomatologia similar com presença de prostração, desconforto respiratório, tremores, pêlos arrepiados, cromodacriorreia, convulsão tônica e tônico-clônica. Na necropsia destes animais não foram observadas alterações macroscópicas, contudo alterações histopatológicas significativas foram encontradas no cérebro, miocárdio, fígado e rins. Os estudos das análise toxicológicas utilizando as três técnicas cromatográficas mostraram que os resultados obtidos na CCD foram adequados para detecção de FAS nas concentrações acima de 1,0 μg do padrão puro e as análises das duas plantas estudadas apresentaram resultados positivos para FAS. As técnicas utilizando CLAE, com dois diferentes detectores, mostraram que quando se utiliza detector de UV o limite de detecção é de 3,75 μg injetado ou de 75 μg/mL e foi possível, também, detectar a presença de FAS nas duas plantas estudadas, porém a técnica não se apresentou adequada para a detecção de intoxicação em tecido animal. O detector de condutividade iônica mostrou limite de detecção de 0,30 μg/mL, isto é, o mais sensível entre os três métodos estudados, e portanto o mais adequado para diagnóstico de intoxicação em tecido animal; no entanto, para identificação do FAS na M. rigida este método não se mostrou adequado devido a interferentes no extrato aquoso e no soro de animais intoxicados. / Sodium fluoroacetate (SFA), also known as \"compound 1080,\" was a rodenticide widely used in the 1940s, but its use was banned in several countries, including Brazil, for its high toxicity and lack of an effective treatment in poisoning. Despite this fact, several cases of poisoning are still registered in humans and animals, because of their misuse as baits, in addition to criminal poisoning. SFA, in the form of natural molecules, can be found in some toxic plants of great importance in Brazil, such as Palicourea marcgravii, and possibly is the active principle of other plants of relevance, such as Mascagnia rigid. Therefore, this study aimed to evaluate the clinical and pathological findings in poisoning caused by the SFA in rats, and compare them with those found in poisoning by the aqueous extract of M. rigid. Moreover, we compared analytical methods for identifying this toxic agent using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) employing two types of detectors: ultraviolet (UV) and ionic conductivity. Therefore, four groups containing six male Wistar rats each, received a single oral dose of 7.0 mg / kg of SFA, an equal volume of distilled water, or 1.6 or 3.2 g / kg of an aqueous extract of M. rigid, and the clinical signs displayed by these animals were evaluated. After euthanasia fragments of brain, heart, liver, kidney, lung, intestine and spleen were collected for histological examination. Both the animals that received SFA and those who received the aqueous extract of M. rigid showed similar symptoms, including prostration, respiratory distress, tremors, shivering, and tonic or tonic-clonic seizures. At necropsy, these animals did not display macroscopic changes. Nevertheless, significant histopathological changes were found in the brain, heart, liver and kidneys. Toxicological analysis using the three chromatographic techniques described above showed that the results obtained in the TLC were suitable for detection of SFA at concentrations above 1.0 μg of pure standard, and analysis of the two plants mentioned showed positive results for SFA. The techniques using HPLC with two different detectors showed that when using the UV detector, the detection limit is of 3.75 μg (injected) or 75 μg / mL, and SFA could also be detected by these methods in the two plants studied, but the technique does not seem suitable for the detection of intoxication in animal tissue samples. The ionic conductivity detector showed a detection limit of 0.30 μg/ mL, therefore the most sensitive of the three methods studied, and hence is the most appropriate for diagnosis of poisoning in animal tissue; however, for the identification of SFA in the M. rigid, this method was not appropriate due to a putative interference in the aqueous extract.

