Fragment-guided approach to incorporating structural information into a CoMFA study: BACE-1 as an example

Alzheimer`s disease is an ultimately fatal neurodegenerative disease, and BACE-1 has become an attractive validated target for its therapy, with more than a hundred crystal structures deposited in the PDB. In the present study, we present a new methodology that integrates ligand-based methods with structural information derived from the receptor. 128 BACE-1 inhibitors recently disclosed by GlaxoSmithKline R&D were selected specifically because the crystal structures of 9 of these compounds complexed to BACE-1, as well as five closely related analogs, have been made available. A new fragment-guided approach was designed to incorporate this wealth of structural information into a CoMFA study, and the methodology was systematically compared to other popular approaches, such as docking, for generating a molecular alignment. The influence of the partial charges calculation method was also analyzed. Several consistent and predictive models are reported, including one with r (2) = 0.88, q (2) = 0.69 and r (pred) (2) = 0.72. The models obtained with the new methodology performed consistently better than those obtained by other methodologies, particularly in terms of external predictive power. The visual analyses of the contour maps in the context of the enzyme drew attention to a number of possible opportunities for the development of analogs with improved potency. These results suggest that 3D-QSAR studies may benefit from the additional structural information added by the presented methodology.

Generation of Cholesterol Carboxyaldehyde by the Reaction of Singlet Molecular Oxygen [O(2) ((1)Delta(g))] as Well as Ozone with Cholesterol

A few years ago, it was reported that ozone is produced in human atherosclerotic arteries, on the basis of the identification of 3 beta-hydroxy-5-oxo-5,6-secocholestan-6-al and 3 beta-hydroxy-5 beta-hydroxy-B-norcholestane-6 beta-carboxaldehyde (ChAld) as their 2,4-dinitrophenylhydrazones. The formation of endogenous ozone was attributed to water oxidation catalyzed by antibodies, with the formation of dihydrogen trioxide as a key intermediate. We now report that ChAld is also generated by the reaction of cholesterol with singlet molecular oxygen [O(2) ((1)Delta(g))] that is produced by photodynamic action or by the thermodecomposition of 1,4-dimethylnaphthalene endoperoxide, a defined pure chemical source of O(2) ((1)Delta(g)). On the basis of (18)O-labeled ChAld mass spectrometry, NMR, light emission measurements, and derivatization studies, we propose that the mechanism of ChAld generation involves the formation of the well-known cholesterol 5 alpha-hydroperoxide (5 alpha-OOH) (the major product of O(2) ((1)Delta(g))-oxidation of cholesterol) and/or a 1,2-dioxetane intermediate formed by O(2) ((1)Delta(g)) attack at the Delta(5) position. The Hock cleavage of 5 alpha-OOH (the major pathway) or unstable cholesterol dioxetane decomposition (a minor pathway, traces) gives a 5,6-secosterol intermediate, which undergoes intramolecular aldolization to yield ChAld. These results show clearly and unequivocally that ChAld is generated upon the reaction of cholesterol with O(2) ((1)Delta(g)) and raises questions about the role of ozone in biological processes.

Evidence that OGG1 glycosylase protects neurons against oxidative DNA damage and cell death under ischemic conditions

7,8-Dihydro-8-oxoguanine DNA glycosylase (OGG1) is a major DNA glycosylase involved in base-excision repair (BER) of oxidative DNA damage to nuclear and mitochondrial DNA (mtDNA). We used OGG1-deficient (OGG1(-/-)) mice to examine the possible roles of OGG1 in the vulnerability of neurons to ischemic and oxidative stress. After exposure of cultured neurons to oxidative and metabolic stress levels of OGG1 in the nucleus were elevated and mitochondria exhibited fragmentation and increased levels of the mitochondrial fission protein dynamin-related protein 1 (Drp1) and reduced membrane potential. Cortical neurons isolated from OGG1(-/-) mice were more vulnerable to oxidative insults than were OGG1(+/+) neurons, and OGG1(-/-) mice developed larger cortical infarcts and behavioral deficits after permanent middle cerebral artery occlusion compared with OGG1(+/+) mice. Accumulations of oxidative DNA base lesions (8-oxoG, FapyAde, and FapyGua) were elevated in response to ischemia in both the ipsilateral and contralateral hemispheres, and to a greater extent in the contralateral cortex of OGG1(-/-) mice compared with OGG1(+/+) mice. Ischemia-induced elevation of 8-oxoG incision activity involved increased levels of a nuclear isoform OGG1, suggesting an adaptive response to oxidative nuclear DNA damage. Thus, OGG1 has a pivotal role in repairing oxidative damage to nuclear DNA under ischemic conditions, thereby reducing brain damage and improving functional outcome. Journal of Cerebral Blood Flow & Metabolism (2011) 31, 680-692; doi:10.1038/jcbfm.2010.147; published online 25 August 2010

