Microglial responses to amyloid β peptide opsonization and indomethacin treatment

BACKGROUND: Recent studies have suggested that passive or active immunization with anti-amyloid β peptide (Aβ) antibodies may enhance microglial clearance of Aβ deposits from the brain. However, in a human clinical trial, several patients developed secondary inflammatory responses in brain that were sufficient to halt the study. METHODS: We have used an in vitro culture system to model the responses of microglia, derived from rapid autopsies of Alzheimer's disease patients, to Aβ deposits. RESULTS: Opsonization of the deposits with anti-Aβ IgG 6E10 enhanced microglial chemotaxis to and phagocytosis of Aβ, as well as exacerbated microglial secretion of the pro-inflammatory cytokines TNF-α and IL-6. Indomethacin, a common nonsteroidal anti-inflammatory drug (NSAID), had no effect on microglial chemotaxis or phagocytosis, but did significantly inhibit the enhanced production of IL-6 after Aβ opsonization. CONCLUSION: These results are consistent with well known, differential NSAID actions on immune cell functions, and suggest that concurrent NSAID administration might serve as a useful adjunct to Aβ immunization, permitting unfettered clearance of Aβ while dampening secondary, inflammation-related adverse events

Epigenetic Differences in Cortical Neurons from a Pair of Monozygotic Twins Discordant for Alzheimer's Disease

DNA methylation [1], [2] is capable of modulating coordinate expression of large numbers of genes across many different pathways, and may therefore warrant investigation for their potential role between genes and disease phenotype. In a rare set of monozygotic twins discordant for Alzheimer's disease (AD), significantly reduced levels of DNA methylation were observed in temporal neocortex neuronal nuclei of the AD twin. These findings are consistent with the hypothesis that epigenetic mechanisms may mediate at the molecular level the effects of life events on AD risk, and provide, for the first time, a potential explanation for AD discordance despite genetic similarities

Altered sphingolipid function in Alzheimer's disease;:a gene regulatory network approach

Sphingolipids (SLs) are bioactive lipids involved in various important physiological functions. The SL pathway has been shown to be affected in several brain-related disorders, including Alzheimer's disease (AD). Recent evidence suggests that epigenetic dysregulation plays an important role in the pathogenesis of AD as well. Here, we use an integrative approach to better understand the relationship between epigenetic and transcriptomic processes in regulating SL function in the middle temporal gyrus of AD patients. Transcriptomic analysis of 252 SL-related genes, selected based on GO term annotations, from 46 AD patients and 32 healthy age-matched controls, revealed 103 differentially expressed SL-related genes in AD patients. Additionally, methylomic analysis of the same subjects revealed parallel hydroxymethylation changes in PTGIS, GBA, and ITGB2 in AD. Subsequent gene regulatory network-based analysis identified 3 candidate genes, that is, SELPLG, SPHK1 and CAV1 whose alteration holds the potential to revert the gene expression program from a diseased towards a healthy state. Together, this epigenomic and transcriptomic approach highlights the importance of SL-related genes in AD, and may provide novel biomarkers and therapeutic alternatives to traditionally investigated biological pathways in AD.</p

Seasonal changes in a sandy beach fish assemblage at Canto Grande, Santa Catarina, South Brazil