A tomografia por emissão de pósitrons associada à tomografia computadorizada (PET/CT) é um dispositivo que combina as características de medicina nuclear (PET) e de radiologia (CT) obtendo imagens metabólicas (PET) e anatômicas sobrepostas (CT). Combinando as duas tecnologias de exames, o exame PET / CT permite aos médicos diagnosticar com maior precisão e identificar o câncer, doenças cardíacas e distúrbios cerebrais. O radiofármaco 18FFAc (fluoroacetato) é promissor para a detecção de tumores primários de próstata e de mama, utilizando a técnica de PET/CT. Estudos recentes mostram a eficácia do 18F-FAc na detecção de tumores que têm baixa captação de 18F-FDG (fluordesoxiglicose). O fluoroacetato é um substrato para a acetil-CoA sintase, enzima que metaboliza ácido fluorocitrato que não é mais metabolizado, levando à inibição da aconitase e do ácido tricarboxílico. O objetivo deste trabalho foi desenvolver um radiofármaco emissor de pósitron, 18F-FAc no IPEN-CNEN/SP em um acordo com o Hospital AC-Camargo / São Paulo. O íon fluoreto (18F-) foi produzido, usando os cíclotrons Cyclone 30 e 18 da IBA localizados no IPEN-CNEN/SP, através da irradiação de água enriquecida em 18O com prótons e dose integrada de 30μAh. A marcação do 18F-FAc foi realizada em um módulo de síntese TRACERlab MXFDG (GE), utilizando kits adquiridos da ABX. O controle de qualidade radioquímico de 18F-FAc foi realizado por cromatografia em camada fina TLC-SG 25 folhas de alumínio em tiras (1,5 x 12 cm ) usando como solvente clorofórmio:metanol (1:1). Para o controle de qualidade radionuclídico, amostras de 18F-FAc e 18F-Fluoreto foram analisadas por espectroscopia de raios-gama. A avaliação dos solventes residuais foi realizada por cromatografia em fase gasosa e a análise de kryptofix foi realizada por TLC utilizando tiras de TLC-SG, metanol:clorofórmio (9:1) como solvente e padrões de kryptofix 2.2.2. Os estudos de biodistribuição foram realizados com 18FFAc injetado em camundongos swiss sadios. Um procedimento reprodutível foi desenvolvido para o preparo do 18F-FAc com um rendimento de marcação de 37% (não corrigido) e 52% (corrigido para o decaimento) e estabilidade de 19 horas. A análise de controle de qualidade mostrou que o produto tinha as exigências adequadas para utilização, com pureza radioquímica superior a 99,9%. Os estudos de biodistribuição em animais sadios mostraram a esperada captação em todos os órgãos medidos com eliminação renal e intestinal. / PET / CT (positron emission tomography / computed tomography) is a device that combines the features of diagnostic nuclear medicine (PET) and Radiology (CT) superimposing metabolic (PET) and anatomical (CT) images. By combining the two technologies examinations, the PET/CT scan allows physicians to diagnose more accurately and identify cancer, heart disease and brain disorders. The radiopharmaceutical 18F-FAc (fluoroacetate) is promising for application in detection of primary tumors of prostate and breast, using PET-CT techniques. Recent studies are showing the efficacy of the 18F-FAc in the detection of tumors that have low uptake of 18F-FDG (fluorodeoxyglucose). The fluoroacetate is a substrate for the enzyme acetyl-CoA synthase that metabolizes acid fluorcitric that, not being metabolized, causes inhibition of aconitase and inhibition of tricarboxylic acid. The aim of this work was to develop a positron emitting radiopharmaceutical, 18F-FAc at IPEN-CNEN/SP in agreement with Hospital AC-Camargo/ São Paulo. The 18F-fluoride ion was produced using the Cyclone 30 and 18 cyclotrons from IBA located at IPEN-CNEN/SP, by irradiating enriched 18O water with protons with integrated dose 30μAh. The labelling of 18F-FAc was performed in a synthesis module TRACERlab MXFDG (GE), using kits purchased from ABX. The radiochemical quality control of 18F-FAc was performed by Thin Layer Chromatography using TLC-SG 25 aluminium sheets strips (1.5 x 12 cm) and chloroform:methanol (1:1) as the solvent. For the radionuclidic quality control, samples of 18F-FAc and 18F-Fluoride were analysed by gama-ray spectroscopy. The evaluation of the residual solvents was performed by gas chromatography and the analysis of kryptofix was performed by TLC using TLC-SG strips, methanol:chloroform (9:1) as solvent and kryptofix 2.2.2 standards. Biodistribution studies were performed with 18F-FAc injected into healthy Swiss mice. A reliable procedure was developed for preparation of 18F-FAc with a labelling yield of 37% (uncorrected) and 52% (corrected for decay) and stability of 19 hours. The quality control analysis showed that the product had the proper requirements for use, with radiochemical purity exceeding 99.9%. The biodistribution studies in healthy animals showed the expected uptake results in all the measured organs with intestinal and renal elimination.

