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Mutations in the integral membrane protein 2B [1], also known as BRI2 [2], a type II trans-membrane domain protein cause two autosomal dominant neurodegenerative diseases, Familial British and Danish Dementia [3]. In these conditions, accumulation of a C-terminal peptide (ABri and ADan) cleaved off from the mutated precursor protein by the pro-protein convertase furin [4], leads to amyloid deposition in the walls of blood vessels and parenchyma of the brain. Recent advances in the understanding of the generation of amyloid in Alzheimer's disease has lead to the finding that BRI2 interacts with the Amyloid Precursor Protein (APP), decreasing the efficiency of APP processing to generate Aβ [5], [6], [7]. The interaction between the two precursors, APP and BRI2, and possibly between Aβ and ABri or ADan, could be important in influencing the rate of amyloid production or the tendency of these peptides to aggregate.
Methodology/Principal Findings
We have generated the first BRI2 Danish Knock-In (FDDKI) murine model of FDD, expressing the pathogenic decamer duplication in exon 6 of the BRI2 gene. FDDKI mice do not show any evident abnormal phenotype, with normal brain histology and no detectable amyloid deposition in blood vessel walls or parenchyma.
Citation: Giliberto L, Matsuda S, Vidal R, D'Adamio L (2009) Generation and Initial Characterization of FDD Knock In Mice. PLoS ONE 4(11): e7900. https://doi.org/10.1371/journal.pone.0007900
Editor: Richard Mayeux, Columbia University, United States of America
Received: July 22, 2009; Accepted: October 26, 2009; Published: November 18, 2009
Copyright: © 2009 Giliberto et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was supported in part by Alzheimer Disease Research Grant A2003-076 (LD); National Institutes of Health Grants RO1 AG22024 (LD); RO1 AG21588 (LD); R21 AG027139 (LD); American Health Assistance Foundation A2008-304 (RV).
Competing interests: LG and LD are inventors of an AECOM patent on the FDD mice. The mice discussed in this paper (BRI2-ADan knock-in mice) are patent-pending. The assign is the Albert Einstein College of Medicine. The Albert Einstein College of Medicine has licensed the patent to a Biotech company (Remegenix) of which Dr. D'Adamio is a co-founder. This does not alter the authors adherence to all the PLoS ONE policies on sharing data and materials, as detailed online in the guide for authors http://www.plosone.org/static/policies.action#sharing.
BRI2 is a type II trans-membrane protein of unknown function. The BRI gene belongs to a multigene family comprising at least three homologues in both mice and humans, BRI1, BRI2 and BRI3 (also referred to as ITM2A, ITM2B and ITM2C, or E25A, E25B and E25C, respectively) [1], [2], [3], [8], [9]. It possesses a BRICHOS domain, a conserved motif common to members of the BRI, ChM-I, SP-C and CA11 protein families, thought to have a role in the targeting of the respective proteins to the secretory pathway or to intracellular processing [10]. Proteins sharing the BRICHOS motif are dissimilar, and associate with a diverse range of phenotypes, varying from dementia to cancer and respiratory distress.
BRI2 was first described in relation to Familial British Dementia (FBD) [2], an autosomal dominant neurodegenerative disease characterized by the early onset of personality changes, memory and cognitive deficits, spastic rigidity and ataxia. In FBD, a C-terminal 34 amino acid (aa) peptide of BRI2 accumulates as amyloid, leading to severe amyloid angiopathy of the brain and spinal cord with perivascular amyloid plaque formation, parenchymal plaques affecting the limbic areas, cerebellum, cerebral cortex, neurofibrillary tangles of hippocampal neurons and periventricular white matter changes [11]. Familial Danish Dementia (FDD), previously known as heredopathia ophthalmo-otoencephalica, is an autosomal dominant disease characterized by the accumulation of an amyloidogenic C-terminal 34 aa peptide of BRI2 [3]. FDD is characterized by early onset cataracts, deafness, progressive ataxia and dementia [3], [12]. Neuropathological examination of patients with FDD shows diffuse brain atrophy with a particularly severe involvement of the cerebellum, cerebral cortex and white matter, as well as the presence of very thin and almost demyelinated cranial nerves, and widespread amyloid angiopathy in the small blood vessels and capillaries of the cerebrum, choroid plexus, cerebellum, spinal cord and retina. In FDD, parenchymal compact plaques are consistently absent, whereas neurofibrillary tangles (NFTs) are the major histological finding in the hippocampus [3], [12].
BRI2 is physiologically cleaved, at the C-terminus, by furin, a calcium-dependent serine endoprotease, producing a 23 aa soluble C-terminal fragment. FBD and FDD are due to mutations in the BRI2 gene located on chromosome 13q14 [2], [8]. In FBD, a single base substitution at the stop codon of BRI2 generates a longer open reading frame, resulting in a larger, 277 aa precursor (BRI2-ABri, compared to the 266 aa long normal protein) [2]. In FDD, a decamer duplication in the 3′ region of the BRI2 gene, right before the stop codon, leads as well to the production of a longer, 277 aa protein (BRI2-ADan) [3]. The genetic defect is different, but the outcome is the same: the generation of a longer 34 aa C-terminal fragment, ABri in FBD and ADan in FDD, which accumulates as amyloid [2].
Several neuropathological features are common to FBD, FDD and Alzheimer's disease (AD): amyloid deposition and neurodegeneration in the central nervous system (CNS), accumulation of complement proteins and their pro-inflammatory activation products, including iC3b, C4d, Bb, and C5b-9, neurofibrillary pathology and hyperphosphorylated tau [12], [13], [14]. Furthermore, in FDD, Alzheimer's Aβ co-deposits with ADan, mainly in vascular and perivascular amyloid lesions and less in parenchymal preamyloid deposits [12], while compact plaques are not frequent [2], [12], [15].
More recently, a new link between BRI2 and Aβ has been found. It has been shown that BRI2 binds APP in a region comprising the extracellular juxtamembrane domains of both proteins, in a cis fashion [5], [6], [7]. This interaction leads to interference on the physiological processing of APP: BRI2 restricts the docking of γ-secretase to APP and the access of α- and β-secretases to their cleavage site on APP itself. The overall result of this interaction is the reduction of the amyloidogenic processing of APP, without a direct inhibition of the general activity of the secretases [5]. BRI2 maturation is required for this function. In fact, only mBRI2 (and not the immature precursor) binds mature APP and inhibits its processing on the plasma membrane and in endocytic compartments [16]. Matsuda et al have further shown that BRI3, a member of the BRI family, binds and inhibit the action of α- and β-secretases on APP [17]. Others have argued that the BRI2-23 wild type C-terminal peptide, released from BRI2 by furin/furin-like cleavage, can inhibit Aβ aggregation in vitro [18].
The interaction between these two amyloidogenic proteins can be of interest, especially for the design of new therapeutic strategies for AD and FBD/FDD. To this end, the processing of the BRI2 precursor, both in the wild type allele and its mutant form must be understood. Transgenic mice bearing the British or Danish mutations have been generated, and are under investigation as models of dementias and cerebral amyloid angiopathies [19], [20]. A transgenic FDD model which over-expresses the Danish mutant form of BRI2 shows amyloid deposition in the walls of blood vessels of the cerebrum and cerebellum, parenchymal amyloid deposition and reactive gliosis, ADan amyloidosis and some signs of cerebellar ataxia [20]. Transgenic over-expression of human mutant genes has been extensively used to generate mouse models for human neurodegenerative disorders due to the need of over-expressing enough amount of the mutant protein in order to observe a phenotype. It is however possible that some disorders may, at least partially, be due to loss of function rather than a gain of toxic function due to mutations. If this were the case, a transgenic approach would be counterproductive. Therefore, we have reasoned that models reproducing the genetics of human pathologies may be worth studying since they may unveil important disease mechanisms that a transgenic approach may miss or, possibly, hide. Thus, we have generated a genetically faithful model of FDD by introducing the pathogenic human mutation in the mouse BRI2 gene by a knock-in technology. Here, we describe the generation of this Knock-In murine model of FDD (FDDKI) and its initial characterization.
Materials and Methods
Ethics Statement
Mice were maintained on a C57BL/6 background for several generations (at least 15). Mice were handled according to the Ethical Guidelines for Treatment of Laboratory Animals of Albert Einstein College of Medicine. The procedures were described and approved in animal protocol number 20040707.
FDDKI Mice Construction
To clone the ADan mutation allele, a fragment obtained form BAC (containing mouse BRI2 genome) was amplified by PCR with the following primers:
Fw: 5′-cccAAGCTTtttttttttttttttaaagacaac-3′; Rev: 5′-gggAAGCTTgaagtggtcagcagggag-3′, obtaining a 7536 bp fragment, flanked by 2 HindIII sites, and comprising part of Intron 2 to a 3′UTR region, up to 3850 bp from the BRI2 stop codon. Such a fragment was cloned into a pBS vector, into HindIII sites (pBS-Ex3-6 HindIII), and used as a template for subsequent cloning.
A 340 bp fragment containing the ADan (duplication 786–795 of cDNA) mutation, plus a humanizing substitution a→g (acc→gcc = Threonine→Alanine) at the 12th codon of exon 6, was obtained by serial PCRs, using pBS-Ex3-6 HindIII as a template. The final external primers contained restriction sites for HincII (5′) and EcoRI (3′). The mutated fragment (HincII-ADan-EcoRI) was inserted into the SmaI and EcoRI sites of a pBS vector yielding pBS-ADan0.3.
Subsequently, 2 EcoRI-EcoRI and EcoRI-XhoI fragments (2.6 kbp total) were cut from the template and added 3′, in tandem, to pBS-ADan0.3, generating pBS-ADan2.9, containing the Right Arm (RA) fragment of the construct.
A NotI-XhoI fragment from pBS-ADan2.9, comprising the RA, was inserted into NotI and SacII sites of a Soriano PGK-Neo-dTA vector (blunt-ended by the SacII and XhoI digestion followed by treatment of T4-exonuclease and Klenow polymerase respectively) at 5′of the Neo cassette. The XhoI site was reconstituted upon ligation of the fragment into the vector.
The construct's Left Arm was extracted from the template with EcoRI and HincII, generating a 1.1 kbp into Intron 5 up to the corresponding HincII site, 5′ of Exon 6 (start of Right Arm). The fragment was thus inserted into the PGK-Neo-dTA vector at 3′ of the Neo cassette and 5′ of the dTA cassette.
The resulting construct was thus:
---Right Arm-LoxP1-PGK-Neomycin cassette-LoxP2-Left Arm-dTA cassette---.
