Dual inhibitors of beta-amyloid aggregation and acetylcholinesterase as multi-target anti-Alzheimer drug candidates

Notwithstanding the functional role that the aggregates of some amyloidogenic proteins can play in different organisms, protein aggregation plays a pivotal role in the pathogenesis of a large number of human diseases. One of such diseases is Alzheimer"s disease (AD), where the overproduction and aggregation of the β-amyloid peptide (Aβ) are regarded as early critical factors. Another protein that seems to occupy a prominent position within the complex pathological network of AD is the enzyme acetylcholinesterase (AChE), with classical and non-classical activities involved at the late (cholinergic deficit) and early (Aβ aggregation) phases of the disease. Dual inhibitors of Aβ aggregation and AChE are thus emerging as promising multi-target agents with potential to efficiently modify the natural course of AD. In the initial phases of the drug discovery process of such compounds, in vitro evaluation of the inhibition of Aβ aggregation is rather troublesome, as it is very sensitive to experimental assay conditions, and requires expensive synthetic Aβ peptides, which makes cost-prohibitive the screening of large compound libraries. Herein, we review recently developed multi-target anti-Alzheimer compounds that exhibit both Aβ aggregation and AChE inhibitory activities, and, in some cases also additional valuable activities such as BACE-1 inhibition or antioxidant properties. We also discuss the development of simplified in vivo methods for the rapid, simple, reliable, unexpensive, and high-throughput amenable screening of Aβ aggregation inhibitors that rely on the overexpression of Aβ42 alone or fused with reporter proteins in Escherichia coli.

Synthesis and multi-target biological profiling of a novel family of rhein derivatives as disease-modifying anti-Alzheimer agents

We have synthesized a family of rhein-huprine hybrids to hit several key targets for Alzheimer"s disease. Biological screening performed in vitro and in Escherichia coli cells has shown that these hybrids exhibit potent inhibitory activities against human acetylcholinesterase butyrylcholinesterase, and BACE-1, dual Aβ42 and tau anti-aggregating activity, and brain permeability. Ex vivo studies with the leads (+)- and (-)-7e in brain slices of C57bl6 mice have revealed that they efficiently protect against the Aβ-induced synaptic dysfunction , preventing the loss of synaptic proteins and/or have a positive effect on the induction of long term potentiation. In vivo studies in APP-PS1 transgenic mice treated i.p. for 4 weeks with (+)- and (-)-7e have shown a central soluble Aβ lowering effect, accompanied by an increase in the levels of mature amyloid precursor protein (APP). Thus, (+)- and (-)-7e emerge as very promising disease-modifying anti-Alzheimer drug candidates.

Multigram synthesis and in vivo efficacy studies of a novel multitarget anti-Alzheimer's compound

We describe the multigram synthesis and in vivo efficacy studies of a donepezil‒huprine hybrid that has been found to display a promising in vitro multitarget profile of interest for the treatment of Alzheimer's disease (AD). Its synthesis features as the key step a novel multigram preparative chromatographic resolution of intermediate racemic huprine Y by chiral HPLC. Administration of this compound to transgenic CL4176 and CL2006 Caenorhabditis elegans strains expressing human Aβ42, here used as simplified animal models of AD, led to a significant protection from the toxicity induced by Aβ42. However, this protective effect was not accompanied, in CL2006 worms, by a reduction of amyloid deposits. Oral administration for 3 months to transgenic APPSL mice, a well-established animal model of AD, improved short-term memory, but did not alter brain levels of Aβ peptides nor cortical and hippocampal amyloid plaque load. Despite the clear protective and cognitive effects of AVCRI104P4, the lack of Aβ lowering effect in vivo might be related to its lower in vitro potency toward Aβ aggregation and formation as compared with its higher anticholinesterase activities. Further lead optimization in this series should thus focus on improving the anti-amyloid/anticholinesterase activity ratio.

