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Pancreatic α- and β-Cell Function and Metabolic Changes during Oral L-Alanine and Glucose Administration: Comparative Studies between Normal, Diabetic and Cirrhotic SubjectsSAKAMOTO, NOBUO, TAMAGAWA, TATSUO, KAKUTA, HIRONOBU, NOMURA, TAKAHIDE, KUNIEDA, TAKEHIDE, SHINODA, HIROSHI, OHARA, KIYOJI, HOTTA, NIGISHI, HATTORI, TADAKAZU 03 1900 (has links)
No description available.
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Avaliação de dose administrada no tratamento de neoplasia ginecológica por ressonância de spin eletrônico com L-alanina / Dose evaluation administered in the treatment of gynecological câncer by eléctron spin resonance with L-alanineRech, Amanda Burg 29 June 2012 (has links)
Neoplasias ginecológicas são os tumores de maior incidência no público feminino. Como alternativa para o tratamento destes tumores existe a radioterapia, em que a sua eficácia depende prioritariamente da correta administração da dose prescrita ao tumor, com acurácia de ±5%, conforme estabelecido pela American Association of Physicists in Medicine (AAPM). Para verificar a administração de doses existem diversas técnicas dosimétricas, entre elas a ressonância de spin eletrônico (RSE) com L-alanina, material tecido-equivalente, que relaciona dose com a quantidade de radicais livres formados pela radiação. Portanto, o objetivo deste estudo é a verificação da dose administrada na região tumoral de pacientes diagnosticadas com câncer ginecológico submetidas à teleterapia, utilizando L-alanina e a RSE. Para comparação entre as modalidades de radioterapia, foi realizada braquiterapia ginecológica com objeto simulador, verificando a dose no reto durante o tratamento, que deve ser inferior a 65% da dose prescrita ao tumor, conforme a International Commission on Radiation Units (ICRU). Para a análise de teleterapia foi construído um aparato, para a inserção vaginal do material radio-sensível, que foi utilizado durante 5 das 25 sessões de tratamento. As doses foram determinadas com o sistema de planejamento do tratamento mediante as imagens da tomografia computadorizada, em conjunto com softwares de planejamento, e a existência de marcadores radiopacos no aparato permitiu a confirmação das doses, conforme referência óssea através de imagens de portal feitas no momento anterior ao tratamento. Após a irradiação foi feita a leitura das amostras com o espectrômetro de RSE, em que a amplitude pico a pico da linha principal do sinal, normalizado pela massa, determinou a dose entregue nas regiões de interesse. Previamente ao estudo in vivo foi efetuada a teleterapia com objeto simulador, nas mesmas condições de tratamentos in vivo, apresentando desvio médio em relação ao planejamento de (0,2 ± 3,5)%, viabilizando o estudo com pacientes. Para a primeira paciente o desvio médio obtido foi de (0,5 ± 3,7)% e para a segunda de (-0,7 ± 2,8)%. Na simulação da braquiterapia o desvio médio observado foi de (-1,3 ± 9,2)%. Por causa do pequeno número amostral de pacientes, esta pesquisa é classificada como um estudo de viabilidade, em que a utilização de L-alanina com a RSE se mostrou eficiente para o objetivo proposto, fazendo com que os resultados da teleterapia estivessem de acordo com o estipulado pela AAPM e na braquiterapia a dose administrada no reto obedecesse ao relatório 38 da ICRU. / Gynecological malignancies are the most incident female tumors. As an alternative treatment for these tumors exist the radiotherapy, which their effectiveness mainly depends on the right administration of the prescribed dose to the tumor, with an accuracy of ±5%, as established by the American Association of Physicists in Medicine (AAPM). There are a lot of dosimetric techniques for dose verification, including electron spin resonance (ESR) with L-alanine, a tissue-equivalent material, which relates dose with the amount of free radicals formed by radiation. Therefore, the aim of this study is to verify the dose administered in the tumor of patients diagnosed with gynecological cancer subjected to external beam radiation therapy using L-alanine and