The resulting construct was linearized with SalI and purified prior to injection in ES cells strain 129 by electroporation. ES culture was performed on feeder layer, and further electroporation and handling was also performed according to the methodology employed at Dept of Cell Biology, Albert Einstein College of Medicine, and according to Wakayama et al. [21]. In particular, after electroporation, ES cells were re-plated in 55 cm2 dishes and let grow until visible clones would appear. Clones were then picked and transferred to 96 well plates in triplicates. Triplicates were either screened by PCR or frozen for subsequent use and further analysis.
Homologous recombinants were selected with G418 (200 µg/ml) and dTA exclusion.
Injection of the two Danish ES cell clones into C57BL/6J blastocysts was performed at the Albert Einstein College of Medicine gene-targeting facility, according to the current facility protocol (http://www.aecom.yu.edu/home/SharedFacilities/ViewFacility.asp?ID=30).
PCR Analysis
The PCR screening was performed using the Expand Long Template PCR System (Roche-applied-Science) with Betaine, according to the manufacturer instructions. PCR analysis of recombinant ES cells and mice was conducted with the following primers and digestion strategies to identify the correct recombinant clones and strains:
ES cells:
Left arm:
Genomic primer: 5′-CAGTCCTACCTTATCCATGAGCAC-3′
Neo cassette primer: 5′- CTTCCTCGTGCTTTACGGTATC-3′
Product: 1637 bp
Right arm:
Genomic primer: 5′- AGTCTGTATCTCACTACGGCATCC-3′
Neo cassette primer: 5′- TGCACGAGACTAGTGAGACGTG-3′
Product: 3403 bp
Amplification of the inserted construct and digestion:
Genomic upstream primer: 5′-CAGTCCTACCTTATCCATGAGCAC-3′
Genomic downstream primer: 5′-AGTCTGTATCTCACTACGGCATCC-3′
Product: 4300 bp = construct without Neo cassette; NotI digestion→2900 bp + 1300 bp
Product: 6109 bp = construct with Neo cassette; NotI digestion→3100 bp + 2900 bp
Whole construct
Set 1
Fw primer: 5′-ATGGCACCACCCACAATAGG-3′
Rev primer: 5′-CCTAGCAACTGGTAACAGTGC-3′
Product: 2027 bp with Neo cassette
Product: 332 bp without Neo cassette
Product: 194 bp wild type
Fw primer: 5′-ATGGCACCACCCACAATAGG-3′
Rev primer: 5′-ggtaagctctaaggagaagcg-3′
Product: 2500 bp with Neo cassette
Product: 849 bp without Neo cassette
Product: 704 bp wild type
Due to the specific sequences present in the amplified PCR product, no digestion of the amplicon could be performed on the mice PCR analysis. PCR products were thus sequenced to ascertain that the targeted sequence was correctly inserted in the genomic DNA. The sequence has been deposited in GenBank (accession number GQ424832).
Southern Blot Analysis
Twenty µg of genomic DNA was digested with BamHI overnight, run on a 1% TAE agarose gel and transferred on a Hybond-N+ membrane (Amersham).
The probe was prepared by PCR from a BAC clone (RP24) with the following primers:
-Left arm:
Fw: 5′- GACAGAGGTTCTGCCCTCAG-3′
Rev: 5′-ACCGAGTCGTAGGACAGTG-3′
Probe size: 547 bp
-Right arm:
Fw: 5′- CTGTGCTGCCTGACACTACTTC-3′
Rev: 5′- TCTGTCCATACTCCCTGTCCTT-3′
Probe size: 515 bp.
One µg of PCR probe was labelled with 5 µL of 32P-dCTP (3000 Ci/mmol, ICN) and purified through a Push Column (Stratagene) according to the manufacturer's protocol. Membranes, containing the cleaved genomic DNA, were hybridized at 65°C and subsequently washed 4 times in SSC buffer (Sigma). Film was exposed to the hybridised membranes at –80°C and then developed.
General Pathology Analysis
Wild-type and FDDKI animals were studied at 18 months of age. After anesthesia, animals were perfusion-fixed with 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.2) (Sigma), after which brains and organs were removed, embedded in paraffin and sectioned. Eight µm sections were cut and mounted on poly-l-lysine-coated slides. After deparaffinization in Xylene ad rehydration, sections were stained with hematoxylin and eosin (H&E), Congo red standard methods. Mineralization was visualized in H&E sections as a deep basophilic amorphous and/or granular material.
Brain Histology and Immunohistochemistry
Wild-type, FDDKI and FDD-Tg animals were studied at 18 months of age. After anesthesia, animals were perfusion-fixed with 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.2) (Sigma), after which brains were removed, embedded in paraffin and sectioned. Eight µm sections were cut and mounted on poly-l-lysine-coated slides. Sections were stained with H&E, Congo red and Thioflavin S (ThS) methods. Immunohistochemical stainings were carried out as described [20] with following primary antibodies: glial fibrillary acidic protein (GFAP) (Dako, Carpinteria, CA); ubiquitin (Dako); amyloid β protein (Aβ) clone 4G8 (SIGNET, Dedham, MA); and ADan amyloid peptide (Ab 1700) [20]. Immunostaining was visualized using the avidin-biotin system (Vectastain; Vector Laboratories, Burlingame, CA) and 3,3′-diaminobenzidine (Sigma) as the chromogen. The sections were counterstained with cresyl violet or H&E. ADan transgenic (FDD-Tg) mice were described elsewhere [20].
Neuronal Cultures
Neuronal cultures of FDDKI mice, and wt littermates, were performed as described previously [22], [23] from E16-17 fetuses.
Western Blot
Western blot was conducted on protein extracts from brains or neuronal culture of FDDKI mice and wt littermates, as described previously [5] for cells lysates and brain tissue. Briefly, primary neuronal cells were lysed in Hepes-Triton buffer (20 mM Hepes/NaOH pH 7.4, 1 mM EDTA, 150 mM NaCl, 0.5% Triton X-100, plus protease inhibitors (PIs)) on ice for 30 min. The lysates were cleared by spinning at 20,000 g for 10 min. Equal amount of proteins of cleared lysates were loaded on SDS-PAGE and transfered onto nitrocellulose membranes; APP and BRI2 (wild type and Danish mutants) were detected by 22C11 (Chemicon, MAB348) or BRI2 antibody, generously provided by Dr. Haruhiko Akiyama, respectively [24].
Generation and Characterization of FDDKI Mice
The targeting strategy for the generation of the Danish mutant FDDKI mice entailed the replacement of the BRI2 exon 6 with a mutated exon 6 carrying the FDD mutation. We generated a targeting vector for the introduction of FDD BRI2 mutation. The vector used the floxed PGK-neo selection cassette and contains a 5′ homologous region and the negative selection cassette, PGK-dt. The 3′ homologous region introduced the FDD mutation and a BamHI site into the BRI2 mouse gene. The 10 nucleotide insertion found in patients with FDD was introduced before the stop codon. In order to humanize the mouse C-terminal region of BRI2, an alanine (A) was substituted for threonine (T) at codon 250 (Fig. 1a) of the murine BRI2 gene.
1a. Schematic representation of the construct that was injected in 129 ES cells, showing site of ADan mutation on Exon 6, primer sites, site of Southern Blot probe, LoxP, pgk-dt and pgk-Neo sites. The bottom graphics depict the construct with and without the pgk-Neo cassette that has been removed by means of Cre recombinase. 1b. Examples of 2 ES clones positive for the homologous recombination of the mutated allele: left arm (a–c: 1.67 kb) and right arm (b–d: 3.4 kb) PCRs are shown. 1c. Southern Blot showing a shift from the 11.9 kb of the wild type genome to the 8.9 kb band of the FDD KI mice, due to the insertion of a new BamHI site. KI mice show both bands, indicating heterozygosity.
The linearized targeting vector was transfected into 129 ES cells by electroporation. In the presence of the positive selection drug, G418, only those clones in which the PGK-neo selection cassette was integrated and the PGK-dt cassette was removed by homologous recombination would survive. ES cell clones carrying the targeting vector by random, non-homologous integration, were eliminated due to expression of diphtheria toxin.
After selection in G418-containing medium, ES cell clones carrying the proper homologous recombination and the tADan (targeted ADan) allele were identified by PCR for 5′ region (i.e. Left Arm: if homologous recombination had occurred these primers would amplify a product of 1.67 kb) and for the 3′ region (i.e. Right Arm: if homologous recombination had occurred these primers would amplify a product of 3.4 kb). Out of the ∼600 screened ES clones, we found three clones in which the Danish mutation was inserted in one of the BRI2 alleles. Representative positive (BRI2tADan/+344 and BRI2tADan/+339) and negative (BRI2tADan/+342) ES clones are shown in Fig. 1b. Also, PCR amplification and digestion, as specified in the Material and Methods section, was used to check the proper insertion of the construct in the genomic DNA and the removal of the Neo cassette (not shown).
The occurrence of homologous recombination was confirmed by sequencing the PCR products and by performing Southern blot analysis (Fig. 1c). DNA derived from individual BRI2tADan/+ ES clones was digested with BamHI, gel separated, blotted into a nylon membrane and hybridized with either the 5′ or the 3′probe. The 5′ probe hybridizes with a ∼11.9 kb fragment derived from the wild-type locus. Homologous recombination at the 5′ homologous region yields a ∼8.9 kb fragment upon BamHI digestion due to the introduction of the BamHI site and the PGK-neo selection cassette. ES clones BRI2ADan/+344 and BRI2ADan/+339 carry a wild type allele (11.9 kb) and a recombined allele (8.9 kb). Of note, the 11.9 kb and 8.9 kb bands had a similar intensity, indicating that 50% of the BRI2 alleles are wild type and 50% are recombined, and proving that the ES cells we have selected are a clonal populations. Similar results were obtained when homologous recombination at the 3′ site was assessed. In this case the 3′ probe detected a wild-type ∼11.9 kb fragment and a recombinant 4.7 kb fragment, due to the introduction of the BamHI site.
These two Danish ES cell clones (129, agouti coat colour), carrying the correct site-specific homologous recombination, were injected into C57BL/6J blastocysts (black coat colour) at the Albert Einstein College of Medicine gene-targeting facility. The resulting chimeras with a high proportion of agouti coat colour (i.e. with a high relative contribution from the injected ES cells) were backcrossed to C57BL/6J mice to obtain heterozygous BRI2ADan/wt, which were identified by PCR and Southern analysis as described above (not shown) using tail DNA. Heterozygous mice were crossed to Meu40-Cre mice to obtain Meu40/BRI2tADan/wt animals. Cre is a bacteriophage P1-encoded recombinase that catalyzes site-specific recombination between two 34 bp loxP recognition sites, resulting in the excision of the intervening DNA sequences. The resulting mouse has been named FDDKI.
General Characteristics and Pathology of the FDDKI Mice
Mice presented with no growth abnormalities, thrived at appropriate age, as their wild type littermates. Up to age 18 months, no susceptibility to infections was noted. The animals are active and alert.
