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Notes: Abstract: Alzheimer's disease (AD) is identified by the accumulation of amyloid plaques, neurofibrillary degeneration, and the accompanying neuronal loss. AD amyloid assembles into compact fibrous deposits from the amyloid β(Aβ) protein, which is a proteo-lytic fragment of the membrane-associated amyloid precursor protein. To examine the effects of amyloid on neuron growth, a hybrid mouse motoneuron cell line (NSC34) exhibiting spontaneous process formation was exposed to artificial “plaques” created from aggregated synthetic Aβ peptides. These correspond to full-length Aβ residues 1–40 (Aβ1–40), an internal β-sheet region comprising residues 11–28 (Aβ11–28), and a proposed toxic fragment comprising residues 25–35 (Aβ25–35). Fibers were immobilized onto culture dishes, and addition of cells to these in vitro plaques revealed that Aβ was not a permissive substrate for cell adhesion. Neurites in close contact with these deposits displayed abnormal swelling and a tendency to avoid contact with the Aβ fibers. In contrast, Aβ did not affect the adhesion or growth of rat astrocytes, implicating a specific Aβ-neuron relationship. The inhibitory effects were also unique to Aβ as no response was observed to deposits of pancreatic islet amyloid poly-peptide fibers. Considering the importance of cell adhesion in neurite elongation and axonal guidance, the antiadhesive properties of Aβ amyloid plaques found in vivo may contribute to the neuronal loss responsible for the clinical manifestations of AD.

Notes: Abstract: A detailed comparative study of RNA transcripts isolated from the neocortex of control and Alzheimer postmortem brains was made to determine whether morphological changes in the chromatin of Alzheimer neurons and glia, which we reported earlier, are accompanied by changes in the products of transcription. A number of parameters were determined including the yields of total and mRNA per gram of tissue, the relative proportions of polyadenylated [poly(A) +] mRNA in the total RNA, the size distribution of the transcripts and the length of their poly(A) tails, and the nature of their in vitro translation products. The levels of endogenous RNase activity were also measured. The effect of the agonal process on the transcript complement was examined by Northern blotting of a cloned human heat-shock cDNA to total human brain RNA. Our results reveal that the yields of total RNA, unadenylated mRNA, and poly(A) tail lengths from Alzheimer neocortex samples do not differ significantly from those of control and non Alzheimer dementia neocortex. On the other hand we find a significant reduction in the levels and proportion of poly(A)+ mRNA in the Alzheimer samples as compared to control brain samples. Quantitative rather than qualitative differences were observed in the in vitro translation products when programmed with control and Alzheimer mRNA. No differences were found in the levels of RNase activity between control and Alzheimer simples. Heatshock mRNA transcripts were detected in brain samples from patients in whom fever was associated with death. The direct correlation of reduced poly(A)+ mRNA and chromatin condensation in Alzheimer neocortex suggests a cause-and-effect relationship. Whether all transcribed genes are affected or only a specific subset has yet to be determined.

Notes: Abstract— The enzyme micrococcal nuclease was used to examine the accessibility of chromatin extracted from brains of 13 patients with senile and presenile dementia of the Alzheimer type. Compared with chromatin extracted from brains of 8 patients without neurological signs or brain pathology and brains of 7 patients with nonAlzheimer dementia, Alzheimer chromatin was less accessible to this enzyme-. Reduced accessibility was reflected by a reduced yield of mononucleosomes in comparison with dinucleosomes and larger oligomers. Both neuronal and glial chromatin were found to be similarly affected. The reduced yield of mononucleosomes from Alzheimer chromatin is not due to their increased breakdown, but is probably related to protein associated with the internucleosomal linker region that retards nuclease action. Dinucleosomes isolated from control and Alzheimer nuclease digests were examined for their protein complement. Three perchloric acid-soluble proteins situated in the histone HI region of sodium dodecyl sulfate (SDS) gels were present in elevated levels in Alzheimer dinucleosomes. These results represent the first example of altered chromosomal proteins associated with a diseased state of the brain.

Notes: Summary Brains of patients with senile dementia of the Alzheimer type and with the dialysis encephalopathy syndrome, exhibit elevated aluminum levels. In Alzheimer's disease and in the experimental aluminum induced encephalopathy, intracellular aluminum is associated with nuclear chromatin. The work reported here was undertaken to test whether chromatin bound aluminum in hippoccampus of the rabbit interferes with nuclear binding of corticosterone-receptor complexes. The results showed that the mean binding of corticosterone decreased from 460±71 fmol/mg DNA in controls, to 343±81 fmol/mg DNA in hippocampal nuclei from aluminum treated rabbits, representing a decrease of 25%. This reduction occurred in the absence of aluminum induced neurofibrillary degeneration and indicates a possible functional consequence of the presence of aluminum on chromatin, and importantly, in the absence of morphological changes.

Notes: [Auszug] Available members of 48 pedigrees (Table 1) were typed with five anonymous DNA markers (D21S16, D21S13, D21S52, D21S1 and D21S11) that constitute three independent loci whose physical and genetic relationships have been previously determined (Table 2)7~9. These data were then assessed using both ...

