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Author Affiliations: University of Antwerp (Drs Gijselinck, Engelborghs, Martin, De Deyn, Kumar-Singh, Van Broeckhoven, and Cruts; Mss Maes, Cuijt, Peeters, and Mattheijssens; and Mr Joris); Neurodegenerative Brain Diseases Group, Department of Molecular Genetics, VIB, and Laboratory of Neurogenetics, Institute Born-Bunge (Drs Gijselinck, Kumar-Singh, Van Broeckhoven, and Cruts; Mss Maes, Cuijt, Peeters, and Matteijssens; and Mr Joris); Memory Clinic and Department of Neurology, ZNA Middelheim (Drs Engelborghs and De Deyn); Laboratories of Neurochemistry and Behavior (Drs Engelborghs and De Deyn), Neurobiology (Dr Cras), and Neuropathology (Dr Martin), Institute Born-Bunge; and Division of Neurology, University Hospital Antwerp (Dr Cras), Antwerp, Belgium.
Frontotemporal lobar degeneration (FTLD) is a neurodegenerative brain disorder that can be accompanied by signs of amyotrophic lateral sclerosis (ALS).
To identify a novel gene for FTLD-ALS.
Genome-wide linkage study in a multiplex family with FTLD-ALS with subsequent fine mapping and mutation analyses.
Memory Clinic of the Middelheim General Hospital.
An extended Belgian family with autosomal dominant FTLD-ALS, DR14, with a mean age at onset of 58.1 years (range, 51-65 years [n = 9]) and mean disease duration of 6.4 years (range, 1-17 years [n = 9]). The proband with clinical FTLD showed typical FTLD pathology with neuronal ubiquitin-immunoreactive inclusions that were positive for the transactivation response DNA-binding protein 43 (TDP-43).
Main Outcome Measure
Linkage to chromosome 9 and 14.
We found significant linkage to chromosome 9p23-q21 (multipoint logarithm of odds [LOD] score = 3.38) overlapping with a known FTLD-ALS locus (ALSFTD2) and nearly significant linkage to a second locus at chromosome 14q31-q32 (multipoint LOD score = 2.79). Obligate meiotic recombinants defined candidate regions of 74.7 megabase pairs (Mb) at chromosome 9 and 14.6 Mb near the telomere of chromosome 14q. In both loci, the disease haplotype segregated in all patients in the family. Mutation analysis of selected genes and copy number variation analysis in both loci did not reveal segregating pathogenic mutations.
Family DR14 provides additional significant evidence for the importance of the chromosome 9 gene to FTLD-ALS and reveals a possible novel locus for FTLD-ALS at chromosome 14. The identification of the underlying genetic defect(s) will significantly contribute to the understanding of neurodegenerative disease mechanisms in FTLD, ALS, and associated neurodegenerative disorders.
Frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) are 2 severe neurodegenerative disorders. Frontotemporal lobar degeneration has a prevalence close to that of Alzheimer disease in individuals younger than 65 years,1 whereas ALS is the most common motor neuron disorder.2 Frontotemporal lobar degeneration and ALS belong to an overlapping clinicopathological spectrum of disorders that might have common genetic etiologies.3 Clinically, patients with FTLD present with behavioral, personality, and language disturbances, while memory impairment and other cognitive dysfunctions occur in a later stage. Signs of ALS or parkinsonism may also occur.4 Conversely, a minority of patients with ALS develop dementia and up to 50% show disturbances in executive functions.5,6 Frontotemporal lobar degeneration is caused by degeneration of neurons in frontal and/or temporal lobes, while ALS results from motor neuron degeneration in the motor cortex, brainstem, and spinal cord. Ubiquitinated transactivation response DNA-binding protein 43 (TDP-43) is a major component of pathological inclusions in both tau-negative FTLD (FTLD-TDP) and ALS.7,8 A positive family history is observed in up to 50% of FTLD patients9 and in 5% to 10% of ALS patients,10- 13 indicating a significant genetic contribution to disease etiology. Mutations causing FTLD were observed in genes that encode the microtubule-associated protein tau (MAPT),14 granulin (GRN),15,16 the valosin-containing protein (VCP),17 and the charged multivesicular body protein 2B (CHMP2B),18 while in ALS patients, mutations in the genes encoding superoxide dismutase 1 (SOD1)19 and TDP-43 (TARDBP)20- 24 and FUS25,26 were found, among other genes. Furthermore, several families segregating both FTLD and ALS are linked to the ALSFTD2 locus at chromosome 9p13-21, but the causal gene remains elusive.27- 32 Mutations in these genes explain about 35% of familial patients, indicating that other genes exist. Herein, we performed a genome-wide linkage scan and mutation analyses in an FTLD-ALS family with TDP-43 pathology but without a GRN mutation.
The proband, patient III-12 (Figure 1 and Figure 2), who had FTLD (subtype frontotemporal dementia [FTD]), was contacted for molecular genetic studies. Detailed information on family history was obtained and family members were asked to participate in the study. Clinical information was collected through family informants and medical records from treating physicians. Informed consent was obtained from all 29 participating family members (aged ≥18 years). The research was approved by the local medical ethics committee of the University of Antwerp, University Hospital Antwerp, and ZNA Middelheim. Blood samples were collected for genomic DNA (gDNA) extractions and Epstein-Barr virus transformations.
Segregation of the 9p23-q21 haplotype in family DR14. Haplotypes are based on a selection of 20 informative short tandem repeat markers at chromosome 9. The black haplotype represents the disease haplotype. Haplotypes for deceased individuals were inferred based on genotype data obtained in their offspring (in parentheses). The disease haplotype was arbitrarily set for I-1. For confidentiality reasons, haplotypes are only shown for patients and obligate carriers. Asterisks indicate individuals for whom DNA was available.
Segregation of the 14q31-q32 haplotype in family DR14. Haplotypes are based on a selection of 15 informative markers at chromosome 14. The black haplotype represents the disease haplotype. Haplotypes for deceased individuals were inferred based on genotype data obtained in their offspring (in parentheses). The disease haplotype was arbitrarily set for I-1. For confidentiality reasons, haplotypes are only shown for patients and obligate carriers. Asterisks indicate individuals for whom DNA was available.
