RESUMO
Because epidemiologic and in vitro evidence conflict, the authors studied the association between smoking and Alzheimer disease (AD) in 46 never, 47 former, and 15 active smokers with AD followed to autopsy. Disease parameters were examined by smoking status and amount smoked in bivariate tests and in multivariate models controlling for age, sex, education, and APOE status. Smoking status was not associated with cognitive or neuropathologic measures. However, active smokers were significantly younger at death and higher levels of smoking were associated with shorter disease duration.
Assuntos
Doença de Alzheimer/mortalidade , Encéfalo/efeitos dos fármacos , Encéfalo/fisiopatologia , Fumar/efeitos adversos , Fumar/epidemiologia , Fatores Etários , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/genética , Doença de Alzheimer/psicologia , Apolipoproteínas E/genética , Encéfalo/patologia , Causalidade , Morte , Progressão da Doença , Escolaridade , Feminino , Genótipo , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Fenótipo , Estudos Prospectivos , Fatores SexuaisRESUMO
We investigated the relationship between the loss of nicotinic acetylcholine receptors (nAChR) and the cognitive decline or neuropathological changes seen in Alzheimer's Disease (AD) and dementia with Lewy bodies (DLB). Midfrontal (MF) cortex of 31 AD, 24 DLB and 11 non-demented controls was examined. Total plaque (TP), neuritic plaque (NP) and neurofibrillary tangle (NFT) counts were obtained. NAChR binding was assayed using 3H-epibatidine [3H-EPI]. Last Blessed Information-Memory-Concentration scores (BIMC), Mini-Mental State Examination (MMSE), Mattis Dementia Rating Scale (DRS) scores were collected. There were no correlations between 3H-EPI binding and TP, NP, NFTs counts in either AD or DLB. Last BIMC, MMSE, DRS scores did not correlate with 3H-EPI binding in AD or DLB. Thus, decline in cognitive function does not correlate with loss of nAChR in DLB or AD at the end of life suggesting that later in these diseases, loss of nAChR binding is not a reliable marker of cognitive function in AD or DLB. Loss of nAChR activity does not appear to be related to plaques or NFTs in AD or DLB.
Assuntos
Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Doença por Corpos de Lewy/metabolismo , Doença por Corpos de Lewy/patologia , Receptores Nicotínicos/metabolismo , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Córtex Cerebral/metabolismo , Córtex Cerebral/patologia , Transtornos Cognitivos/metabolismo , Transtornos Cognitivos/patologia , Humanos , Emaranhados Neurofibrilares/metabolismo , Emaranhados Neurofibrilares/patologiaRESUMO
We wished to determine the influence of the apolipoprotein E (Apo-E) genotype on the loss of high affinity nicotinic acetylcholine receptor (nAChR) binding in Alzheimer's disease (AD). The interaction between epsilon4 allele gene dose and cholinergic loss in AD remains controversial. We have demonstrated that nicotinic binding is significantly lost in AD. Tissue from the midfrontal (MF) cortex of 7 subjects with no epsilon4 allele copies (epsilon-/epsilon-) (mean death age 75.1 +/ 10.4 years) was compared to MF cortex of 14 subjects heterozygous for the epsilon4 allele (epsilon4/epsilon-) (mean death age 81.4 +/- 7.3 years) and MF cortex of 10 subjects homozygous for the epsilon4 allele (epsilon4/epsilon4) (mean death age 79.6 +/- 5.0 years). All subjects were autopsy confirmed AD (using NIA and CERAD criteria) and met NINCDS-ADRDA clinical criteria for probable or possible AD. Nicotine AChR binding was assayed using the high affinity nicotinic agonist 3H-epibatidine ([3H]-Epi). Apo-E genotype was determined in blood samples or in post-mortem tissue. The mean age at death was not significantly different among the groups (p = 0.19). There was no difference in mean [3H]-Epi total binding among the three groups (6.7 +/- 4.6, 6.1 +/- 2.4, and 6.0 +/- 1.0 fmol/mg protein for epsilon-/epsilon-, epsilon4/epsilon-, and epsilon4/epsilon4 respectively. We conclude that the presence or absence of the Apo-E4 genotype does not influence the loss of high affinity nAChR in AD.