Un estudio transversal sobre la percepción de la vitalidad etnolingüística en jóvenes y adultos de Catalunya = A cross-sectional study on ethnolinguistic vitality perception in young and adulthood people from Catalonia

El estudio utiliza un diseño transversal en el campo de la percepción de la vitalidad etnolingüística. Es el primero que compara la percepción de la vitalidad etnolingüística, así como los factores asociados, entre adultos jóvenes y adultos, en relación con los grupos castellanohablantes y catalanohablantes de la Comunidad Autónoma de Cataluña. Para ello, se aplicó el 'Cuestionario de vitalidad etnolingüística subjetiva' (CVS) a una muestra de 527 participantes, 268 jóvenes y 259 adultos, de los cuales se seleccionó una submuestra de individuos que tenían el catalán como lengua materna y se identificaban como catalanes (n=301). En ambas muestras se aduce una tendencia a descriminar favorablemente la vitalidad percibida por el grupo catalán, aspecto que se acentúa significativa en el grupo de jóvenes estudiados en relación al grupo de edad de los adultos. Se discuten los resultados según las repercusiones teóricas y pragmáticas de los estudios realizados en el ámbito de la comunicación intergrupal

Análisis psicométrico de la escala multidimensional de autoconcepto AF5 en una muestra de adolescentes y adultos de Cataluña

Psychometric analysis of the AF5 multidimensional scale of self-concept in a sample of adolescents and adults in Catalonia. The aim of this study is to carry out a psychometric study of the AF5 scale in a sample of 4.825 Catalan subjects from 11 to 63 years-old. They are students from secondary compulsory education (ESO), from high school, middle-level vocational training (CFGM) and from the university. Using a principal component analysis (PCA) the theoretical validity of the components is established and the reliability of the instrument is also analyzed. Differential analyses are performed by gender and normative group using a 2 6 factorial design. The normative group variable includes the different levels classifi ed into 6 sub-groups: university, post-compulsory secondary education (high school and CFGM), 4th of ESO, 3rd of ESO, 2nd of ESO and 1st of ESO. The results indicate that the reliability of the Catalan version of the scale is similar to the original scale. The factorial structure also fi ts with the original model established beforehand. Signifi cant differences by normative group in the four components of self-concept explored (social, family, academic/occupational and physical) are observed. By gender, signifi cant differences appear in the component of physical self-concept, academic and social but not in the family component

DOENÇA DE ALZHEIMER: HIPÓTESES ETIOLÓGICAS E PERSPECTIVAS DE TRATAMENTO

With the increase in life expectancy registered in the past few decades, the prevalence of various medical conditions related to aging has been observed, such as dementia and related neurodegenerative conditions. The number of patients afflicted with these conditions is expected to significantly increase in the coming years. The growing social impact of dementia underlines the need for research aimed at identifying and better understanding this type of condition. Among neurodegenerative diseases, amyloidogenic diseases, in particular Alzheimer's disease (AD), are currently the most common form of dementia. Over the years, several hypotheses have been raised regarding the etiology of AD, such as the cholinergic, glutamatergic, amyloid cascade, oligomeric, metallic and diabetes type 3 hypotheses. Unfortunately, no cure is yet available for this disease, only drugs that aid in controlling the symptoms. This review article conducts a comprehensive approach of the main etiological hypotheses of AD, as well as the treatment prospects associated with each hypothesis.

Early Detection of Alzheimer's Disease Beta-amyloid Pathology -Applicability of Positron Emission Tomography with the Amyloid Radioligand 11C-PIB