A postmortem neuropathological study was performed on the brain of the proband (Figure 1 and Figure 2). After 3 months' fixation in 10% buffered formalin, the brain was embedded in paraffin. Sections of 5 μm were prepared from 3 frontocortical regions and Brodmann area 9, the olfactory bulb and tracts, cingulate gyrus, corpus callosum, superior temporal gyrus, hippocampus, parahippocampal gyrus, lateral occipitotemporal gyrus, occipital cortex with area striata, cerebellum, midbrain, and pons with locus coeruleus. The sections were examined using routine histopathological methods and immunostained using antibody AT8 against hyperphosphorylated tau (Innogenetics, Zwijnaarde, Belgium), 4G8 against residues 17 through 24 of amyloid β (Signet, Dedham, Massachusetts), 3F4 against prion protein (Signet), antiglial fibrillary acidic protein (Dako, Glostrup, Denmark), CD68 (Dako), antiubiquitin (Dako), anti-p62 (Progen Biotechnik), rabbit TDP-43 antisera (Proteintech Group Inc, Chicago, Illinois), and phospho-TDP-43 against 409/410 serines of TDP-43.7 Antigen retrieval and staining were performed as described.33
We performed a genome-wide scan using 425 short tandem repeat (STR) markers, with a mean intermarker distance of 8 cM as described.34 For fine mapping of regions with a multipoint logarithm of odds (LOD) score above 1.9, additional STR markers were selected from the Marshfield genetic map (http://research.marshfieldclinic.org/genetics) or from the Microsatellites track in the University of California–Santa Cruz Human Genome Browser (http://genome.ucsc.edu), based on Tandem Repeats Finder.35 Primers were designed with an algorithm implemented in a polymerase chain reaction (PCR) multiplexer program (primers are available from the authors upon request). Polymerase chain reaction amplification reactions and analyses were performed as described.34
Genes were selected for mutation analysis on gDNA or complementary DNA (cDNA) of 2 patients (III-9 and III-12) (Figure 1 and Figure 2) and 2 married-in control individuals. Epstein-Barr virus–transformed lymphoblasts were treated with 100-μg/mL cycloheximide or with dimethyl sulfoxide for 4 hours prior to RNA isolation. Total RNA was isolated using the Ribopure Kit (Ambion; Applied Biosystems, Foster City, California) and first-strand cDNA was synthesized using the SuperScript III First-Strand Synthesis System for reverse transcriptase–PCR kit (Invitrogen, Carlsbad, California). Primers were designed using Primer 3 (available from the authors upon request).36 Standard PCRs on gDNA amplified exons and exon-intron boundaries with optimized conditions. Amplified cDNA of the entire open reading frame was analyzed for aberrantly sized transcripts by agarose gel electrophoresis. Amplicons of both gDNA and cDNA were purified, sequenced in both directions using BigDye Terminator Cycle Sequencing kit, version 3.1 (Applied Biosystems), and analyzed on an ABI3730 DNA Analyzer (Applied Biosystems). Sequences were analyzed using novoSNP.37 Variations segregating with the disease were tested in at least 90 neurologically healthy control individuals.
Oligonucleotide-based array comparative genomic hybridization (CGH) was performed in the proband and a control individual not carrying the disease haplotype using 385K Tiling arrays (Nimblegen, Madison, Wisconsin) of chromosomes 9 and 14, with a mean probe spacing of 255 base pairs (bp) and 200 bp, respectively. Data were analyzed with SignalMap (Nimblegen). Copy number variations (CNVs) were validated by quantitative real-time PCR using SYBR Green assays on the ABI Prism 7900HT Sequence Detection System (Applied Biosystems) as described previously.38 Confirmed CNVs were tested for segregation in family DR14 using quantitative PCR.
The statistical power of family DR14 to obtain genome-wide significant linkage was assessed using SLINK,39,40 assuming autosomal dominant inheritance with age-dependent penetrance and phenocopy rates. A cumulative risk curve was calculated based on the mean age at onset (58.1 years), and 9 liability classes for disease penetrance were defined with a maximum penetrance of 90%.
Two-point and multipoint parametric linkage analyses were performed using MLINK and LINKMAP from the Linkage package, version 5.2,41 or Simwalk, version 2.91,42 for genome-wide multipoint parametric linkage analysis. All patients with FTLD, dementia, and/or ALS were considered affected, and the phenotypes of individuals I-1, I-2, II-6, and II-7 were considered unknown (Figure 1 and Figure 2). Parameters were the same as in the simulation study and marker allele frequencies were set equally.
At age 57 years, patient III-12 presented with marked behavioral disturbances and personality changes, including agitation, irritability, and compulsive behavior, and experienced forgetfulness. His brain computed tomographic scan showed moderate frontal atrophy. One year later, the patient's personality disturbances had progressively worsened and the patient showed mild impairment of short- and long-term memory, impaired concentration, and impaired verbal fluency. His Mini-Mental State Examination score was 28 of 30. Brain-perfusion single-photon emission computed tomography showed hypoperfusion in the frontotemporal region at both sides, and on brain magnetic resonance imaging severe frontotemporal atrophy was observed (eFigure). At age 59 years, the patient presented with severe wandering (away from home), and his cognitive state had severely declined, as illustrated by a Mini-Mental State Examination score of 3 of 30. He did not show muscle weakness, fasciculations, or hyperactive reflexes. The patient received a diagnosis of familial FTLD (subtype FTD)4 and died at age 60 years.
Macroscopic examination of the autopsied brain demonstrated a generalized cortical atrophy, which was most prominent in the frontal lobe resulting in dilatation of frontal (and temporal) horns of the lateral ventricles. On microscopic examination, marked neuronal loss was observed mostly in cortical layers I and II of the frontal cortical regions, including the prefrontal cortex (Figure 3A), but it was also observed in the cortex of the gyrus temporalis superior and the CA1 segment of the hippocampus. Mild microspongiosis and astrocytic gliosis were present in the cortical layers I and II of the prefrontal cortex and the deep cortical layers, respectively. Except for some very rare neurofibrillary tangles in the hippocampus, AT8 staining did not reveal neurofibrillary tangles or Pick bodies in any other region analyzed (Figure 3D), and in most regions 4G8 staining showed some focal accumulations of diffuse or senile amyloid β plaques (Figure 3E), but amyloid angiopathy was absent. These observations, accompanied by corpora amylacea in these regions, most likely represent age-dependent phenomena. Immunoreactivity with the antiprion antibody was negative.
Brain pathology of proband III-12 of family DR14. A, Hematoxylin-eosin staining showing marked neuronal loss and microspongiosis of the superficial cortical layers in the frontal cortical region that was most affected (asterisk); layers II/III of this region were further analyzed by immunohistochemistry (B-H). B, Small cytoplasmic inclusions were relatively common (arrows) (ubiquitin staining). C, Cytoplasmic inclusions were rim shaped (lower right arrow), and very often also well-formed, large, globular aggregates were present in the cytoplasm (upper left arrow) (ubiquitin staining). AT8 reactivity was absent in these areas (D), and very rarely, age-related amyloid plaques were observed (E, amyloid β). F, Small cytoplasmic inclusions were p62-positive (arrows). G, Transactivation response DNA-binding protein 43 (TDP-43) reactivity was also evident in cytoplasmic inclusions (arrows) as well as in parenchyma with a fine granular staining pattern. Characteristic clearing of the nuclei of inclusion-bearing neurons is visible. H, Phospho-TDP-43 also stained the rimlike or small granular cytoplasmic inclusions (arrows) as well as fine granules in the parenchyma.
Ubiquitin staining frequently demonstrated a small number of rim-shaped, granular, or occasionally large globular ubiquitin-positive neuronal cytoplasmic inclusions in the superficial cortical layers, especially of the frontal neocortex (Figures 3B and C). In addition, small ubiquitin-positive neuritic profiles were observed in the affected neocortical layers. Immunoreactivity of p62 was observed in a subset of neuronal cytoplasmic inclusions but never in the nucleus or neurites (Figure 3F). Immunostaining with TDP-43 and phospho-TDP-43 antibodies also showed a strong presence of different neuronal cytoplasmic inclusions in the superficial cortical layers with clearing of nuclei of inclusion-bearing neurons as well as short neuritic profiles or dot-like structures (Figures 3G and H). Although the observed FTLD-TDP pathology did not completely fit into one of the described FTLD-TDP subtypes,43,44 the presence of neuronal cytoplasmic inclusions without neuronal intranuclear inclusions was more typical for type III.43
Family DR14 was ascertained through its proband, individual III-12, who initially had been referred to our Molecular Diagnostic Unit for genetic testing of FTLD genes. No mutations were observed in MAPT, GRN, VCP, or CHMP2B. Informed consent was obtained for further genetic testing, and mutations were also excluded in the presenilin genes PSEN1 and PSEN2; exons 16 and 17 of the amyloid precursor protein gene (APP); the coding exon 2 of PRNP; TARDBP; and the intraflagellar transport 74 homologue gene (IFT74).29
Genealogical analysis identified a 4-generation family that showed indications of autosomal dominant transmission of dementia (Figure 1 and Figure 2). In total, we collected DNA from 29 family members, including 3 patients in generation III and 11 at-risk individuals in both generation III and IV. The mean age at onset was 58.1 years (SD, 4.2 years [n = 9]; range, 51-65 years) and mean disease duration was 6.4 years (SD, 4.9 years; range 1-17 years). Two patients (III-2 and III-12) (Figure 1 and Figure 2) had dementia specified as FTLD (subtype FTD), while patient III-10 had a diagnosis of ALS. The clinical characteristics of the 9 patients are summarized in Table 1. Of individuals I-1, I-2, II-6, and II-7, no or very limited clinical information was available. In individuals II-6 and II-7, some suggestive symptoms pointed toward an (unconfirmed) ALS phenotype, including swallowing problems and loss of muscle strength.