Assuntos
Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Apolipoproteínas E/genética , Lobo Frontal/metabolismo , Neurônios/metabolismo , Receptores Nicotínicos/genética , Acetilcolina/metabolismo , Idoso , Apolipoproteína E4 , Apolipoproteínas E/metabolismo , Colina O-Acetiltransferase/metabolismo , Lobo Frontal/patologia , Lobo Frontal/fisiopatologia , Dosagem de Genes , Frequência do Gene , Genótipo , Humanos , Neurônios/patologia , Receptores Nicotínicos/metabolismo , Sinapses/genética , Sinapses/metabolismoRESUMO
HEK293 cells were stably transfected with the cDNAs encoding full-length human neuronal nicotinic acetylcholine receptor (nAChR) subunit combinations alpha3beta2 or alpha4beta2. [(3)H]-(+/-)Epibatidine ([(3)H]-(+/-)EPI) bound to membranes from A3B2 (alpha3beta2) and A4B2.2 (alpha4beta2) cells with K(d) values of 7.5 and 33.4 pM and B(max) values of 497 and 1564 fmol/mg protein, respectively. Concentration-dependent increases in intracellular free Ca(2+) concentration were elicited by nAChR agonists with a rank order of potency of EPI>1,1-dimethyl-4-phenylpiperazinium (DMPP)>nicotine (NIC)=suberyldicholine (SUB)>cytisine (CYT)=acetylcholine (ACh) for A3B2 cells and EPI>CYT=SUB=NIC=DMPP>ACh for A4B2.2 cells. Antagonists of nAChRs blocked NIC-induced responses with a rank order of potency of d-tubocurarine (d-Tubo)=mecamylamine (MEC)>dihydro-beta-erythroidine (DHbetaE) in A3B2 cells and MEC=DHbetaE>d-Tubo in A4B2.2 cells. Whole-cell patch clamp recordings indicate that the decay rate of macroscopic ACh-induced currents is faster in A3B2 than in A4B2.2 cells and that A3B2 cells are less sensitive to ACh than A4B2.2 cells. ACh currents elicited in alpha3beta2 and alpha4beta2 human nAChRs are maximally potentiated at 20 and 2 mM external Ca(2+), respectively. Our results indicate that stably expressed alpha3beta2 and alpha4beta2 human nAChRs are pharmacologically and functionally distinct.
Assuntos
Receptores Nicotínicos/metabolismo , Northern Blotting , Western Blotting , Cálcio/metabolismo , Linhagem Celular , Estimulação Elétrica , Eletrofisiologia , Humanos , Rim/metabolismo , Ligantes , Membranas/efeitos dos fármacos , Membranas/metabolismo , Agonistas Nicotínicos/farmacologia , Técnicas de Patch-Clamp , RNA/biossíntese , RNA/isolamento & purificação , Ensaio Radioligante , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/genética , Proteínas Recombinantes/químicaRESUMO
In this study, the mechanism of nicotine-induced hippocampal acetylcholine (ACh) release in awake, freely moving rats was examined using in vivo microdialysis. Systemic administration of nicotine (0.4 mg kg(-1), s.c.) increased the levels of ACh in hippocampal dialysates. The nicotine-induced hippocampal ACh release was sensitive to the pretreatment of neuronal nicotinic acetylcholine receptor (nAChR) antagonists mecamylamine (3.0 mg kg(-1), s.c.) and dihydro-beta-erythrodine (DHbetaE; 4.0 mg kg(-1), s.c.) as well as systemic administration of the dopamine (DA) D1 receptor antagonist SCH-23390 (R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-benzaz epine; 0.3 mg kg(-1), s.c.). Local perfusion of mecamylamine (100 microM), DHbetaE (100 microM) or SCH-23390 (10 microM) through microdialysis probe did not increase basal hippocampal ACh release. Hippocampal ACh release elicited by systemic administration of nicotine (0.4 mg kg(-1), s.c.) was antagonized by local perfusion of SCH-23390 (10 microM), but not by MEC (100 microM) or DHbetaE (100 microM). Direct perfusion of nicotine (1 mM, but not 0.1 mM) increased hippocampal ACh levels; however, this effect was relatively insensitive to blockade by co-perfusion of either mecamylamine (100 microM) or SCH-23390 (10 microM). These results suggest that nicotine-induced hippocampal ACh release occurs by two distinct mechanisms: (1) activation of nAChRs outside the hippocampus leading to DA release and subsequent ACh release involving a permissive DA synapse, and (2) direct action of nicotine within the hippocampus leading to ACh release via non-DA-ergic mechanism.
