O presente trabalho teve como objetivo estimar a concentração de Phe em 22 amostras de sopas desidratadas instantâneas, por serem úteis na diversificação do cardápio de fenilcetonúricos. Foi analisada a concentração de glutamato monossódico (GMS) por ser uma provável fonte de N não protéico (NNP) que pode resultar em concentrações protéicas superestimadas. A concentração de proteína real estimada foi realizada após precipitação da proteína com TCA 10%, seguida da análise do N pelo método de Kjeldahl, o qual foi convertido para proteína por um fator de conversão (Fc) adequado. A legislação Brasileira estabelece um Fc de 5,75 para proteínas vegetais, 6,25 para proteínas da carne e misturas de proteínas e 6,38 para proteínas lácteas. A concentração de GMS foi determinada por método enzimático com eletrodo sensível a amônia. A concentração de proteína bruta (N totalxFc) variou entre 6,05 e 21,51%, tendo sido estes valores, na maioria das vezes, similares aos declarados no rótulo, indicando que os fabricantes utilizam o N totalxFc para expressar o conteúdo protéico. A concentração protéica real estimada foi baixa, variando entre 1,28 e 16,31%. A concentração de NNP teve uma variação de 0,33 a 1,27g/100g de amostra, representando de 11,10 a 81,33% do NT presente. A concentração de GMS variou entre 1,01 e 7,86g/100g de amostra, sendo que o N proveniente deste realçador de sabor contribuiu com 2,53 a 47,71% na quantidade total de N. A diferença entre a concentração de proteína bruta e real estimada se deve à presença de NNP, na forma de GMS. Com base nos valores protéicos reais estimados, foram calculados os teores de Phe que variaram entre 51,16 e 652,24mg de Phe/100g de amostra. Assim, recomenda-se que todos os alimentos adicionados de realçadores de sabor sejam analisados quanto à concentração de proteína real para que a Phe seja corretamente estimada. / The aim of this work was to estimate the concentration of Phe in 22 samples of commercially available dehydrated soups, as they are useful to add variety to the diet for phenilketonurics. The monosodium glutamate (MSG) contents had been analyzed as it is a likely source of non protein N (NPN) that might result in overestimated protein contents. The true protein content was accomplished after protein precipitation with 10% TCA and followed by N analysis according to the Kjeldahl method, which was converted to protein by a suitable conversion factor (Fc). The Brazilian legislation establishes a Fc of 5,75 for vegetables proteins, 6,25 for meat and blended proteins and 6,38 for milk proteins. The MSG concentration was determined by an enzymatic method employing an ammonia gas-sensitive electrode. The crude protein content (total NxFc) varied from 6,05 to 21,51% and were similar, in most cases, to those stated on the label, showing that manufacturers use total NxFc to express the protein content. Nevertheless, the true protein content was low, varying from 1,28 to 16,31%. The NPN concentration varied from 0,33 to 1,27g/100g of sample, which represents from 11,10 to 81,33% of the existing total N. The MSG concentration varied from 1,01 to 7,86g/100g of sample; the N arose from this flavor enhancer gives about 2,53 to 47,71% of the total quantity of N. The difference between the crude protein and true protein contents is due to the presence of MSG-like NPN. The Phe concentrations were calculated in accordance with the true protein values and varied from 51,16 to 652,24 mg/100g of sample. Thus, we recommend the analysis of all flavor-enhancer-added foods, in order to get reliable results for Phe estimation from the protein contents.

