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Ashes of War
Eberron is a fantasy world of intrigue built upon pulp action and dark adventure. The great nations of Khorvaire have only recently accepted peace after a century of conflict. It is a place of magic, where the advances of the Dragonmarked Houses have brought everyday magic to the people. It is a world of adventure, where the mysterious ruins of the giants of Xen'drik and relics closer to home beckon. Finally, it is a place of secrets, with shadowy organizations and spy networks working at cross purposes.
Welcome to Eberron!
Creation Myth
Across Eberron, scholars have found the same creation myth echoed in various forms. While some Vassals of the Host have a few creation myths involving Onatar or Aueron, most of the faith accepts the common creations story: The story of the Progenitor Dragons.
Before creation, the three great Progenitors existed: Siberys, the Dragon Above, Khyber, the Dragon Below, and Eberron, the Dragon Between. Siberys and Khyber each took part in the initial creation. Stories tell of Siberys putting the stars in the sky and Khyber devouring them; others claim that the two split the creation of the planes, their light or dark nature a reflection of their Progenitor. In all cases, some great difference forced the two Progenitors to fight each other over creation.
The battle was endless in the timeless era before the world. But at the end, Khyber slew Siberys, scattering his remains across reality. Then Khyber sought to slay Eberron. Every attack Eberron met with a feint; when Khyber lunged, Eberron twisted out of the way. Eventually, Eberron trapped Khyber within her folds, containing all that was evil within herself. And thus the world was formed.
According to the tales, dragons generated from the drops of Siberys' blood hitting Eberron while the rest of life on Eberron sprung directly from her. As for Khyber… the fiends and other dark things came forth from the Dragon Below.
A Brief Known History
The University of Wynarn separates the history of Eberron into four separate ages: The Age of Demons, Age of Giants, Age of Monsters, and the Current Age.
Age of Demons
For a time unrecorded, fiends from Khyber, lead by their god-like Overlords, ruled over Eberron's surface. It took a coalition of the dragons and couatls to bind the evil away from the world. According to the records from various Serpent Cults, the discovery of the Draconic Prophesy and the sacrifice of the feathered serpents were the catalyst for the dragons ascendancy over the fiends.
Thus ended the Age of Demons.
Age of Giants
The Titans of Xen'drik were a primordial civilization 80,000 years before the modern age. With the help of the dragons, they reached the pinnacle of arcane arts. The secrets of elemental binding and the creation of the warforged supposedly originated within the dark continent. However, when an invasion by creatures from the Plane of Dreams culminated with the utilization of a magic weapon beyond comprehension, the giant's civilization began to falter. When a rebellion of their elven slaves caused the giants to contemplate another "final solution," the dragons intervened, destroying the giants and cursing Xen'drik in the process. After 40,000 years of rule, all that was left of the giant civilization was ruin.
Thus ended the Age of Giants.
Age of Monsters
Elves from Xen'drik, fleeing its destruction, eventually settled in Aerenal where they delved into the secrets of life, eventually creating the Undying Court and their deathless counsellors. While human civilization was still in a primitive state on Sarlona, the goblin races of the Dhakaani Empire formed the first major nation during the Age of Monsters. Dhaakan spread across Khorvaire, subjugating or limiting the expansion of developing gnomish, dwarven, and orcish cultures. Elves and hobgoblins also came into conflict over an elven colony established in the south of the land (the eventual location of Valenar). It is also during this time that the great dragon Vvaraak taught the ways of druidism to the orcs of the Shadow Marshes. But after 30,000 years, another planar invasion, this time from the Realm of Madness, weakened and eventually destroyed Dhakaan.
Thus ended the Age of Monsters.
The Current Age
The current age saw the genesis of the Dragonmarks, first amongst the elves and then the halflings of the Talenta Plains. During this time, Lhazaar the Explorer lead a colony of humans from Sarlona as part of a large human diaspora to Khorvaire. As humans spread over the lands, more dragonmarks emerged amongst the common races; with them came the more sporadic and dangerous aberrant dragonmarks. With the destruction of the Mark of Death by the elves and dragons, and the War of the Mark between those of aberrant marks and those of the "pure" marks, the remaining houses came together to formerly organize the Dragonmarked Houses, the first in a long step of their eventual dominance of the Khorvaire economy.
As the Dragonmark Houses unified, so did the different human nations. Karrn the Conqueror, from the area that would eventually bear his name, tried first 2000 years ago. However, it was not until his descendant Galifar I that the Five Nations would be united and the Kingdom of Galifar born. For almost 1000 years Galifar was united. But with the Last War and the mysterious and destructive Day of Mourning, unity is no more. Now, a continent faces the slow process of rebuilding, trying to make peace, and keeping a wary eye out for danger.
The future awaits!