Assuntos
Acetilcolina/metabolismo , Hipocampo/efeitos dos fármacos , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Animais , Benzazepinas/farmacologia , Di-Hidro-beta-Eritroidina/farmacologia , Antagonistas de Dopamina/farmacologia , Relação Dose-Resposta a Droga , Hipocampo/metabolismo , Masculino , Mecamilamina/farmacologia , Antagonistas Nicotínicos/farmacologia , Perfusão , Ratos , Ratos Sprague-Dawley , Receptores Dopaminérgicos/efeitos dos fármacos , Receptores Dopaminérgicos/fisiologia , Sinapses/efeitos dos fármacosRESUMO
The loss of neocortical synapses that occurs in Alzheimer's disease (AD) has been shown to correlate with cognitive decline. In addition, marked losses in the cholinergic system in AD, specifically choline acetyltransferase (ChAT) activity and high affinity presynaptic neuronal nicotinic cholinergic receptors (nAChRs), have also been described. We hypothesized that in AD, the loss of [3H]-ligand binding to nAChRs, which are largely presynaptic, would correlate with changes in two other presynaptic markers: synaptophysin (Syn), a measure of synaptic density, and ChAT activity. The midfrontal (MF) cortex of 36 autopsy confirmed (NIA and CERAD criteria) AD patients (mean death age +/- SD 80.1 +/- 8.4 years) who met NINDS-ADRDA criteria for a clinical diagnosis of probable or possible AD, and 11 nondemented controls (mean death age +/- SD 77.9 +/- 8.0) were examined. Synapse counts were quantified by a dotimmunobinding assay for Syn. ChAT activity was assessed by standard biochemical assays. Nicotinic cholinergic receptor binding was assayed using the high affinity nicotinic agonist [3H]-(+/-)-epibatidine ([3H]-EPI). The mean +/- SD Syn in AD (83.4 +/- 31.9 arbitrary units (AU)/mg protein) was significantly lower than controls (126.1 +/- 19.9, p = 0.0003; t-test). The mean ChAT activity in AD (139.0 +/- 75.6 nmol ACh/hr/100 mg protein) was significantly lower than controls (219.6 +/- 70.8, p = 0.004). The mean [3H]-EPI total binding in AD (6.2 +/- 2.8 fmol/mg protein) was significantly lower than controls (14.8 +/- 3.2; p < 0.0001). Syn correlated with [3H]-EPI binding in AD (r = 0.48, p = 0.006; Pearson) but ChAT did not (r = -0.20, p = 0.34). We conclude that loss of high affinity nAChR binding correlates with loss of synapses in AD. The lack of correlation between [3H]-EPI binding and ChAT activity suggests that the targeted receptor populations may not be located exclusively on cholinergic neurons.