La phénylcétonurie (PCU est une maladie métabolique génétique causée par une déficience d'activité phénylalanine hydroxylase (PAH). Une hypomyélinisation du cerveau a été documentée chez les patients non traités, mais sa pathophysiologie reste floue. Nous avons investigué l'influence de la phénylalanine (Phe), phénylpyruvate (PP) et phénylacétate (PA) sur les oligodendrocytes. Nous avons premièrement montré dans un modèle murin de PCU que le nombre d'oligodendrocytes n'était pas différent dans le corps calleux entre animaux PCU et sains. Ensuite, en utilisant la technique des co-cultures myélinisantes nous avons pu déterminer que Phe, PP et PA n'ont pas d'effet direct sur la synthèse des gaines de myéline. Ces données indiquent que ces trois composés n'exercent probablement pas de rôle direct dans l'hypomyélinisation du système nerveux central constatée dans le cadre de la PCU. Ces données suggèrent donc des mécanismes d'action indirects. De plus, nous avons investigué la faisabilité d'un modèle de thérapie génique pour la PCU. Celui-ci implique la transduction ex vivo d'hépatocytes ou cellules souches mésenchymateuses par un vecteur lentiviral puis leur implantation dans le foie de l'organisme receveur. Phenylketonuria (PKU) is a metabolic genetic disease characterized by deficient phenylalanine hydroxylase (PAH) enzymatic activity. Brain hypomyelination has been reported in untreated patients, but its mechanism remains unclear. We therefore investigated the influence of phenylalanine (Phe), phenylpyruvate (PP), and phenylacetate (PA) on oligodendrocytes. We fisrt showed in a mouse model of PKU that the number of oligodendrocytes is not different in corpus callosum sections from adult mutants or from control brains. Then, using enriched oligodendroglial cultures, we detected no cytotoxic effect of high concentrations of Phe, PP, or PA. Finally, we analyzed the impact of Phe, PP, and PA on the myelination process in myelinating cocultures using both an in vitro index of myelination, based on activation of the myelin basic protein (MBP) promoter, and the direct quantification of myelin sheaths by both optical measurement and a bioinformatics method. None of these parameters was affected by the increased levels of Phe or its derivatives. Taken together, our data demonstrate that high levels of Phe, such as in PKU, are unlikely to directly induce brain hypomyelination, suggesting involvement of alternative mechanisms in this myelination defect. Moreover, we investigated the feasibility of a gene therapy for phenylketonuria. This project involved the ex vivo transduction of hepatocytes and mesenchymal stem cells with lentivirus vector and the engraftment of these cells in the liver's recipient.

Previous research has shown a relationship between prenatal teratogenic exposure and impaired cognitive functioning. However, data regarding the long-term outcome of prenatal teratogenic exposure are minimal. The present study investigated the long-term neuropsychological functioning (specifically attention and memory) of adults prenatally exposed to alcohol or phenylalanine, and examined whether there was evidence to suggest that there are effects specific to individual teratogens. Using a battery of attention and memory measures the performance of 17 adults diagnosed with Fetal Alcohol Syndrome (FAS) and 13 adults with Maternal Phenylketonuria Syndrome (MPKUS) was assessed. In order to identify the pattern of deficits associated with prenatal teratogenic exposure, an age and CA and IQ matched control group was assessed. Attention was broadly assessed using Mirsky et al.'s (1991) neuropsychological model of attention. The memory and learning tests administered included a number of well standardized measures of verbal learning, verbal and visual recall, delayed recall, and recognition. Paired comparisons between the FAS group and age and CA and IQ matched controls indicated a unique pattern of attention and memory deficits consistent with previous research with children and adolescents. Specifically, adult individuals with FAS appear to have deficits in acquisition of new material, delayed recall of verbal material and in response inhibition. Paired comparisons between the MPKUS group and CA and IQ matched controls indicated that the pattern of attention and memory deficits seen in adults with MPKUS is difficult to distinguish when the effect of IQ is removed. A randomized block design using IQ as the blocking variable and group (FAS, MPKUS, or Controls) as the treatment variable was utilized to examine the question of whether the two prenatal teratogen groups differ from one another and from Controls in terms of attention and memory ability. Ten blocks of three participants (FAS, MPKUS and Control) matched on IQ were formed. The randomized block analyses revealed few differences between the groups and failed to reveal a number of the differences found in the paired comparisons between the prenatal teratogen groups and the CA and IQ matched Control group. Possible reasons for these differences are discussed.