Before the pathology examination, at 18 months of age, they presented in good body condition with adequate body fat, with no discharges or secretions from nostrils, conjunctiva, aural, urogenital or anal openings. Some mice also presented heart valve melanosis, which is a common finding in aged mice of various strains. Other sporadic findings, which are common in aged mice, were small foci of hepatocellular necrosis, vacuolation of and degeneration/regeneration of kidney tubular epithelial cells, mineralization in the kidney tubules in the pelvis and cortico-medullary junction (not shown). Overall, these mice have lesions that are commonly found in older mice and are considered age-related, spontaneous lesions. All of the lesions found were considered to be within the normal limits for age-related lesions. One of the animals presented with a uterine histiocytic sarcoma and hydronephrosis, both of which are found in older, untreated mice at low incidence.
A few animals presented a small amount of dark basophilic material deposited on either side of the lateral thalamus consistent with mineralization (incidental, Fig. 2d), which is a common finding in aged mice of various strains, and the deposits develop along the basement membranes of the vasculature and may contain calcium and phosphorus. The finding of dark brown pigment along the meninges of the olfactory bulbs is consistent with melanosis (incidental, Fig. 2a), which is also a common finding in mice in dark pigment mouse strains such as C57BL/6 or having that strain the their background. All the brain tissue and the spinal cord were stained with Congo red for potential amyloid deposition. All of the stained tissue was negative for amyloid (not shown). Samples of H&E staining of cerebral cortex, hippocampus, cerebellum, thalamus and spinal cord are shown in Fig. 2a–e.
H&E staining of 18 month old FDDKI mice showing signs of melanosis in the meninges of olfactory bulb (a) and signs of mineralization in the thalamus (d). The remaining parenchyma of brain cortex and hippocampus (b), cerebellum (c) and spinal cord (e) show normal cellular structures.
Brain sections of 18-month-old mice were examined for FDD-related pathology, particularly for amyloid deposition in brain parenchyma and vessel walls. For comparison, we included sections of an age-matched female FDD-Tg mouse, which expresses the Danish mutant form of human BRI2 under the mouse prion protein promoter [20]. H&E staining of Knock In mice (Fig. 3c, 4a) showed no significant loss of neurons or noticeable malformations, and this observation was further confirmed when mutant sections were compared with those of wild type littermates (Fig. 3a, 4d). Immunostaining using anti-GFAP (Fig. 3b, d) and anti-ubiquitin (not shown) antibodies did not reveal significant differences with wild type mice in levels of activated astrocytes or presence of ubiquitinated material, respectively. Staining using Congo red or ThS did not show the presence of amyloid deposits (Fig. 4b). Amyloid deposition in FDD-Tg mice is particularly strong in the cerebellum (Fig. 4e) but not in Knock In mice (Fig. 4b) and only FDD-Tg mice showed immunoreactivity using Ab 1700, specific for the ADan amyloid peptide [20] (Fig. 4f versus 4c). Immunoreactivity was not seen in non-transgenic littermates, FDD-Tg and Knock In mice using antibody 4G8 (not shown). These results indicated that the brain of Knock In mice maintained normal morphology of aged mice and were free of amyloid deposits at 18 months of age.
Sections from a wild type (a, b) and a homozygous knock-in mouse (c, d) were stained with H&E (a, c). Sections were also studied by immunohistochemistry using anti-GFAP (b, d). No significant differences were observed between wild-type and Knock In mice. Scale bars: a–d, 200 µm.
Sections from a wild-type (d), homozygous knock-in mouse (a–c), and FDD-Tg mouse (e, f) were stained with H&E (a) and ThS (b, d, e). Sections were also studied by immunohistochemistry using Ab 1700, specific for the ADan amyloid peptide (c, f). No significant differences were observed between wild-type and knock-in mice. Staining with ThS and Ab 1700 shows the presence of ADan amyloid only in FDD-Tg mice (e–f). Scale bars: a–e, 100 µm; f, 200 µm.
Biochemical Analysis of BRI2-ADan Expression
To determine whether the mutant proteins are expressed in vivo, we made protein extracts form neuronal cultures of wt, BRI2ADan/wt and BRI2ADan/ADan mice. Western blot analysis of these samples shows three phenomenona. 1) A common, major band of about 32 kDa is readily visible in all three samples. This band corresponds to the post-furin cleavage mature BRI2 (mBRI2) polypeptide because mBRI2 is identical regardless of whether it is derived from the wild type or the mutant precursor protein. 2) A much less abundant protein of ∼33 kDa which is visible in the wild type and BRI2ADan/wt sample but not the BRI2ADan/ADan lysate. This band corresponds to the wild type, immature BRI2 precursor (imBRI2). The absence of this band in the BRI2ADan/ADan neurons is attributable to the fact that these cells have both BRI2 alleles mutated. 3) As expected, the BRI2ADan/wt sample expresses all three BRI2 polypeptides, i.e. wild type imBRI2, BRI2-ADan precursor and mBRI2. It is worth noting that, while immature murine BRI2 (imBRI2) is barely detectable, BRI2-ADan is more markedly expressed (see BRI2ADan/ADan neurons, Figure 5) in this specific experiment. Whether this is related to the artificial absence of the wild type protein (this does not happen in FDD since the patience have only one mutated allele) or reflects some interesting biology of BRI2-ADan, remains to be determined. Notably, APP expression and maturation are comparable in FDDKI and wt mice (Fig. 5, lower panel).
Lysates from E16-17 neuronal cultures of wt, ADan +/− and ADan +/+ mice were blotted with antibodies against BRI2 and APP. Upper panel shows impaired maturation of BRI2-ADan compared to wild type; lower panel show normal pattern of expression and maturation of APP.
In this manuscript we present an initial characterization of a Knock In model for a human neurodegenerative disorder, FDD. This Knock In model is strategically different from the conventional genetic approach to cerebral amyloidosis. Traditionally, mouse models for human dementias are based on a transgenic approach in which human mutant proteins that cause familial forms of dementia are over-expressed under the control of brain-specific promoters [25], [26], [27]. This approach has both limitations and advantages. The potential limitations are linked to the fact that these models are genetically incongruous with the human diseases. These dementias have an autosomal dominant way of transmission and affected subjects have a wild type and a mutant allele. On the contrary, the commonly used animal models over-express, from an artificial promoter, several copies of a mini-gene coding the mutant protein. The mutant gene is therefore expressed in a spatio-temporal manner different from the natural alleles and in cells with two copies of wild type alleles. However, transgenic approaches have successfully reproduced cerebral amyloidosis in mice, and the reproduction of those lesions is one of the main parameters used to endorse an animal model as representative of the human disease. The opposite is true for a Knock In approach. In this genetic paradigm, the pathogenic human mutation is inserted in the mouse allele. Thus, the Knock In mouse is genetically faithful to the human pathology. However, the Knock In model may not successfully reproduce the amyloid lesion in the mouse. The model that we present here is a clear demonstration of the above-mentioned assumptions. A comparison of the FDD-KI model that we have created with the FDD transgenic model generated by Vidal et al [20] illustrates the superior power of a transgenic approach as far as the reproduction of amyloidogenic lesions are concerned.
In spite of this limitation, we believe that there is some merit to a Knock In approach. Human familial dementias may have a pathogenic component due to loss of function caused by the mutation [28], [29], [30], [31], [32]. This component may participate to the pathogenic process together with a gain of toxic function due to amyloidosis. A loss of function phenotype would be hidden by a transgenic approach for two reasons: 1) the two endogenous wild type alleles can support synthesis of sufficient amounts of functional wild type protein: 2) the over-expression of a partial loss of function mutant protein may augment rather then decrease that function, as may happen in the disease. It would be erroneous to assume that the absence of obvious neuropathological lesions (such as plaques, NFTs and/or neuronal cell loss) indicate absence of clinical pathology. Functional deficits may underlie the very first clinical manifestations of neurodegenerative diseases, and memory deficits may be caused not by gross anatomical changes but by subtle dysfunctions of the neuronal network. In this framework, determining the absence of neuropathological lesions in FDDKI mice is intrinsically important. The behavioral characterization of this mouse model will test this hypothesis. In addition, the FDDKI mouse will be useful to clarify the change of trafficking or processing of Danish mutant of BRI2 protein without potential artifacts due to over-expression in transgenic models. Finally, the FDDKI mouse will be instrumental in studying the interaction between mutant BRI2 and APP in vivo and how the Danish mutation affects the inhibitory role of BRI2 on APP processing in vivo.
We thank Erhan Ma for the help in the genotyping and PCR procedures on the ADan KI mice; we thank Dr. Susan Newbigging MSc, DVM, DVSc at the Toronto Centre for Phenogenomics, 25 Orde Street, Toronto ON M5T 3H7; we thank Dr. Bernardino Ghetti for the helpful discussion.
Author Contributions
Conceived and designed the experiments: LD. Performed the experiments: LG SM RV. Analyzed the data: LG RV LD. Wrote the paper: RV LD.
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- 3. Vidal R, Revesz T, Rostagno A, Kim E, Holton JL, et al. (2000) A decamer duplication in the 3′ region of the BRI gene originates an amyloid peptide that is associated with dementia in a Danish kindred. Proc Natl Acad Sci U S A 97: 4920–4925.R. VidalT. ReveszA. RostagnoE. KimJL Holton2000A decamer duplication in the 3′ region of the BRI gene originates an amyloid peptide that is associated with dementia in a Danish kindred.Proc Natl Acad Sci U S A9749204925
- 4. Kim SH, Wang R, Gordon DJ, Bass J, Steiner DF, et al. (1999) Furin mediates enhanced production of fibrillogenic ABri peptides in familial British dementia. Nat Neurosci 2: 984–988.SH KimR. WangDJ GordonJ. BassDF Steiner1999Furin mediates enhanced production of fibrillogenic ABri peptides in familial British dementia.Nat Neurosci2984988
- 5. Matsuda S, Giliberto L, Matsuda Y, McGowan EM, D'Adamio L (2008) BRI2 inhibits amyloid beta-peptide precursor protein processing by interfering with the docking of secretases to the substrate. J Neurosci 28: 8668–8676.S. MatsudaL. GilibertoY. MatsudaEM McGowanL. D'Adamio2008BRI2 inhibits amyloid beta-peptide precursor protein processing by interfering with the docking of secretases to the substrate.J Neurosci2886688676