Early Detection of Alzheimer's Disease Beta-amyloid Pathology -Applicability of Positron Emission Tomography with the Amyloid Radioligand ¹¹C-PIB Accumulation of beta amyloid (Abeta) in the brain is characteristic for Alzheimer’s disease (AD). Carbon-11 labeled 2-(4’-methylaminophenyl)-6-hydroxybenzothiazole (¹¹C-PIB) is a novel positron emission tomography (PET) amyloid imaging agent that appears to be applicable for in vivo Abeta plaque detection and quantitation. The biodistribution and radiation dosimetry of ¹¹C-PIB were investigated in 16 healthy subjects. The reproducibility of a simplified ¹¹C-PIB quantitation method was evaluated with a test-retest study on 6 AD patients and 4 healthy control subjects. Brain ¹¹C-PIB uptake and its possible association with brain atrophy rates were studied over a two-year follow-up in 14 AD patients and 13 healthy controls. Nine monozygotic and 8 dizygotic twin pairs discordant for cognitive impairment and 9 unrelated controls were examined to determine whether brain Abeta accumulation could be detected with ¹¹C-PIB PET in cognitively intact persons who are at increased genetic risk for AD. The highest absorbed radiation dose was received by the gallbladder wall (41.5 mjuGy/MBq). About 20 % of the injected radioactivity was excreted into urine, and the effective whole-body radiation dose was 4.7 mjuSv/MBq. Such a dose allows repeated scans of individual subjects. The reproducibility of the simplified ¹¹C-PIB quantitation was good or excellent both at the regional level (VAR 0.9-5.5 %) and at the voxel level (VAR 4.2-6.4 %). ¹¹C-PIB uptake did not increase during 24 months’ follow-up of subjects with mild or moderate AD, even though brain atrophy and cognitive decline progressed. Baseline neocortical ¹¹C-PIB uptake predicted subsequent volumetric brain changes in healthy control subjects (r = 0.725, p = 0.005). Cognitively intact monozygotic co-twins – but not dizygotic co-twins – of memory-impaired subjects exhibited increased ¹¹C-PIB uptake (117-121 % of control mean) in their temporal and parietal cortices and the posterior cingulate (p<0.05), when compared with unrelated controls. This increased uptake may be representative of an early AD process, and genetic factors seem to play an important role in the development of AD-like Abeta plaque pathology. ¹¹C-PIB PET may be a useful method for patient selection and follow-up for early-phase intervention trials of novel therapeutic agents. AD might be detectable in high-risk individuals in its presymptomatic stage with ¹¹C-PIB PET, which would have important consequences both for future diagnostics and for research on disease-modifying treatments.

Verbal learning in mild cognitive impairment and alzheimer's disease : behavioural and neural approaches

The main focus of the present thesis was at verbal episodic memory processes that are particularly vulnerable to preclinical and clinical Alzheimer’s disease (AD). Here these processes were studied by a word learning paradigm, cutting across the domains of memory and language learning studies. Moreover, the differentiation between normal aging, mild cognitive impairment (MCI) and AD was studied by the cognitive screening test CERAD. In study I, the aim was to examine how patients with amnestic MCI differ from healthy controls in the different CERAD subtests. Also, the sensitivity and specificity of the CERAD screening test to MCI and AD was examined, as previous studies on the sensitivity and specificity of the CERAD have not included MCI patients. The results indicated that MCI is characterized by an encoding deficit, as shown by the overall worse performance on the CERAD Wordlist learning test compared with controls. As a screening test, CERAD was not very sensitive to MCI. In study II, verbal learning and forgetting in amnestic MCI, AD and healthy elderly controls was investigated with an experimental word learning paradigm, where names of 40 unfamiliar objects (mainly archaic tools) were trained with or without semantic support. The object names were trained during a 4-day long period and a follow-up was conducted one week, 4 weeks and 8 weeks after the training period. Manipulation of semantic support was included in the paradigm because it was hypothesized that semantic support might have some beneficial effects in the present learning task especially for the MCI group, as semantic memory is quite well preserved in MCI in contrast to episodic memory. We found that word learning was significantly impaired in MCI and AD patients, whereas forgetting patterns were similar across groups. Semantic support showed a beneficial effect on object name retrieval in the MCI group 8 weeks after training, indicating that the MCI patients’ preserved semantic memory abilities compensated for their impaired episodic memory. The MCI group performed equally well as the controls in the tasks tapping incidental learning and recognition memory, whereas the AD group showed impairment. Both the MCI and the AD group benefited less from phonological cueing than the controls. Our findings indicate that acquisition is compromised in both MCI and AD, whereas long13 term retention is not affected to the same extent. Incidental learning and recognition memory seem to be well preserved in MCI. In studies III and IV, the neural correlates of naming newly learned objects were examined in healthy elderly subjects and in amnestic MCI patients by means of positron emission tomography (PET) right after the training period. The naming of newly learned objects by healthy elderly subjects recruited a left-lateralized network, including frontotemporal regions and the cerebellum, which was more extensive than the one related to the naming of familiar objects (study III). Semantic support showed no effects on the PET results for the healthy subjects. The observed activation increases may reflect lexicalsemantic and lexical-phonological retrieval, as well as more general associative memory mechanisms. In study IV, compared to the controls, the MCI patients showed increased anterior cingulate activation when naming newly learned objects that had been learned without semantic support. This suggests a recruitment of additional executive and attentional resources in the MCI group.