With an assumed hypothetical marker with 4 equifrequent alleles and a disease allele frequency of 0.01%, simulation analyses indicated an average maximum LOD score of 2.45 at recombination fraction (θ) 0.04. The mean maximum false-positive LOD score was 2.26 (θ = 0.5).
Multipoint LOD scores were calculated for markers of the genome-wide linkage scan (Figure 4). At 3 loci, we obtained a LOD score above 1.9, which is compatible with suggestive linkage45: one at chromosome 9 with a maximal LOD score of 2.71 between D9S1121 and D9S270, one at chromosome 14 with the highest LOD score of 2.61 between D14S302 and D14S611, and one at chromosome 11 with a maximal LOD score of 1.99 between D11S4464 and D11S4410.
Multipoint logarithm of odds (LOD) scores of the genome-wide linkage scan. The curves represent multipoint LOD scores under a dominant inheritance model with age-dependent penetrance. The dark horizontal line indicates the cutoff value of 1.9 for suggestive linkage.
At chromosome 9, 31 additional STR markers were analyzed, in an interval of 80.7 cM (85.9 Mb) between D9S2169 and D9S253 (eTable 1). A maximal 2-point LOD score of 2.82 was obtained with D9S171 (θ = 0.0), and a maximal multipoint LOD score of 3.38 was calculated in the region between D9S1833 and D9S1121 (Table 2). Haplotypes were reconstructed for all individuals, and segregation analysis defined a candidate region of 64.6 cM (74.7 Mb) between D9S235 and D9S257 at 9p23-q21. Obligate meiotic recombinants defining the minimal interval were observed in patient III-9 (Figure 1).
At chromosome 14, 17 additional STR markers were genotyped in an interval of 41.8 cM (17.9 Mb) between D14S1005 and the 14q telomere (deTable 2). The marker D14S302 reached the highest 2-point LOD score of 2.45, and a maximal multipoint LOD score of 2.79 was obtained between D14S1030 and D14S987 (Table 3). Based on an obligate meiotic recombination in patient II-5 between D14S1015 and D14S973, the disease haplotype was delineated to a region of a maximal length of 33.2 cM (14.6 Mb) at 14q31-q32 between D14S1015 and the 14q telomere (Figure 2). Individual II-7 carried only part of the haplotype owing to a telomeric recombinant between D14S985 and D14S1051. However, his disease status was unclear.
At chromosome 11, 11 additional STR markers were analyzed in an interval of 42.0 cM (21.0 Mb) between D11S897 and D11S4112. MFD316 reached the maximal 2-point LOD score of 1.73, and the highest multipoint LOD score of 2.18 was obtained in the interval between MFD316 and D11S4464 (Table 4). Haplotype analysis showed a disease haplotype with a maximal length of 11.0 cM (7.0 Mb) at 11q23-q25 between D11S1998 and D11S933, defined by obligate meiotic recombinants in patients II-5 and III-3.
Because of the significant or nearly significant linkage at chromosomes 9p23-q21 and 14q31-q32, we prioritized genes in these 2 loci. Positional candidate genes were selected from the National Center for Biotechnology Information Reference Sequence (RefSeq) database of protein-coding genes. The chromosome 9p23-q21 locus in DR14 harbored 265 genes. Because this region did not reduce the known ALSFTD2 locus at 9p13-p21, we based the selection of genes on published segregation data in chromosome 9–linked FTLD-ALS families with a multipoint LOD score above 3.0.27,28,30- 32 We defined a minimal region of 6.96 Mb between D9S16928 and D9S180531 that segregated in all these families and contained 27 protein-coding genes. Based on functions, protein homology and interactions with known FTLD genes, we analyzed the 17 best candidate genes by sequencing of the complete coding sequence of cDNA from lymphoblasts treated with or without cycloheximide, allowing the detection of putatively degraded aberrant transcripts. These 17 genes included MOBKL2B, C9orf72, ACO1, DDX58, TOPORS, NDUFB6, DNAJA1, SMU1, B4GALT1, BAG1, CHMP5, AQP3, NOL6, UBE2R2, UBAP2, WDR40A, and UBAP1. We did not identify novel variants segregating on the disease haplotype.
The candidate region at chromosome 14q31-q32 harbored 121 protein-coding RefSeq genes. Sixty-six genes located in the region between D14S1015 and D14S1051defined in individual II-7 (Figure 2) were prioritized for exon-based genomic sequencing. We identified 9 novel variants segregating on the disease haplotype; however, all were present in at least 1 control individual (Table 5). The Gly191Ala variant in YY1 and the IVS5-43G>C variant in SERPINA9 were found in only 1 of 300 and 1 of 360 control individuals with age at last examination of 83 and 77 years, respectively; no positive familial history of dementia was described.
We performed high-density array CGH at chromosomes 9 and 14 that covered the complete candidate regions. In the chromosome 9 locus, we identified 2 large CNVs (chromosome 9: 24492891-24508926 and chromosome 9: 29082732-29087816) covered by 64 and 20 CGH probes, respectively. Only the deletion of the second region was confirmed in the proband by 6 quantitative PCR fragments that demonstrated a deleted region of at least 5273 bp (chromosome 9: 29082677-29087949) (data not shown). This deletion did not segregate with disease in DR14 and represented a polymorphism, because it was also present in individuals not carrying the disease haplotype and a frequent CNV had previously been reported at this position (chromosome 9: 29082445-29088195).46 The chromosome 14 locus did not show the presence of a large CNV except for a known CNV region in the immunoglobulin heavy chain gene family cluster near the 14q telomere.
