Human amyloid precursor-like protein 1--cDNA cloning, ectopic expression in COS-7 cells and identification of soluble forms in the cerebrospinal fluid

Amyloid precursor-like protein 1 (APLP1) represents an integral membrane type 1 protein of unknown function which was originally cloned from a mouse cDNA library on the basis of sequence similarity with the Alzheimer's amyloid precursor protein (APP). Here we report on the molecular cloning and expression of the human APLP1 (hAPLP1). hAPLP1 consists of 650 amino acids, displays 89% identity on the amino acid level to its mouse homologue and has a calculated molecular mass of 72 kDa. hAPLP1 synthesized in a cell-free system displays an apparent molecular mass of approximately 80 kDa in SDS-containing gels and becomes N-glycosylated when the in vitro translation is performed in the presence of microsomes. The hAPLP1 cDNA was also expressed ectopically in COS-7 cells and the protein expression was analyzed by immunoprecipitation and western blotting. We have demonstrated that hAPLP1 represents a novel glycoprotein which carries both N- and O-linked glycans. Moreover, hAPLP1 undergoes limited proteolysis which results in the secretion of the carboxy-terminal truncated molecule into the cells conditioned medium. Examination of cells transfected with hAPLP1 cDNA by confocal laser microscopy reveals an intense perinuclear and Golgi staining, a pattern resembling the subcellular distribution of APP. Using a novel hAPLP1-specific antiserum, we identified soluble hAPLP1 in the human cerebrospinal fluid, which suggests that secretion of hAPLP1 from brain cells also takes place in vivo.     

Plasma Amyloid-Beta Levels in a Pre-Symptomatic Dutch-Type Hereditary Cerebral Amyloid Angiopathy Pedigree: A Cross-Sectional and Longitudinal Investigation

Plasma amyloid-beta (Aβ) has long been investigated as a blood biomarker candidate for Cerebral Amyloid Angiopathy (CAA), however previous findings have been inconsistent which could be attributed to the use of less sensitive assays. This study investigates plasma Aβ alterations between pre-symptomatic Dutch-type hereditary CAA (D-CAA) mutation-carriers (MC) and non-carriers (NC) using two Aβ measurement platforms. Seventeen pre-symptomatic members of a D-CAA pedigree were assembled and followed up 3–4 years later (NC = 8; MC = 9). Plasma Aβ1-40 and Aβ1-42 were cross-sectionally and longitudinally analysed at baseline (T1) and follow-up (T2) and were found to be lower in MCs compared to NCs, cross-sectionally after adjusting for covariates, at both T1(Aβ1-40: p = 0.001; Aβ1-42: p = 0.0004) and T2 (Aβ1-40: p = 0.001; Aβ1-42: p = 0.016) employing the Single Molecule Array (Simoa) platform, however no significant differences were observed using the xMAP platform. Further, pairwise longitudinal analyses of plasma Aβ1-40 revealed decreased levels in MCs using data from the Simoa platform (p = 0.041) and pairwise longitudinal analyses of plasma Aβ1-42 revealed decreased levels in MCs using data from the xMAP platform (p = 0.041). Findings from the Simoa platform suggest that plasma Aβ may add value to a panel of biomarkers for the diagnosis of pre-symptomatic CAA, however, further validation studies in larger sample sets are required.     

Generation effects and source memory in healthy older adults and in adults with dementia of the Alzheimer type.

Recognition and source memory were explored in healthy older adults, adults diagnosed with very mild dementia of the Alzheimer type (DAT), and adults diagnosed with mild DAT. Two sentence-completion tasks were used. In Task 1, half of the sentences were completed (clozed) by the participant, and half by the experimenter. In Task 2, half were participant clozed, and half were participant read (already clozed). Recognition of the cloze words and accuracy of categorizing them as participant generated or experimenter generated (Task 1) and participant generated or participant read (Task 2) were measured (source discriminalion). Contrary to previous reports, the DAT groups showed the generation effect, that is, better recognition for participant-generated words than experimenter-generated words (Task 1) or read words (Task 2). Source discrimination was disproportionately impaired in the DAT groups. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Inhibition of platelet activation by the Alzheimer's disease amyloid precursor protein