Expression Profile of Rat Hippocampal Neurons Treated with the Neuroprotective Compound 2,4-Dinitrophenol: Up-Regulation of cAMP Signaling Genes

2,4-Dinitrophenol (DNP) is classically known as a mitochondrial uncoupler and, at high concentrations, is toxic to a variety of cells. However, it has recently been shown that, at subtoxic concentrations, DNP protects neurons against a variety of insults and promotes neuronal differentiation and neuritogenesis. The molecular and cellular mechanisms underlying the beneficial neuroactive properties of DNP are still largely unknown. We have now used DNA microarray analysis to investigate changes in gene expression in rat hippocampal neurons in culture treated with low micromolar concentrations of DNP. Under conditions that did not affect neuronal viability, high-energy phosphate levels or mitochondrial oxygen consumption, DNP induced up-regulation of 275 genes and down-regulation of 231 genes. Significantly, several up-regulated genes were linked to intracellular cAMP signaling, known to be involved in neurite outgrowth, synaptic plasticity, and neuronal survival. Differential expression of specific genes was validated by quantitative RT-PCR using independent samples. Results shed light on molecular mechanisms underlying neuroprotection by DNP and point to possible targets for development of novel therapeutics for neurodegenerative disorders.

O polimorfismo da apolipoproteina e em três grupos étnicos e suas influências sobre os níveis lipídicos e a doença de Alzheimer

O gene da apolipoproteina E (APOE) possui três alelos com freqüências polimórficas. Esta apolipoproteína possui um importante papel no metabolismo de lipídeos, crescimento e regeneração neuronal, e parece estar relacionada com a doença de Alzheimer. No entanto, a magnitude destas influências difere de acordo com a população estudada, sugerindo uma interação genótipo/ambiente. No presente trabalho, foram estudadas seis tribos indígenas sul-americanas (n=186), 100 negróides e 466 caucasóides de Porto Alegre. Destes últimos, 343 foram investigados quanto à associação com níveis lipídicos e 23 quanto à associação com doença de Alzheimer. Todas as amostras foram amplificadas pela reação em cadeia da polimerase (PCR) e clivadas com a enzima de restrição Hha I. Os genótipos foram identificados após separação dos fragmentos de restrição por eletroforese em gel de agarose a 4% corado com brometo de etídeo. O presente estudo teve os seguintes objetivos específicos: 1)Determinar as freqüências gênicas e genotípicas da APOE nas populações negróides e caucasóides de Porto Alegre e de seis tribos indígenas da América do Sul; 2)Verificar se as associações entre os alelos da APOE e lipídeos séricos descritas em caucasóides também ocorrem em populações indígenas brasileiras; 3)Investigar a influência do polimorfismo do gene APOE em pacientes com hipercolesterolemia e hipertrigliceridemia, bem como em indivíduos normais da população de Porto Alegre e 4)Determinar a distribuição dos alelos da APOE em uma amostra de pacientes com Doença de Alzheimer.