We identified a multigenerational autosomal dominant FTLD-ALS family, family DR14, of whom the proband presented clinically with FTD and neuropathologically with FTLD-TDP. We showed significant linkage to chromosome 9p23-q21 overlapping with the known ALSFTD2 locus and nearly significant linkage to a novel locus at chromosome 14q31-q32. At both loci, the disease haplotype segregated in all patients (Figure 1 and Figure 2). Also, an asymptomatic individual in generation III, aged 76 years at last examination, carried the complete chromosome 9 disease haplotype, possibly suggesting reduced penetrance of the mutation. In addition, the current ages of asymptomatic at-risk individuals were within or below the wide age at onset range (individual data not shown for confidentiality). Therefore, recombinants in these individuals were not accounted for to define the candidate regions. Alternatively, because the asymptomatic carrier of the chromosome 9 haplotype did not carry the chromosome 14 disease haplotype, the chromosome 9 mutation is possibly insufficient to cause the disease, implicating a digenic effect by which the disease will only be fully expressed if mutant genes at both loci are present in the same individual. The latter is supported by the observation that all affected individuals carry both the chromosome 9 and 14 disease haplotypes. Another potential explanation could be that a mutation in a gene in the one locus has a modifying effect on the disease-causing mutation at the other. Alternatively, 1 of the 2 loci might be a false-positive finding due to coincidental segregation even if both loci reached a near equal level of genetic significance. The slight difference in the maximum LOD scores is not essential, because they largely depend on the allele frequencies in the control population. Owing to accumulating evidence for an FTLD-ALS locus at 9p23-q21 and because family DR14 has a similar clinicopathological phenotype than other chromosome 9–linked families,27- 32 one might reason that the chromosome 14 linkage is more likely false.27- 32,47
At chromosome 9, in a minimal region shared between chromosome 9–linked families,27,28,30- 32 we excluded in 17 genes the presence of simple mutations, exonic insertions or deletions, and detectable altered splicing by transcript analysis. Currently, all 27 RefSeq genes located in this minimal region have been analyzed for mutations on gDNA and/or cDNA in 1 or more families.28,29,31,32 In addition, we excluded large CNVs in the complete DR14 candidate region. However, mutations might still have been missed because most candidate genes were either exclusively screened on gDNA or in families not conclusively linked to chromosome 9.29 Also, a noncoding mutation in 1 of the screened RefSeq genes or a mutation in a non-RefSeq gene or outside an annotated gene cannot be excluded. Furthermore, the potential of phenocopies within these families and subsequent false meiotic recombinants makes it possible that the disease gene is actually located outside the delineated minimal candidate region. Alternatively, although the increasing number of families with similar phenotypes linked to chromosome 9p supported the idea that 1 common gene might be responsible for the occurrence of both FTLD and ALS, the possibility remains that the FTLD and ALS symptoms are the result of 1 larger genomic rearrangement affecting 2 different disease genes. The array CGH applied here could only detect CNVs of at least 1 kb, while inversions or small insertions or deletions could not be detected at all. Performing other techniques, including macrorestriction mapping, would provide more information on the presence of more complex rearrangements.
In the chromosome 14q31-q32, locus 66 genes were excluded for the presence of simple mutations in the coding exons and exon/intron boundaries. We selected these genes because of their localization within the candidate region delineated by a centromeric recombinant in patient II-5 and a telomeric recombinant in individual II-7 (Figure 2). In the latter patient, the disease status was uncertain, though it suggested ALS. Therefore, the remaining 55 RefSeq genes between the telomeric recombinant and the 14q telomere might still carry the mutation. Large CNVs were excluded in the complete candidate region by array CGH. Other types of mutations or mutations outside the screened sequences are still possible. Also at chromosome 11q23-q25, we observed suggestive linkage where we did not yet prioritize genes for mutation analysis because of its lower significance.
In conclusion, additional mutation analyses in both loci are needed to identify the underlying genetic etiology of the disease in family DR14. Although we did not reduce the known ALSFTD2 locus, this family provided supportive evidence for an important contribution of the chromosome 9 gene to FTLD and/or ALS and a novel disease-linked locus at chromosome 14q31-q32. We expect that the disease gene(s), once identified, will significantly contribute to understanding the neurodegenerative disease mechanisms in FTLD, ALS, and related diseases.
Correspondence: Marc Cruts, PhD, Neurodegenerative Brain Diseases Group, VIB–Department of Molecular Genetics, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium (firstname.lastname@example.com).
Accepted for Publication: October 8, 2009.
Author Contributions: Dr Cruts had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. Study concept and design: Gijselinck, Van Broeckhoven, and Cruts. Acquisition of data: Gijselinck, Engelborghs, Maes, Cuijt, Peeters, Mattheijssens, Joris, Cras, Martin, De Deyn, Kumar-Singh, Van Broeckhoven, and Cruts. Analysis and interpretation of data: Gijselinck, Engelborghs, Maes, Cuijt, Joris, Cras, Martin, De Deyn, Kumar-Singh, Van Broeckhoven, and Cruts. Drafting of the manuscript: Gijselinck, Van Broeckhoven, and Cruts. Critical revision of the manuscript for important intellectual content: Gijselinck, Engelborghs, Maes, Cuijt, Peeters, Mattheijssens, Joris, Cras, Martin, De Deyn, Kumar-Singh, Van Broeckhoven, and Cruts. Obtained funding: Van Broeckhoven. Administrative, technical, and material support: Gijselinck, Engelborghs, Maes, Cuijt, Peeters, Mattheijssens, Joris, Cras, Martin, De Deyn, and Van Broeckhoven. Study supervision: Van Broeckhoven and Cruts.
Financial Disclosure: None reported.
Funding/Support: This research was funded in part by the Special Research Fund of the University of Antwerp, the Fund for Scientific Research Flanders, the Institute for Science and Technology–Flanders, the Methusalem Excellence Grant of the Flemish Government, the Interuniversity Attraction Poles program P6/43 of the Belgian Science Policy Office, the Stichting Alzheimer Onderzoek, and the Association for Frontotemporal Dementias. Drs Gijselinck and Engelborghs hold postdoctoral fellowships from the Fund for Scientific Research Flanders.
Additional Contributions: We are grateful to the DR14 family members for their cooperation. We further acknowledge the contribution of the personnel of the VIB Genetic Service Facility and the Biobank of the Institute Born-Bunge. We also thank Masato Hasegawa, PhD, for donating the phospho-TDP-43 antibody.
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Created in this Craftsy class
Jewelry Workshop: Bead, Wrap, Chain & Etch taught by Max Goodman
Build a broad skill base for getting started making beautiful, timeless jewelry. This is the perfect class for beginners to test many different types of techniques, from beading to wire wrapping and beyond.
Other projects made as part of this class:
JewelryFlower Bracelet
JewelryFirst pearl knotting project
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nice work!
04/14/2014 Flag
Thank you. It's my first piece with loops that are different sizes and very few are round and centered. But I'm proud that I finished it and I will definitely wear it.
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Well most of us here find eating the higher fats, keeps us satisfied for much longer, so of course eat less.
What are your fat sources?
Starvation mode is a CW thing, when you are actually starving and not eating.
Not sure on PCOS and fasting, but fasting I feel great.
But I would not attempt to try until fully fat adapted, or you will probably get hungry and not feel great.
I tried it last year, but was not ready and it made me binge.
I am up to nearly 40 hours of a half fast (bulletproof coffee and bone broth only) and I still feel good. Hopefully someone will PCOS will answer this for you soon.


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Child of Eden Screenshots for Xbox 360On: Xbox 360, PS3
Child of Eden is a multi-sensory shooter that will send players diving into a kaleidoscopic matrix of synchronized music and mind-blowing visuals that will usher forth a landmark game experience.
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8Out of 10Back to game info
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Rez creator Mizuguchi wants to start new game project this year or next
Is thinking about what's next and what's the future experience.
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Rez and Child of Eden creator left studio last year, report claims.
No downloadable Child of Eden for now, says Ubisoft
No downloadable release for Child of Eden is currently planned, according to Ubisoft's senior VP of sales and marketing Tony Key.
New Kinect bundle revealed, includes Child of Eden
Microsoft has announced that it will be launching a new Kinect for Xbox 360 bundle, which will be arriving in stores as soon as next week.


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What is Google Looking For?
When people ask me conceptually about the effect of certain elements on search engines, I often respond, "If you were a search engine, what would you think of it?" It's a hard question to answer if you don't know what they are looking for. Here's the low-down on search engines from the eyes of Google. .
Google is a Business
First and foremost, Google is a business. They do what they do because it helps them turn a profit. The fact that the majority of people view Google as a benevolent organization stems from great user experiences. And it's true–they allow us to use their great products for free in exchange for our usage data.