ESR. For comparison between the radiotherapy modalities was performed gynecological brachytherapy in phantom, verifying the dose to the rectum during treatment, which must be less than 65% of the prescribed dose to the tumor, according to the International Commission on Radiation Units (ICRU). For teletherapy analysis was constructed an apparatus for vaginal insertion of the radio-sensitive material, which was used in 5 of the 25 treatment fractions. The doses were determined with the treatment planning system with the computed tomography images, together with the planning software, and the existence of radiopaque markers in the apparatus allowed the dose confirmation, according to the bone reference through portal images made before treatment. After the irradiation the samples was read with the ESR spectrometer, wherein the peak to peak amplitude of the signal main line, normalized by weight, determined the dose delivered into the regions of interest. Prior to the in vivo study was performed the teletherapy with phantom, under the same conditions of in vivo treatments, presenting mean deviation to the planning dose of (0.2 ± 3.5)%, enabling the study with patients. For the first patient, the mean deviation obtained was of (0.5 ± 3.7)% and for the second one of (-0.7 ± 2.8)%. In the brachytherapy simulation the mean deviation observed was of (-1.3 ± 9.2)%. Due to the small sample of patients, this research is classified as a feasibility study, which the use of L-alanine with the ESR was efficient for the objective proposed, making the teletherapy results consistent with the stipulated by the AAPM, and in the brachytherapy the dose to the rectum obey the ICRU report 38.
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Cellular transport and secretion of the cyanobacterial neurotoxin BMAA into milk and egg : Implications for developmental neurotoxicityAndersson, Marie January 2015 (has links)
The cyanobacterial amino acid β-N-methylamino-L-alanine (BMAA) is a neurotoxin implicated in the etiology of neurodegenerative diseases. Cyanobacteria are cosmopolitan organisms present in various environments. BMAA can cause long-term neurodegenerative alterations in rats exposed during the neonatal period, a period that corresponds to the last trimester and the first few years of life in humans. As BMAA has been reported to be bioaccumulated in the aquatic food chain and detected in mussels, crayfish and fish used for human consumption, the main aim of this thesis has been to investigate the final step in the mammalian food-chain, i.e. the transfer of BMAA into breast milk. Autoradiographic imaging and mass spectrometry analysis showed an enantiomer-selective uptake of BMAA and that the neurotoxin was transferred from lactating mice and rat, via the milk, to the brain of the nursed pups. The results show that transport of BMAA may be disproportional to dose. In addition, BMAA was found present both as free amino acid and tightly associated to proteins in rat brains. Surprisingly, however, no association to milk proteins was found. In vitro studies of murine (HC11) and human (MCF7) mammary epithelial cells suggest that BMAA can pass the human mammary epithelium into milk. Additional transport studies on human intestinal, glioblastoma and neuroblastoma cells showed that L-BMAA was consistently favored over D-BMAA and that the transport was mediated by several amino acid transporters. We also demonstrated that egg-laying quail transfer BMAA to its offspring by deposition in the eggs, particularly in the yolk but also in the albumen. Furthermore, comparative analysis of carboxyl- and methyl-labeled [14C]-BMAA suggested that BMAA was not metabolized to a large degree. Altogether, the results indicate that BMAA can be transferred from mothers, via the milk, to the brain of nursed human infants. Determinations of BMAA in mothers’ milk and cows’ milk are therefore warranted. We also propose that birds’ eggs could be an additional source of BMAA exposure in humans. It might therefore be of concern that mussels are increasingly used as feed in commercial egg production.