O cuidador familiar do idoso com alzheimer: percepções e sentimentos

The Brazilian Constitution maintains that care for elderly people is a responsibility shared by the state, the family and the society. The politics for the elderly corroborate this understanding and treats home as a privileged place for elderly care taking. This determines the participation of the familiar as a caregiver, but highlights the lack of strategic assistance for the needs of the relative caregiver who feels helplessly and unattended in their responsibility for elderly homecare. In recent years , despite the recently pursuit for health and life quality, there is an increasing incidence of elderly patients with dementia diseases that lead to disability, the most common among then is the Alzheimer´s disease. This disease affects seriously and irreversibly cognition, memory and independence of the elderly, making it dependent on others to perform basic activities of daily life, for all his life. The present study aims to evaluate the perceptions and feelings of family caregivers of elders with Alzheimer on the role of caregiver. This is a qualitative study conducted with family caregivers of seniors with Alzheimer´s, caregivers linked to the group of the Specialized Care Center of the Elderly´s Health, located in Natal / RN. Through semi-structured interview research sought to investigate the perceptions of family caregivers on the role of caregiver, the feelings and the changes in the caregiver´s life since they assumed this role. The data were organized into categories and units of semantic analysis and analyzed using thematic content analysis by Bardin. The reports originated three categories: the perception of the role of caregiver, feelings related to the caregving and consequences of the caregiver role. Perceptions of caregivers of elderly from the requirement of dedication to the care generates losses in personal and professional life for the familiar who assumes this responsibility. The lack of family and social support, aggravates the burden of care for the dependent elderly. Public health politics for the elderly recognize the importance and needs of family caregivers, but not enough to provide support and meet the needs and assist them in supporting their limitations. The research results show the urgent need to take measures to assist the caregivers of seniors with Alzheimer, recognizing them as an action of promotion quality of life and health of the elderly and protection the health of the caregiver

Efeito do treinamento com pesos na apatia, funções cognitivas frontais e funcionalidade motora em pacientes com doença de Alzheimer

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeitos do treinamento com pesos nos sintomas depressivos e variáveis metabólicas em pacientes com doença de Alzheimer

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo de Jatropha gossypifolia e J. multifida (Euphorbiaceae) aplicando métodos analíticos in silico e de desreplicação, visando a detecção e elucidação in situ dos constituintes micromoleculares com atividade acetilcolinesterásicas e antioxidante

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Alterações oxidativas em portadores da doença de Alzheimer: correlação com os estágios da doença e o quadro cognitivo dos pacientes

A sobrevida da população tem aumentado de forma progressiva e atribui-se à melhoria da qualidade dos serviços de saúde e das condições gerais de vida, sendo marcante, sobretudo nos países industrializados. Este aumento da expectativa de vida repercute no aumento da incidência de doenças comuns em idades avançadas. O envelhecimento é considerado fator de risco para o desenvolvimento de doenças neurodegenerativas como a doença de Alzheimer (DA). Uma das teorias para a patogênese da DA postula que a neurodegeneração é o resultado de alterações no metabolismo oxidativo com acometimento do tecido cerebral vulnerável. O fato de o envelhecimento ser um fator de risco na DA, reforça também a hipótese da participação dos radicais livres, pois os efeitos de sua ação podem se acumular durante anos. A DA é uma doença insidiosa e progressiva e caracteriza-se clinicamente por uma perda progressiva da memória e de outras funções cognitivas, além de mudanças comportamentais e sociais. O déficit de memória é o principal e usualmente o primeiro sintoma da DA, com comprometimento especialmente da retenção e recordação de novas informações. O presente trabalho visa avaliar se ocorrem alterações no metabolismo oxidativo detectáveis no sangue de pacientes com a DA, e se estas podem ser relacionadas com os diferentes estágios da doença e com o quadro cognitivo dos pacientes. Foram avaliados 30 pacientes com a doença de Alzheimer e 28 indivíduos no grupo controle atendidos na Unidade de Ensino e Assistência de Fisioterapia e Terapia Ocupacional da Universidade do Estado do Pará (UEAFTO-UEPA). O metabolismo oxidativo foi avaliado através da medida da capacidade antioxidante total equivalente ao Trolox (TEAC) e da mensuração dos níveis das substâncias que reagem ao ácido tiobarbitúrico - TBARS. Também foram realizados testes neuropsicológicos em todos os sujeitos participantes do estudo. Não foram observadas correlações significativas no desempenho dos testes neuropsicológicos com os níveis de TBARS e TEAC nos pacientes com a DA. Observou-se que a capacidade antioxidante total (CAOT) estava significativamente diminuída nos pacientes com a DA em comparação com o grupo controle independentemente do estágio da doença, mostrando uma possível relação entre a CAOT e a DA. Na avaliação de TBARS houve uma tendência para maiores concentrações nos pacientes com DA do que no grupo controle, porém a diferença não foi estatisticamente significativa, apenas a fase moderada foi significativa quando comparada com o grupo controle. A avaliação da peroxidação lipídica através dos níveis de TBARS provavelmente não seria um biomarcador adequado para a doença de Alzheimer.