A tomografia por emissão de pósitrons associada à tomografia computadorizada (PET/CT) é um dispositivo que combina as características de medicina nuclear (PET) e de radiologia (CT) obtendo imagens metabólicas (PET) e anatômicas sobrepostas (CT). Combinando as duas tecnologias de exames, o exame PET / CT permite aos médicos diagnosticar com maior precisão e identificar o câncer, doenças cardíacas e distúrbios cerebrais. O radiofármaco 18FFAc (fluoroacetato) é promissor para a detecção de tumores primários de próstata e de mama, utilizando a técnica de PET/CT. Estudos recentes mostram a eficácia do 18F-FAc na detecção de tumores que têm baixa captação de 18F-FDG (fluordesoxiglicose). O fluoroacetato é um substrato para a acetil-CoA sintase, enzima que metaboliza ácido fluorocitrato que não é mais metabolizado, levando à inibição da aconitase e do ácido tricarboxílico. O objetivo deste trabalho foi desenvolver um radiofármaco emissor de pósitron, 18F-FAc no IPEN-CNEN/SP em um acordo com o Hospital AC-Camargo / São Paulo. O íon fluoreto (18F-) foi produzido, usando os cíclotrons Cyclone 30 e 18 da IBA localizados no IPEN-CNEN/SP, através da irradiação de água enriquecida em 18O com prótons e dose integrada de 30μAh. A marcação do 18F-FAc foi realizada em um módulo de síntese TRACERlab MXFDG (GE), utilizando kits adquiridos da ABX. O controle de qualidade radioquímico de 18F-FAc foi realizado por cromatografia em camada fina TLC-SG 25 folhas de alumínio em tiras (1,5 x 12 cm ) usando como solvente clorofórmio:metanol (1:1). Para o controle de qualidade radionuclídico, amostras de 18F-FAc e 18F-Fluoreto foram analisadas por espectroscopia de raios-gama. A avaliação dos solventes residuais foi realizada por cromatografia em fase gasosa e a análise de kryptofix foi realizada por TLC utilizando tiras de TLC-SG, metanol:clorofórmio (9:1) como solvente e padrões de kryptofix 2.2.2. Os estudos de biodistribuição foram realizados com 18FFAc injetado em camundongos swiss sadios. Um procedimento reprodutível foi desenvolvido para o preparo do 18F-FAc com um rendimento de marcação de 37% (não corrigido) e 52% (corrigido para o decaimento) e estabilidade de 19 horas. A análise de controle de qualidade mostrou que o produto tinha as exigências adequadas para utilização, com pureza radioquímica superior a 99,9%. Os estudos de biodistribuição em animais sadios mostraram a esperada captação em todos os órgãos medidos com eliminação renal e intestinal. / PET / CT (positron emission tomography / computed tomography) is a device that combines the features of diagnostic nuclear medicine (PET) and Radiology (CT) superimposing metabolic (PET) and anatomical (CT) images. By combining the two technologies examinations, the PET/CT scan allows physicians to diagnose more accurately and identify cancer, heart disease and brain disorders. The radiopharmaceutical 18F-FAc (fluoroacetate) is promising for application in detection of primary tumors of prostate and breast, using PET-CT techniques. Recent studies are showing the efficacy of the 18F-FAc in the detection of tumors that have low uptake of 18F-FDG (fluorodeoxyglucose). The fluoroacetate is a substrate for the enzyme acetyl-CoA synthase that metabolizes acid fluorcitric that, not being metabolized, causes inhibition of aconitase and inhibition of tricarboxylic acid. The aim of this work was to develop a positron emitting radiopharmaceutical, 18F-FAc at IPEN-CNEN/SP in agreement with Hospital AC-Camargo/ São Paulo. The 18F-fluoride ion was produced using the Cyclone 30 and 18 cyclotrons from IBA located at IPEN-CNEN/SP, by irradiating enriched 18O water with protons with integrated dose 30μAh. The labelling of 18F-FAc was performed in a synthesis module TRACERlab MXFDG (GE), using kits purchased from ABX. The radiochemical quality control of 18F-FAc was performed by Thin Layer Chromatography using TLC-SG 25 aluminium sheets strips (1.5 x 12 cm) and chloroform:methanol (1:1) as the solvent. For the radionuclidic quality control, samples of 18F-FAc and 18F-Fluoride were analysed by gama-ray spectroscopy. The evaluation of the residual solvents was performed by gas chromatography and the analysis of kryptofix was performed by TLC using TLC-SG strips, methanol:chloroform (9:1) as solvent and kryptofix 2.2.2 standards. Biodistribution studies were performed with 18F-FAc injected into healthy Swiss mice. A reliable procedure was developed for preparation of 18F-FAc with a labelling yield of 37% (uncorrected) and 52% (corrected for decay) and stability of 19 hours. The quality control analysis showed that the product had the proper requirements for use, with radiochemical purity exceeding 99.9%. The biodistribution studies in healthy animals showed the expected uptake results in all the measured organs with intestinal and renal elimination.

