Síntese de potencial inibidor de acetilcolinesterase para tratamento da Doença de Alzheimer

A doença de Alzheimer (DA) é a forma mais comum de demência, representando cerca de 80% dos casos. A DA é caracterizada por um processo de declínio progressivo e irreversível das funções cognitivas e da memória, que se estende para a desorganização do comportamento. Atualmente, 46,8 milhões de pessoas em todo o mundo foram diagnosticadas com demência. Embora vários fatores tenham sido implicados na DA, sua etiologia ainda não é completamente conhecida. Do ponto de vista neuropatológico, é observado no cérebro de indivíduos com DA atrofia cortical difusa, presença de grande número de placas senis, emaranhados neurofibrilares, processo inflamatório e perda neuronal. A progressão dos sintomas está associada a mudanças estruturais nas sinapses colinérgicas em certas regiões do cérebro, que consequentemente, apresentam neurotransmissão colinérgica reduzida. Os vários eventos patológicos interligados contribuem para o avanço da doença e direcionam diversas pesquisas na busca por tratamentos multialvos com base no processo multifatorial de DA. Assim o presente trabalho descreve a síntese de derivados híbridos dual binding site de donepezila-tacrina (fármacos inibidores de acetilcolinesterase), com potencial para agir em dois alvos terapêuticos pela (i) inibição da acetilcolinesterase em ambos os sítios ativo e periférico, como demonstrado pelos estudos de modelagem molecular, e (ii) na agregação do peptídeo A? neurotóxico induzido pela acetilcolinesterase, na tentativa de interromper a progressão da doença. A estratégia sintética envolveu a condensação da 5,6-dimetóxiindanona com a unidade 4-piperidinil carbaldeído, a qual forneceu o intermediário 5,6- dimetóxindan-1-ona-4-piperidinil-metileno-1-[(4-cloroquinolin-2-il)metil], seguido de redução da dupla ligação, gerada na reação de condensação anterior, e substituição do átomo de cloro-quinolina por amino para obtenção do produto final, ou manutenção da função olefina, seguido de substituição do átomo de cloro-quinolina por azido ou amino, gerando cinco híbridos estruturalmente correlacionados. Os híbridos foram testados em ensaio de inibição de acetilcolinesterase e butirilcolinesterase pelo método de Ellman, e o híbrido insaturado, contendo a função amino-quinolina foi o mais ativo da série com IC50 na faixa de nanomolar (0,014 ?M). Futuramente, os intermediários da reação e produto final serão submetidos ao ensaio de inibição da agregação do peptídeo A? neurotóxico pelo método da tioflavina T. Neste trabalho, também são descritos os testes de predição in vitro para permeação pela barreira hematoencefálica, bem como sua absorção intestinal, pelo método PAMPA.

Activity and biomass of small benthic biota in the central Arctic Ocean

Sediment samples collected during the expedition "Arctic Ocean '96" with the Swedish ice-breaker ODEN were investigated to estimate for the first time heterotrophic activity and total microbial biomass (size range from bacteria to small metazoans) from the perennially ice-covered central Arctic Ocean. Benthic activities and biomass were evaluated analysing a series of biogenic sediment compounds (i.e. bacterial exoenzymes, total adenylates, DNA, phospholipids, particulate proteins). In contrast to the very time-consuming sorting, enumeration and weight determination, analyses of biochemical sediment parameters may represent a useful method for obtaining rapid information on the ecological situation in a given benthic system. Bacterial cell numbers and biomass were estimated for comparison with biochemically determined biomass data, to evaluate the contribution of the bacterial biomass to the total microbial biomass. It appeared that bacterial biomass made up only 8-31% (average of all stations = 20%) of the total microbial biomass, suggesting a large fraction of other small infaunal organisms within the sediment samples (most probably fungi, yeasts, protozoans such as flagellates, ciliates or amoebae, as well as a fraction of small metazoans). Activity and biomass values determined within this study were generally extremely low, and often even slightly lower than those given for other deep oceanic regions, thus characterizing the seafloor of the central Arctic Ocean as a "benthic desert". Nevertheless, some clear trends in the data could be found, e.g. generally sharply decreasing values within the sediment column, a vague tendency for declining values with increasing water depth of sampling stations, and also differences between various Arctic deep-sea regions.