Notes: Abstract The yields of total and poly(A) RNA were examined in rabbit forebrains during an experimentally induced aluminum encephalopathy. Rabbits (35 day old) were injected intracranially with 13 μmole Al lactate and sacrificed 1, 3, 7, 10, or 12 days later. IRNA yields (total RNA minus transfer RNA) were not significantly altered during the encephalopathy. Poly(A) RNA yields, assayed by oligo(dT)-cellulose fractionation and by a [3H]poly(U) hybridization assay on IRNA, were increased significantly by the end of the asymptomatic stage of the encephalopathy (7 days post-Al injection). The increase in messenger RNA population may represent either a compensatory response to cell damage induced by aluminum or the accumulation of messager RNA for proteins directly related to the expression of aluminum toxicity.

Notes: Abstract BC200 RNA is a polyadenylated 200 nucleotide primate brain-specific transcript with 80% homology to the left monomer of the human Alu family of repetitive elements. Whether this transcription product contributes anything to normal brain gene function or is a residue of post transcriptional processing of brain heterogeneous nuclear RNA (hnRNA) is uncertain. However, the high abundance, tissue-specific expression and nucleotide sequence characteristics of BC200 RNA suggests that the generation of this small RNA is associated with some brain cell function. Sustained levels of the BC200 RNA transcript may be indicative of a genetically competent and normally functioning cerebral neocortex. In this investigation, we have measured the abundance of the BC200 RNA transcript in total RNA isolated from 18 temporal neocortices (Brodman area 22) of brains with no pathology and those affected with neurodegenerative disease. Neocortices were examined from 3 neurologically normal brains, 5 non-Alzheimer dernented [NAD; 3 Huntingtons chorea (HC), 1 amyotrophic lateral sclerosis (ALS) and 1 dementia unclassified] and 10 Alzheimer disease (AD) affected brains. Our results indicate a strong BC200 presence in both the normal brains and NAD affected neocortices, but a 70 per cent reduction in BC200 signal strength in AD afflicted brains. These results may be related to the observation that Alzheimer brains exhibit marked deficits in the abundance of neuron-specific DNA transcripts; these deficits are consistent with the idea that AD is characterized by an impairment in the primary generation of brain gene transcription products.

Notes: Abstract The etiology of some, if not all, cases of Alzheimer's disease is linked to a mutation in the proximal portion of the long arm of chromosome 21∶21q11.2 → 21q22.2. While the functional consequences of the mutation are unknown, we speculate that one consequence of the mutation is loss of the natural barriers and intracellular ligands for aluminum. As a result, aluminum gains access to several brain sites including the nuclear compartment in certain neurons of the central nervous system. Both sporadic and familial Alzheimer's disease are associated with an increased compaction of DNA within chromatin as measured by physical shearing and resistance to digestion by micrococcal nuclease and DNase I. There is also an increase in linker histone Hlo content on dinucleosomes released by light (3–5% ASN) micrococcal nuclease digestion, and an increase in the affinity of histone Hlo for DNA as measured by a salt elution technique. The change in enzyme accessibility to chromatin also involves the 5′ promoter region of at least one physiologically important gene: the gene which codes for the low molecular weight moiety of neurofilament (NF-L). The conformation change involving the 5′ regulator region probably reduces transcription because the pool size of the mRNA coding for NF-L is reduced to 14% of age matched control in cerebral grey matter. Reduced transcription may account for many disorders in cellular metabolic processes including the regulation of phosphorylation, calcium homeostasis, free radical metabolism, proteolysis and neurotransmitter metabolism. The experimental evidence indicates that one important toxic action of aluminum in Alzheimer's disease neocortex is to increase the binding of histones, particularly Hlo, to DNA which results in increased compaction of chromatin and reduced transcription. The supporting evidence includes: (1) A statistically reliable correlation between the aluminum to DNA ratio on intermediate euchromatin and the amount of highly condensed heterochromatin found in a given preparation from Alzheimer affected neocortex (Crapperet al., 1980). (2) A nine-fold increase in aluminum content in Alzheimer's disease in the di- and tri- nucleosome fraction released by light micrococcal nuclease digestion of nuclei from cerebral grey matter compared to age matched controls. Compared to age matched control dinucleosomes, the Alzheimer affected dinucleosomes contain an increased abundance of the linker histone Hlo and an increased proportion of DNA containing the promoter region of the gene coding for NF-L. (3) A reduction in abundance to 14% of control mRNA coding for NF-L in Alzheimer affected neocortex (Crapper McLachlanet al., 1988). (4) In vitro evidence that Alzheimer linker histones bind more tightly to DNA than control and that aluminum added to nuclei,in vitro, extracted from normal control brain, enhances DNA-protein binding of Hl and Hlo at concentrations found in the Alzheimer affected chromatin (Lukiwet al., 1987). (5) Application of a band retardation assay indicates that aluminum,in vitro, selectively binds human Hlo to a 300 bp human ALU DNA fragment from a crude extract of 5% per chloric acid soluble proteins. (6) Aluminum experimentally applied to rabbit CNS induces a marked reduction in NF-L mRNA in anterior horn cells (Mumaet al., 1988). We therefore conclude that aluminum plays a major role in the pathogenesis of Alzheimer's disease. Further understanding of the role of aluminum in Alzheimer's disease requires a detailed investigation of the precise sites of co-ordination of this trivalent metal within chromatin.