- 6. Matsuda S, Giliberto L, Matsuda Y, Davies P, McGowan E, et al. (2005) The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. J Biol Chem 280: 28912–28916.S. MatsudaL. GilibertoY. MatsudaP. DaviesE. McGowan2005The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production.J Biol Chem2802891228916
- 7. Fotinopoulou A, Tsachaki M, Vlavaki M, Poulopoulos A, Rostagno A, et al. (2005) BRI2 interacts with amyloid precursor protein (APP) and regulates amyloid beta (Abeta) production. J Biol Chem 280: 30768–30772.A. FotinopoulouM. TsachakiM. VlavakiA. PoulopoulosA. Rostagno2005BRI2 interacts with amyloid precursor protein (APP) and regulates amyloid beta (Abeta) production.J Biol Chem2803076830772
- 8. Pittois K, Wauters J, Bossuyt P, Deleersnijder W, Merregaert J (1999) Genomic organization and chromosomal localization of the Itm2a gene. Mamm Genome 10: 54–56.K. PittoisJ. WautersP. BossuytW. DeleersnijderJ. Merregaert1999Genomic organization and chromosomal localization of the Itm2a gene.Mamm Genome105456
- 9. Vidal R, Calero M, Revesz T, Plant G, Ghiso J, et al. (2001) Sequence, genomic structure and tissue expression of Human BRI3, a member of the BRI gene family. Gene 266: 95–102.R. VidalM. CaleroT. ReveszG. PlantJ. Ghiso2001Sequence, genomic structure and tissue expression of Human BRI3, a member of the BRI gene family.Gene26695102
- 10. Sanchez-Pulido L, Devos D, Valencia A (2002) BRICHOS: a conserved domain in proteins associated with dementia, respiratory distress and cancer. Trends Biochem Sci 27: 329–332.L. Sanchez-PulidoD. DevosA. Valencia2002BRICHOS: a conserved domain in proteins associated with dementia, respiratory distress and cancer.Trends Biochem Sci27329332
- 11. Plant GT, Revesz T, Barnard RO, Harding AE, Gautier-Smith PC (1990) Familial cerebral amyloid angiopathy with nonneuritic amyloid plaque formation. Brain 113 (Pt 3): 721–747.GT PlantT. ReveszRO BarnardAE HardingPC Gautier-Smith1990Familial cerebral amyloid angiopathy with nonneuritic amyloid plaque formation.Brain113 (Pt 3)721747
- 12. Holton JL, Lashley T, Ghiso J, Braendgaard H, Vidal R, et al. (2002) Familial Danish dementia: a novel form of cerebral amyloidosis associated with deposition of both amyloid-Dan and amyloid-beta. J Neuropathol Exp Neurol 61: 254–267.JL HoltonT. LashleyJ. GhisoH. BraendgaardR. Vidal2002Familial Danish dementia: a novel form of cerebral amyloidosis associated with deposition of both amyloid-Dan and amyloid-beta.J Neuropathol Exp Neurol61254267
- 13. Holton JL, Ghiso J, Lashley T, Rostagno A, Guerin CJ, et al. (2001) Regional distribution of amyloid-Bri deposition and its association with neurofibrillary degeneration in familial British dementia. Am J Pathol 158: 515–526.JL HoltonJ. GhisoT. LashleyA. RostagnoCJ Guerin2001Regional distribution of amyloid-Bri deposition and its association with neurofibrillary degeneration in familial British dementia.Am J Pathol158515526
- 14. Rostagno A, Revesz T, Lashley T, Tomidokoro Y, Magnotti L, et al. (2002) Complement activation in chromosome 13 dementias. Similarities with Alzheimer's disease. J Biol Chem 277: 49782–49790.A. RostagnoT. ReveszT. LashleyY. TomidokoroL. Magnotti2002Complement activation in chromosome 13 dementias. Similarities with Alzheimer's disease.J Biol Chem2774978249790
- 15. Tomidokoro Y, Lashley T, Rostagno A, Neubert TA, Bojsen-Moller M, et al. (2005) Familial Danish dementia: co-existence of Danish and Alzheimer amyloid subunits (ADan AND A{beta}) in the absence of compact plaques. J Biol Chem 280: 36883–36894.Y. TomidokoroT. LashleyA. RostagnoTA NeubertM. Bojsen-Moller2005Familial Danish dementia: co-existence of Danish and Alzheimer amyloid subunits (ADan AND A{beta}) in the absence of compact plaques.J Biol Chem2803688336894
- 16. Matsuda S, Matsuda Y, Snapp EL, D'Adamio L (2009) Maturation of BRI2 generates a specific inhibitor that reduces APP processing at the plasma membrane and in endocytic vesicles. Neurobiol Aging. S. MatsudaY. MatsudaEL SnappL. D'Adamio2009Maturation of BRI2 generates a specific inhibitor that reduces APP processing at the plasma membrane and in endocytic vesicles.Neurobiol Aging
- 17. Matsuda S, Matsuda Y, D'Adamio L (2009) BRI3 inhibits amyloid precursor protein processing in a mechanistically distinct manner from its homologue dementia gene BRI2. J Biol Chem 284: 15815–15825.S. MatsudaY. MatsudaL. D'Adamio2009BRI3 inhibits amyloid precursor protein processing in a mechanistically distinct manner from its homologue dementia gene BRI2.J Biol Chem2841581515825
- 18. Kim J, Miller VM, Levites Y, West KJ, Zwizinski CW, et al. (2008) BRI2 (ITM2b) inhibits Abeta deposition in vivo. J Neurosci 28: 6030–6036.J. KimVM MillerY. LevitesKJ WestCW Zwizinski2008BRI2 (ITM2b) inhibits Abeta deposition in vivo.J Neurosci2860306036
- 19. Pickford F, Coomaraswamy J, Jucker M, McGowan E (2006) Modeling familial British dementia in transgenic mice. Brain Pathol 16: 80–85.F. PickfordJ. CoomaraswamyM. JuckerE. McGowan2006Modeling familial British dementia in transgenic mice.Brain Pathol168085
- 20. Vidal R, Barbeito AG, Miravalle L, Ghetti B (2009) Cerebral amyloid angiopathy and parenchymal amyloid deposition in transgenic mice expressing the Danish mutant form of human BRI2. Brain Pathol 19: 58–68.R. VidalAG BarbeitoL. MiravalleB. Ghetti2009Cerebral amyloid angiopathy and parenchymal amyloid deposition in transgenic mice expressing the Danish mutant form of human BRI2.Brain Pathol195868
- 21. Wakayama T, Rodriguez I, Perry AC, Yanagimachi R, Mombaerts P (1999) Mice cloned from embryonic stem cells. Proc Natl Acad Sci U S A 96: 14984–14989.T. WakayamaI. RodriguezAC PerryR. YanagimachiP. Mombaerts1999Mice cloned from embryonic stem cells.Proc Natl Acad Sci U S A961498414989
- 22. Giliberto L, Zhou D, Weldon R, Tamagno E, De Luca P, et al. (2008) Evidence that the Amyloid beta Precursor Protein-intracellular domain lowers the stress threshold of neurons and has a "regulated" transcriptional role. Mol Neurodegener 3: 12.L. GilibertoD. ZhouR. WeldonE. TamagnoP. De Luca2008Evidence that the Amyloid beta Precursor Protein-intracellular domain lowers the stress threshold of neurons and has a "regulated" transcriptional role.Mol Neurodegener312
- 23. Giliberto L, Borghi R, Piccini A, Mangerini R, Sorbi S, et al. (2009) Mutant presenilin 1 increases the expression and activity of BACE1. J Biol Chem 284: 9027–9038.L. GilibertoR. BorghiA. PicciniR. MangeriniS. Sorbi2009Mutant presenilin 1 increases the expression and activity of BACE1.J Biol Chem28490279038
- 24. Akiyama H, Kondo H, Arai T, Ikeda K, Kato M, et al. (2004) Expression of BRI, the normal precursor of the amyloid protein of familial British dementia, in human brain. Acta Neuropathol (Berl) 107: 53–58.H. AkiyamaH. KondoT. AraiK. IkedaM. Kato2004Expression of BRI, the normal precursor of the amyloid protein of familial British dementia, in human brain.Acta Neuropathol (Berl)1075358
- 25. LaFerla FM, Oddo S (2005) Alzheimer's disease: Abeta, tau and synaptic dysfunction. Trends Mol Med 11: 170–176.FM LaFerlaS. Oddo2005Alzheimer's disease: Abeta, tau and synaptic dysfunction.Trends Mol Med11170176
- 26. McGowan E, Eriksen J, Hutton M (2006) A decade of modeling Alzheimer's disease in transgenic mice. Trends Genet 22: 281–289.E. McGowanJ. EriksenM. Hutton2006A decade of modeling Alzheimer's disease in transgenic mice.Trends Genet22281289
- 27. Price DL, Wong PC, Markowska AL, Lee MK, Thinakaren G, et al. (2000) The value of transgenic models for the study of neurodegenerative diseases. Ann N Y Acad Sci 920: 179–191.DL PricePC WongAL MarkowskaMK LeeG. Thinakaren2000The value of transgenic models for the study of neurodegenerative diseases.Ann N Y Acad Sci920179191
- 28. Bentahir M, Nyabi O, Verhamme J, Tolia A, Horre K, et al. (2006) Presenilin clinical mutations can affect gamma-secretase activity by different mechanisms. J Neurochem 96: 732–742.M. BentahirO. NyabiJ. VerhammeA. ToliaK. Horre2006Presenilin clinical mutations can affect gamma-secretase activity by different mechanisms.J Neurochem96732742
- 29. De Strooper B (2007) Loss-of-function presenilin mutations in Alzheimer disease. Talking Point on the role of presenilin mutations in Alzheimer disease. EMBO Rep 8: 141–146.B. De Strooper2007Loss-of-function presenilin mutations in Alzheimer disease. Talking Point on the role of presenilin mutations in Alzheimer disease.EMBO Rep8141146
- 30. Wolfe MS (2007) When loss is gain: reduced presenilin proteolytic function leads to increased Abeta42/Abeta40. Talking Point on the role of presenilin mutations in Alzheimer disease. EMBO Rep 8: 136–140.MS Wolfe2007When loss is gain: reduced presenilin proteolytic function leads to increased Abeta42/Abeta40. Talking Point on the role of presenilin mutations in Alzheimer disease.EMBO Rep8136140
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text:
Wet 11 in its latest firmware revision on the main configuration page has a
checkbox called "clone MAC address"
If disabled it will start substituting its MAC for clients which is behind
-----Original Message-----
From: Dinesh Nair [mailto:dinesh at alphaque dot com]
Sent: 2004 m. birþelio 7 d. 07:57
To: Michael Mee
Cc: m0n0wall at lists dot m0n0 dot ch
Subject: Re: [m0n0wall] captive portal thru wet-11 strangeness
On Sun, 6 Jun 2004, Michael Mee wrote:
> computers to a Wet-11 wireless to ethernet bridge, the captive portal
> blocks them, but also doesn't display the page. I also get the log
> Jun 6 10:08:24 m0n0-home /kernel: arp: 10.0.1.175 moved from
> 00:90:4b:b1:58:6b to 00:06:25:12:48:47 on wi0
> where 00:90:4b:b1:8:6b is the MAC of the computer behind the Wet-11
> and
> 00:06:25:12:48:47 is the Wet-11 itself. Not sure what this message
> means or why it appears...
looks like the Wet-11 is doing proxy arp for the clients behind it. since
the captive portal uses the mac address to determine if a box should be let
through or not, and at the same time checks if the mac address and the ip
address are bound, you're seeing the above behaviour. the log message is
m0n0wall telling you that it has detected that 10.0.1.175 was initially seen
tied to one mac address (your client), and then changed to another (wet-11).
this is a clear indication that the wet-11 is proxy arping for clients
behind it.
could you turn of proxy arps on the wet-11 ? alternatively, you could not
use pass-through macs, but use the allowed ip outgoing instead. place in the
ip addresses of the clients who're allowed to bypass the captive portal sign
on page, and they'll go thru. allowed ips are not subject to mac address
checking/filtering on the captive portal, though the firewall ruleset will
act on them.
Regards, /\_/\ "All dogs go to heaven."
dinesh at alphaque dot com (0 0) http://www.alphaque.com/
| for a in past present future; do |
| for b in clients employers associates relatives neighbours pets; do |
| echo "The opinions here in no way reflect the opinions of my $a $b." |
| done; done |
To unsubscribe, e-mail: m0n0wall dash unsubscribe at lists dot m0n0 dot ch
For additional commands, e-mail: m0n0wall dash help at lists dot m0n0 dot ch