A fenilcetonúria (PKU) é caracterizada por uma deficiente atividade da fenilalanina hidroxilase (PAH), que converte fenilalanina em tirosina. Consequentemente, pacientes fenilcetonúricos apresentam níveis aumentados de fenilalanina no sangue e em tecidos. A fenilalanina atinge níveis tóxicos no cérebro e, assim, pode levar a deficiência intelectual grave se não tratada. Hoje em dia, os pacientes fenilcetonúricos são diagnosticados através de programas de triagem neonatal e são colocados em tratamento imediatamente. O tratamento da PKU é baseado em uma dieta restrita em fenilalanina juntamente com uma mistura de aminoácidos, que visa reduzir a ingestão de fenilalanina ao mesmo tempo em que fornece todos os outros aminoácidos e nutrientes essenciais. Apesar de eficiente, o tratamento é extremamente difícil de seguir e, portanto, os pacientes ainda apresentam altos níveis de fenilalanina e seus problemas associados, tais como estresse oxidativo, distúrbios motores e cognitivos. Ainda, a dieta em si é tão restritiva que a completa adesão pode levar a problemas nutricionais como obesidade e perturbações hormonais. A fim de abordar estas questões na PKU, é necessário compreender os mecanismos pelos quais a fenilalanina perturba homeostasia, bem como propor novas estratégias de tratamento para a doença. Portanto, esta tese teve como objetivo verificar o estado metabólico basal e em exercício aeróbico em pacientes fenilcetonúricos, além de avaliar possíveis benefícios do treinamento físico em camundongos PAHenu2. Para tanto, foram avaliados composição basal, estado nutricional, taxa metabólica basal, bem como a resposta ventilatória e bioquímica a uma sessão de exercício aeróbico em pacientes fenilcetonúricos em comparação com controles pareados, por bioimpedância elétrica e calorimetria indireta, respectivamente. Foram encontrados resultados similares entre pacientes e controles para todos os marcadores antropométricos e nutricionais, mostrando, assim, que os pacientes fenilcetonúricos têm a mesma composição corporal e perfil metabólico que controles. Além disso, os níveis de fenilalanina não se modificaram imediatamente após o exercício em comparação com a condição de repouso, demonstrando assim que o exercício aeróbico é seguro para pacientes fenilcetonúricos. No modelo animal de PKU, este estudo avaliou os efeitos crônicos do exercício aeróbico voluntário no cérebro de camundongos PAHenu2. Apesar de correrem menores distâncias do que os controles, os camundongos PKU apresentaram melhoras em parâmetros de estresse oxidativo no cérebro, embora os níveis de fenilalanina no sangue e no cérebro permanecessem inalterados. Os animais PKU apresentaram habilidades motoras e de equilíbrio deficientes em comparação com os controles, enquanto o exercício não afetou esses parâmetros comportamentais. Adicionalmente, os aminoácidos gliconeogênicos e os relacionados com o ciclo da ureia foram encontrados em níveis inferiores no plasma dos animais PKU que se exercitaram em comparação com os sedentários. Por outro lado, os camundongos selvagens não apresentaram nenhuma alteração causada pelo exercício. Assim, os fenilcetonúricos não apresentam distúrbios em marcadores metabólicos tanto em repouso como durante exercício aeróbico. Por conseguinte, os fenilcetonúricos devem ser encorajados a praticar exercício para, possivelmente, beneficiarem-se das adaptações positivas geradas pelo treinamento físico. No entanto, o pequeno número de pacientes avaliados em nossos estudos destaca a necessidade de mais pesquisas para descrever o exercício mais adequado para pacientes com PKU. / Phenylketonuria (PKU) is characterized by poor phenylalanine hydroxylase (PAH) activity, which acts converting phenylalanine into tyrosine. Consequently, PKU patients show increased levels of phenylalanine in the blood and tissues. Phenylalanine reaches toxic levels in the brain and therefore can lead to severe mental retardation, if untreated. Nowadays, PKU patients are diagnosed through newborn screening programs and are put on treatment immediately. PKU treatment is based on a phenylalanine-restricted diet along with an amino acid mixture, which aims to reduce the intake of phenylalanine while providing other essential amino acids and nutrients. Despite efficient, the treatment is extremely hard to follow so that PKU patients show high levels of phenylalanine and related issues such as oxidative