So how do they make their money? Since they give away all their products for free, how can they fund the development? Many of you are whispering the answer right now…PPC. Google made 98% of their revenue last year from pay-per-click advertising. That means there is a disconnect somewhere – how does providing excellent user experience help Google make money? Easy. Make the user experience for all of your users so awesome that they will want to spend all of their time using your products. The more time you spend on Google software, the higher the chance you're going to click on an ad, which gives them cash.
Google Mimics Your Behavior
In order to provide the best user experience, the Goog wants to know how you use the web. They track usage data from everyone who has Google Analytics installed, from your Gmail box, and from information about how you search. It all belongs to them. They use it for the purpose of making your experience faster, cleaner, and more pleasant…and possibly for taking over the world.
How does that reflect in SEO? Take, for instance, the idea that links which are further up the page pass more authority. They pass more authority because the further up a link is on the page, the more of a chance that the users are going to have to click on it. A link in the footer or sidebar where advertisements traditionally reside isn't going to get much play; therefore, the links will be worth less to the target domain.
Google Wants to Rid the World of Spam
My guess is that with all of Google's spider overhead (servers, power, maintenance, storage, etc), plus the lost opportunity cost of crawling and indexing new, fresh content elsewhere, Google's time is probably worth a few thousand dollars per second. Spam wastes everyone's time.
Spam takes up valuable space in Google's index. It forces them to update their algorithm over 500 times per year. 500. And just in case you missed that, there are 365 days in a year, so the algorithm is updated every 16 – 18 hours (approximately). Spam makes the user experience poor. For these reasons, spam is the antithesis of Google's mission.
Next time you are considering a tactic, ask yourself:
- Does it provide bad user experience?
- Does it waste Google's time?
If you answered yes to either of these two questions, you can rest assured that one of this year's 500 updates will send you packing.
So right !! For the little time that I have been on the internet staying away from spam can only help you. If just everybody would litsen and quit trying to beat the system how nice would that be. Thanks for putting the word out No Spam !!!!
Good info AJ. I think there is disconnect between the way business owners view the web vs. their website. We know businesses spend a lot of money and time to give a good customer experience in the store, they should approach the web the same way.
Oh, and I'm sure that Google is working on ways to take over the world right now.
hey a.j., do you have a reference for the "98% of revenue" stat?
[…] every time Google makes a change. From the beginning, we've always asked ourselves: "What is Google's goal?" Google's goal is to provide users the most relevant and best search results possible. So […]


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Star Tracks: Monday, May 16, 2016 42 years, 2,191 covers and 55,436 stories from PEOPLE magazine's history for you to enjoy
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People Top 5
LAST UPDATE: Tuesday February 10, 2015 01:10PM EST
PEOPLE Top 5 are the most-viewed stories on the site over the past three days, updated every 60 minutes
- January 25, 1999
- Vol. 51
- No. 3
Brave Hearts
Brits Rejoice at Prince Edward's Choice, a Commoner with Uncommon Good Sense. But Can She Handle the Royal Glare?
Then she reached the door of the offices of R-JH (for Rhys-Jones Harkin), the public relations company she cofounded in 1997, and discovered that, inexplicably, it wouldn't open. As reporters watched, Rhys-Jones fumbled with the lock before ringing the bell and, a hint of panic in her voice, implored through the intercom: "Will somebody let me in?" (Somebody did.)
You may have to get used to the cold shoulder, Sophie. Cracking the code to surviving in a family strewn with bitter marital baggage is not going to be easy. The Windsors' sink-or-swim approach toward outsiders (daughters-and sons-in-law, that is) was not something they picked up in charm school. "I do think the treatment that Diana and Fergie received from the royal household contributed to the breakup of their marriages," says royals expert Judy Wade. "You're not a person when you marry into the royal family. You're a chattel." So much for fairy-tale marriages.
If anyone has a sporting chance of getting it right, though, it just might be Sophie. In the five years she has dated the Queen's youngest son, this down-to-earth daughter of a tire salesman (Christopher) and a homemaker (Mary), who turns 34 on Jan. 20, has gone riding with the Queen at Sandringham, had regular sleepovers at Buckingham Palace and learned the difference between a full curtsy and a bob. Still, when the couple announced their engagement on Jan. 6 in the garden at St. James's Palace—following her demure "Yes, please" response to Edward's proposal—Rhys-Jones admitted to some trepidation over her new role. Then she added, "I think I am ready for it."
Time will tell. "There's so much intense media scrutiny, especially in the beginning," says British Press Association royals reporter Peter Archer. "It's a real test." Even more so since Edward, seventh in line to the throne, is the only one of the Queen's four children to not yet bungle a marriage (see box, page 89).
For Sophie, there's also the Princess Diana lookalike problem. Described as "Di-dentical!" by British tabs from the moment she appeared at Edward's side in 1993, Rhys-Jones is constantly likened to the late Princess (though at about 5'5" she couldn't possibly fill the void left by the 5'10" Di). Hoping to quash the comparisons, Sophie reportedly made a request to the Queen recently that, when she and Edward marry, she not be referred to as the Princess Edward, the graceless title she's slated for. (Her concern will be moot if, as most royal experts predict, the Queen makes Edward a duke and Sophie gets the title of duchess.)
Despite the downsides, Sophie seems delighted with her choice of life partner. "I've never seen her so happy," says Murray Harkin, her friend and business partner. Others say her prince is equally smitten and has been from the start. "If I ever could have chosen somebody who would be perfect for Edward," says Malcolm Cockren, chairman of Ardent Productions, Edward's TV production company, "it would be Sophie."
Those who knew Rhys-Jones back in Brenchley, Kent, the village 40 miles southeast of London where she grew up, would hardly have predicted it. A bright but unremarkable student, Sophie, along with her brother David (now 35 and an insurance agent), attended local private schools—thanks to extra money her mother earned taking on home typing jobs—before enrolling in a secretarial course at "West Kent College. Marion Vellino, now 50, taught Rhys-Jones shorthand and typing. "I was very fond of Sophie—she was a lively young lady," Vellino remembers. "But if you had told me she was going to marry a prince, I would have said, 'Pull the other one!' "
If former classmates don't recall her as a knockout—"All of us have said how surprised we are at how she has blossomed," says Louisa Feldhaus, now a Sussex homemaker—men always noticed her. "She was a very attractive girl," says retired farmer Michael Noakes, 56, who frequented the Halfway House, a Brenchley pub Sophie worked at when she was 18. "I wished I was 20 years younger myself. And she pulled a good pint."
At age 18, Rhys-Jones moved to London, where she landed a series of odd jobs—and beaus who didn't last—before leaving the city for stints in Switzerland and Australia. Returning to London in 1992, she entered the public relations field. She was helping put the likes of Thomas the Tank Engine and Mr. Blobby (Britain's answer to Barney) on the publicity map when she was hired to promote an August 1993 real-tennis (an old-fashioned form of the game) charity event supported by Edward. "Nobody knew that there was an attraction between them that day," recalls real-tennis champion Sally Jones. "But Edward always had an eye for a bright girl—and Sophie is bright, sparky and fun."
Shortly after, the prince and the PR girl began dating—riding at Windsor Castle, windsurfing, playing tennis or barbecuing together. Friends knew it was serious when the low-profile Edward, who uses "Edward Windsor" as his business name, complained to Britain's press commission when he and Sophie were snapped by photogs exchanging a chaste peck on the grounds of Balmoral in 1994. By 1995, Sophie was abandoning her flatmate in West Kensington several nights a week to bunk at Buckingham Palace, where—to the dismay of the Archdeacon of York, who denounced the Queen for setting an immoral example—she was given a room adjoining Edward's. "If they couldn't be together there, they wouldn't have had any privacy at all," notes Majesty magazine editor-in-chief Ingrid Seward, author of a 1995 biography of the prince.