The use of positron emission tomography (PET) for imaging in oncology has grown rapidly in recent years. 2-[18F]-fluorodeoxyglucose (FDG) is the most common tracer of PET, although drawbacks exist. Radiolabeled 1-[11C]-acetate (C-AC) is a simple probe for evaluation of perfusion, anabolism (lipogenesis) and catabolism (oxidative metabolism) in all living tissues. This study explored the potential of AC PET in head and neck cancer, benign and malignant lymph nodes in prostate cancer and normal distribution.  In head and neck cancer, C-AC PET detected more primaries and lymph node metastases than FDG PET. The mean primary tumor volumes delineated by C-AC was 51% larger than that of FDG before radiotherapy (RT). Both FDG and C-AC PET tumor volumes must be carefully validated before used in clinical routine. Baseline tumor clearance rate (kmono) was higher in complete responders (CR) than that in partial responders (PR). kmono tended to correlate inversely with FDG SUV at baseline. Radiosensitive tumors might rely predominantly on oxidative metabolism for their biogenetic needs. kmono increased in PR during RT. The potential reversibility of impaired kmono in radioresistant tumors imply that treatment targeting the intermediary metabolism might improve the outcome. Tumor relative perfusion index (rF) and kmono were coupled in CR throughout the RT, but not in PR. Dynamic C-AC PET provides a new non-invasive method to simultaneously evaluate the tumor oxidative metabolism and perfusion which link the RT response in patients by a single tracer injection. In prostate cancer, elevated C-AC accumulation is common in benign inguinal lymph nodes, probably due to increased lipogenesis rather than lymphatic drainage. CT Hounsfield unit of benign nodes was lower than that of metastases, suggesting that density measurement using CT might improve the specificity of nodal staging of prostate cancer. A novel tracer 2-[18F]-fluoroacetate (F-AC) was synthesized and used for dynamic PET-CT imaging in animals. Compared with C-AC PET-CT, F-AC showed prolonged blood retention, no detectable trapping in myocardium and salivary glands, rapid excretion from liver to bile and urine and de-fluorination resulting in intensive skeletal activity. F-AC does not mimic the normal physiologic path of C-AC and appears to be of little use for assessment of perfusion, intermediary metabolism or lipogenesis.

Fluorotelomer alcohols (FTOHs, F(CF2)nCH2CH2OH) are emerging contaminants in the environment. Biodegradation of 6:2 and 8:2 FTOHs has been intensively studied using soils and activated sludge. However, little is known about the bacteria responsible for biotransformation of FTOHs. This study deciphered factors affecting biodefluorination of FTOHs and their metabolites, and developed three effective FTOH-degrading consortia. Two alkane-degrading Pseudomonas strains (P. oleovorans and P. butanovora) can defluorinate 4:2, 6:2 and 8:2 FTOHs, with a higher degree of defluorination for 4:2 FTOH. According to the identified metabolites, P. oleovorans transformed FTOHs via two pathways I and II. Pathway I led to formation of x:2 ketone (x = n-1), x:2 sFTOH and perfluorinated carboxylic acids (PFCAs). Pathway II resulted in the formation of x:3 polyfluorinated acid and relatively minor shorter-chain PFCAs. Conversely, P. butanovora transformed FTOHs by pathway I only. Mycobacterium vaccae JOB5 (a C1-C22alkane-degrading bacterium) and P. fluorescens DSM 8341 (a fluoroacetate-degrading bacterium) can transform 6:2 FTOH via both pathways I and II with the formation of odd-numbered short-chain PFCAs. In the presence of dicyclopropylketone or formate, P. oleovorans transformed 6:2 FTOH six times faster and produced odd-numbered PFCAs. P. butanovora, utilized both pathways I and II in the presence of lactate, and it also produced odd-numbered PFCAs. Unlike P. oleovorans, P. fluorescens DSM 8341 could slightly convert 5:3 polyfluorinated acid (a key metabolite during 6:2 FTOH degradation, [F(CF2)5CH2CH2COOH]) to 4:3 acid and PFPeA via one-carbon removal pathways. Three FTOH-degrading consortia transformed FTOHs, with enhanced removal of FTOHs in the presence of n-octane. A higher copy number of alkB gene was found to correspond to better removal of FTOHs, suggesting that alkane-degrading bacteria might be the key degraders in the enrichments. The three enrichment cultures showed a similar microbial community structure. This is the first study reporting that pure strains of alkane- and fluoroacetate-degrading bacteria can bio-transform FTOHs via different or preferred transformation pathways to remove multiple –CF2– groups from FTOHs to form shorter-chain PFCAs, and to other perfluorinated acids. The results of this study also suggest that enhanced FTOH biodegradation is possible through co-substrate addition and/or using enrichment cultures.