The amyloid precursor protein (APP) of Alzheimer's disease is abundantly expressed in the platelet alpha-granule where its role remains unclear. This study describes a novel function for APP in regulating human platelet activation. Preincubation of platelet-rich plasma with recombinant secreted APP (sAPP) isoforms dose-dependently inhibited platelet aggregation and secretion induced by ADP or adrenaline. Similarly, sAPP potently inhibited low-dose thrombin-induced activation in washed platelet suspensions, indicating that the activity does not require plasma cofactors. There were no functional differences between sAPP forms with or without the Kunitz protease inhibitor domain or derived from either alpha- or beta-secretase cleavage. In fact, the N-terminal cysteine-rich region of APP (residues 18-194) was as effective as the entire sAPP region in the inhibition of platelet activation. The inhibitory activity of sAPP correlated with a significant reduction in the agonist-induced production of the arachidonic acid (AA) metabolites thromboxane B2 and prostaglandin E2. However, sAPP did not affect AA-induced platelet aggregation or secretion, indicating the enzymatic conversion of AA was not inhibited. The addition of a threshold dose of AA reversed the sAPP-inhibition of agonist-induced platelet activation. This suggests that sAPP decreases the availability of free AA, although the mechanism is not yet known. These data provide evidence that the release of sAPP upon platelet degranulation may result in negative feedback regulation during platelet activation.     

Aging, source, and decision criteria: When false fame errors do and do not occur.

Two experiments investigated the influence of decision criteria on source memory performance of older adults and younger adults. Experiment 1 used the false fame paradigm, which encourages people to use relatively loose decision criteria when making what are, in essence, source judgments. Consistent with previous research, older adults made more false fame errors than younger adults. Experiment 2 was identical to Experiment 1 except that the fame judgments were made with the traditional source task format that encourages relatively stringent decision criteria when making source judgments: Possible sources were listed, and participants categorized names in terms of their source. In contrast to Experiment 1, older adults reduced their false fame errors to the level of younger adults. Encouraging older adults to use relatively stringent decision criteria when making source discriminations can reduce age differences in source misattributions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Phonological Blocking During Picture Naming in Dementia of the Alzheimer Type.

Individuals with dementia of the Alzheimer type (DAT; n = 53, ages 55-91), healthy older adults (n = 75, ages 59-91), and younger adults (n = 24, ages 18-24) performed a word-primed picturenaming task. Word primes were neutral (ready), semantically or phonologically related, or unrelated to the correct picture name. AH groups produced equivalent unrelated-word interference and semantic priming effects in response latencies. However, analysis of errors revealed a DAT-related increase of phonological blocking. The results suggest that picture-naming errors in DAT are due, at least in part, to a breakdown in access to phonological representations of object names as a consequence of reduced inhibitory control over other highly active alternatives. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Copper-binding amyloid precursor protein undergoes a site-specific fragmentation in the reduction of hydrogen peroxide

The extracellular domain of transmembrane Abeta amyloid precursor protein (APP) has a Cu(II) reducing activity upon Cu(II) binding associated with the formation of a new disulfide bridge. The complete assignment of the disulfide bond revealed the involvement of cysteines 144 and 158 around copper-binding histidine residues. The vulnerability of APP-Cu(I) complexes to reactive oxygen species was elaborated as a site-specific and random fragmentation of APP in a time-dependent manner and at low concentrations of H2O2. Analysis of the specific reaction revealed the generation of C-terminal polypeptides, containing the Abeta domain. APP catalyzed the reduction of H2O2 and oxidation of Cu(I) to Cu(II) in a "peroxidative" reaction in vitro. The resulting bound copper-hydroxyl radical intermediate [APP-Cu(II)(.OH)] then likely participated in a Fenton type of reaction with radical formation as a prerequisite for protein degradation. Evidence from two observations suggests that the reaction takes place in two phases. Bathocuproine, a trapping agent for Cu(I), abolished the initial fragmentation, and chelation of Cu(II) by DTPA (diethylenetriaminepentaacetic acid) interrupted the reaction cascade induced by H2O2 at later stages. Consequently, the results suggest that a cytotoxic gain-of-function of APP-Cu(I) complexes might result in a perturbation of free radical homeostasis. What significance such a perturbation may have for the pathogenesis of Alzheimer's disease remains to be determined.     

Inhibition in the processing of garden-path sentences.

The Hartman and Hasher (1991) garden-path sentence completion task has been used in several studies to assess the efficiency of the deletion function of inhibition (e.g., L. Hasher, R. Zacks, & C. P. May, 1999), with results suggesting that younger adults are efficient at suppressing once relevant but no longer appropriate information, whereas older adults generally are not (e.g., M. Hartman & L. Hasher, 1991; L. Hasher, M. B. Quig, & C. P. May, 1997; C. P. May & L. Hasher, 1998). An alternative interpretation of patterns of access to relevant and no-longer-relevant sentence endings focuses on the difficulty of selecting final words for sentence frames and on integration effects in implicit memory (M. Hartman, 1995). This alternative is considered and found wanting on the basis of both new and old empirical data. On the basis of present data and related findings, it is concluded that the task does measure inhibitory efficiency. (PsycINFO Database Record (c) 2010 APA, all rights reserved)