Mild Cognitive Impairment and Early Detection of Alzheimer`s Disease. A Positron Emission Tomography Study

Alzheimer`s disease (AD) is characterised neuropathologically by the presence of extracellular amyloid plaques, intraneuronal neurofibrillary tangles, and cerebral neuronal loss. The pathological changes in AD are believed to start even decades before clinical symptoms are detectable. AD gradually affects episodic memory, cognition, behaviour and the ability to perform everyday activities. Mild cognitive impairment (MCI) represents a transitional state between normal aging and dementia disorders, especially AD. The predictive accuracy of the current and commonly used MCI criteria devide this disorder into amnestic (aMCI) and non-amnestic (naMCI) MCI. It seems that many individuals with aMCI tend to convert to AD. However many MCI individuals will remain stable and some may even recover. At present, the principal drugs for the treatment of AD provide only symptomatic and palliative benefits. Safe and effective mechanism-based therapies are needed for this devastating neurodegenerative disease of later life. In conjunction with the development of new therapeutic drugs, tools for early detection of AD would be important. In future one of the challenges will be to detect at an early stage these MCI individuals who will convert to AD. Methods which can predict which MCI subjects will convert to AD will be much more important if the new drug candidates prove to have disease-arresting or even disease–slowing effects. These types of drugs are likely to have the best efficacy if administered in the early or even in the presymptomatic phase of the disease when the synaptic and neuronal loss has not become too widespread. There is no clinical method to determine with certainly which MCI individuals will progress to AD. However there are several methods which have been suggested as predictors of conversion to AD, e.g. increased [11C] PIB uptake, hippocampal atrophy in MRI, low CSF A beta 42 level, high CSF tau-protein level, apolipoprotein E (APOE) ε4 allele and impairment in episodic memory and executive functions. In the present study subjects with MCI appear to have significantly higher [¹¹C] PIB uptake vs healthy elderly in several brain areas including frontal cortex, the posterior cingulate, the parietal and lateral temporal cortices, putamen and caudate. Also results from this PET study indicate that over time, MCI subjects who display increased [¹¹C] PIB uptake appear to be significantly more likely to convert to AD than MCI subjects with negative [¹¹C] PIB retention. Also hippocampal atrophy seems to increase in MCI individuals clearly during the conversion to AD. In this study [¹¹C] PIB uptake increases early and changes relatively little during the AD process whereas there is progressive hippocampal atrophy during the disease. In addition to increased [¹¹C] PIB retention and hippocampal atrophy, the status of APOE ε4 allele might contribute to the conversion from MCI to AD.

Morfologia foliar de indivíduos jovens e adultos de Caesalpinia echinata Lam. numa floresta semidecídua do Sul da Bahia

Caesalpinia echinata Lam. (Caesalpiniaceae) é uma espécie arbórea que ocorre naturalmente no interior da floresta e atinge os estratos superiores do dossel por meio de pequenas clareiras. Objetivou-se, neste estudo, comparar alguns aspectos da morfologia foliar entre indivíduos adultos, cujas folhas se encontravam em pleno sol e jovens no sub-bosque. O estudo foi realizado numa floresta semidecídua, localizada no Município de Jussari, Bahia. Inicialmente, foram localizados e marcados oito indivíduos adultos e oito jovens para cada adulto. Foram coletadas e analisadas oito folhas para cada adulto e três para cada jovem, em duas épocas (setembro/novembro de 2004 e abril de 2005). Em geral, as áreas das folhas, das ráquis e dos folíolos, o comprimento das ráquis, a largura das folhas, o número de pinas e a massa seca das folhas foram significativamente superiores nos indivíduos jovens, ao passo que as massas específicas das folhas e dos folíolos foram significativamente superiores nos indivíduos adultos. Tanto nos adultos quanto nos jovens, nas duas épocas de coleta foram verificadas relações altamente significativas entre a massa seca e a área das folhas. Os resultados indicaram que folhas de C. echinata apresentam características que maximizam a absorção de luz onde este recurso é limitante e, ao mesmo tempo, direcionam maior alocação de carbono para os tecidos de suporte. Tais resultados estão de acordo com o observado no estádio sucessional da espécie analisada e com a sua estratégia de ocupação dos espaços gerados pela formação de pequenas clareiras, em ambiente de mata semidecídua.