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Avaliação de dose administrada no tratamento de neoplasia ginecológica por ressonância de spin eletrônico com L-alanina / Dose evaluation administered in the treatment of gynecological câncer by eléctron spin resonance with L-alanineAmanda Burg Rech 29 June 2012 (has links)
Neoplasias ginecológicas são os tumores de maior incidência no público feminino. Como alternativa para o tratamento destes tumores existe a radioterapia, em que a sua eficácia depende prioritariamente da correta administração da dose prescrita ao tumor, com acurácia de ±5%, conforme estabelecido pela American Association of Physicists in Medicine (AAPM). Para verificar a administração de doses existem diversas técnicas dosimétricas, entre elas a ressonância de spin eletrônico (RSE) com L-alanina, material tecido-equivalente, que relaciona dose com a quantidade de radicais livres formados pela radiação. Portanto, o objetivo deste estudo é a verificação da dose administrada na região tumoral de pacientes diagnosticadas com câncer ginecológico submetidas à teleterapia, utilizando L-alanina e a RSE. Para comparação entre as modalidades de radioterapia, foi realizada braquiterapia ginecológica com objeto simulador, verificando a dose no reto durante o tratamento, que deve ser inferior a 65% da dose prescrita ao tumor, conforme a International Commission on Radiation Units (ICRU). Para a análise de teleterapia foi construído um aparato, para a inserção vaginal do material radio-sensível, que foi utilizado durante 5 das 25 sessões de tratamento. As doses foram determinadas com o sistema de planejamento do tratamento mediante as imagens da tomografia computadorizada, em conjunto com softwares de planejamento, e a existência de marcadores radiopacos no aparato permitiu a confirmação das doses, conforme referência óssea através de imagens de portal feitas no momento anterior ao tratamento. Após a irradiação foi feita a leitura das amostras com o espectrômetro de RSE, em que a amplitude pico a pico da linha principal do sinal, normalizado pela massa, determinou a dose entregue nas regiões de interesse. Previamente ao estudo in vivo foi efetuada a teleterapia com objeto simulador, nas mesmas condições de tratamentos in vivo, apresentando desvio médio em relação ao planejamento de (0,2 ± 3,5)%, viabilizando o estudo com pacientes. Para a primeira paciente o desvio médio obtido foi de (0,5 ± 3,7)% e para a segunda de (-0,7 ± 2,8)%. Na simulação da braquiterapia o desvio médio observado foi de (-1,3 ± 9,2)%. Por causa do pequeno número amostral de pacientes, esta pesquisa é classificada como um estudo de viabilidade, em que a utilização de L-alanina com a RSE se mostrou eficiente para o objetivo proposto, fazendo com que os resultados da teleterapia estivessem de acordo com o estipulado pela AAPM e na braquiterapia a dose administrada no reto obedecesse ao relatório 38 da ICRU. / Gynecological malignancies are the most incident female tumors. As an alternative treatment for these tumors exist the radiotherapy, which their effectiveness mainly depends on the right administration of the prescribed dose to the tumor, with an accuracy of ±5%, as established by the American Association of Physicists in Medicine (AAPM). There are a lot of dosimetric techniques for dose verification, including electron spin resonance (ESR) with L-alanine, a tissue-equivalent material, which relates dose with the amount of free radicals formed by radiation. Therefore, the aim of this study is to verify the dose administered in the tumor of patients diagnosed with gynecological cancer subjected to external beam radiation therapy using L-alanine and ESR. For comparison between the radiotherapy modalities was performed gynecological brachytherapy in phantom, verifying the dose to the rectum during treatment, which must be less than 65% of the prescribed dose to the tumor, according to the International Commission on Radiation Units (ICRU). For teletherapy analysis was constructed an apparatus for vaginal insertion of the radio-sensitive material, which was used in 5 of the 25 treatment fractions. The doses were determined with the treatment planning system with the computed tomography images, together with the planning software, and the existence of radiopaque markers in the apparatus allowed the dose confirmation, according to the bone reference through portal images made before treatment. After the irradiation the samples was read with the ESR spectrometer, wherein the peak to peak amplitude of the signal main line, normalized by weight, determined the dose delivered into the regions of interest. Prior to the in vivo study was performed the teletherapy with phantom, under the same conditions of in vivo treatments, presenting mean deviation to the planning dose of (0.2 ± 3.5)%, enabling the study with patients. For the first patient, the mean deviation obtained was of (0.5 ± 3.7)% and for the second one of (-0.7 ± 2.8)%. In the brachytherapy simulation the mean deviation observed was of (-1.3 ± 9.2)%. Due to the small sample of patients, this research is classified as a feasibility study, which the use of L-alanine with the ESR was efficient for the objective proposed, making the teletherapy results consistent with the stipulated by the AAPM, and in the brachytherapy the dose to the rectum obey the ICRU report 38.