Biochemical investigation of multicorer sediment profile PS74/109-2

To detect and track the impact of large-scale environmental changes in a the transition zone between the northern North Atlantic and the central Arctic Ocean, and to determine experimentally the factors controlling deep-sea biodiversity, the Alfred- Wegener-Institute for Polar and Marine Research (AWI) established the deep-sea long-term observatory HAUSGARTEN, which constitutes the first, and until now only open-ocean long-term station in a polar region. Virtually undisturbed sediment samples have been taken using a video-guided multiple corer (MUC) at 13 HAUSGARTEN stations along a bathymetric (1,000 - 4,000 m water depth) and a latitudinal transect in 2,500 m water depth as well as two stations at 230 and 1,200 m water depth within the framework of the KONGHAU project. Various biogenic sediment compounds were analyzed to estimate the input of organic matter from phytodetritus sedimentation, benthic activities (e.g. bacterial exoenzymatic activity), and the total biomass of the smallest sediment-inhabiting organisms (size range: bacteria to meiofauna).

Biochemical investigation of multicorer sediment profile PS74/113-2

To detect and track the impact of large-scale environmental changes in a the transition zone between the northern North Atlantic and the central Arctic Ocean, and to determine experimentally the factors controlling deep-sea biodiversity, the Alfred- Wegener-Institute for Polar and Marine Research (AWI) established the deep-sea long-term observatory HAUSGARTEN, which constitutes the first, and until now only open-ocean long-term station in a polar region. Virtually undisturbed sediment samples have been taken using a video-guided multiple corer (MUC) at 13 HAUSGARTEN stations along a bathymetric (1,000 - 4,000 m water depth) and a latitudinal transect in 2,500 m water depth as well as two stations at 230 and 1,200 m water depth within the framework of the KONGHAU project. Various biogenic sediment compounds were analyzed to estimate the input of organic matter from phytodetritus sedimentation, benthic activities (e.g. bacterial exoenzymatic activity), and the total biomass of the smallest sediment-inhabiting organisms (size range: bacteria to meiofauna).

Biochemical investigation of multicorer sediment profile PS74/118-2

To detect and track the impact of large-scale environmental changes in a the transition zone between the northern North Atlantic and the central Arctic Ocean, and to determine experimentally the factors controlling deep-sea biodiversity, the Alfred- Wegener-Institute for Polar and Marine Research (AWI) established the deep-sea long-term observatory HAUSGARTEN, which constitutes the first, and until now only open-ocean long-term station in a polar region. Virtually undisturbed sediment samples have been taken using a video-guided multiple corer (MUC) at 13 HAUSGARTEN stations along a bathymetric (1,000 - 4,000 m water depth) and a latitudinal transect in 2,500 m water depth as well as two stations at 230 and 1,200 m water depth within the framework of the KONGHAU project. Various biogenic sediment compounds were analyzed to estimate the input of organic matter from phytodetritus sedimentation, benthic activities (e.g. bacterial exoenzymatic activity), and the total biomass of the smallest sediment-inhabiting organisms (size range: bacteria to meiofauna).

Neurotoxicity : identifying and controlling poisons of the nervous system : summary.

"April 1990"--p.4 of cover.

Effects of foods and drugs on the development and function of the nervous system : methods for predicting toxicity /

Symposium organized by the Office of Health Affairs, Food and Drug Administration.

Reducing and recycling mercury switch thermostats and vehicle components : a report to Governor Blagojevich and the Illinois General Assembly.

Over the last two years, Governor Blagojevich and the Illinois General Assembly have enacted a number of laws to reduce the health risks associated with the use and disposal of consumer, household and commercial products that contain mercury. These products include thermometers, switches, electrical relays and scientific instruments used in schools. The purpose of these laws is to reduce mercury releases into the environment. Mercury is a strong neurotoxin that can be harmful to the health of humans and wildlife. Mercury exposure poses a particular risk to young children and pregnant women because mercury may inhibit the development of the brain and nervous system. This report presents recommendations for improving efforts to reduce and recycle mercury components that are found in thermostats and motor vehicles. Illinois EPA prepared the report in response to Public Act 93-0964. In drafting the report, Illinois EPA conducted research on mercury reduction and recycling programs in other states, reviewed technical studies and consulted with officials in the private and public sectors.

Recycling mercury vehicle switches in Illinois.