stress, motor and cognitive disturbances. Moreover, the diet itself is so restrictive that adhering to it may lead to nutritional problems like obesity and hormonal disruptions. In order to tackle these issues in PKU, it is necessary to understand the mechanisms in which phenylalanine disturbs homeostasis as well as propose new treatment strategies for the disease. Therefore, this thesis aimed to evaluate metabolic markers in rest and exercise in PKU patients, as well as to verify the possible benefits of exercise in the brain of PAHenu2 mice. For that, we evaluated basal body composition, nutritional status, basal metabolic rate, as well as ventilatory and biochemical response to an aerobic exercise session in PKU patients and matched-controls using electrical bioimpedance analysis and indirect calorimetry, respectively. The groups did not differ for anthropometric and nutritional markers, thus showing that PKU patients have the same body and metabolic profiles as controls. Moreover, phenylalanine levels were not modified immediately after exercise in comparison to rest condition, thus proving that aerobic exercise is safe for PKU patients. In the animal model of PKU, we evaluated the effects of voluntary training in the brain of PAHenu2 mice. Despite running less distances than the controls, the PKU mice showed improved oxidative stress parameters although phenylalanine levels in the blood and in brain remained unchanged. PKU animals showed poor motor and balance skills than controls while exercise did not affect these behavioral markers. In addition, gluconeogenic and urea cycle-related amino acids were found in lower levels in the plasma of the exercised PKU animals in comparison to the sedentary PKU group. On the other hand, wild-type mice did not show any of those changes. Taken together, we conclude that PKU patients do not show disturbed metabolism in rest and during aerobic exercise. Therefore, PKU patients have to be encouraged to exercise in order to possibly benefit from long-term exercise-related adaptations. Nevertheless, the small number of patients evaluated in our studies highlights the need of further research to describe the most suitable exercise for PKU patients.

A fenilcetonúria (PKU) é caracterizada por uma deficiente atividade da fenilalanina hidroxilase (PAH), que converte fenilalanina em tirosina. Consequentemente, pacientes fenilcetonúricos apresentam níveis aumentados de fenilalanina no sangue e em tecidos. A fenilalanina atinge níveis tóxicos no cérebro e, assim, pode levar a deficiência intelectual grave se não tratada. Hoje em dia, os pacientes fenilcetonúricos são diagnosticados através de programas de triagem neonatal e são colocados em tratamento imediatamente. O tratamento da PKU é baseado em uma dieta restrita em fenilalanina juntamente com uma mistura de aminoácidos, que visa reduzir a ingestão de fenilalanina ao mesmo tempo em que fornece todos os outros aminoácidos e nutrientes essenciais. Apesar de eficiente, o tratamento é extremamente difícil de seguir e, portanto, os pacientes ainda apresentam altos níveis de fenilalanina e seus problemas associados, tais como estresse oxidativo, distúrbios motores e cognitivos. Ainda, a dieta em si é tão restritiva que a completa adesão pode levar a problemas nutricionais como obesidade e perturbações hormonais. A fim de abordar estas questões na PKU, é necessário compreender os mecanismos pelos quais a fenilalanina perturba homeostasia, bem como propor novas estratégias de tratamento para a doença. Portanto, esta tese teve como objetivo verificar o estado metabólico basal e em exercício aeróbico em pacientes fenilcetonúricos, além de avaliar possíveis benefícios do treinamento físico em camundongos PAHenu2. Para tanto, foram avaliados composição basal, estado nutricional, taxa metabólica basal, bem como a resposta ventilatória e bioquímica a uma sessão de exercício aeróbico em pacientes fenilcetonúricos em comparação com controles pareados, por bioimpedância elétrica e calorimetria indireta, respectivamente. Foram encontrados resultados similares entre pacientes e controles para todos os marcadores antropométricos e nutricionais, mostrando, assim, que os pacientes fenilcetonúricos têm a mesma composição corporal e perfil metabólico que controles. Além disso, os níveis de fenilalanina não se modificaram imediatamente após o exercício em comparação com a condição de repouso, demonstrando