The Queen's decision to sanction what amounted to cohabitation under her roof—license she gave to neither Diana or Fergie—no doubt stemmed from her determination that future wives of Windsor see what they were getting into. A girlfriend in the house also helped quell rumors that Edward, despite his own string of romantic attachments, was gay. (The rumors began circulating just before the prince took a production job with Andrew Lloyd Webber's Really Useful Theatre Company in 1988. Edward publicly denied them in 1990 as being "so unfair to me and my family. I am not gay.")
The cozy quarters at the palace also helped familiarize the new recruit with the royal fold. "She was terrified when she first met the Queen," Sophie's friend Judy Ledger recalls. "But the Queen has been wonderful to her. And she gets on well with Prince Philip because she likes his dry sense of humor." As for Edward, unlike his brother Charles, he has taken care to make sure the woman he loves doesn't feel overwhelmed. "Sophie said he would call to make sure she hadn't had problems after she went out in public," Ledger says. "That's one of the things I think will make the relationship succeed—they have a good friendship, and they talk."
That's not all that bodes well. Sophie and Edward are both devoted to their work. His six-year-old Ardent Productions, which specializes in historical documentaries, is expected to turn a profit this year, and R-JH, at which Sophie plans to continue working after her marriage, has amassed a roster of major clients, including posh jewelers Boodle & Dunthorne and Thomas Goode, purveyors of silver and glassware. "She's very media-aware, since that's what her job is," an observer points out. "And that's most of the battle [in the royal family] nowadays."
At the moment, everything about Prince Edward's future wife makes news: her evolving style (she has gone from Sloane Ranger skirts and scarves to smart, conservative suits by designer Tomasz Starzewski), her relatively low-key wedding plans (she and Edward hope to marry in early summer at St. George's Chapel at Windsor Castle) and, of course, her wedding dress. "If I was doing it, I would make it sleek, not fussy," says Jon Moore, chief designer at the Hardy Amies fashion firm. "I usually say to brides, if they can lay their hands on a tiara somewhere, it's a good opportunity to wear one. There might be someone in the family who could lend Sophie one!"
Once the newlyweds move into their home, Bagshot Park—Edward's 87-acre Surrey estate, now undergoing a $3 million renovation—the hoopla should die down. And while Sophie's ordinary roots have some pundits spelling doom ("Every time one of the royals marry a commoner, they get it wrong," sniffs James Whitaker, royals commentator for Britain's Mirror), veteran royals photographer Jayne Fincher, for one, predicts smooth sailing. "I've photographed Sophie on a number of occasions," she says, "and she is more levelheaded than Diana. She is a nice, unassuming, good fun, sensible girl. They will be fine."
Kim Hubbard
Simon Perry, Nina Biddle, Joanna Blonska and Ellen Lieberman in London
- Contributors:
- Simon Perry,
- Nina Biddle,
- Joanna Blonska,
- Ellen Lieberman.
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Featured Golf News
Kiawah Island's Ocean Course Will Be No Beach Vacation
Unless Tiger Woods, Phil Mickelson or one of the 20 PGA professionals in the field shoots three straight 63s establishing a 10-stroke lead, it's likely the most intriguing story prior to the final round of the 94th PGA Championship will be the venue.
The Ocean Course at Kiawah Island, 20 miles south of Charleston on the shores of South Carolina's Lowcountry, would be worthy of superlatives even if it hadn't been built quite so quickly. That it was designed, shaped, seeded and groomed for major international competition in just 25 months - less if you take into account the time lost clearing up and recreating the coastline after Hurricane Hugo blasted through two months after construction began - surely makes it one of the most remarkable courses in the world.
Architect Pete Dye dug his first bucket-load of soil, sand and salt from the dunes and marshes on the eastern end of the island in July 1989, a few months following the PGA of America's decision to move the 1991 Ryder Cup from the original venue - PGA West in Palm Springs, Calif. - to the East Coast in order to avoid the desert heat and so European audiences could tune in during prime time.
Dye and his wife Alice, whose idea it was to raise the fairways to, or above, dune level - thus subjecting players to the full brunt of offshore winds from the south and west, got the job done in good time, and the tantalizing layout definitely played its part in producing the most exciting finish in Ryder Cup history. A contest that came down to the final putt on the final hole of the final singles match would obviously have created a great deal of excitement wherever the matches were played - even the Belfry (the 14-14 tie that happened there in 1989 wasn't quite as enthralling as the finish in the '91 matches). But the spectacular ocean setting at Kiawah certainly added to the drama of the occasion.
It would be fair to say the Ocean Course has flown under the radar somewhat since the unfortunately-named "War by the Shore" 21 years ago. Yes, it is now part of a Golf Digest top-20 resort thanks to the opening of the highly acclaimed Sanctuary Hotel in 2004, and is ranked by the same magazine as America's toughest - and 26th best - course. Yes, it was featured in the movie "Bagger Vance" and was the first course made available for gamers on worldgolftour.com. Yes, it hosted a "Shell Wonderful World of Golf" match between Annika Sorenstam and Dottie Pepper in 1996. And yes, it has hosted two World Cups (1997, 2003), one PGA Professional National Championship (2005) and a Senior PGA Championship (2007, won by Denis Watson).
But after what happened in 1991 only the very highest rankings and most prestigious events are going to move Kiawah's needle and make it part of a global conversation again.
The ocean views, huge sandy waste areas, undulating fairways and greens, and exposure to the wind characterize this endlessly captivating course. There are 18 distinct, well-defined holes, any one of which can ruin a player's scorecard and his hopes of winning; these are 18 quite separate tales, romantic or tragic, coming together to create a grueling and utterly exhausting compilation.
Because the holes are so singular and easily recognizable, many regard the course as more suitable for the thrill of 18-hole match play than the steady grind of 72-hole stroke play. Also working against it is the fact that Tuesday's torrential rain and the isolated/scattered thunderstorms forecast for every day of the tournament will soften the ground, making the course play more like a regular PGA Tour stop than a firm-and-fast major championship venue.
The combination of soft fairways and immoderate length - 7,676 yards, which makes it the longest course ever used for a major - suggests the group of potential winners might well be reduced to only the mightiest of hitters. But then they said that about a lengthened Augusta National in 2002 and '05 since when Mike Weir, Zach Johnson and Trevor Immelman have each won a green jacket.
The Ocean Course is a tough test even when calm, but as with most courses in this day and age it will need a little help from the heavens to defend itself against the best players in the world and all their made-to-measure, turbo-charged equipment. Watson won the '07 Senior PGA with a four-round total of 9-under 279 in pleasant but gusty conditions. If it blows hard this week, very few, if any, in the field will get remotely close to that number. But with yielding greens and the forecast 5-10mph breezes, several players might equal or surpass it.
The question though is who will best take advantage of a course that has lost its sting and go the lowest.
Until last week, the reigning PGA champion Keegan Bradley, whose victory at Atlanta Athletic Club 12 months ago came at the first major he ever played in, was having a slightly disappointing 2012. In his second, third and fourth major championship appearances, he finished T27, T68, and T34, and he had missed three cuts elsewhere. It hadn't been a disastrous second-year on the PGA Tour for the Vermont native by any means, but after two wins and a fifth-place finish in the FedEx Cup standings in 2011, he was justified in expecting a little better.