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Redistribution of Hepatocyte Chloride During L-Alanine UptakeWang, Kening, Wondergem, Robert 01 September 1993 (has links)
We used ion-sensitive, double-barrel microelectrodes to measure changes in hepatocyte transmembrane potential (Vm), intracellular K+, Cl-, and Na+ activities (aik, aCliand aNai), and water volume during l-alanine uptake. Mouse liver slices were superfused with control and experimental Krebs physiological salt solutions. The experimental solution contained 20 μml-alanine, and the control solution was adjusted to the same osmolality (305 mOsm) with added sucrose. Hepatocytes also were loaded with 50 m m tetramethylammonium ion (TMA+) for 10 min. Changes in cell water volume during l-alanine uptake were determined by changes in intracellular, steady-state TMA+ activity measured with the K+ electrode. Hepatocyte control Vm was -33±1 mV. l-alanine uptake first depolarized Vm by 2±0.2 mV and then hyperpolarized Vm by 5 mV to-38±1 mV (n = 16) over 6 to 13 min. During this hyperpolarization, aNaiincreased by 30% from 19±2 to 25±3 m m (P < 0.01), and aKidid not change significantly from 83±3 m m. However, with added ouabain (1 m m) l-alanine caused only a 2-mV increase in Vm, but now aKidecreased from 61±3 to 54±5 m m (P < 0.05). Hyperpolarization of Vm by l-alanine uptake also resulted in a 38% decrease of aClifrom 20±2 to 12±3 m m (P < 0.001). Changes in Vm and VCl - Vm voltage traces were parallel during the time of l-alanine hyperpolarization, which is consistent with passive distribution of intracellular Cl- with the Vm in hepatocytes. Added Ba2+ abolished the l-alanineinduced hyperpolarization, and aCliremained unchanged. Hepatocyte water volume during l-alanine uptake increased by 12±3%. This swelling did not account for any changes in ion activities following l-alanine uptake. We conclude that hepatocyte aKiis regulated by increased Na+-K+ pump activity during l-alanine uptake in spite of cell swelling and increased Vm due to increased K+ conductance. The hyperpolarization of Vm during l-alanine uptake provides electromotive force to decrease aCli. The latter may contribute to hepatocyte volume regulation during organic solute transport.
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ARRESTED AND CHAINED: The role of AmiB and AmiC in Pseudomonas aeruginosa daughter cell separationAl-Saigh, Sarra 10 1900 (has links)
<p>Peptidoglycan (PG) remodelling and cell division are two important cellular processes that are the major target of antibiotics. Due to rising resistance, the need for new antibiotics today has never been greater. Therefore it is important to fill the gaps in our understanding of these two important processes in order to discover new and promising antibiotic targets. Peptidoglycan synthesis and remodelling is a highly coordinated event that involves a wide number of enzymes and processes which are not well understood. N-acetylmuramoyl-L-alanine amidases, whose function is to cleave the amide linkage between the stem peptides and the lactyl moiety of N-acetylmuramic acid, is a major class of PG-active proteins. Their role in daughter cell separation during cell division is well established in <em>Escherichia coli</em> however little is known about it in other systems. Using enzymatic assays we characterize AmiC as a novel amidase in <em>Pseudomonas aeruginosa. </em>Through mutational analysis and microscopy we show that AmiB and AmiC are required for daughter cell separation. A deletion of both enzymes results in a cell chaining phenotype with abnormal cell morphology. Transmission electron microscopy reveals that the double mutant is arrested at the septal peptidoglycan separation step. In addition to cell chaining, the ∆<em>amiB/amiC</em> mutant exhibits a significant increase in susceptibility to antibiotics. We also demonstrate that the LysM motif of AmiB is not required for its role in cell separation. Furthermore, the <em>amiB</em> mutant has significantly shorter cells than the wildtype indicating an additional role for the enzyme in the cell. Lastly, through a novel bioinformatics strategy we identify PA5047 as a potential PG amidase.</p> / Bachelor of Science (BSc)
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Neurotoxicity and Degenerative Disorders: Studies of β-N-methylamino-L-alanine (BMAA)-induced Effects in SH-SY5Y Cells using Immunohistochemistry (IHC)Robbani, Elin January 2017 (has links)