Assuntos
Doença de Alzheimer/metabolismo , Receptores Nicotínicos/metabolismo , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/patologia , Doença de Alzheimer/psicologia , Biomarcadores , Encéfalo/metabolismo , Encéfalo/patologia , Compostos Bicíclicos Heterocíclicos com Pontes/metabolismo , Colina O-Acetiltransferase/metabolismo , Transtornos Cognitivos/etiologia , Humanos , Agonistas Nicotínicos/metabolismo , Piridinas/metabolismo , Valores de Referência , Sinapses/ultraestrutura , Sinaptofisina/metabolismoRESUMO
Human embryonic kidney (HEK293) cells were transfected with cDNA encoding the human beta4 neuronal nicotinic acetylcholine (ACh) receptor subunit in pairwise combination with human alpha2, alpha3 or alpha4 subunits. Cell lines A2B4, A3B4.2 and A4B4 were identified that stably express mRNA and protein corresponding to alpha2 and beta4, to alpha3 and beta4 and to alpha4 and beta4 subunits, respectively. Specific binding of [3H]epibatidine was detected in A2B4, A3B4.2 and A4B4 cells with Kd (mean +/- S.D. in pM) values of 42 +/- 10, 230 +/- 12 and 187 +/- 29 and with Bmax (fmol/mg protein) values of 1104 +/- 338, 2010 +/- 184 and 3683 +/- 1450, respectively. Whole-cell patch-clamp recordings in each cell line demonstrated that (-)nicotine (Nic), ACh, cytisine (Cyt) and 1, 1-dimethyl-4-phenylpiperazinium iodide (DMPP) elicit transient inward currents. The current-voltage (I-V) relation of these currents showed strong inward rectification. Pharmacological characterization of agonist-induced elevations of intracellular free Ca++ concentration revealed a distinct rank order of agonist potency for each subunit combination as follows: alpha2beta4, (+)epibatidine (Epi) > Cyt > suberyldicholine (Sub) = Nic = DMPP; alpha3beta4, Epi > DMPP = Cyt = Nic = Sub; alpha4beta4, Epi > Cyt = Sub > Nic > DMPP. The noncompetitive antagonists mecamylamine and d-tubocurarine did not display subtype selectivity. In contrast, the Kb value for the competitive antagonist dihydro-beta-erythroidine (DHbetaE) was highest at alpha3beta4 compared with alpha2beta4 or alpha4beta4 receptors. These data illustrate that the A2B4, A3B4.2 and A4B4 stable cell lines are powerful tools for examining the functional and pharmacological properties of human alpha2beta4, alpha3beta4 and alpha4beta4 neuronal nicotinic receptors.
Assuntos
Receptores Nicotínicos/química , Compostos Bicíclicos Heterocíclicos com Pontes/metabolismo , Cálcio/metabolismo , Linhagem Celular , Humanos , Potenciais da Membrana/efeitos dos fármacos , Agonistas Nicotínicos/farmacologia , Antagonistas Nicotínicos/farmacologia , Técnicas de Patch-Clamp , Piridinas/metabolismo , RNA Mensageiro/genética , Ensaio Radioligante , Receptores Nicotínicos/efeitos dos fármacos , Proteínas Recombinantes , Relação Estrutura-AtividadeRESUMO
Nicotine, the prototypical agonist for neuronal nicotinic acetylcholine receptors (NAChR), nonselectively activates NAChR limiting its use in elucidating the function of NAChR subtypes. SIB-1765F is a subtype selective NAChR agonist that displaces [3H]-nicotine binding with an IC50 of 4.6 nM and [3H]-cytisine binding with an IC50 of 12.2 nM which is 2000- to 6000-fold lower than its displacement of [3H]-QNB or [125I]-alpha-bungarotoxin. SIB-1765F did not inhibit human or rat cholinesterases or the uptake of [3H]-DA in synaptosomal preparations. SIB-1765F mimicked (-)-nicotine in stimulating [3H]-DA release from rat striatal and olfactory tubercle slices, with EC50 values of 99.6 and 39.6 microM, respectively. Such stimulation was sensitive to mecamylamine and DH beta E. SIB-1765F also released endogenous DA in the striatum and the nucleus accumbens as measured by in vivo microdialysis. SIB-1765F was less efficacious than (-)-nicotine at stimulating [3H]-NE release from rat hippocampal slices; in contrast, SIB-1765F increased [3H]-NE release from rat thalamic and cortical slices with efficacies approaching those of (-)-nicotine. Similar to (-)-nicotine and (+/-)-epibatidine, subcutaneous administration of SIB-1765F increased the turnover rate of dopamine ex vivo both in the striatum and olfactory tubercles in a mecamylamine-sensitive manner. Because the release of striatal DA and hippocampal NE appears to be regulated by distinct NAChR, differential effects of SIB-1765F on striatal DA and hippocampal NE release supports the NAChR subtype selectivity of SIB-1765F compared to (-)-nicotine. This is further demonstrated by observations showing that SIB-1765F has a higher affinity for h alpha 4 beta 2 NAChR relative to h alpha 4 beta 4 NAChRs in displacing [3H]-epibatidine binding and increasing cytosolic CA+2 concentration in cell lines stably expressing h alpha 4 beta 2 or h alpha 4 beta 4.