On Sunday, however, the 26-year-old New Englander won the WGC-Bridgestone Invitational in Akron with a brilliant six-birdie-no-bogey 64. And with that, Bradley thrust himself into the reckoning for a successful defense of the Wanamaker Trophy, something only Tiger Woods has achieved (twice - '99 and '00; '06 and '07) since the tournament became a stroke-play event in 1958.
Woods himself is the clear favorite and, at 10-1 odds, a very attractive bet considering he tops the PGA Tour money list, FedEx Cup standings, U.S. Ryder Cup Team standings, PGA Tour scoring average standings, and has the best GIR and accuracy stats on the PGA Tour from 175 to 200 yards, a distance from which might hit as many as nine approaches per round this week. Woods's record in inclement weather has never been terribly impressive, but if he can avoid the storms and whatever wind does show up, he may be difficult to beat. The strategic options of Dye's typically thought-provoking layout will appeal to the world No. 2, who united sound course management and decision-making with superior ball-striking in his 14 major championship victories.
Sitting one place ahead of Woods in the world rankings, Luke Donald still remains major-less after 36 attempts and a total of 55 weeks as the world No. 1. The consensus is the Englishman doesn't possess the firepower to hit Kiawah's four lengthy par-5s in two or threaten birdies at the five brutishly long par-4s. But the Copperhead Course at the Innisbrook Resort in Florida, where he won this year's Transitions Championship, and the West Course at Wentworth, where he has won the last two BMW Championships, aren't exactly short. Indeed, who's to say Donald won't be to Kiawah Island what Weir, Johnson and Immelman were to Augusta? And with the desire to emulate the Great Britain Olympics team performing so well in London, Donald has extra motivation in capturing the title that may yet justify his world ranking to the doubters.
Ernie Els might be a new, old golfer following his Open Championship win and earn the title that pushes him one step closer to a career Grand Slam, and what about Martin Kaymer, the last man to win the PGA Championship on a Pete Dye design? The German won in 2010 at Whistling Straits, which shares several characteristics with the Ocean Course. Kaymer wasn't in the best of form entering the 2010 tournament, but he went on a tear after his victory winning his next two events and his first of the 2011 season, eventually climbing to the top spot in the world golf rankings.
The form book might mean very little at a place like the Ocean Course, to which some in the field might take an instant dislike while others feel inspired by its many unconventional challenges. On Sunday, the winner will hoist the trophy and be able to say he beat what the PGA of America claims is the strongest field of the year. But it remains to be seen if he can master the Ocean Course and beat Old Man Dye.
Tony Dear is an Englishman living in Bellingham, Wash. In the early 1990s he was a member of the Liverpool University golf team which played its home matches at Royal Liverpool GC. Easy access to Hoylake made it extremely difficult for him to focus on Politics, his chosen major. After leaving Liverpool, he worked as a golf instructor at a club just south of London where he also made a futile attempt at becoming a 'player.' He moved into writing when it became abundantly clear he had no business playing the game for a living. A one-time golf correspondent of the New York Sun, Tony is a member of the Golf Writers Association of America, the Pacific Northwest Golf Media Association and the Golf Travel Writers Association. He is a multi-award winning journalist, and edits his own website at www.bellinghamgolfer.com.
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Gov. Bobby Jindal made a visit to two southeast Louisiana parishes Friday to hand out millions of dollars in federal grants for hurricane protection and other recovery projects.
St. John the Baptist Parish received $8.7 million, and Plaquemines Parish got $2.4 million for hurricane recovery and hazard mitigation, Jindal announced during his "Louisiana Working Tour."
"Right after Gustav, one of the commitments I made then was that we weren't going to create the kind of bureaucracy the state did in 2005, " Jindal said. "We weren't going to create a Road Home program, we weren't going to create all these rules and red tape, but instead, the second the state got recovery dollars, we'd give those directly to the parish."
St. John's $8.7 million is part of three grants that were awarded to the parish during a River Region Chamber of Commerce luncheon at the St. John Civic Center in LaPlace.
|Governor Bobby Jindal video July 31, 2009
St. John Parish President Bill Hubbard said the money also will go toward infrastructure improvements and flood prevention.
Hubbard already has one project in mind.
Hubbard, who last year asked Jindal for money to start a project that would use flap valves to prevent storm surges from flooding neighborhoods said last September's wishes could be a reality.
Several streets throughout the parish were flooded after Hurricane Ike, so much so that exits on Interstate 10 were closed because of the rising water.
Plans are to install the valves on main drainage canals near the interstate. The valves, which would let water flow in one direction, would cost about $4 million, Hubbard said.
He said the parish will hold town hall meetings, the first of which will be held Thursday, to get public input on how the money will be spent.
A.J. "Pete" Landry of LaPlace said he's glad Jindal gave money directly to the parishes.
"We have the capability to use it in the areas where we know what our priorities are, " Landry said. "We can put it to work where our needs really are."
Jindal spent the latter part of the afternoon at the Belle Chasse Public Library in Plaquemines Parish addressing the Plaquemines Association of Business and Industry, during which he signed a $2.4 million ceremonial check to parish officials for two grants for hurricane recovery and hazard mitigation.
The first grant, totaling $1.92 million, will aid the parish's recovery efforts for housing and infrastructure damage from Hurricanes Gustav and Ike. The second, a $491,404 grant, will support local projects that will help prevent damage from future hurricanes.
"It certainly allows us to help rebuild some of that damage from Gustav and Ike, but even more importantly, it allows the parish to start getting ready for future storms, " Jindal said. "I'm praying for a quiet storm season. We don't need a busy storm season."
. . . . . . .
Victoria St. Martin can be reached at firstname.lastname@example.com or 985.652.0952.C.J. Lin can be reached at email@example.com or 504.826.3796.


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It's been too long since we wrote anything here, we need to change that.
We haven't added any new apps to the market lately, but at the moment we are working on a small project for iOS. For this project we have "recruited" three awesome testers. Here's a video of them testing the first prototype of the app:


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Carol Burnett has a saying about cotton candy: "It's fluff, it's nothing, but sometimes cotton candy tastes really good!" If you look into a trash can at any carnival you will see a half-eaten poof of cotton candy - someone thought they wanted it but one can only take so much. "Grease" is spun sugar: It is silly, and it sends a controversial message, but it is beloved, and hey, sometimes you just want some.
Director Brian Swasey offers a "Grease" smorgasbord of treats. His choreography is organic and appears almost improvisational. He has incorporated songs from two different Broadway versions, as well as songs written just for the 1978 film. By enlisting some of Maui's finest singers and dancers, his "Grease" works.
What has always worked in "Grease" is Rizzo. How can you not love Rizzo? Lia Krieg shines in the role, and at times, carries the entire production. Krieg is one of the finest singers on the island, and her "There Are Worse Things I Could Do" is a chicken-skin moment. Rueben Carrion is surprising as usual in the role of Rizzo's steady guy, Kenickie. Carrion generally appears in four to five plays per year, and with Kenickie he has added another layer to his skill set, especially when singing lead on "Greased Lightening."
Article Photos
The vocals of Eric Peterson's Danny and Jacqui Sherwood's Sandy help Maui OnStage's "Grease" soar. Performances continue through March 11 at the Historic Iao Theater.
JACK GRACE photo
Of course, "Grease" just falls flat on its face without a quality Danny and Sandy. Eric Peterson is probably the only person on Maui who could pull off playing Danny. His vocal range alone makes it possible for Swasey's "Grease" to soar. If Danny can't sing extremely well, you've got bad cotton candy. Jacqui Sherwood as Sandy is a sugar cube wrapped in pretty paper, tied with a bow, emitting a song when you touch the ribbon. Don't misunderstand me, this is not sarcasm; she just happens to be Marlo Thomas' "That Girl." Sherwood has a chameleon side, too. She alters her voice from sweet to rock-ish and her Sandy thinks, acts, reacts and evolves constantly before your eyes.