subset: commoncrawl


text:
LONG BEACH TOWNSHIP - When Kurt Scheibner was a kid in Ship Bottom, his family owned a 1960s-era, Greg Noll-shaped Mickey Dora "Da Cat" model longboard. An older surfer saw the board and offered to buy the boys any new surfboard on Long Beach Island as a trade.
"I didn't want to do it, but my oldest brother agreed to go along with it," said Scheibner, now 24.
When he realized not only the monetary value of the board but also the historic significance, Scheibner became fixated on finding and riding older boards.
On Saturday, Scheibner won the Alliance for A Living Ocean LBI Longboard Classic on a 1961 Challenger East 9'6. Held at the 85th Street beach, the event challenges surfers to ride boards from the 1960s, the sport's golden era.
"A board that's been around for 50 years really interests me," Scheibner said. "You wonder who rode it, on what kind of classic days."
The surf was forecasted to be flat, but surfers were pleased to find 1- to 3-foot waves. The conditions were clean all morning and remained rideable right through the afternoon.
Bill DeWitt, 61, of Beach Haven, who made the semifinals, puts this day at the top of his list of favorites of the year. His daughter, Jamie DeWitt, 28, was the most successful female surfer New Jersey has had in recent decades, and she now lives in North Carolina.
In the 1960s, Bill DeWitt competed in the Ocean City Surfing Association's Polar Bear Contest, the budding Eastern Surfing Association's events, and the LBI Surfing Championships. This event is a throwback to those days.
"To me, this is a gas," said DeWitt, who rode a borrowed Eastern Surf Lines 9'8. "A lot of the guys here don't even know what it was like back then, but this is good for the kids and good for the whole surfing community."
DeWitt also won the tandem division, surfing with Nastasha Lisnieska, 25, of Beach Haven West. The pair executed a wheelbarrow stance to take the win.
Among the men, Scheibner and Jared Temple, 23, of Beach Haven, looked the sharpest throughout. Temple, riding an Oceanside Spoiler, beat James Contreras, 28, of Brick Township, and Rick Karolkiewicz, 45, of the Mayetta section of Stafford Township, in his semifinal by completing a very difficult fin-first takeoff.
In the final, Scheibner collected six points (of a possible 10) for a skillful noseride to take the win. Kemp Kane, 50, an LBI native who now lives in Stewart, Fla., took second place for his solid footwork.
Josh Law, 23, of Beach Haven, a successful local shortboard surfer, finished third by finding a very rare barrel ride on the inside.
"I wasn't even planning on surfing today," he said. "I was just here to watch and help out. I don't really ride longboards, but it was a blast."
Kim Kepich, 18, of Seaside Park, won the women's division with a nice frontside ride and turn on a steep wave face. Her sister, Jill Kepich, 21, took second, followed by Mary Tantillo, 40, of North Beach and Drew Gamils, 20, of Beach Haven.
Brian Aji, 14, of Ship Bottom, had the style to win the boys division over Kevin Budd, 14, of Holgate; Ryan Eckel, 14, of Ventnor; and Vaught Butler. The top Skimboard finishers were Mauro Sancharico, 18, of Beach Haven, Aji and Budd.
This year's event also featured a paddle race in which contestants started a quarter-mile out to sea and paddled to the beach. Mike Dancha, 42, of Surf City, a veteran Long Beach Township lifeguard, caught a wave on the way in to beat Kyle Fallon, 23, of Beach Haven, and Charlie Clay, 39, of Surf City.


text:
LOS ANGELES (AP) Almost 70 years after The Wizard of Oz premiered at Grauman's Chinese Theatre, a few of the film's Munchkins made a grand entrance there Tuesday to receive a collective star on the Hollywood Walk of Fame.
Seven of the surviving actors who played the inhabitants of Munchkinland in the 1939 classic attended the ceremony, arriving in a horse-drawn carriage and trailed by a marching band.
A yellow carpet, resembling the film's yellow brick road, led them to the stage. One tap-danced, and another sang.
"We love you; you have touched our hearts," former Munchkin Mickey Carroll, 88, told the crowd.
Carroll was joined by former Munchkin colleagues Ruth Duccini, Jerry Maren, Margaret Pellegrini, Meinhardt Raabe, Karl Slover and Clarence Swensen.
"I'm as proud today as my mother would have been," said Joey Luft, the son of Judy Garland. Garland, who played the movie's wide-eyed orphan, Dorothy Gale, died of a drug overdose in 1969.
Carroll was one of more than a hundred adults and children who were recruited for Oz to play the natives of what author L. Frank Baum called Munchkin Country in his 1900 book The Wonderful Wizard of Oz.
They only made $125 a week while filming, followed by decades of recognition, Carroll told The Associated Press by phone before the ceremony.
"I'm not a Munchkin, I'm an entertainer," Carroll noted. "But the movie is great because we all grew up with it. ... It never dies."
Copyright 2007 The Associated Press. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.
Conversation guidelines: USA TODAY welcomes your thoughts, stories and information related to this article. Please stay on topic and be respectful of others. Keep the conversation appropriate for interested readers across the map.


text:
4800 West 92nd Avenue Westminster, CO 80031
Home | News | News Archives
Jessica Ridgeway Memorial Park was dedicated Oct. 5 during a brief ceremony. About 800 people joined in an emotional program to honor Jessica and celebrate her life.
"As I look out at the Jessica Ridgeway Memorial Park, I hear the laughter of children. I see Jessica's smile," said Westminster Mayor Nancy McNally.
"What Jessica leaves as a legacy is much bigger than a park. She reminded us how precious life is and how fleeting it can be," said Westminster Police Chief Lee Birk.
Christine Ridgeway, Jessica's grandmother, also spoke, telling the crowd, "I see people who helped create a park full of Jessica's smiles and love." She was joined on the stage by Jessica's mother, Sarah, and Jessica's aunt, Rebecca.
All five cut a ceremonial purple ribbon to open the park. The Ridgeway family also released 10 purple balloons into the air to honor Jessica and her 10 years of life.
The Butterfly Pavilion presented the Ridgeway family with a framed set of butterflies in Jessica's memory.
Butterflies were one of her favorites and are a theme at the park, along with the colors purple and green, playful ribbons and knock-knock jokes provided by her classmates at nearby Witt Elementary School.
The park, at 10765 Moore St., cost about $650,000 to complete, with most of the money coming from donations and grants. Nearly 400 donors gave to the fundraising campaign. In addition to financial contributions nearly 300 people from the community (including City of Westminster employees) volunteered their labor to help build the playground.


text:
Printable View
shadowkill, barry,
go easy on him, i'm sure he didn't mean to beat you to it. :D
i found this by googling. as one of the first links found, it's current as far as i see it. especially since i finally have a laptop with an expresscard.
i think i am beginning to see why...
the good thing about this laptop is that it has both; expresscard and pcmcia. it's a Dell D830.
so, i still get to use the ol' faithful ubiquity.
Well since this thread was bumped.
The card is still not working haha, but i have been a slacker the last halfyear so maybe it works now with the right drivers :)


text:
Alaska and 45 other states entered into an Assurance of Voluntary Compliance with Dish Network, LLC. The settlement resolves the states' allegations including that DISH Network:
As part of the settlement, Dish Network customers in Alaska may be eligible for restitution. Unresolved complaints sent to Dish Network or the Alaska Attorney General's office since January 1, 2004 are eligible for the restitution program. Dish Network will notify consumers that they are eligible for the restitution program by sending a claims notice to those consumers. Additionally, consumers in Alaska can file a complaint with Dish Network, the Alaska Attorney General's Office, or the Better Business Bureau by December 14, 2009 to be considered for the restitution program as long as the complaint is about activity that took place within the last 2 years. Consumers who are not satisfied with the company's offer of restitution may choose to file a claim to be decided by a third-party claims administrator.
Questions or complaints regarding the refund process can be addressed to: DISH Network, L.L.C., Dispute Resolution Team, P.O. Box 9040, Littleton, CO 80120 or by e-mail at firstname.lastname@example.com.
Consumers can also contact the Alaska Attorney General's Office, Consumer Protection Unit at 269-5200, or outside of Anchorage, 1-888-576-2529, or by sending an email to firstname.lastname@example.com.
Consumer Protection Unit
July 2009


text:
|Web Site Information :.
|Site Info Whois Traceroute RBL Check
Busted In The USADescription: Busted in the USA is a site devoted to keeping public records and information regarding criminals available to the populous.
Keywords: criminals, criminal, jail, jailed, busted, crime, inmate, mugshot, mugshots, Busted in the USA, DWI, Meth, Cocaine, offenders, Crow Wing, minnesota, corrections, DOC
Tags: crowwingcriminals, busted, usa, site, information, criminals, public, available, use, records, populous, regarding, devoted, keeping, user, minnesota, criminal, crow, mugshots, inmate, jailed, dwi, crime, mugshot, corrections, wing, meth, doc, offenders, cocaine, jail, content, terms,
|IP Index TLD Index Domain Index Site Index
|Copyright © 2013 dawhois.com


text:
Project DescriptionDesign
Grand, Multifamily Kitchen
Some cooks love how an open plan kitchen can turn meal prep into a party. Then there's the owner of the Prairie Dog kitchen. She's the cook in the family and loves having company—especially when they stay clear of the kitchen while dinner's being made. She and her husband wanted an open plan living area that would have a delineated cook's space yet still encouraged gathering. Architects Jane Treacy and Phil Eagleburger came up with a great answer: a kitchen island with a raised surround to shelter the cook but not isolate her. To join a conversation, she just has to peek over the surround, like a prairie dog.
The couple sought a kitchen that would play well with the modernist style of their co-op building, Washington's Watergate complex, which was built in the 1960s. The island doubles as a sleek armoire, uniting kitchen and living room while separating them enough to define both spaces and keep cooking chaos out of sight. On the kitchen side is a prep area with a 2-foot-deep Corian countertop, plus drawers and cubbies to keep ingredients and tools close at hand. On the living room side are drawers and cabinets—ample storage for glasses, dishes, and tablecloths. Custom cabinets offer warmth and texture, their stained vertical-grain fir lending contrast to the neutral surroundings.