Submitted by Jordan (jordanbiblio@gmail.com) on 2017-10-04T14:12:15Z No. of bitstreams: 1 DISS_2015_Geovanny Bruno Gonçalves Dias.pdf: 1405701 bytes, checksum: 24fbe33b137779f7f8668a0dfcb69dd5 (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2017-10-04T14:12:38Z (GMT) No. of bitstreams: 1 DISS_2015_Geovanny Bruno Gonçalves Dias.pdf: 1405701 bytes, checksum: 24fbe33b137779f7f8668a0dfcb69dd5 (MD5) / Made available in DSpace on 2017-10-04T14:12:38Z (GMT). No. of bitstreams: 1 DISS_2015_Geovanny Bruno Gonçalves Dias.pdf: 1405701 bytes, checksum: 24fbe33b137779f7f8668a0dfcb69dd5 (MD5) Previous issue date: 2015-03-03 / CAPES / CNPq / Do grupo das plantas que causa morte súbita em animais de interesse pecuário no Brasil, sete delas contém monofluoroacetato de sódio (MFA) como princípio tóxico. MFA causa bloqueio do ciclo de Krebs e consequente acúmulo de citrato nos tecidos. A movimentação dos animais intoxicados desencadeia os sinais clínicos e a morte com evolução superaguda. Uma das alternativas preconizadas para prevenção da intoxicação é a ingestão de doses não tóxicas de plantas que contém MFA. Sugere-se que a resistência ocorra devido à multiplicação de microrganismos que expressam o gene fluoroacetato dehalogenase e degradam o MFA no rúmen. Este estudo teve por objetivo verificar se E. faecalis EF09, JQ661270.1 e E. faecalis EF08, JQ661271.1, isoladas do rúmen de bovinos, degradam MFA in vivo e podem ser usadas para detoxificação de plantas que contém MFA. Para a realização deste experimento, seis ovinos com idade média de dois anos, fêmeas, entre 35-40 kg, de raça Santa Inês, foram divididos em dois grupos com três animais cada. O Grupo Inoculação recebeu durante três dias seguidos o inóculo de E. faecalis de 30 ml dividido em três doses de 10 ml. O Grupo Controle recebeu água destilada durante o mesmo período. Os animais de ambos os grupos foram mantidos em descanso por 48 horas e após esse período, o MFA foi fornecido em uma dose de 1,5 mg/Kg/PV por via oral para todos os ovinos. Os sinais clínicos, frequência cardíaca e respiratória foram obtidos em repouso a cada uma hora, e após 10 minutos de movimentação decorridas seis horas da intoxicação. Todos os animais de ambos os grupos morreram entre três e oito horas após o fornecimento de MFA. Os resultados indicam que os animais não desenvolveram resistência ao MFA após a inoculação com E. faecalis. / From the group of plants that cause sudden death in livestock in Brazil, seven of which contains sodium fluoroacetate (MFA) as toxic principle. MFA cause blockage of the Krebs cycle and subsequent accumulation of citrate in the tissues. The movement of animals that are intoxicated triggers the clinical signs and cause the death with acute evolution. One envisaged alternative to preventing poisoning is the ingestion of nontoxic doses of plants containing the MFA. It is suggested that the resistance occurs due to proliferation of microorganisms that express the MFA degrading gene fluoroacetate dehalogenase and that the MFA is degrade in the rumen. This study aimed to determine whether E. faecalis FY09, JQ661270.1 and FY08, JQ661271.1, isolated from bovine rumen, degrade MFA in vivo and can be used for detoxification of plants containing MFA. To carry out this experiment, six sheep with an average age of two years old, female, between 35-40 kg, Santa Ines, were divided into two groups of three animals each. The inoculation Group received for three consecutive days, E. faecalis - 30 ml inoculum divided into three doses of 10 ml. The control group received distilled water for the same period. The animals of both groups were kept at rest for 48 hours and after this period, the MFA is provided in a dose of 1.5 mg / kg / Live Weight orally to all sheep. Clinical signs, heart and respiratory rate were obtained at rest every hour, and after 10 minutes of elapsed drive six hours of poisoning. All animals in both groups died between three and eight hours after delivery of MFA. The results indicate that the animals have not developed resistance to MFA after inoculation with E. faecalis.

Diss. (sammanfattning) Umeå : Umeå universitet, 2010.