The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.

Efeito dos ácidos graxos ômega-3 sobre o relaxamento-dependente do endotélio em coelhos hipercolesterolêmicos

OBJETIVO: Estudar o efeito dos ácidos graxos ômega-3 sobre o relaxamento-dependente do endotélio, o colesterol plasmático, as LDL, VLDL, HDL, triglicérides e a peroxidação lipídica das partículas de LDL-nativas, oxidadas e da parede arterial. MÉTODOS: Coelhos da raça Nova Zelândia foram submetidos a dieta enriquecida com colesterol (0,5%) e gordura de coco (2%), por 30 dias e separados em grupo hipercolesterolemia (H) e ômega-3 (O-3), sendo administrado ao O-3 ácidos graxos ômega-3 na dose de 300mg/kg/dia, durante 15 dias, através de gavagem. O colesterol plasmático, triglicérides, LDL-colesterol, VLDL e HDL-colesterol foram medidos através de kits enzimáticos e os resultados expressos em mg/dl. As LDL foram obtidas por ultracentrifugação e oxidadas através da exposição ao Cu++. A peroxidação lipídica das LDL e da parede da aorta foi mensurada pela dosagem do malondialdeido (MDA). A função endotelial foi avaliada por curvas de concentração-efeito obtidas pela acetilcolina e nitroprussiato, após contração com norepinefrina. RESULTADOS: Houve aumento do colesterol plasmático e das VLDL, sem interferência nos níveis de LDL e HDL, no O-3. Observou-se redução significante dos triglicérides. Verificou-se aumento significante do teor de MDA nas LDL-nativas e oxidadas, assim como na parede arterial. O relaxamento-dependente do endotélio foi significativamente menor no O-3. CONCLUSÃO: A administração de ácidos graxos ômega-3 na dosagem de 300/mg/kg/dia, a coelhos hipercolesterolêmicos aumentou o colesterol e as VLDL plasmáticas, enquanto reduziu os triglicérides. O relaxamento-dependente do endotélio foi menor que no grupo H.

Motivação para o desporto escolar: estudo em alunos do 2º e 3º ciclo de escolaridade numa Escola Básica do concelho de Fafe

Relatório de estágio de mestrado em Ensino de Educação Física nos Ensinos Básico e Secundário

Metodologia para o ensino da química no 3.º ciclo do ensino básico: Química para todos: um estudo centrado na estrutura da matéria, transformações químicas e físicas, Lei de Lavoisier e velocidade das reações

Dissertação de mestrado em Ciências – Formação Contínua de Professores (área de especialização em Física e Química)

Síntese de novas tieno[3,2-b]piridinas com potencial atividade biológica contendo 1,2,3-triazoles 1,4-dissubstituídos obtidos por reação "click" de ciclioadição azida-alquino catalisada por Cu(I)-CuAAC

Dissertação de mestrado em Química Medicinal

Role of sirtuin 3 on mitochondrial dynamics in Huntington's disease striatal cells

Dissertação de mestrado em Genética Molecular

24-h urinary free cortisol from mid-pregnancy to 3-months postpartum: gender and parity differences and effects

Background: Pregnancy and postpartum have been associated to several physiological changes;however, empirical evidence was almost exclusively obtained in primiparous women and few studies focus on hormonal changes in men and second-time parents. The main aim of this study is to examine 24-h urinary free cortisol from mid-pregnancy to 3-months postpartum, comparing women/men and first/second-time parents.Methods: Twenty-six women and 22 men (N = 48) were recruited from an antenatal obstetric unit in Porto, Portugal. 24-h urinary free cortisol was measured at the 2nd and 3rd trimester and at 3-months postpartum. Repeated measures analyses of variance were conducted, in order to analyze 24-h urinary free cortisol patterns of change over this period. Gender and parity were included in the analyses as potential modifiers, in order to compare women and men, and first-and second-time parents.Results: An increase from the 2nd to the 3rd trimester (p = .006) and a decrease from the 3rd trimester to 3-months postpartum (p = .005) were reported in all parents’ 24-h urinary free cortisol. The interaction effects for Time * Gender (p = .03) and Time * Parity (p = .02) were found. Women and first-time parents revealed higher levels, while men and second-time parents showed lower 24-h urinary free cortisol levels at the 2nd trimester than at 3-months postpartum.Conclusions: Findings appear to clarify the direction, as well as, the timing, gender and parity extension of 24-h urinary free cortisol changes from mid-pregnancy to 3-months postpartum.The same pattern of change in all parents’ 24-h urinary free cortisol from mid-pregnancy to 3-months postpartum is consistent with the proposed role of hormones in preparation to parenting.