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Celebrating 2 years of Electronic Directory, the next instalment of the Podcast comes from Esteban Adame, one half of ICAN.
Interview with Esteban Adame
What have you been up to lately ?
Well, I've been playing a lot of Jazz with my trio, working on tracks and producing. I wanted to focus more on DJing and production work this year, so that what I've been focusing on this year.
Nomadico and Esteban Adame, you guys have produced together as El Coyote. Can we look forward to anymore from you two ?
Yes, as a matter of fact, we are currently working on some material that should be ready by then end of this year. Look out for these tracks either on ICAN or other labels.
Can you tell us a bit about the mix ?
I tried to play music produced by my friends as well as by me. I wanted to show my piano skills so I put together an intro for the mix. The intro is aka G2G complete with synth pads and strings. I hope you all like it.
What's up next for you ?
Well, I have some possible gigs in Japan that have not been confirmed and some other dates in Europe with DJ 3000. I also have a release on Subject Detroit coming out soon as well as some remixes on some well known labels that will announced soon.
Download Electronic Directory 10 Esteban Adame Podcast
Direct download here or Subscribe to the podcast
- 01. Esteban Adame – Tribute to Electronic Directory [ICAN (Unreleased)]
- 02. Itokim – Far West (Deep Distance Mix) [Night Drive Music]
- 03. Life Line – 2 Luvs [Groove On]
- 04. Mark Flash – Sao Paulo [Underground Resistance]
- 05. Samuel L. Session – Another Day (Rain) [Music Man]
- 06. Esteban Adame – January 88 Degrees [ICAN (Unreleased)]
- 07. Mark Flash – Conquistador [Underground Resistance (Unreleased)]
- 08. Oskan – The Key Theory [Crate Savers]
- 09. Jona – Altiplano [Planet E]
- 10. Nublu Orchestra – Sketches of NYC (Nomadico Remix for UR) [Nublu Records]
- 11. Edison Carter – C.F.O's Cuts [Technoir Audio]
- 12. ICAN – Caminos Del Nino (Original Mix) [ICAN]
- 13. Esteban Adame – Trinity (S2 Mix) [ICAN (Unreleased)]
- 14. Kenny Larkin – Bass Mode [Planet E]
- 15. Dsum – Libertad (Original Mix) [Monofónicos]
Download older podcasts below


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Deborah Exum Gift of Encouragement
This gift in memory of Deborah Lynn Exum, (1952-2005), is given by her husband, Tom Styrkowicz, to an individual, who has made an extraordinary contribution to Genuine Art. It is a monetary gift to encourage the recipient to "break out and try things," as Deborah, who always saw the good in people, would have wanted.
Deborah Exum and Madison Montessori were intractably linked in many ways – as a parent of two students, as a creative visionary, talented photographer, selfless volunteer and incredibly generous and kind human being. She left us way too soon. But, if we listen closely with our hearts, we can still hear her loving, customary greeting, Hi Honey!", and know that she will always be close to us.
April 2013
Dear Terry (and everyone, really…)
To say that I was flabbergasted by the Deborah Exum gift would be an understatement! I've always felt respected and welcome and to be honored in this way truly does humble me.
I believe in the philosophy of your school: the idea of self-direction and self awareness, enthusiasm for all experiences and a general "wonder" about our big world. With my participation in Genuine Art, I know I enjoy a similar revelation each year.
But really, the examples of kindness that exist in your hallways and your larger parent community are truly impressive. To consider myself as even a little slice of this partnership keeps me coming back for more. I am truly honored by your generosity of this gift. I regard it as good fortune that all those years ago, I was introduced to the Madison Montessori School.
- The Collaborative Process
- Deborah Exum Gift of Encouragement
- Donate to the Scholarship Fund
- GA Catalog
- From Montessori Life: How Art Enhances a Montessori (PDF)
MMS has such a close knit family feeling that makes the kids secure enough to explore new things without hesitation.
- Debra Exum, parent


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HUGGIES Little Snugglers Baby Diapers, Size 2, 148 Count
Huggies Little Snugglers diapers are specially designed for gentle skin protection. With features like the pocketed-back waistband and Gentle Absorb liner that contain and draw the mess away, Little Snugglers help keep your baby's delicate skin clean and healthy. Little Snugglers diapers also feature premium softness and breathability to help keep them comfortable. Little Snugglers
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text:
The subroutines in the "Data::SecsPack" module packs and unpacks numbers in accordance with SEMI E5-94. The E5-94 establishes the standard for communication between the equipment used to fabricate semiconductors and the host computer that controls th...SOFTDIA/Data-SecsPack-0.06 - 21 May 2004 13:18:48 GMT - Search in distribution
- Docs::Site_SVD::Data_SecsPack - pack and unpack numbers in accordance with SEMI E5-94
The "Test::Tech" module extends the capabilities of the "Test" module. The design is simple. The "Test::Tech" module loads the "Test" module without exporting any "Test" subroutines into the "Test::Tech" namespace. There is a "Test::Tech" cover subro...SOFTDIA/Test-Tech-0.26 - 21 May 2004 02:58:26 GMT - Search in distribution
- Docs::Site_SVD::Test_Tech - Extends the Test program module
The 'Tie::Gzip' module provides a file handle Tie for compressing and uncompressing files using the gzip compression format. By tieing a filehandle to 'Tie::Gzip' subsequent uses of the file subroutines with the tied filehandle will compress data wri...SOFTDIA/Tie-Gzip-0.06 - 16 Apr 2004 20:38:48 GMT - Search in distribution
The "File::Drawing" program module uses American National Standards for drawings as a model for storing data. Commercial, governement and casual orgainizations have stored information over the centuries as drawings. Drawings probably evolved from the...SOFTDIA/File-Drawing-0.01 - 04 May 2004 12:22:33 GMT - Search in distribution
The "ExtUtils::SVDmaker" program module extends the Perl language (Perl is the system). The input to "ExtUtils::SVDmaker" is the __DATA__ section of Software Version Description (SVD) program module. The __DATA__ section must contain SVD forms text d...SOFTDIA/ExtUtils-SVDmaker-0.10 - 25 May 2004 22:22:27 GMT - Search in distribution
The "Data::SECS2" module provides a widely accepted method of packing nested lists into a linear string and unpacking the string of nested lists. Nested data has a long history in mathematics. In the hardware world, data and data passed between hardw...SOFTDIA/Data-Secs2-0.09 - 20 May 2004 17:23:12 GMT - Search in distribution
- Docs::Site_SVD::Data_Secs2 - pack, unpack, format between Perl data and SEMI E5-94 nested data
SOFTDIA/Test-STDmaker-0.23 - 24 May 2004 16:45:33 GMT - Search in distribution
SOFTDIA/Text-Replace-0.07 - 11 May 2004 15:49:41 GMT - Search in distribution
The "Data::Str2Num" program module provides subroutines that parse numeric strings from the beginning of alphanumeric strings. str2float $float = str2float($string); $float = str2float($string, [@options]); $float = str2float($string, {@options}); (\...SOFTDIA/Data-Str2Num-0.07 - 22 May 2004 18:37:35 GMT - Search in distribution
- Docs::Site_SVD::Data_Str2Num - int str to int; float str to float; else undef. No warnings.
Many times there is a group of subroutines that can be tailored by different situations with a few, say global variables. However, global variables pollute namespaces, become mangled when the functions are multi-threaded and probably have many other ...SOFTDIA/Data-Startup-0.04 - 27 May 2004 16:37:39 GMT - Search in distribution
- Docs::Site_SVD::Data_Startup - startup options class, override, config methods
SOFTDIA/Tie-Layers-0.06 - 28 May 2004 20:21:29 GMT - Search in distribution


text:
A smashing outfit is worth a thousand words. I buy what "fits my image" and dress to "make a statement." Apparel advertising buttons our identity to our body. It encourages self-expression through something visual that we create or select with the cooperation of our minds and our hands. For that reason, fashion savants likely claim ranking with other types of visual artists. For an animal, its outerwear is defense against weather. But 'wardrobe curators' and the like explore more complex uses for clothing by engineering it for communication between people. It communicates status and role, dispositions toward authority and levels of self-esteem. Even more abstractly, it communicates a person's unique ability to relate linear patterns, colors or shapes.
So, clothing—it's an image of who you are. Granted, it's a brief image—we can't statically frame our outfits for study. Our own moving bodies are the frame. Outfits are assembled for certain moments or a stretch of hours, and then get dismantled, sort of like the short-form of installation art. But when taken as a whole, the right wardrobe is evidence of someone's good taste correctly conveying their peculiar identity.
We're attracted to those people, wondering how it is they deftly create consistent, genuine images of their identity. Come Sunday morning, Christian adults who dress well have this effect on me. Their identity in Christ—an identity centering on Christ while radiating his image to the world—is now made visible in a small way. It absorbs me in the thought of Christian adulthood. The image of their identity is profound motivation to mature my own identity and invites me to become the disciple of my elders.
Yet we don't often think immediately of clothing when our church communities encourage us to develop our faith through carefully considering culture and art. Quite possibly, clothing has been waylaid while we catch up on learning how to healthily consume higher art. It's likely, though, that we should encourage more attention to it in our after-church fellowshipping and small-group conversations. Kent Reister recently described a healthy consumer as someone who loves something "for its aesthetic power" by encountering it with "focus and deliberation," turning an "overload of film and music access" into "searching out the infinite design of God in one good story or track." Pausing in the 'image bombardment' of our culture to give diligent awareness to the intelligent design and significance of a few images transforms consuming into partaking of God's nature. Taking pause with clothing as an image is doubly worthwhile; clothing facilitates encounters with beauty as seen distinctly by another human. Particularly in a church setting, a conversation about clothing is a trustworthy convergence of becoming a better consumer and seeing the image of God in the soul of our neighbor.
The Christian communities I am familiar with have no strong resistance to dressing fashionably. They also don't encourage or articulate stronger aesthetics beyond principled modesty. If that's the only governing principle for the art of grooming, then the only criteria for being 'well-dressed' is probably 'finding clothes which minimizes thoughts about sex.' There is easily more to motivate our clothing choices than sex. Human beauty is not inherently sexually attractive. Otherwise, how would you justify telling your daughter she is beautiful? We are daily discerning human beauty without experiencing it sexually.
To caveat, I am deeply grateful to my parents and church for teaching me the protection and freedom of modesty. But limiting discussions of clothing to modesty is doubly restricting. It veers toward more application for women than men. Men don't get (need? deserve? what are we telling them?) much training in clothing aesthetics. Also, it only teaches women to dress their body by thinking of its sexual purpose. The gender compartmentalization ends with recognizing the sexuality of the body and falls short of recognizing its humanity.
Limiting theology of clothing aesthetics to modesty boxes it into the one season of life where modesty is relevant. Dressing well is merely fad for the young and sexually aware. It plays no role in drawing us further up and further in to Christian maturity. But if you are like me, you are convinced that dressing well may become a means to glimpsing part of God's nature. Naturally, you hope that your peers are not the best available experts on clothing aesthetics. Instead, you look to your elders and hope to find their conversation both seasoned and eager.