Copyright © 2004 Coastal Education and Research Foundation (CERF).Neste trabalho realizaramse amostragens, com uma rede de praia, de modo a estudar a comunidade de peixes de substrato arenoso na enseada de Canto Grande, Santa Catarina, Brasil. As amostragens realizaramse em intervalos de 3 horas durante períodos de 24 h, numa base bimensal, entre Abril de 1996 e Fevereiro de 1997. Verificouse existir uma variação sazonal no número de espécies, densidade de peixes e biomassa, tendo os valores mais elevados ocorrido em Fevereiro (38 espécies, 257.6 peixes 1000 mˉ², 2286.4 g 1000 mˉ²). Recolheuse um total de 67 espécies, pertencentes a 56 géneros e a 33 famílias, sendo a comunidade dominada por sete espécies pertencentes a três famílias: Atherinella brasiliensis (Atherinidae); Brevoortia pectinata, Harengula clupeola e Sardinella brasiliensis (Clupeidae); Anchoviella lepidontostole, Cetengraulis edentulus e Lycengraulis grossidens (Engraulidae). Tanto a diversidade de espécies (H′) como a equitabilidade (J′) foram médias a elevadas ao longo do ano devido à baixa dominância. A maior mudança na estrutura da comunidade ocorreu entre os meses de Inverno (Julho e Agosto) e as outras estações. Nenhuma das espécies dominantes pode ser classificada como residente. Os principais predadores foram Pomatomus saltator (Inverno) e Trichiurus lepturus (Verão). A maior parte das espécies observadas foram ou peixes juvenis ou espécies pelágicas de pequeno tamanho e fortemente gregárias.ABSTRACT: A shallow-water fish assemblage, over a soft, sandy bottom, at Canto Grande, Santa Catarina, Brazil, was sampled with a beach seine. Sampling was undertaken at 3 h intervals over 24 h on a bimonthly basis between April 1996 and February 1997. There was a seasonal variation in the number of species, density of fishes and biomass with the highest values in February (38 species, 257.6 fish 1000 mˉ², 2286.4 g 1000 mˉ²). A total of 67 species, belonging to 56 genera and 33 families were collected and the assemblage was dominated by seven species belonging to three families: Atherinella brasiliensis (Atherinidae); Brevoortia pectinata, Harengula clupeola and Sardinella brasiliensis (Clupeidae); Anchoviella lepidontostole, Cetengraulis edentulus and Lycengraulis grossidens (Engraulidae). Species diversity (H′) and equitability (J′) were medium to high throughout the year due to the low dominance. The largest change in the assemblage structure occurred between winter months (July and August) and the other seasons. None of the dominant species can be classified as a resident. Main predators were Pomatomus saltator (winter) and Trichiurus lepturus (summer). Most of the species observed were either juvenile fish or small pelagic and strongly gregarious species

Targeted methylation profiling of single laser-capture microdissected post-mortem brain cells by adapted limiting dilution bisulfite pyrosequencing (LDBSP)

A reoccurring issue in neuroepigenomic studies, especially in the context of neurodegenerative disease, is the use of (heterogeneous) bulk tissue, which generates noise during epigenetic profiling. A workable solution to this issue is to quantify epigenetic patterns in individually isolated neuronal cells using laser capture microdissection (LCM). For this purpose, we established a novel approach for targeted DNA methylation profiling of individual genes that relies on a combination of LCM and limiting dilution bisulfite pyrosequencing (LDBSP). Using this approach, we determined cytosine-phosphate-guanine (CpG) methylation rates of single alleles derived from 50 neurons that were isolated from unfixed post-mortem brain tissue. In the present manuscript, we describe the general workflow and, as a showcase, demonstrate how targeted methylation analysis of various genes, in this case, RHBDF2, OXT, TNXB, DNAJB13, PGLYRP1, C3, and LMX1B, can be performed simultaneously. By doing so, we describe an adapted data analysis pipeline for LDBSP, allowing one to include and correct CpG methylation rates derived from multi-allele reactions. In addition, we show that the efficiency of LDBSP on DNA derived from LCM neurons is similar to the efficiency obtained in previously published studies using this technique on other cell types. Overall, the method described here provides the user with a more accurate estimation of the DNA methylation status of each target gene in the analyzed cell pools, thereby adding further validity to this approach

Analysis of the putative role of CR1 in Alzheimer’s disease: Genetic association, expression and function

Chronic activation of the complement system and induced inflammation are associated with neuropathology in Alzheimer's disease (AD). Recent large genome wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) in the C3b/C4b receptor (CR1 or CD35) that are associated with late onset AD. Here, anti-CR1 antibodies (Abs) directed against different epitopes of the receptor, were used to localize CR1 in brain, and relative binding affinities of the CR1 ligands, C1q and C3b, were assessed by ELISA. Most Abs tested stained red blood cells in blood vessels but showed no staining in brain parenchyma. However, two monoclonal anti-CR1 Abs labeled astrocytes in all of the cases tested, and this reactivity was preabsorbed by purified recombinant human CR1. Human brain-derived astrocyte cultures were also reactive with both mAbs. The amount of astrocyte staining varied among the samples, but no consistent difference was conferred by diagnosis or the GWAS-identified SNPs rs4844609 or rs6656401. Plasma levels of soluble CR1 did not correlate with diagnosis but a slight increase was observed with rs4844609 and rs6656401 SNP. There was also a modest but statistically significant increase in relative binding activity of C1q to CR1 with the rs4844609 SNP compared to CR1 without the SNP, and of C3b to CR1 in the CR1 genotypes containing the rs6656401 SNP (also associated with the larger isoform of CR1) regardless of clinical diagnosis. These results suggest that it is unlikely that astrocyte CR1 expression levels or C1q or C3b binding activity are the cause of the GWAS identified association of CR1 variants with AD. Further careful functional studies are needed to determine if the variant-dictated number of CR1 expressed on red blood cells contributes to the role of this receptor in the progression of AD, or if another mechanism is involved