Amygdala, hippocampus and six cortical gyri were examined for the Lewy body (LB) degeneration and Alzheimer's disease (AD) type changes in 45 patients with Parkinson's disease (PD). For detection of LBs, the brain areas were stained with an antibody against alpha-synuclein. The extent of neuropathological lesions was investigated in relation to cognitive dysfunction and apolipoprotein E (apoE) epsilon4 allele dosage. At least one cortical LB was found in 95% of cases (43/45). Furthermore, 40% of cases (18/45) had histological findings of definite AD (CERAD class C). Those PD cases with the apoE epsilon4 allele had a significantly greater number of cortical LBs than those without the apoE epsilon4 allele, but this was statistically significant only in precentral, angular and temporal gyri. The LB density correlated better with the number of plaques than with the density of tangles. The number of LBs in several cortical areas correlated significantly with the cognitive impairment. In stepwise linear regression analysis, the number of LBs in the cingulate gyrus and the amount of tangles in the temporal cortex remained statistically significant. When the CERAD class C was excluded, the correlation between cognitive decline and the number of LBs in cortical areas became even more pronounced. A stepwise linear regression analysis in these cases found the number of LBs in the frontal gyrus to be the statistically most significant predictor of cognitive impairment. This study shows, for the first time, that in PD, alpha-synuclein-positive cortical LBs are associated with cognitive impairment independent of AD-type pathology.

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Semliki Forest virus (SFV) is a mosquito-transmitted pathogen of small rodents, and infection of adult mice with SFV4, a neurovirulent strain of SFV, leads to lethal encephalitis in a few days, whereas mice infected with the avirulent A7(74) strain remain asymptomatic. In adult neurons, A7(74) is unable to form virions and hence does not reach a critical threshold of neuronal damage. To elucidate the molecular mechanisms of neurovirulence, we have cloned and sequenced the entire 11,758-nucleotide genome of A7(74) and compared it to the highly neurovirulent SFV4 virus. We found several sequence differences and sought to localize determinants conferring the neuropathogenicity by using a panel of chimeras between SFV4 and a cloned recombinant, rA774. We first localized virulence determinants in the nonstructural region by showing that rA774 structural genes combined with the SFV4 nonstructural genome produced a highly virulent virus, while a reciprocal recombinant was asymptomatic. In addition to several amino acid mutations in the nonstructural region, the nsp3 gene of rA774 displayed an opal termination codon and an in-frame 21-nucleotide deletion close to the nsp4 junction. Replacement in rA774 of the entire nsp3 gene with that of SFV4 reconstituted the virulent phenotype, whereas an arginine at the opal position significantly increased virulence, leading to clinical symptoms in mice. Completion of the nsp3 deletion in rA774 did not increase virulence. We conclude that the opal codon and amino acid mutations other than the deleted residues are mainly responsible for the attenuation of A7(74) and that the attenuating determinants reside entirely in the nonstructural region.Semliki Forest virus (SFV) is an enveloped positive-stranded RNA virus of the family Togaviridae. The SFV prototype was isolated in 1942 from a pool of mosquitoes in Uganda (40). The A7(74) strain (4) is a derivative of an SFV strain isolated from mosquitoes in Mozambique in 1959 (26). Several strains of SFV, such as L10 (4) and SFV4 (24), cause lethal encephalitis in mice of all ages, leading to death of the animals in a few days (3, 11), whereas A7(74) infection of adult mice is asymptomatic per se but leads to axonal demyelination mediated by CD8 ϩ T cells (2). However, the severity of A7(74) infection is strictly age dependent, being lethal for neonatal mice less than 2 weeks old (8, 27), probably because the strain is capable of virion formation in propagating neurons, unlike in mature neurons (27).SFV nonstructural proteins (nsP) are translated as a polyprotein (nsP1234) from the genomic 42S RNA, and they form essential components of viral RNA replication and transcription complexes (14). The nsP1 protein is a methyl-and guanylyltransferase (1, 18), whereas the nsP2 is a proteinase (45) and nucleoside triphosphatase (33). The nsp3 gene product is a phosphoprotein, and it has been proposed to function together with nsP1 in anchoring the replication complex proteins to cytoplasmic membrane structures (30, 31). In Sindbis virus (SIN), p123 a...