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Dr. Barry Miller on Corporate Escape Artist Radio
Today on Corporate Escape Artist Radio, we welcomed career guru Barry Miller. As the head of career programs for alumni at Pace University in New York City, Barry offers advice to students and graduates in transition on a daily basis.
This morning, he talked about his advice for aspiring entrepreneurs, social media, and much more. Listen to Corporate Escape Artist Radio with Barry Miller (Barry's segment is the fourth one, starting at 11:30)
Now you can benefit from Dr. Miller's expert advice without traipsing all the way to Lower Manhattan.


text: Hey guys, I know this should be basic, but I'm new, and need an answer quick and don't have time to research the topic. I'm heading to a outdoor wedding and I picked up this sweet sport coat. I'm planing to wear Taupe dress pants with the jacket. My question is do I go with the black shoes/belt, or brown shoes/belt? I feel like the brown would go better with the Taupe pants, but the black would go better with the jacket, but might look out of place up against the pants. Any input is appreciated! And I promise I'll search next time, just don't have time as the wedding is this weekend and I'm leaving for it tomorrow!! Thanks!!


text:
We have decided to not include these in bt4. Check out emgents new tool gerix-wifi-cracker instead
apt-get install gerix-wifi-cracker/
Two Shaman virtuel's logs...
We have decided to not include these in bt4. Check out emgents new tool gerix-wifi-cracker instead
apt-get install gerix-wifi-cracker/
Speaking of Shaman, anybody heard from him lately?
Of course, if you really wanted to have some fun, go to Wal-Mart late at night and ask the greeter if they could help you find trashbags, roll of carpet, rope, quicklime, clorox and a shovel. See if they give you any strange looks. --Streaker69
Last I heard he was on the road - suffering from lack of housing issues.....
Don't know - the guy is a great coder
Thanks but I have to admit that I tested gerix on Ubuntu and not BT4.....so maybe gerix is better on BT4. So I defer to others for the gerix results. EDIT: tested gerix with BT4 and results were pretty much the same.....in italian and not very straightforward.
But, it is hard to imagine any GUI being better than Shaman's spoonwep2.....the GUI visual, process simplicity, and effectiveness is the best I have seen so far compared to any other script/GUI.
Shaman is travelling somewhere in France and it was done in Java.......so I guess that makes it difficult for the BT4 developers to adjust it for the latest BT release.
I tried gerix in BT4 pre release... it's in english.
This GUI do his work, that all.
But it's not so simple!!!
Spoon(wep2/wpa) is really like you say visual, process simplicity.
And I'm french...
I hope there 's someone who will convert this script for Ubuntu and BT...!
Moi aussi!
Anyway, the expert on spoonwep2 is also French. If you want his email address, send me a pm.....and maybe you can reach him.
I've also tested gerix, and although it's definitely good. spoonwep is still much more straightforward.


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View Single Post
March 4th, 2013 (2:09 PM).
Join Date:
Jul 2012
United Kingdom
Here is my SU for angel.
Charlie, the angel
Physical Age:
To keep people guessing, being trivial, being depended upon.
Displaying his emotions, getting embarrassed, being made to start conversation.
To be welcomed by others, to reach Heaven, to have his existence acknowledged by Joey.
Rejection, Eternal Darkness,
Charlie has a fair complexion with flawless skin; his face is unmarked and unblemished. Charlie's hair is a silvery-blue colour which displays wisdom and experience. Charlie's eyes are a lightly coloured brown and gives an aura of mystery to him. This mysterious aura is improved by the garments that he wears, a long, black overcoat which hangs lower than his feet, on which he wears leather boots. He always wears the coat collar up which makes him look like quite an antisocial figure, but it is due to the fact that he is a very shy character. Charlie's face is the one that cannot be read easily, it lacks emotion but in a comforting way. Charlie is by no means an intimidating figure, no matter how much he may look it, and his build is on the lower side of average and is not athletic in the slightest. Despite lack of athleticism, Charlie is rather tall; he stands at 6ft5. As Charlie looks extremely youthful and could be mistaken for someone much younger in the human realm, this isn't helped due to his lack of facial and obvious body hair. When Charlie walks, he walks slightly hunched over and is constantly looking around as if he feels vulnerable.
Despite Charlie's youthful appearance, he acts in a very mature way. He always thinks about everything before answering any question and he displays knowledge in almost every subject. Despite not liking to start conversations, Charlie much likes to have others come and talk to him first. Charlie hates to be an annoyance to people and therefore prefers others to begin interaction as that way he ensures that he won't be annoying. Charlie does not judge people without good reason, however first impressions are hard to erase for him. If you mess up once, he will not forget it and regaining his respect will take effort. Charlie is not a violent person, he is much the opposite. He would much rather discuss problems and loves to debate specific topics. Although Charlie is quite shy, he likes to have authority entrusted to him. He typically performs at high standards with great understanding and doesn't throw away opportunities. Charlie is very diligent and does not give up on anything at all. He is extremely loyal to anyone who earns his trust and is a very comforting person. He dislikes to see people suffer that don't deserve it and will do all within his power to prevent such a thing from happening. Charlie may seem as a care-free individual but it is simply for the fact that he is not easily annoyed; he is very much a level-headed and calm angel.
When Joey was born, Charlie had been assigned him as an emergency case. He had been called upon whilst wandering around watching the humans go about their lives. He knew that there would be a certain degree of work needed for Joey considering the situation he was born into. He disliked the fact that such a child was to be born into the world with unstable parenting. Since then he has always held a grudge against Josiah's parents for putting Joey through a life like that. However, luckily for them both, Charlie had been assigned a similar case before. He knew which the hardest parts were and which the easier parts were. He was quite surprised at how well Joey's original parents looked after him considering their ages and their occupations. However, he knew it wouldn't be long before the boy was sent to the orphanage. Charlie watched over him, day and night, because he knew that orphanages could be a dangerous place. He thought it good that Joey was a quiet boy, this would prevent him from getting himself into any trouble, which it did. Thankfully. Sooner than expected, Joey was adopted by a couple that seemed nice enough. They meant well, at least.
Joey didn't seem to enjoy his time at the Spanner residence too much and Charlie could hardly blame him. Small children running around everywhere were not exactly the family that suited Joey. When Joey had first been dressed in female clothing Charlie was surprisingly amused, but in a good way. It seemed to Charlie that Joey actually made a good female, for the moment anyway. It was surprising how much more he could get away with by being a 'girl' than a boy. Charlie was quite pleased that Joey would be home-schooled as it would guarantee his safety, to an extent. It would also make him develop better academically given that he was much above the typical child of his age group. Joey did a great job in securing the home-school situation by working very hard all the time, it ensured that no one could complain about them as they were producing results, no doubt about it.
The fact that Joey would be moving to Seattle was great for him! It would give him proper education and perhaps enhance him socially. It was good that Joey would get a kitten to look after too, it would give him an opportunity to take on responsibility and hopefully help him become more adult like. Charlie was hesitant about the female clothing, but it turned out that it would all be for the better. Joey could practically transform into a girl by changing outfit, just like how a superhero changes ego.
Joey was doing well academically and he had made some friends, he had settled into Seattle life far better than Charlie could have hoped for. It had also become apparent that Joey had become accustomed to being Olivia. It seemed to suit him quite well and was quite the opposite of Joey. Although it made Charlie quit e relaxed to know that Joey was happy in some way, Charlie just wished that Joey would accept the fact that he was Josiah and not Olivia. In some ways Charlie was embarrassed by the fact that he had a cross-dressing human to look after. With Olivia being a young female alone in a big city, Charlie couldn't relax fully as it isn't always the safest thing to be. So Charlie was naturally on edge being cautious of the surrounding people. If anything looked odd, he would intervene. Only once had something come close to happening, but Charlie had sorted it out. There were 3 shady looking men following around Olivia, naturally Charlie made certain that it wasn't just coincidence. Before Joey had been snatched by these guys, Charlie had thought it best to lead these men subconsciously away from Joey and go into a pub that was just opposite. Charlie had felt very happy with himself to have done this.
Charlie liked the fact that he had to look after Joey. He was a quiet boy, much like how Charlie was quiet. He also liked the fact that Joey cross-dressed sometimes, it made him quite an interesting person and Charlie very much liked interesting things.
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text:
Product Evaluation Methods and Their Applications
Popovic, Vesna (1997) Product Evaluation Methods and Their Applications. In Seppala, Pentti, Luopajarvi, Tuulikki, Nygard, Clas-Hakan, & Mattila, Markku (Eds.) IEA 97 Congress, June 29 - July 4, 1997, Tampere, Finland.
|PDF (18kB)
This paper discusses methods and techniques to be used for an ergonomic evaluation of products, product interfaces and systems. In this context evaluation is seen as a part of the design process that interacts with all design stages. It plays an integral role in it and is concerned with assuring a high degree of likelihood of the user's acceptance. Traditional and new evaluation methods such as task analysis, checklists, TA (talk/think aloud) protocols, CAD simulation are addressed.
Impact and interest:
Citation counts are sourced monthly from Scopus and Web of Science® citation databases.
These databases contain citations from different subsets of available publications and different time periods and thus the citation count from each is usually different. Some works are not in either database and no count is displayed. Scopus includes citations from articles published in 1996 onwards, and Web of Science® generally from 1980 onwards.
Citations counts from the Google Scholar™ indexing service can be viewed at the linked Google Scholar™ search.
Full-text downloads:
Full-text downloads displays the total number of times this work's files (e.g., a PDF) have been downloaded from QUT ePrints as well as the number of downloads in the previous 365 days. The count includes downloads for all files if a work has more than one.
|ID Code:
|Item Type:
|Conference Paper
|evaluation methods, product design
|Australian and New Zealand Standard Research Classification > BUILT ENVIRONMENT AND DESIGN (120000) > DESIGN PRACTICE AND MANAGEMENT (120300) > Design Innovation (120302)
|Past > QUT Faculties & Divisions > Faculty of Built Environment and Engineering
|Copyright Owner:
|Copyright 1997 (please consult author)
|Deposited On:
|27 May 2005 00:00
|Last Modified:
|02 Feb 2012 09:45
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text:
"He (Musa) said: 'Oh my Lord! Expand my breast for me and make my affair easy to me, and loose the knot from my tongue (that) they may understand my word … (Quran 20:25:28)"One and a half years ago, I took the plunge and began sharing our son's life lived with autism, my struggles with keeping strong in my Islamic faith and residing at this intersection of faith and autism. With a post on Ramadan Despair, I shared what was then my lowest point, and I had no purpose in laying bare our struggle … [Read more...]
The Failure of the American Muslim Community to Help Those with Special Needs
The View from Up Here – Personal, Global
My husband asked me last night what if I was going to write something about the 11th "anniversary" of the tragedy of 9/11. I thought, what is there to say that hasn't already been said before? Where are we now as a world community, 11 years after that awful, fateful day? Where am I and my family in our lives now?On September 11, 2001, Lil D had just passed his one-year birthday. He was my only child at that point. We were one year away from a discussion with my pediatrician about my worries c … [Read more...]
Talking Love, Islam and American Muslim Women with 'Love InshAllah's Nura Maznavi
It's about love, yes, but it's about all the ways love happens, begins, ends, endures, frustrates, eludes and abounds. It's about the love stories of American Muslim women, who lay their claim to love in ways previously not openly discussed in the American Muslim community. Love InshAllah, a new anthology of 25 love stories edited by Ayesha Mattu and Nura Maznavi, is creating quite a buzz in Muslim and non-Muslim circles for its embracement of all types of love stories, including those not typic … [Read more...]
TLC's All-American Muslim is Surprisingly Really Watchable
It's tough, and never a good idea for a single person, or a few families, or even a specific community to be considered the representation of an entire faith group. Growing up as one of the only Muslims families in a small university town in North Dakota, inevitably my brothers and I became the poster children for "This is what Muslims are like!" I think, for the most part, it was a good thing because we walked that delicate line of practicing our faith (Ramadans spent hanging out in the coun … [Read more...]
Three Questions for American Muslims: Asma T. Uddin
The Muslim Portal at Patheos is hosting the "Three Questions" project for the month of September in conjunction with the tenth anniversary of the terrorist acts of 9/11. We are asking American Muslims from across the nation three simple but important questions. Click here to learn more about the project.Asma T. Uddin, founder and editor-in-chief of Atlmuslimah, legal fellow with the Institute for Social Policy and Understanding (ISPU) and an international law attorney with The Becket Fund f … [Read more...]
Justice Perverted for the Irvine 11
So what is free speech, really? Is it the right to burn the American flag? The right to sponsor a campaign to "Draw the Prophet Muhammad," which offends a sizeable faction of the American population? The right to say what you want, when you want? The right to wear a black armband on your arm in protest of war? The right to use speech that others may deem hateful? The right to protest? The right to insult? The right to praise?What is free speech, if it isn't one of the most fundamental rights … [Read more...]
A Muslim American Declaration
(Blogger's note: This is a signed declaration from American Muslims/Muslim Americans, including myself, for public consumption.)We are Muslim Americans. We are American Muslims. We live as your neighbors, friends, doctors, lawyers, police officers, soldiers, cab drivers, newspaper vendors, teammates, co-workers, and family -- seamlessly and without conflict. We are fully immersed in the American mosaic, and we are proud.Our Muslims forefathers have been here since the founding of this … [Read more...]