assim que o exercício aeróbico é seguro para pacientes fenilcetonúricos. No modelo animal de PKU, este estudo avaliou os efeitos crônicos do exercício aeróbico voluntário no cérebro de camundongos PAHenu2. Apesar de correrem menores distâncias do que os controles, os camundongos PKU apresentaram melhoras em parâmetros de estresse oxidativo no cérebro, embora os níveis de fenilalanina no sangue e no cérebro permanecessem inalterados. Os animais PKU apresentaram habilidades motoras e de equilíbrio deficientes em comparação com os controles, enquanto o exercício não afetou esses parâmetros comportamentais. Adicionalmente, os aminoácidos gliconeogênicos e os relacionados com o ciclo da ureia foram encontrados em níveis inferiores no plasma dos animais PKU que se exercitaram em comparação com os sedentários. Por outro lado, os camundongos selvagens não apresentaram nenhuma alteração causada pelo exercício. Assim, os fenilcetonúricos não apresentam distúrbios em marcadores metabólicos tanto em repouso como durante exercício aeróbico. Por conseguinte, os fenilcetonúricos devem ser encorajados a praticar exercício para, possivelmente, beneficiarem-se das adaptações positivas geradas pelo treinamento físico. No entanto, o pequeno número de pacientes avaliados em nossos estudos destaca a necessidade de mais pesquisas para descrever o exercício mais adequado para pacientes com PKU. / Phenylketonuria (PKU) is characterized by poor phenylalanine hydroxylase (PAH) activity, which acts converting phenylalanine into tyrosine. Consequently, PKU patients show increased levels of phenylalanine in the blood and tissues. Phenylalanine reaches toxic levels in the brain and therefore can lead to severe mental retardation, if untreated. Nowadays, PKU patients are diagnosed through newborn screening programs and are put on treatment immediately. PKU treatment is based on a phenylalanine-restricted diet along with an amino acid mixture, which aims to reduce the intake of phenylalanine while providing other essential amino acids and nutrients. Despite efficient, the treatment is extremely hard to follow so that PKU patients show high levels of phenylalanine and related issues such as oxidative stress, motor and cognitive disturbances. Moreover, the diet itself is so restrictive that adhering to it may lead to nutritional problems like obesity and hormonal disruptions. In order to tackle these issues in PKU, it is necessary to understand the mechanisms in which phenylalanine disturbs homeostasis as well as propose new treatment strategies for the disease. Therefore, this thesis aimed to evaluate metabolic markers in rest and exercise in PKU patients, as well as to verify the possible benefits of exercise in the brain of PAHenu2 mice. For that, we evaluated basal body composition, nutritional status, basal metabolic rate, as well as ventilatory and biochemical response to an aerobic exercise session in PKU patients and matched-controls using electrical bioimpedance analysis and indirect calorimetry, respectively. The groups did not differ for anthropometric and nutritional markers, thus showing that PKU patients have the same body and metabolic profiles as controls. Moreover, phenylalanine levels were not modified immediately after exercise in comparison to rest condition, thus proving that aerobic exercise is safe for PKU patients. In the animal model of PKU, we evaluated the effects of voluntary training in the brain of PAHenu2 mice. Despite running less distances than the controls, the PKU mice showed improved oxidative stress parameters although phenylalanine levels in the blood and in brain remained unchanged. PKU animals showed poor motor and balance skills than controls while exercise did not affect these behavioral markers. In addition, gluconeogenic and urea cycle-related amino acids were found in lower levels in the plasma of the exercised PKU animals in comparison to the sedentary PKU group. On the other hand, wild-type mice did not show any of those changes. Taken together, we conclude that PKU patients do not show disturbed metabolism in rest and during aerobic exercise. Therefore, PKU patients have to be encouraged to exercise in order to possibly benefit from long-term exercise-related adaptations. Nevertheless, the small number of patients evaluated in our studies highlights the need of further research to describe the most suitable exercise for PKU patients.