Espectrometria de massas aplicada aos estudos de biossíntese de alcalóides de Senna spectabilis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudos computacionais e sintéticos visando o planejamento racional de novos agentes anticolinesterásicos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Association between oxidative stress and nutritional status in the elderly

O envelhecimento é um processo dinâmico e progressivo que se caracteriza pela ocorrência de modificações morfológicas, bioquímicas, funcionais e psicológicas do organismo. O objetivo do presente artigo é fornecer conceitos atualizados sobre estresse oxidativo, abordando sua importância no envelhecimento, assim como o estado nutricional e suplementação com antioxidantes (substâncias que evitam ou atenuam a oxidação de substratos oxidáveis, como lipídeos, proteínas, carboidratos e o ácido desoxirribonucleico) na população geriátrica. Evidências sugerem que existe uma relação inversa entre estresse oxidativo e estado nutricional em indivíduos idosos. Embora seja comprovado o aumento do estresse oxidativo nas doenças crônicas associadas ao envelhecimento, como doença de Parkinson e doença de Alzheimer, não há, até o momento, evidências clínicas consistentes que comprovem a eficiência da suplementação com antioxidantes contra o estresse oxidativo. Nesse contexto, a suplementação não é recomendada. Por outro lado, idosos devem ser encorajados a ingerir alimentos antioxidantes, como, por exemplo, frutas e vegetais. A manutenção do peso dentro da faixa de normalidade (índice de massa corpórea entre 23 e 28 kg/m2) também deve ser estimulada.

A percepção da doença de Alzheimer pelos estudantes da F.F.C. Júlio de Mesquita Filho da UNESP, Marília

Alzheimer’s disease (AD) accounts for approximately 56% of all cases of dementia. The present study aimed to report the perception of AD by students of Physiotherapy, Speech Therapy and Occupational Therapy from Faculty of Philosophy and Science “Julio de Mesquita Filho” – UNESP – Marília, with regard to the symptoms, diagnosis and treatment, between 2011 and 2012. The questionnaire with open and closed questions was administered to 101 students, using as a criterion for exclusion those who refused to answer the survey and / or not signed the informed consent form. After the questionnaire application, the answers have undergone quantitative analysis, checking which information and concepts about Alzheimer’s disease these students acquired during the undergraduate years. The data shows that although students have knowledge about the signs and symptoms of the disease, few mention the importance of family, the caregiver and the relationship of the disease with dependence that commonly reaches this population.

Análise do equilíbrio e da qualidade de vida de idosos com Alzheimer e a influência na qualidade de vida do Cuidador

With the growing aging population will be an increase of chronic degenerative diseases such as dementia. Among the various forms of dementia Alzheimer’s disease (AD) is the most prevalent. In individuals with AD, there is a loss in the processing of sensory information, which may aggravate the imbalance and falls. As the disease progresses, the individual lose the ability to function independently, becoming dependent on a caregiver. This study aimed to analyze the balance of the mental state and quality of life of individuals with AD, to determine whether a correlation exists between these variables and analyze the influence on quality of life of caregivers. This study was conducted with thirty individuals (82.86 ± 9.07 years) with AD, both sexes, and their caregivers. The evaluation of the balance was accomplished by the Scale of Functional Balance of Berg (EEFB), the cognitive function for the Mini-exam of the Mental State (MEEM), and the quality of life (QV) for the scale “life Quality - Disease of Alzheimer “ (QdV - DA) that is composed for three versions: patient, caregiver and family The data were analyzed by coefficient of correlation of Spearman. The balance analyses (EEFB=32,17 ± 13,26 points) shows increased in the risk of falls in the elderly and negative correlation (R = - 0,55, p <0,01) with age and good correlation with MEEM (R=0,63 p <0,01). Already in relation of the MEEM and QV, can observed correlation between the familiar version and the MEEM ((R=0,40 p=0,02). In Relation the versions of the QV questionnaire, found significant correlation among: QdV-DA patient X caregiver (R=0,41 p=0,02), QdV-DA patient X family (R=0,40 p=0,03). In this way we can conclude that the individuals with DA, appraised in this study, present a deficit in the balance, so much related with the age as with to the cognitive decline, and the greater the cognitive decline worse the impression of caring about the QOL of their family, and still, that the worsening in the quality of the patient’s life contemplates in a worsening in the quality of your caregiver’s life.