"July 2008"

Retinal glutamate transporter activity persists under simulated ischemic conditions

Elevated extracellular concentrations of the neurotransmitter glutamate are neurotoxic and directly contribute to CNS damage as a result of ischemic pathologies. However, the main contributors to this uncontrolled rise in glutamate are still unconfirmed. It has been reported that the reversal of high-affinity glutamate transporters is a significant contributing factor. Conversely, it has also Peen observed that these transporters continue to take up glutamate, albeit at a reduced saturation concentration, under ischemic conditions. We sought to determine whether glutamate transporters continue to remove glutamate from the extracellular space under ischemic conditions by pharmacologically modulating the activity of high-affinity retinal glutamate transporters during simulated ischemia in vitro. Retinal glutamate transporter activity was significantly reduced under these ischemic conditions. The suppression of retinal glutamate transporter activity, with the protein kinase C inhibitor chelerythrine, significantly reduced ischemic glutamate uptake and enhanced retinal neurodegeneration. These findings imply a limited but protective role for retinal glutamate transporters under certain ischemic conditions, suggesting that pharmacological enhancement of high-affinity glutamate transporter activity may reduce tissue damage and loss of function resulting from toxic extracellular glutamate concentrations. (C) 2004 Wiley-Liss, Inc.

Factors that influence the prevalence of acaricide resistance and tick-borne diseases

This manuscript provides a summary of the results presented at a symposium organized to accumulate information on factors that influence the prevalence of acaricide resistance and tick-borne diseases. This symposium was part of the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), held in New Orleans, LA, USA, during August 10-14, 2003. Populations of southern cattle ticks, Boophilus microplus, from Mexico have developed resistance to many classes of acaricide including chlorinated hydrocarbons (DDT), pyrethroids, organ ophosphates, and formamidines (amitraz). Target site mutations are the most common resistance mechanism observed, but there are examples of metabolic mechanisms. In many pyrethroid resistant strains, a single target site mutation on the Na+ channel confers very high resistance (resistance ratios: >1000x) to both DDT and all pyrethroid acaricides. Acetylcholine esterase affinity for OPs is changed in resistant tick populations. A second mechanism of OP resistance is linked to cytochrome P450 monooxygenase activity. A PCR-based assay to detect a specific sodium channel gene mutation that is associated with resistance to permethrin has been developed. This assay can be performed on individual ticks at any life stage with results available in a few hours. A number of Mexican strains of B. microplus with varying profiles of pesticide resistance have been genotyped using this test. Additionally, a specific metabolic esterase with permethrin-hydrolyzing activity, CzEst9, has been purified and its gene coding region cloned. This esterase has been associated with high resistance to permethrin in one Mexican tick population. Work is continuing to clone specific acetylcholinesterase (AChE) and carboxylesterase genes that appear to be involved in resistance to organophosphates. Our ultimate goal is the design of a battery of DNA- or ELISA-based assays capable of rapidly genotyping individual ticks to obtain a comprehensive profile of their susceptibility to various pesticides. More outbreaks of clinical bovine babesisois and anaplasmosis have been associated with the presence of synthetic pyrethroid (SP) resistance when compared to OP and amidine resistance. This may be the result of differences in the temporal and geographic patterns of resistance development to the different acaricides. If acaricide resistance develops slowly, herd immunity may not be affected. The use of pesticides for the control of pests of cattle other than ticks can affect the incidence of tick resistance and tick-borne diseases. Simple analytical models of tick- and tsetse-bome diseases suggest that reducing the abundance of ticks, by treating cattle with pyrethroids for example, can have a variety of effects on tick-bome diseases. In the worst-case scenario, the models suggest that treating cattle might not only have no impact on trypanosomosis but could increase the incidence of tick-bome disease. In the best-case, treatment could reduce the incidence of both trypanosomosis and tick-bome diseases Surveys of beef and dairy properties in Queensland for which tick resistance to amitraz was known were intended to provide a clear understanding of the economic and management consequences resistance had on their properties. Farmers continued to use amitraz as the major acaricide for tick control after the diagnosis of resistance, although it was supplemented with moxidectin (dairy farms) or fluazuron, macrocyclic lactones or cypermethrin/ chlorfenvinphos. (C) 2004 Published by Elsevier B.V.

Alcoholic neurobiology: Changes in dependence and recovery

This article presents the proceedings of a symposium held at the meeting of the International Society for Biomedical Research on Alcoholism (ISBRA) in Mannheim, Germany, in October, 2004. Chronic alcoholism follows a fluctuating course, which provides a naturalistic experiment in vulnerability, resilience, and recovery of human neural systems in response to presence, absence, and history of the neurotoxic effects of alcoholism. Alcohol dependence is a progressive chronic disease that is associated with changes in neuroanatomy, neurophysiology, neural gene expression, psychology, and behavior. Specifically, alcohol dependence is characterized by a neuropsychological profile of mild to moderate impairment in executive functions, visuospatial abilities, and postural stability, together with relative sparing of declarative memory, language skills, and primary motor and perceptual abilities. Recovery from alcoholism is associated with a partial reversal of CNS deficits that occur in alcoholism. The reversal of deficits during recovery from alcoholism indicates that brain structure is capable of repair and restructuring in response to insult in adulthood. Indirect support of this repair model derives from studies of selective neuropsychological processes, structural and functional neuroimaging studies, and preclinical studies on degeneration and regeneration during the development of alcohol dependence and recovery from dependence. Genetics and brain regional specificity contribute to unique changes in neuropsychology and neuroanatomy in alcoholism and recovery. This symposium includes state-of-the-art presentations on changes that occur during active alcoholism as well as those that may occur during recovery-abstinence from alcohol dependence. Included are human neuroimaging and neuropsychological assessments, changes in human brain gene expression, allelic combinations of genes associated with alcohol dependence and preclinical studies investigating mechanisms of alcohol induced neurotoxicity, and neuroprogenetor cell expansion during recovery from alcohol dependence.