The aim of this study was to evaluate the National Neonatal Screening Program in Sergipe State in Brazil Northeastern (PNTN/SE) for phenylketonuria (PKU). It was performed a cross-sectional study. Variables assessed were: phenylalanine blood concentrations at filter paper collected from the heel of 43.449 children (PKUneo); blood phenylalanine concentrations obtained by venipuncture in the children with abnormal PKUneo; children s age in the different program phases from January 2007 to June 2008; and the coverage in 2007. The suspected children were selected when PKUneo were above the cut-off level of 5 mg/dL. Furthermore, these children were classified by the venous concentration of phenylalanine in according to the literature, thereby obtaining the prevalence of hyperphenylalaninemy (HPA) and phenylketonuria from January 2007 to June 2008. The cases diagnosed before 2007 were not analysed. Finally, we verified the venous concentrations of phenylalanine at those children on dietetic treatment for the disease as much as the amount of phenylalanine present on their diet. The children s age at PKUneo collection was 107 days (MDP), the age when the assay was done was 2813 days and at the venous collection in the diagnosis confirmation was 5317 days. Twelve children were called based on the PKUneo cut-off. From these, the concentrations of phenylalanine collected by venipuncture were normal in five children, one child was classified as hyperphenylalaninemy and five as PKU with the prevalence of 1/43449 and 1/8690, respectively. The treatment for PKU began with 5112 days. The coverage of PNTN/SE/2007 was 78.93%, besides, 11% of the Sergipe´s children that have private health care. In conclusion, PNTN/SE presented satisfactory coverage, PKU and hyperphenylalaninemy prevalences compatible with the literature and adequate cut-off. On the other hand, the collection of PKUneo is late and the onset of treatment is delayed. / O objetivo deste trabalho foi avaliar o Programa Nacional de Triagem Neonatal no Estado de Sergipe no Nordeste do Brasil (PNTN/SE) para a fenilcetonúria (PKU). Foi realizado um estudo transversal. As variáveis estudadas foram: concentrações de fenilalanina no sangue coletado em papel-filtro do calcanhar de 43.449 crianças (PKUneo); concentrações de fenilalanina no sangue coletado por punção venosa realizada nas crianças convocadas após resultado de PKUneo alterado; idade das crianças nas diferentes fases do PNTN/SE, no período entre janeiro de 2007 a junho de 2008, e a cobertura do programa no ano de 2007. As crianças suspeitas foram selecionadas quando apresentavam concentrações de PKUneo acima do ponto de corte de 5 mg/dL. Além disso, classificamos estas crianças segundo as concentrações venosas de fenilalanina de acordo com a literatura, calculando, assim, a prevalência de PKU e da hiperfenilalaninemia (HPA) no período de janeiro de 2007 a junho de 2008, não utilizando os casos diagnosticados antes de janeiro de 2007 e depois de junho de 2008. Por fim, foram acompanhadas as concentrações venosas de fenilalanina das crianças classificadas como fenilcetonúricas e hiperfenilalaninêmicas em tratamento dietético, assim como a quantidade de fenilalanina ingerida na alimentação. A idade das crianças, na coleta do PKUneo, foi de 107 dias (MDP), na realização do ensaio foi de 2813 dias e na coleta para confirmação do diagnóstico foi de 5317 dias. Foram convocadas doze crianças após resultado de PKUneo alterado, das quais cinco tiveram concentrações venosas normais de fenilalanina, uma foi classificada como hiperfenilalaninêmica e cinco como fenilcetonúricas com prevalência de 1/43449 e 1/8690, respectivamente. A terapia nas cinco crianças com PKU foi iniciada com 5112 dias. A cobertura do PNTN/SE em 2007 foi de 79%, não sendo considerados nesse resultado os 11% da população coberta por planos privados de saúde. Deste modo, o PNTN/SE apresentou no período estudado cobertura satisfatória, prevalências de PKU e HPA compatíveis com àquelas encontradas na literatura e ponto de corte adequado. Em contrapartida, a coleta do PKUneo é tardia e o início do tratamento é demorado.