Jonna Ahn as Marty gets her own show-stealing moment with "Freddy My Love," one of the best production numbers of the show. Danielle Allaire is fantastic via multiple characters, especially as Cha Cha DiGregorio. This is Allaire's Maui debut of sorts, and I look forward to seeing her in future productions.
In addition to Carrion, "Grease" allows several familiar Maui faces to step out of their usual roles. Aly Cardinalli plays the socially inept Doody, ironically pretending to dance badly at the Rydell High dance, saying, "Don't talk to me, I'm trying to count!" Another great dancer, Caleb Rhodes, plays Sonny, the sex-obsessed tough guy. Mamie Gesen, usually cast as sweet and innocent, plays the smoking, drinking, dropout Frenchy. And Patty Silva, last seen in sexy "Chicago" attire, plays the frumpy, constantly eating wallflower, Jan. Also of note is Jerry Eiting's stellar performance as the teen idol who croons "Beauty School Dropout." The well-known and popular Eiting gets a big laugh upon making his entrance, clad in a white tux as he comes in from the back out the house.
The ensemble cast shares in creating the highlight of the show, "Born To Hand Jive," (sung by Mark Bolden) in what seems like a never-ending dance workout. The energy and enthusiasm of the young cast is infectious, and the packed opening-night crowd cheered wildly at its finish.
The "Grease" band is another reason why the audience taps and claps along. John Zangrado on saxophone adds that extra something that most orchestras lack on Maui; it's refreshing to hear some horns in a musical as opposed to just electric keyboards. Other "Grease" musicians include Anne Durham, keyboard; Perry Gragas, percussion; Eric Molina, bass; and Dave Anthony, guitar.
Filling out the the Rydell High alumni are two current King Kekaulike High School students Marissa Godinez as Patty Simcox, "the Little Lulu of Rydell High," and Trevor Natividad as Eugene, the class nerd. James Natividad plays Roger, "king of the mooners," Laura Bloom Farber plays "old lady Lynch," the lone teacher in the Broadway version, and Steven Dascoulias plays the lecherous disc jockey Vince Fontaine.
The music of "Grease" is essentially its selling point. By the looks of the nearly sold-out show of all ages, singing along to "Grease is the Word" at curtain call, Maui is going to have a run on cotton candy. Make your reservations in advance because it appears Maui OnStage has found another "Cats" and "Chicago."
* "Grease" continues through March 11. Book, music and lyrics by Jim Jacobs and Warren Casey. Directed by New York guest director/choreographer Brian Swasey. Performances are Fridays and Saturdays at 7:30 p.m. and Sundays 3 p.m. at the Historic Iao Theater. Tickets are $40 and $22; keiki 12 and younger are $15. Dinner packages available with Cafe O'Lei and Bistro Casanova. Call the Maui OnStage box office at 242-6969 or visit mauionstage.com.


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Bheki Mseleku
Perhaps the most telling attribute of South African-born Bheki Mseleku, a pianist-saxophonist-vocalist-composer, is that he doesn't read music. This, in the age of the well-studied but pedestrian jazz musician, is something to smile about. His third release, Meditations, is not a jazz record per se but an eclectic journey through musical idioms: jazz, indigenous African, experimental. His piano playing alludes to the great McCoy Tyner, and his haunting tenor saxophone—played simultaneously with the piano—aspires to an Eastern aesthetic. "Meera-Ma" (Divine Mother) sounds like a celestial daydream, and on "Meditation Suite," Mseleku's regal piano is interspersed with his vocal incantations—African scatting, if you will—and despite its 32-minute length, it never loses its resonance. It's like a good three-hour movie. (Verve)
Picks and Pans Review: Meditations
updated 01/23/1995 AT 01:00 AM EST
•originally published 01/23/1995 AT 01:00 AM EST


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Michael Hackney
Mi historia
With a degree in computer graphics and a degree in graphic design, Michael Hackney has 5 years experience as a graphic designer. He has strong print design skills, and an intermediate knowledge of web design; he writes semantic XHTML, and CSS. Michael currently works full-time as an administrative assistant and print designer. Additionally, he is a freelance web designer for small organizations.
Experience in using Dreamweaver, (X)HTML, CSS, Flash, Photoshop, Illustrator, InDesign, Alias Maya, 3D Studio Max, Word, Excel, PowerPoint, CSO, and Datatel Comfortable in Windows and Macintosh environments
Web Design, Semantic (X)HTML, Adobe Suite, Branding & IdentityCuéntanos algo sobre ti


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BEAT THE WIND CONTEST - Show us how you challenge the power of the wind with your windiest selfie – all tricks allowed! The most windblown image is awarded with a brand-new Spitz wind jacket. Upload your image to Facebook or Instagram using the hashtag #beatthewind
Competition runs until Sept 28 23:59 CET. A Haglöfs representative jury will announce the winner by Sept 30, 2016. By participating you agree that any images posted under the hashtag #beathewind may be used freely by Haglöfs in any channel.
Full terms & conditions available here
Don't let the weather get you down. Rely on Icon GT Shoe when the temperature is falling. The shoe provides a good comfort, grip and stability with sustainable materials. A good bonus is that the shoe also keeps your feet dry and warm, see more aboutROC ICON GT
Photographer Anna Lovehed
Johanna Davidsson who received our scholarship 2015 will now in November carry out the world's longest solo expedition to the South Pole without any assistance or depots. Johanna will start her skiing adventure to the South Pole from Gould Bay coast which will mean a distance of 1400 km that will take about 2 months. Nobody has ever done this alone earlier. Johanna wants to be the first.
Beat the elements as JohannaDiscover Spitz Jacket
An iconic jacket famous for beating the elements, shielding you from the wind, cold and rain. Meet its challenger: The Northern Wind.
Check it out


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Tax Compliance
AviorData is a tax engine tackles the tax preparation and filing process of tobacco, fuel, beverage, and other related tax reporting for distributors, manufacturers, and retailers.
AviorData automates the complex process of importing, processing, and filing of indirect taxes to local, county, state, and federal jurisdictions across the entire U.S. and Canada. It identifies and calculates liabilities for financial transactions involving fuel, alcohol and tobacco. AviorData integrates with any back-end financial system to seamlessly determine the tax consequences of transactions in real time, with no manual intervention.
In using AviorData, tax filers improve efficiency, reduce costs, and mitigate risks associated with fuel, alcohol and tobacco taxes. Our on-demand reporting services provide the most robust coverage and flexibility necessary to address tobacco tax complexity and rate change of a variety of tax types.
AviorData tax automation engine generates signature-ready tax returns from the customer transactions data, enabling distributors and other filers to automate the entire filing process. AviorData imports and transforms data from the back-office systems that track alcohol & tobacco transactions. Using customizable rules that embody local, state, and federal tax laws; AviorData determines applicable taxes generating schedules and returns for all major reporting jurisdiction in the U.S. and Canada.
AviorData automates the end-to-end tax filing process eliminating labor-intensive procedures. Distributors gain the ability to use electronic filing, adopt industry best practices, and scale with tax law and standards changes.
AviorData Tax Engine integrates with most of accounting systems including Quickbooks, Microsoft Dynamics, Sage ERP, PeachTree, Atrex, NetSuite, Intacct, Epicor, Infor Syteline, Epicor and many others.
- Phone client support
- Dedicated Software Programmer
- User training and support
- Experienced technical support team
- All queries answered within 48 hours, guaranteed