The cyanobacterial neurotoxin β-N-methylamino-L-alanine (BMAA), a non-protein amino acid, first attracted attention in correlation to reports of high incidence of the unusual neurological disease amyotrophic lateral sclerosis/Parkinsonism-dementia (ALS/PDC) among the people of Guam in the South Pacific Ocean. Experimental studies have revealed that BMAA causes neuronal cell death. The neurotoxin is suggested to act via excitotoxicity through interaction with glutamatergic receptors. More importantly, BMAA is suggested to misincorporate in the synthesis of proteins, and contribute to protein misfolding and/or deleterious aggregation, which are hallmarks of several neurodegenerative disorders. A selective uptake of BMAA in the rat neonatal hippocampus can interfere with brain development, causing learning and memory impairments in adult rats. The aim of the present study was to investigate the effects of BMAA in human neuroblastoma SH-SY5Y cells. These cells were exposed to BMAA (10 μM, 50 μM, 100 μM or 500 μM) for 72 hours, and the expression of five selected proteins, including heat shock protein-27 (HSP-27), lysosomal associated membrane protein-1 (LAMP-1), CCAAT-enhancer-binding protein homologous protein (CHOP), Golgi associated plant pathogenesis related protein-2 (GLIPR-2), and glucose regulated protein-78 (GRP-78). They were carried out with immunohistochemistry (IHC). Results revealed an increased expression of all selected proteins, which indicates an uptake and shows the effects of BMAA in the cell cultures. Taken together, BMAA caused cellular stress, including endoplasmic reticulum (ER) stress that is correlated with HSP-27, LAMP-1, CHOP, GLIPR-2, and GRP-78. Further studies are needed in order to support the results. The experiments require being repeated using the same biomarkers as well as a combination of them with other biomarkers to elucidate the effects of BMAA.
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Estudo de altas pressões por espectroscopia Raman no cristal de Nitrato de L-Alanina / Study of high pressures by Raman spectroscopy on L-Alanine Nitrate crystalSoares, Rilleands Alves 18 April 2017 (has links)
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Previous issue date: 2017-04-18 / In this work nitrate L-Alanine crystals (LAN) were prepared by slow evaporation technique at room temperature and characterization by X-ray diffraction and Raman scattering at room temperature and high pressures in the amino acid salt crystals LAN. The crystals grew after six months of rest. The growth solution was acidic with pH 1.0. With the XRD pattern of the material and the Rietveld analysis, it was found that at room temperature LAN crystallizes in orthorhombic space group (P212121) with four molecules per unit cell. The refinement of the quality parameters were satisfactory, with Rp = 5.52%, 7.73% and S = 1.4. The results of Raman scattering with hydrostatic pressure variation between 0 GPa and 7.4 GPa, show that the LAN crystal undergoes a phase transition between 3.5 and 4.1 GPa, as suggested by the changes in the vibrational modes, both In the low wave number as in the internal mode regions. The correlation between the configuration of hydrogen bonding at ambient pressure and the occurrence of phase transition with pressure variation was proposed. After decompression at ambient pressure, the original spectra were recovered indicating that the phase transitions are reversible. / Este trabalho descreve o preparo dos cristais de nitrato de L-Alanina (NLA) pela técnica de evaporação lenta do solvente à temperatura ambiente e realizadas medidas de caracterização por difração de raios X e espalhamento Raman à temperatura ambiente e a altas pressões em cristais do sal de aminoácido de NLA. Os cristais cresceram após seis meses de repouso. A solução de crescimento era ácida com pH 1,0. Com o difratograma do material e a análise pelo método Rietveld, constatou-se que à temperatura ambiente o NLA cristaliza-se numa estrutura ortorrômbica (P212121) com quatro moléculas por célula unitária. Os parâmetros de qualidade do refinamento foram satisfatórios, com Rp = 5,52 %, Rwp = 7,73 % e S = 1,4. Os resultados de espalhamento Raman com variação da pressão hidrostática entre 0 GPa e 7,4 GPa, sugerem que o cristal de NLA, sofre uma transição de fase entre 3,5 e 4,1 GPa, evidenciando pelas mudanças nos modos vibracionais, tanto no número de onda baixo como nas regiões de modos internos. Foi proposta a correlação entre as ligações de hidrogênio à pressão ambiente e a ocorrência de transição de fase com variação de pressão. Após a descompressão à pressão ambiente, os espectros originais foram recuperados indicando que as transições de fase são reversíveis.