OBJECTIVE-A functional polymorphism leucine 7 proline in the human neuropeptide Y (NPY) gene leading to increased NPY release from sympathetic nerves is associated with traits of metabolic syndrome. Although hypothalamic NPY neurons play an established role in promoting positive energy balance, the role of NPY colocalized with norepinephrine in sympathetic nervous system and brain noradrenergic neurons remains obscure.RESEARCH DESIGN AND METHODS-To clarify the role of NPY in noradrenergic neurons, we generated a transgenic mouse overexpressing NPY under dopamine-␤-hydroxylase promoter and characterized the metabolic phenotype of the OE-NPY D␤H mouse.RESULTS-NPY levels are increased by 1.3-fold in adrenal glands and 1.8-fold in the brainstem but not in the hypothalamus in OE-NPY D␤H mice. They display increased white adipose tissue mass and cellularity and liver triglyceride accumulation without hyperphagia or increased body weight. Hyperinsulinemia and impaired glucose tolerance develop by the age of 6 months in the OE-NPY D␤H mice. Furthermore, circulating ghrelin is significantly increased in comparison with wild-type mice.CONCLUSIONS-The present study shows that even a moderate increase in NPY levels in noradrenergic neurons leads to disturbances in glucose and lipid metabolism. The OE-NPY D␤H mouse is an interesting new model to investigate the pathophysiology of some key components of the cluster of abnormalities characterizing the metabolic syndrome. Diabetes 57: [1517][1518][1519][1520][1521][1522][1523][1524][1525] 2008 N europeptide Y (NPY) plays a well-established role in the hypothalamic control of body energy balance. It is one of the key components of the interconnected orexigenic network, which is upregulated in states of negative energy balance. NPY is a very potent orexigenic peptide, and when chronically administered into the central nervous system, it leads to increased food intake, weight gain, and adiposity (1-3). NPY-induced obesity is not only due to hyperphagia; centrally administered NPY also promotes white adipose tissue (WAT) lipid storage, inhibits brown adipose tissue thermogenesis, and induces hyperinsulinemia and hypercorticosteronemia (4 -6). Increased hypothalamic NPY is also an essential feature of leptin-deficient obesity, which is attenuated by genetic depletion of NPY (7). Although there is plenty of evidence on the key role of NPY in the regulation of energy balance in rodents, the evidence is scarce in humans and based mainly on the functional leucine 7 proline (L7P) polymorphism of the preproNPY associated with alterations in glucose and lipid metabolism and development of atherosclerosis (8).In addition to the NPY neurons of the hypothalamus, NPY is widely distributed in the central and peripheral nervous system. In the periphery, NPY is co-stored and co-released in postganglionic sympathetic neurons and chromaffin cells of adrenal medulla with norepinephrine (NE) (9,10). In the brain, NPY is also colocalized with NE in noradrenergic neurons in the medulla and the brain...

We evaluated the association of HSV-1, HHV-6, and VZV with Alzheimer's disease (AD) and Parkinson's disease (PD). Brain specimens for viral DNA polymerase chain reaction represented 34 patients with AD, 40 with PD, and 40 controls. One AD patient (2.9%) was positive for HSV-1 DNA, 88.2% for HHV-6 DNA, and 26.5% for VZV DNA; 17.5% of PD patients were HSV-1 DNA-positive and 75% HHV-6-positive, whereas 40% had VZV DNA. Twenty-five percent of the controls were positive for HSV-1 DNA, 87.5% for HHV-6, and 27.5% for VZV. HSV-1, VZV, or HHV-6 DNA in brains was no additional risk factor for AD.

SUMMARYHerpes simplex virus type 1 and a fluorescein-labelled lectin (wheat germ agglutinin) were selectively transported to nerve cell bodies located in the inner compartment of a two-chamber tissue culture system after the application of virus or lectin to the neuritic processes in the outer culture compartment. Taxol, which stabilizes and alters intracellular arrangements of microtubules, and nocodazole, which disrupts microtubules, both inhibited this retrograde axonal transport of viral particles and lectin. The transport was also inhibited by erythro-9-3-(2-hydroxynonyl)adenine (EHNA), which blocks ATPases. However, EHNA was also an effective inhibitor of infection with the virus in non-neuronal cells (GMK AH-1). The nature of the action(s) of EHNA on neuritic transport of the virus is therefore less clear.The pathogenesis of herpes simplex virus (HSV) infections involves axonal transport of virus and viral subunits by sensory neurons in the anterograde as well as the retrograde direction. Neurites of dissociated dorsal root ganglion cells have the capacity to internalize the virus (Ziegler & Herman, 1980), presumably by fusion of the viral envelope with the neuritic plasma membrane (Lycke et al., 1984). Since nucleocapsids are detectable in neurites exposed to HSV and are seen around nuclear pores in HSV-infected cell lines (Batterson et al., 1983), we assume that they are involved in retrograde axonal transport of the virus. The mechanisms of this axonal transport of HSV remain to be clarified.We now report the effects on neuritic transport of HSV of three substances capable of inhibiting intracellular transport of endosomes, namely nocodazole, which causes disruption of microtubules, and taxol, which promotes assembly of microtubules (Herman & Albertini, 1984); the third drug, erythro-9-3-(2-hydroxynonyl)adenine (EHNA) (Bouchard et al., 1981) is supposed to inhibit the function of the microtubule-associated ATPase dynein and can inhibit the retrograde movement of organelles in axons. EHNA is considered to have less effect on anterograde axonal transport (Forman et al., 1983).We have used a two-chamber culture system allowing us to infect the neuritic extensions of rat sensory neurons cultured in vitro, without exposing the cell bodies to the virus. For evaluation of the results the neuritic transfer of HSV was compared with the transport of wheat germ agglutinin (WGA).The two-chamber system employed has been described previously (Ziegler & Herman, 1980). Briefly, 35 mm collagen-coated plates were scratched on the bottom, fitted in the centre with an 8 mm cloning cylinder kept in place by silicone high vacuum grease and seeded in the cloning cylinder with trypsin-dissociated dorsal root ganglion (DRG) cells. The cells were obtained by dissecting eight to ten embryos of 16 to 17 day pregnant Sprague-Dawley rats. Trypsinization was performed for 30 min at 37 °C using 0.25~ trypsin (porcine pancreas, type 2, Sigma) in a Ca z÷-and MgZ+-free Hanks' buffer pH 7.2. For the first 2 days, the cells were cultured ...