Assuntos
Canais Iônicos/agonistas , Agonistas Nicotínicos/farmacologia , Piridinas/farmacologia , Pirrolidinas/farmacologia , Animais , Cálcio/metabolismo , Corpo Estriado/metabolismo , Dopamina/metabolismo , Masculino , Microdiálise , Norepinefrina/metabolismo , Condutos Olfatórios/metabolismo , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Receptores Nicotínicos/metabolismoRESUMO
In the present investigation, anti-nociceptive effects of neuronal nicotinic acetylcholine receptor (NAChR) ligands, (+)- and (-)-nicotine, cytisine, methylcarbamylcholine (MCC), dimethylphenylpiperazinium iodide (DMPP), and (+/-)-epibatidine were evaluated in the rat tail-flick assay both after subcutaneous (s.c.) and intracerebroventricular (i.c.v.) administration. The pharmacology of the tail-flick response to NAChR ligands after s.c. and i.c.v. routes was similar. Epibatidine was the most potent ligand examined with a longer duration of action than any other agonist. (-)-Nicotine was more active than (+)-nicotine indicating stereospecificity. ICV administration studies indicated an apparent partial agonist activity for (+)-nicotine in the tail-flick response. Tail-flick responses to NAChR agonists are independent of opioid and muscarinic pathways and appear to be mediated both by central and peripheral NAChR recognition sites. Central administration of MCC activates both NAChR and muscarinic anti-nociceptive mechanisms. Studies employing the alpha-adrenergic receptor alkylating agent, phenoxybenzamine or the noradrenergic neurotoxin, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4), suggested that the NAChR-noradrenergic and NAChR-serotoninergic interactions play an important role in the tail-flick response. Studies employing a selective alpha-bungarotoxin-sensitive NAChR receptor antagonist, methyllycaconitine (MLA), suggested a minimal role for these receptors in the tail-flick response. The biochemical studies also indicated that a sub-population of NAChR receptors are located pre-synaptically on noradrenergic and/or serotoninergic pathways in the hippocampus.
Assuntos
Aconitina/análogos & derivados , Neurônios/efeitos dos fármacos , Agonistas Nicotínicos/farmacologia , Antagonistas Nicotínicos/farmacologia , Medição da Dor/métodos , Receptores Nicotínicos/efeitos dos fármacos , Aconitina/farmacologia , Animais , Cromatografia Líquida de Alta Pressão , Eletroquímica , Estudos de Avaliação como Assunto , Injeções Intraventriculares , Injeções Subcutâneas , Ligantes , Masculino , Norepinefrina/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Virtually all microorganisms require iron for growth. The paucity of iron in surface ocean water (approximately 0.02-1.0 nM (refs 1, 2)) has spurred a lively debate concerning iron limitation of primary productivity, yet little is known about the molecular mechanisms used by marine microorganisms to sequester iron. Terrestrial bacteria use a siderophore-mediated ferric uptake system. A siderophore is a low-molecular-mass compound with a high affinity for ferric ion which is secreted by microorganisms is response to low-iron environments; siderophore biosynthesis is regulated by iron levels, with repression by high iron. Although open-ocean marine microorganisms (such as phytoplankton and bacteria) produce siderophores, the nature of these siderophores has not been investigated. We report here the first structure determination, to our knowledge, of the siderophores from an open-ocean bacterium, alterobactin A and B from Alteromonas luteoviolacea. A. luteoviolacea is found in oligotrophic and coastal waters. Alterobactin A has an exceptionally high affinity constant for ferric ion. We suggest that at least some marine microorganisms may have developed higher-affinity iron chelators as part of an efficient iron-uptake mechanism which is more effective than that of their terrestrial counterparts.