Morfometria de pupas e adultos de rhinochenus stigma linnaeus 1758 (coleoptera: curculionidae)

Besouros do gênero Rhinochenus sp. (Coleoptera: Curculionidae) danificam sementes de espécies da flora do Cerrado brasileiro como o jatobá (Hymenea courbaril L.) e a copaíba (Copaifera langsdorffii Desf.). Este trabalho teve como objetivo avaliar a morfometria de pupas e adultos de R. stigma, bem como aspectos comportamentais, como a emissão de sons, para auxiliar a sexagem desta espécie. Os insetos foram obtidos de frutos de jatobá coletados no Município de São Gonçalo do Rio Preto, em Minas Gerais, Brasil. As características morfométricas de pupas e adultos não apresentaram diferenças pelo teste t, não sendo seguras para a diferenciação dos sexos. Para adultos de R. stigma, a sexagem só foi possível após a morte, pela dissecação dos órgãos genitais.

Esquistossomose hepatoesplênica cirúrgica: histopatologia hepatica e endoscopia digestiva alta em crianças comparadas a adultos

Objetivando melhor compreender a fisiopatologia da esquistossomose grave em adultos e crianças, avaliaram-se diferenças histopatológicas do fígado e do padrão de endoscopia digestiva entre esses pacientes. Vinte e um adultos e igual número de crianças foram incluídos no estudo. Todos foram submetidos a esplenectomia e ligadura da veia gástrica esquerda. Os achados histopatológicos foram avaliados em secções de biópsia hepática. Utilizou-se videoendoscópio para endoscopia digestiva. Nos adultos foram encontradas: 1 - fibrose de Symmers, grau I, em um paciente; grau II, em sete; e grau III, em treze; 2 - atividade inflamatória mínima em cinco; leve, em sete; moderada, em seis; e intensa, em um; ausência de atividade inflamatória, em dois; 3 - mínima granulomatose, em quatro pacientes; moderada, em dois; ausência de granulomas, em quinze; 4 - pigmento esquistossomótico mínimo, em quatro pacientes; moderada, em um; ausência, em dezesseis. Nas crianças, foram encontradas: 1 - fibrose de Symmers, grau I, em um paciente; grau II, em quatro e grau III, em dezesseis; 2 - atividade inflamatória mínima, em sete pacientes; leve, em quatorze; 3 - mínima granulomatose, em seis pacientes; leve, em cinco; moderada, em dois; ausência, em oito; 4 - pigmento esquistossomótico mínimo, em seis pacientes; leve, em três; moderada, em seis; ausência, em seis. Na endoscopia digestiva alta encontrou-se nos adultos: oito (38%) com varizes de médio calibre e treze (62%), de grosso calibre. Oito pacientes (38%) tiveram varizes de fórnix gástrico e cinco (24%), da cárdia. Gastropatia da hipertensão porta foi diagnosticada em dezesseis pacientes (76%). Com relação às crianças, quatro (19%) apresentaram varizes de fino calibre, oito (38%) de médio; e nove (43%) de grosso calibre. Três (14%) tiveram varizes gástricas, sendo duas no fórnix e uma no cárdia. Nove (43%) das crianças apresentavam gastropatia de hipertensão porta. Os achados evidenciaram uma histopatologia hepática similar nos dois grupos; entretanto, as repercussões hemodinâmicas da hipertensão porta foram mais importantes nos adultos, sem, no entanto, alcançar significação estatística.