Background: Specialized adult care of phenylketonuria (PKU) patients is of increasing importance. Adult outpatient clinics for inherited errors of metabolism can help to achieve this task, but experience is limited. Ten years after establishment of a coordinated transition process and specialised adult care for inherited metabolic diseases, adult PKU care was evaluated with respect to metabolic control, therapy satisfaction, life satisfaction, sociodemographic data, economical welfare as well as pregnancy outcome. Methods: All PKU patients transferred from paediatric to adult care between 2005 and 2015 were identified. A retrospective data analysis and a cross-sectional survey in a sub-cohort of 30 patients including a questionnaire for assessing quality of life (FLZm) were performed as a single-centre investigation at the metabolic department of the University Hospital Leipzig, Germany. For statistical analysis, Mann-Whitney-U-test, t-test for independent samples, ANOVA and chi square test were used as appropriate. Results: 96 PKU patients (56 females/40 males; median age 32 years, range 18–62) were included. In the last 3-year period, 81 % of the transferred patients still kept contact to the adult care centre. Metabolic control was stable over the evaluation period and dried blood phenylalanine concentrations mostly remained within the therapeutic range (median 673.0 μmol/l, range 213.0–1381.1). Sociodemographic data, economical welfare and life satisfaction data were comparable to data from the general population. However, differences could be revealed when splitting the cohort according to time of diagnosis and to management during childhood. 83 % of the PKU adults were satisfied with the transition process and current adult care. 25 completed pregnancies were supervised; three newborns, born after unplanned pregnancy, showed characteristic symptoms of maternal PKU syndrome. Conclusions: Continuous care for adult PKU patients in a specialized outpatient clinic is successful, leading to good to satisfactory metabolic control and social outcomes. Uninterrupted good metabolic treatment throughout childhood and adolescence positively influences educational, professional and economic success in later life. Further effort in specialized paediatric and adult metabolic care is needed to prevent loss of follow-up and to support the recommended life-long treatment and/or care.

Exogenously supplied organoselenium compounds, capable of propagating a selenium redox cycle, were shown to supplement natural cellular defenses against oxidants generated during biological activity. Phenylaminoalkyl selenides were developed in our laboratory as novel substrate analogs for the enzyme dopamine beta-monooxygenase. Recently, phenylaminoalkyl selenides were found to protect plasmid DNA and Molecular beacons from oxoperoxynitrate – mediated damage by scavenging this oxidant and forming the corresponding selenoxides as the sole selenium – containing products. Rate constants were determined for the reactions of the phenylaminoalkyl selenoxides with GSH at physiological pH and 25 degrees C. The kinetic data obtained in current and previous research was subsequently used in a MatLab simulation, which showed the feasibility of selenium redox cycling by GSH in the presence of a cellular oxidant, oxoperoxynitrate. Loblolly pine (LP, Pinus taeda) is the primary commercial species in southern forests covering 11.7 million hectares. Somatic embryogenesis (SE) is an effective technique to implement production of high value genotypes of LP. SE is a multi-step process, which includes initiation of somatic embryo (SME) growth from tree tissue, maintenance and multiplication of early stage SMEs and the maturation / germination phase. In this work, we isolated a substance from stage 2 or 3 LP female gametophyte (FG) tissue that stimulates early stage SME growth, and characterized this substance as citric acid on the basis of 1H NMR and mass spectrometry. We then demonstrated that topical application of citric acid to SMEs stimulates embryo colony growth at p = 0.05 for 3 of the 5 genotypes tested. Phenylketonuria (PKU) is an autosomal recessive disorder caused by an impaired conversion of L-phenylalanine (L-Phe) to L-tyrosine (L-Tyr). A novel assay based on enzymatic - colorimetric methodology (ECA) was developed in order to detect elevated concentrations of L-Phe in undeproteinized plasma of PKU patients via continuous spectrophotometric detection. We report here that L-Phe concentrations in undeproteinized plasma measured using our ECA were comparable to those determined on an amino acid analyzer based on Pearson correlation coefficients and a Bland and Altman comparison.