Assuntos
Compostos Férricos/metabolismo , Bactérias Aeróbias Gram-Negativas/química , Oligopeptídeos/química , Peptídeos Cíclicos/química , Sideróforos/química , Sequência de Aminoácidos , Bactérias Aeróbias Gram-Negativas/metabolismo , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Dados de Sequência Molecular , Oligopeptídeos/metabolismo , Peptídeos Cíclicos/metabolismo , Conformação Proteica , Sideróforos/metabolismo , Espectrometria de Massas de Bombardeamento Rápido de ÁtomosRESUMO
A controlled trial was designed to compare both the long-term efficacy and the safety of fenbufen and indomethacin in patients with osteoarthritis. One hundred and ten patients of both sexes (aged 33 to 79 years) who had both subjective and objective (including radiological) evidence of osteoarthritis, and who met the minimal entry criteria, were enrolled into the studies. Patients were randomly assigned to receive either fenbufen capsules b.i.d. (600-1000 mg/day) or identical appearing indomethacin capsules b.i.d. (750-125 mg/day). Thirty-seven fenbufen and 26 indomethacin patients completed twelve months of treatment. Both fenbufen and indomethacin provided statistically nd clinically significant improvement at months 1, 3, 6, 9 and 12. There were no significant differences in improvement between the two treatment groups. Fenbufen-treated patients, however, reported fewer occurrences (4) of severe drug-related adverse experiences than indomethacin-treated patients (20). Significantly fewer headaches were reported with fenbufen and the number of patients terminating treatment because of adverse experiences was significantly greater with indomethacin. Fenbufen, in this patient population, provided a superior ratio of benefits to risk for the long-term treatment of osteoarthritis.U
Assuntos
Anti-Inflamatórios/uso terapêutico , Indometacina/uso terapêutico , Osteoartrite/tratamento farmacológico , Fenilbutiratos , Propionatos/uso terapêutico , Adulto , Idoso , Ensaios Clínicos como Assunto , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Distribuição AleatóriaAssuntos
Anti-Inflamatórios/uso terapêutico , Indometacina/uso terapêutico , Osteoartrite/tratamento farmacológico , Fenilbutiratos , Propionatos/uso terapêutico , Adulto , Idoso , Método Duplo-Cego , Feminino , Humanos , Indometacina/efeitos adversos , Masculino , Pessoa de Meia-Idade , Propionatos/efeitos adversos , Distribuição AleatóriaRESUMO
A six-month, double-blind study was conducted to compare pirprofen (Rengasil), 800 mg per day, to aspirin, 3.6 Gm per day, in patients with active, definite, or classical rheumatoid arthritis. Data from 33 patients, 17 in the pirprofen group and 16 in the aspirin group, were analyzed for efficacy while data from 37 patients were analyzed for safety. In the pirprofen group, four of seven variables analyzed were significantly improved over baseline at the terminal visit; no statistically significant improvement was seen at the terminal visit in the aspirin group, although statistically significant improvement was achieved at a number of interim visits. Between-treatment comparisons indicated that pirprofen was statistically superior to aspirin with respect to the number of swollen joints at the terminal visit and average grip strength after six weeks of treatment. There were no serious signs of toxicity in either treatment group.