Aspergillazines A-E: novel heterocyclic dipeptides from an Australian strain of Aspergillus unilateralis

Biological and chemical pro ling of an Australian strain of the fungus Aspergillus unilateralis (MST-F8675), isolated from a soil sample collected near Mount Isa, Queensland, revealed a complex array of metabolites displaying broad chemotherapeutic properties. Noteworthy among these metabolites were a unique series of highly modified dipeptides aspergillazines A-E, incorporating a selection of unprecedented and yet biosynthetically related heterocyclic systems. Co-occurring with the aspergillazines was the recently described marine-derived fungal metabolite trichodermamide A (cf. penicillazine), whereas re-fermentation of A. unilateralis in NaCl (1%) enriched media resulted in co-production of the only other known example of this structure class, the marine-derived fungal metabolite trichodermamide B. Further investigation of A. unilateralis returned the known terrestrial fungal metabolite viridicatumtoxin as the cytotoxic and antibacterial principle, together with E-2-decenedioic acid, ferulic acid, (7E,7'E)-5,5'-diferulic acid and (7E,7'E)-8,5'-diferulic acid. The aromatic diacids have previously been reported from the chemical and enzymatic (esterase) treatment of plant cell wall material, with their isolation from A. unilateralis being their first apparent reported occurrence as natural products. Structures for all metabolites were determined by detailed spectroscopic analysis and, where appropriate, comparison to literature data and/or authentic samples.

Cytochrome P450-mediated metabolism of haloperidol and reduced haloperidol to pyridinium metabolites

Haloperidol ( HP) has been reported to undergo cytochrome P450 (P450)-mediated metabolism to potentially neurotoxic pyridinium metabolites; however, the chemical pathways and specific enzymes involved in these reactions remain to be identified. The aims of the current study were to (i) fully identify the cytochrome P450 enzymes capable of metabolizing HP to the pyridinium metabolite, 4-(4-chlorophenyl)- 1-(4-fluorophenyl)-4-oxobutylpyridinium (HPP+), and reduced HP (RHP) to 4-(4-chlorophenyl)- 1-(4-fluorophenyl)-4-hydroxybutylpyridinium (RHPP+); and (ii) determine whether 4-(4-chlorophenyl)- 1-(4-fluorophenyl)-4-oxobutyl-1,2,3,6-tetrahydropyridine (HPTP) and 4-(4-chlorophenyl)1-( 4-fluorophenyl)-4-hydroxybutyl-1,2,3,6-tetrahydropyridine (RHPTP) were metabolic intermediates in these pathways. In vitro studies were conducted using human liver microsomal preparations and recombinant human cytochrome P450 enzymes (P450s 1A1, 1A2, 1B1, 2A6, 2B6, 2C9, 2C19 2D6, 2E1, 3A4, 3A5, and 3A7) expressed in bicistronic format with human NADPH cytochrome P450 reductase in Escherichia coli membranes. Pyridinium formation from HP and RHP was highly correlated across liver preparations, suggesting the same enzyme or enzymes were responsible for both reactions. Cytochrome P450s 3A4, 3A5, and 3A7 were the only recombinant enzymes which demonstrated significant catalytic activity under optimized conditions, although trace levels of activity could be catalyzed by NADPHP450 reductase alone. NADPH-P450 reductase-mediated activity was inhibited by reduced glutathione but not catalase or superoxide dismutase, suggesting O-2-dependent oxidation. No evidence was obtained to support the contention that HPTP and RHPTP are intermediates in these pathways. K-m values for HPP+ (34 +/- 5 mu M) and RHPP+ (64 +/- 4 mu M) formation by recombinant P450 3A4 agreed well with those obtained with human liver microsomes, consistent with P450 3A4 being the major catalyst of pyridinium metabolite formation in human liver.