A avaliação da fluência oral em crianças do 3º ano do ensino básico com e sem necessidades educativas especiais: um estudo exploratório quantitativo

Dissertação de mestrado em Educação Especial (área de especialização em Dificuldades de Aprendizagem Específicas)

Infant’s psychophysiological profile and temperament at 3 and 12 months

This study is intended to analyze (1) differences in infant temperament at 3 and 12 months according to infants' psychophysiological profiles: "withdrawn" "extroverted", and "underaroused" and (2) changes in infant temperament from 3 to 12 months, namely according to the infant psychophysiological profile and the quality of mother-infant interaction. Ninety-four 8-week-old infants were assessed using the Neonatal Behavioral Assessment Scale (NBAS, Brazelton & Nugent, 1995) and the Alarm Distress Baby Scale (ADBB, Guedeney & Fermanian, 2001). Saliva samples were collected at 8-12 weeks old, both before and after a routine inoculation for cortisol reactivity measurement. Mother-infant interaction was evaluated at 12-16 weeks, using the Global Rating Scales (GRS, Murray, Fiori-Cowley, Hooper, & Cooper, 1996). Mothers' reports on infant temperament at 3 and 12 months were collected using the Infant Behavior Questionnaire (IBQ, Rothbart, 1981). Significant differences in mothers' perception of infant temperament were found at both 3 and 12 months in infants with distinct psychophysiological profiles. Stability was observed in most of the temperament's dimensions from 3 to 12 months old; still, there were changes in mothers' perception of infant temperament in terms of level of distress, cuddliness, sadness and approach. Infant psychophysiological profile and mother-infant interaction both influence with the pattern of those changes. The results corroborate that infant's characteristics early in life as well as contextual factors influence with mothers' perception of infant temperament and with changes across the 1st year of life.

Anxiety and depression symptoms in women and men from early pregnancy to 3-months postpartum: parity differences and effects

This study aimed to investigate both anxiety and depression symptoms from early pregnancy to 3-months postpartum, comparing women and men and first and second-time parents. Methods: A sample of 260 Portuguese couples (N=520), first or second-time parents, recruited in an Obstetrics Out-patients Unit, filled in the State-Anxiety Inventory (STAI-S) and the Edinburgh Post-Natal Depression Scale (EPDS) at the 1st, 2nd and 3rd pregnancy trimesters, childbirth, and 3-months postpartum. Results: A decrease in anxiety and depression symptoms from early pregnancy to 3-months postpartum was found in both women and men, as well as in first and second-time parents. Men presented less anxiety and depression symptoms than women, but the same pattern of symptoms over time. Second-time parents showed more anxiety and depression symptoms than first-time parents and a different pattern of symptoms over time: an increase in anxiety and depression symptoms from the 3rd trimester to childbirth was observed in first-time parents versus a decrease in second-time parents. Limitations: The voluntary nature of the participation may have lead to a selection bias; women and men who agreed to participate could be those who presented fewer anxiety and depression symptoms. Moreover, the use of self-report symptom measures does not give us the level of possible disorder in participants. Conclusions: Anxiety and depression symptoms diminish from pregnancy to the postpartum period in all parents. Patterns of anxiety and depression symptoms from early pregnancy to 3-months postpartum are similar in women and men, but somewhat different in first and second time parents. Second-time parents should also be considered while studying and intervening during pregnancy and the postpartum.

Mother's stress, mood and emotional involvement with the infant: 3 months before and 3 months after childbirth

Adverse effects of maternal anxiety and depression are well documented, namely on the foetus/child behaviour and development, but not as much attention has been given to the mother's emotional involvement with the offspring. To study mother's prenatal and postpartum stress, mood and emotional involvement with the infant, the State-Trait Anxiety Inventory, the Edinburgh Postnatal Depression Scale and the Mother-to-Infant Bonding Scale were filled in and cortisol levels were measured, 3 months before and 3 months after childbirth, in a sample of 91 Portuguese women. From pregnancy to the postpartum period, mother's cortisol levels, anxiety and emotional involvement toward the child decrease. No significant change was observed regarding mother's depression. Mother's depression predicted a worse emotional involvement before childbirth, while mother's anxiety predicted a worse emotional involvement with the infant after childbirth. Additionally, pregnant women with a worse emotional involvement with the offspring are at risk of poorer emotional involvement with the infant and higher anxiety and depression at 3 months postpartum. It should be given more attention to mother's poor emotional involvement with the offspring during pregnancy, as it interferes with her emotional involvement with the infant and her psychological adjustment 3 months after childbirth.