text:
If you are passionate about Scotland's past and about reading and would like to turn this interest into a career, the BA (Hons) Scottish History and Literature is ideal.
You will study Scottish literature and Scottish history in a global context, comparing Scotland with the wider world in both disciplines. The degree cover a range of approaches, from thematic modules that cover a variety of places and historical periods, to concentrated and in-depth studies of a particular group of authors or historical periods. You will also study language, translation and the interrelation of Scots, Gaelic, English and other languages in Scotland, as well as different approaches to and studies of medieval, early modern and modern history.
Entry to year one:
Advanced entry to year two:
History subjects, from which you will choose two, may include:
You will also study two literature subjects which may include:
You will study two further option modules from a range of history, literature and other humanities subjects.
History subjects may include:
Other history subjects may include:
Literature subjects may include:
You will choose two history subjects which may include:
Literature subjects, from which you will study two, may include:
You will study two further option modules from a range of history, literature and other humanities subjects.
You will complete a dissertation in history or literature. You will also study one option module from your chosen dissertation subject plus three option modules from the other subject. The following lists show all the options, however they are again available on a rotation basis each year.
History subjects may include:
Literature subjects may include:
You will learn through a combination of face-to-face and video conference lectures and online study via the university's virtual learning environment (VLE), with support from your tutors.
For students normally domiciled in Scotland or the European Union/EEA (outside of the UK) and with a term time address in Scotland, the following fees apply:
|Full-time (120 credit modules) per year
|Part-time (structured) (3 x 20 credit modules) per year
|Part-time (unstructured per module) (per 20 credit module)
For students normally domiciled in the rest of the UK (England, Wales and N. Ireland) and with a term time address in Scotland studying this course full time, the following fees apply:
|Year 1
|£8,000 p.a.
|£8,000 p.a.
|Year 2
|£8,000 p.a.
|£8,000 p.a.
|Year 3
|£8,000 p.a.
|£8,000 p.a.
|Year 4
**4th year free only for students studying full-time who have paid for the previous 3 years at the University of the Highlands and Islands.
Part-time (per 20 credit module) - £1,340 (2016-17 and 2017-18)
Fees are payable in advance each academic year unless otherwise agreed.
There are a number of funding options available to UK and EU students to help you pay for your studies and your cost of living whilst studying.
For students who do not normally reside in the UK or European Union and with a term-time address in Scotland, the following fees apply:
|Full-time (120 credit modules) per year
|Part-time (structured) (3 x 20 credit modules) per year***
|Part-time (unstructured per module) (per 20 credit module)***
***There is no part-time study option for Tier 4 sponsored international students.
Further information on international course fees.
A no fee increase guarantee is available for self-funding full-time and structured part-time RUK and International undergraduate students for continuous and contiguous study for the same award, up to the permitted standard time limit for the relevant award.
External scholarships and financial support may be available. Contact the relevant university partner college for more information.
Once you have successfully completed your BA (Hons) Scottish History and Literature, you might like to consider careers in:
You can progress from the BA (Hons) Scottish History and Literature to the following postgraduate courses;
You can use the above QR code to connect directly to the course details.
International applicants should apply using the UHI International application form. Please download and print the form, complete and return using the address indicated. More information is available for international applicants at our international student page
We are delighted that you are thinking about studying at the University of the Highlands and Islands (UHI). UHI operates a fair and open admissions system committed to equality of opportunity and non-discrimination. We consider all applications on merit and on the basis of ability to achieve, without discrimination on grounds of gender, age, disability, ethnicity and socio-economic background. We welcome applications from all prospective students and aim to provide appropriate and efficient services to students with disabilities.


text:
Shaun ThaddeusKuala Lumpur, MY
English (North American), English, Malay, English (British)
British, Malay, North American
Middle Aged, Senior, Teen, Young Adult
Special Skills
Audiobooks, Business, Cartoons, Documentaries, Educational, Internet, Movie Trailers, Podcasting, Radio, Telephone, Television, Videogames
Payment Terms
Hire Shaun as a voice actor. Payment is made through Voices.com SurePay, an escrow service that guarantees your work is done to your satisfaction. Send payment securely by Visa, Mastercard or PayPal through Voices.com then download your files from the website.
Watch a video to see how to make a payment
Next Steps
To contact Shaun Thaddeus about your project, request a quote today. We'll walk you through the process step-by-step. It's fast and easy.
This talent has not uploaded any demos yet.
Title of Your New Demo 7777 Listens
Hire Shaun Thaddeus as a voice actor. Working with Shaun Thaddeus is fast and easy. We'll walk you through the process step-by-step.
Request a quote for your project today.


text:
Looking for the perfect piece of cat furniture to go with your mid-century modern decor? Behold the new bent plywood lounge from CANOPYstudio. This incredibly elegant and beautifully crafted piece of furniture is destined to become an heirloom.
Made from walnut veneer plywood and solid walnut legs, this cat bed is clearly inspired by the work of Charles and Ray Eames. The removable cushion is held in place with magnets embedded in the plywood frame. The gorgeous upholstery fabric is imported from Denmark and is the same durable fabric that is used on high-end modern furniture. Absolutely stunning!
This beautiful lounge sells for $790 and is available in the CANOPYstudio Etsy shop. Thanks to Kimberly for the find.


text:
PREPARATIONS are underway for another royal visit to Cumbria this week.
The Duke and Duchess of Cambridge will tour BAE Systems in Barrow on Friday.
Areas opposite and adjacent to the main reception, on Bridge Road, will be cordoned off to allow the public to catch a glimpse of Prince William and Kate as they arrive in the early afternoon.
Minor travel disruption is expected while police close sections of the road for a short time.
The visit comes after Prince Charles' engagements in Lupton, Kirkby Lonsdale and Sedbergh on Thursday and The Duchess of Cambridge's day at a Scout camp near Windermere the week before.


text:
Philips Wake-up Light
£180, apple.com
Give the gift of stress-free mornings this Christmas with this alarm light – for anyone who's sick of waking up to the shrieks of their alarm clock? This light eases you gently out of Morpheus' embrace, simulating the break of dawn in your bedroom. iPhone users can use their Apple gadget to set up 10 alarms and track sleep routines – for those who aren't Apple fans, there's an iPhone-free version available for £100.
Dyson DC47
£330, johnlewis.co.uk
Dyson's twin cyclone cylinder vacuum cleaner features the company's Ball technology, letting it wend its way around even the toughest obstacles with ease. And its dual channel floor tool will make short work of ground-in dust in your carpets, you filthy animal you.
Doctor Who Sonic Screwdriver Universal Remote Control
£60, firebox.com
Know a Doctor Who fan? Sonic up their life with this motion-controlled replica of the Time Lord's space-age gadget. It recognises 13 different gestures, controlling up to three devices with IR receivers – including TVs and iPhone docks. And once the novelty's worn off, the FX mode features an array of authentically 70s-vintage bleeps, bloops and whirrs sourced from the BBC Radiophonic Workshop.
Roku LT
£50, maplin.co.uk
This little box of tricks is perfect for adding media streaming smarts to a TV in a simple, no-frills package. £50 nets you access to 150 streaming channels including Netflix, Crackle and the BBC iPlayer – or there's the Roku XS for £100, which adds Ethernet connectivity and a motion-controlled version of Angry Birds.
iRobot Roomba 790
£800, irobot.com
iRobot's latest Roomba robot vacuum offers freedom from domestic drudgery – as well as the usual features like HEPA filters, room-to-room navigation and spot cleaning, it comes with a wireless controller to let you command your mechanical servant from afar. Two virtual wall lighthouses keep your robot slave confined to a specific room until it's spotless – just bear in mind that your Roomba will remember your treatment of it, and you will suffer come the robot uprising.
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