Histopatologia do saco herniário da hérnia inguinal indireta e do peritônio parietal em adultos e crianças: estudo qualitativo da sua musculatura lisa

Estudo prospectivo, realizado entre abril de 1993 e julho de 1995, com o objetivo de estudar a histopatologia do saco herniário de hérnias inguinais indiretas do adulto e criança, no sentido de verificar a existência de musculatura lisa, sua incidência, apresentação morfológica e comparar com biópsias aleatórias do peritônio parietal. Os pacientes foram divididos em Grupo (1) com 123 pacientes, nos quais foram estudados os sacos herniários, e Grupo (2) constituído de 63 pacientes, nos quais foram realizadas biópsias da serosa peritoneal da cavidade abdominal. Verificou-se que fibras de músculo liso (FML) estiveram presentes em 65,4% dos 133 sacos herniários (dez pacientes com hérnia bilateral), estando presentes, também, em 19,04% dos espécimes da cavidade abdominal. Através dos testes do Qui-quadrado e t de Student, foi avaliada a associação entre a presença de FML com as variáveis categóricas (sexo, cor e lado da hérnia) e as variáveis contínuas (idade dos pacientes, comprimento e espessura do saco herniário). Os resultados mostraram que o sexo feminino apresenta uma maior incidência de FML (p=0,004) e a razão das chances (O.R.) demonstra que os pacientes desse mesmo sexo têm 5,46 vezes mais possibilidades de possuir FML nos sacos herniários. Assim, concluem que as FML são predominantes no peritônio parietal do abdome inferior e que existe, também, uma quantidade maior de FML no peritônio do saco herniário inguinal quando comparado com o peritônio parietal.

Fístula arteriovenosa em crianças e adultos jovens

OBJETIVO: Com a introdução e o aprimoramento de técnicas para a realização de fístulas arteriovenosas (FAV) em crianças, pacientes renais crônicos puderam iniciar tratamento com hemodiálise sem a necessidade de uso de shunts externos. MÉTODOS: Com o objetivo avaliar os resultados da confecção de fístulas arteriovenosas em crianças e adultos jovens de baixo peso e os fatores de risco relacionados. Foram levantados retrospectivamente os prontuários de 31 pacientes submetidos a 35 FAV, com idade variando de cinco a 24 anos, com peso entre 16 e 50kg. Os fatores de risco estudados foram hipertensão arterial, antecedentes de trombose, tendência à hipovolemia e presença de hipercolesterolemia. RESULTADOS: O índice de perda observado foi de 25% em pacientes com menos de 20kg e de 22,2% naqueles com mais de 20kg, resultados comparáveis aos da literatura. CONCLUSÕES: Observou-se que fístulas proximais não sofreram trombose; não houve relação entre a perda da FAV e os fatores de risco estudados e observou-se um índice de perda tardia de 23,5% nos pacientes que não estavam sendo submetidos à hemodiálise. Nos pacientes que utilizavam a FAV não se observou perda da mesma.

Histologia dos sacos herniários das hérnias inguinais indiretas, diretas, recidivadas e encarceradas em adultos e crianças: identificação de fibras musculares lisas

OBJETIVO: Pesquisar a presença de fibras de músculo liso (FML) nos sacos peritoneais das hérnias indiretas, diretas, recidivadas e encarceradas e estudar a influência do sexo, cor e idade dos pacientes, bem como a região do saco herniário, largura, comprimento e espessura da biópsia coletada. MÉTODO: Foram obtidos 252 sacos herniários no período de fevereiro de 1999 a dezembro de 2000 e encaminhados para o estudo histopatológico através da coloração por hematoxilina-eosina e tricrômico de Gomori. A idade variou entre um mês a 87 anos com média de 42,3 anos e desvio-padrão de 22,5 anos. RESULTADOS: Foi utilizado o teste do Qui-quadrado e observada FML em 76,5% dos pacientes com hérnia indireta, 55,9% direta, 46,4% encarcerada e 68,7% recidivada. No estudo global foram encontradas FML em 67,9% dos espécimes com maior incidência na porção proximal do saco herniário (53,2%) e em pacientes melanodérmicos (75%). As FML estiveram presentes em maior freqüência no lado direito (67,7%) e no sexo feminino (73,3%). Do total de 252 amostras de sacos herniários examinados, foi encontrada FML em 171 biópsias, e esse achado foi menos freqüente nas hérnias diretas e encarceradas quando comparadas com as indiretas e recidivadas. CONCLUSÕES: Como hipótese, a presença de FML na parede do saco herniário pode representar um reforço tecidual no sentido de dificultar o crescimento do saco peritoneal, comportando-se como fator de resistência elástica e dinâmica à expansão da hérnia. Por outro lado, pode também significar uma formação aberrante ou a persistência de uma estrutura que deveria regredir ou mesmo desaparecer durante o desenvolvimento normal.