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Detection, transfer and role of an environmentally spread neurotoxin (BMAA) with focus on cyanobacteria and the Baltic Sea regionBerntzon, Lotta January 2015 (has links)
β-N-methylamino-L-alanine (BMAA) is one of the more recently discovered bioactive compounds produced by cyanobacteria. BMAA is a non-protein amino acid reported present in human brain tissues of patients deceased from a neurodegenerative disease, such as Alzheimer´s disease or amyotrophic lateral sclerosis (ALS). This observation in combination with its neurotoxic effects in eukaryotes (in vivo and in vitro) and its potential to incorporate into (human) proteins, causing protein aggregation, suggests BMAA as a possible causative environmental agent for neurodegenerative diseases. Due to the ubiquitous nature of cyanobacteria with a wide occurrence in both aquatic and terrestrial environments, BMAA could be globally spread. Hence, investigations of a possible coupling between BMAA and neurodegeneration are urgently needed as well as to identify sources of BMAA in Nature. The aim of this thesis was to examine the potential occurrence of BMAA in bloom forming cyanobacteria of the Baltic Sea and its possible transfer to other organisms of this ecosystem. Of importance was also to reveal any likely routes for human BMAA exposure in the Baltic Sea region and to further investigate BMAA as a triggering agent for neurodegenerative diseases. Acknowledged difficulties of analysing BMAA in biological samples also inferred method development as part of the experimental studies. Investigating the role of BMAA in its producers was another purpose of the thesis, which may be crucial for future management of BMAA-producing cyanobacteria. By screening natural populations of the major filamentous bloom forming cyanobacteria of the Baltic Sea, we discovered the presence of BMAA throughout the entire summer season of two consecutive years, using a highly specific analytical method (liquid chromatography-tandem mass spectrometry; LC-MS/MS). BMAA was found to bioaccumulate in zooplankton and fish, as well as in mussels and oysters from the Swedish west coast. To improve the understanding of BMAA analyses in natural samples, the formation of carbamate adducts in the presence of bicarbonate was examined. Using two derivatization techniques in combination with LC-MS/MS, we could show that BMAA detection was not hindered by carbamate formation. Exogenously added BMAA inhibited nitrogen fixation in the model cyanobacterium Nostoc sp. PCC 7120, which was also hampered in growth and displayed signs of nitrogen starvation. Finally, BMAA was detected in cerebrospinal fluid in three of 25 Swedish test individuals, and represents the first confirmation of BMAA in the human central nervous system using LC-MS/MS as the primary analytical method. However, the association of BMAA to neurodegenerative diseases could not be verified as BMAA was present in both control individuals (two) and in one ALS-patient. Nevertheless, the finding of a known neurotoxic compound in the human central nervous system is alarming and potential consequences should be investigated. The discovery of the neurotoxic compound BMAA in Baltic Sea organisms, and in the central nervous system of humans potentially consuming fish from this ecosystem is concerning and warrants continued investigations of BMAA occurrence and human exposure. Further knowledge on the function and regulation of BMAA may help in developing strategies aiming to minimise human exposure. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 3: Manuscript.</p>
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Terahertz Time-Domain Spectroscopy and Imaging of Biological Compounds and TissuesTaulbee, Anita R. 11 August 2009 (has links)
No description available.
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