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In Alzheimer's disease (AD), rod-like cofilin aggregates (cofilin–actin rods) and thread-like inclusions containing phosphorylated microtubule-associated protein (pMAP) tau form in the brain (neuropil threads), and the extent of their presence correlates with cognitive decline and disease progression. The assembly mechanism of these respective pathological lesions and the relationship between them is poorly understood, yet vital to understanding the causes of sporadic AD. We demonstrate that, during mitochondrial inhibition, activated actin-depolymerizing factor (ADF)/cofilin assemble into rods along processes of cultured primary neurons that recruit pMAP/tau and mimic neuropil threads. Fluorescence resonance energy transfer analysis revealed colocalization of cofilin-GFP (green fluorescent protein) and pMAP in rods, suggesting their close proximity within a cytoskeletal inclusion complex. The relationship between pMAP and cofilin–actin rods was further investigated using actin-modifying drugs and small interfering RNA knockdown of ADF/cofilin in primary neurons. The results suggest that activation of ADF/cofilin and generation of cofilin–actin rods is required for the subsequent recruitment of pMAP into the inclusions. Additionally, we were able to induce the formation of pMAP-positive ADF/cofilin rods by exposing cells to exogenous amyloid-β (Aβ) peptides. These results reveal a common pathway for pMAP and cofilin accumulation in neuronal processes. The requirement of activated ADF/cofilin for the sequestration of pMAP suggests that neuropil thread structures in the AD brain may be initiated by elevated cofilin activation and F-actin bundling that can be caused by oxidative stress, mitochondrial dysfunction, or Aβ peptides, all suspected initiators of synaptic loss and neurodegeneration in AD.
Alzheimer's disease (AD) is a progressive, degenerative dementia histopathologically characterized by neurofibrillary tangles of tau protein and amyloid-β (Aβ) plaques (Goedert and Spillantini, 2006; Haass and Selkoe, 2007). In early stages of AD, hyperphosphorylated microtubule-associated protein (pMAP) tau forms striated thread-like structures in neurites, so-called "neuropil threads" (Velasco et al., 1998; Augustinack et al., 2002), that correlate with cognitive decline and comprise >85% of end-stage cortical tau pathology (Velasco et al., 1998; Braak et al., 2006; Giannakopoulos et al., 2007).
Tau, like other MAPs, stabilizes neuronal microtubules (MTs) and facilitates MT dynamics through its phosphorylation and dephosphorylation (Timm et al., 2003) (for review, see Garcia and Cleveland, 2001). Although normal adult neurons exhibit low levels of tau phosphorylation, neurons of AD brain and other tau-related neurodegenerative diseases show high levels of tau phosphorylation at both physiological and pathological disease-specific residues. This tau hyperphosphorylation prevents binding and stabilization of MTs and causes abnormal translocation of tau from axonal MT tracks to neuropil thread inclusions, dendritic processes, and cell bodies in which it accumulates and aggregates (Terry, 1998; Garcia and Cleveland, 2001). The phosphorylation of tau at Ser262 in the microtubule-binding domain is one of the earliest markers of AD neuropathology, readily detected in "pretangle" neuropil threads (Augustinack et al., 2002).
Another prominent feature widespread in the AD brain is abnormal aggregates of the actin-associated protein cofilin, which forms punctate and rod-like linear arrays through the neuropil (Minamide et al., 2000). Neuronal cofilin plays important roles in learning and memory pathways by modulating actin-rich dendritic spine architecture (Hotulainen et al., 2009) (for review, see Bamburg and Bloom, 2009). The activity of cofilin and related protein actin-depolymerizing factor (ADF) is negatively regulated by phosphorylation of the conserved Ser3 by LIM (Lin-11, Isl-1, and Mec-3) and other kinases and reactivated on its dephosphorylation by slingshot or chronophin phosphatases (Huang et al., 2008) (for review, see Bamburg and Bloom, 2009), allowing it to actively bind and sever filamentous actin (F-actin), thus regulating actin turnover (Carlier et al., 1997; Bamburg and Bloom, 2009).
ADF/cofilin–actin rods comparable with those observed in the AD brain are inducible in neuronal cell culture through inhibition of mitochondrial ATP generation and other neurodegenerative stimuli such as oxidative stress or exposure to Aβ peptides (Minamide et al., 2000; Maloney et al., 2005; Davis et al., 2009). Since actin dynamics in neurons are purported to use ∼50% of total cellular ATP (Bernstein and Bamburg, 2003), ADF/cofilin–actin rods have been proposed to represent an early neuroprotective mechanism during times of transient stress since virtually all ADF/cofilin is sequestered into nondynamic polymers of ADF/cofilin–actin, inhibiting actin turnover and thereby preserving ATP (Bernstein et al., 2006). Although mitochondrial dysfunction has been linked to AD (Smith et al., 2005; Wang et al., 2009), the relationship between mitochondrial dysfunction, the generation of tau inclusions, and their relationship to cofilin aggregates remains elusive.
In this study, we aimed to determine the effects of mitochondrial dysfunction on cellular pMAP/tau distribution compared with ADF/cofilin–actin rod distribution (Minamide et al., 2000; Huang et al., 2008). Using primary neuronal cell culture models, we demonstrate that cytoskeletal rods containing ADF/cofilin sequester and bind pMAP. The resulting striated pMAP-positive rods bear striking resemblance to neuropil threads observed in postmortem AD brain labeled with the same pMAP antibody. This process may well represent an early pathogenic event in AD leading to synaptic loss and neurodegeneration.
Materials and Methods
Antibodies and reagents.
Mouse monoclonal antibodies are actin (1A4; Dako), β-actin (Abcam), β(III)-tubulin (Abcam), tau phosphorylated at Ser202/Thr205 (AT8; Pierce), and Ser262/356 (12E8; Elan) (Seubert et al., 1995). The monoclonal antibody 12E8, raised against Ser262-phosphorylated tau, is known to cross- react with other phosphorylated MAPs such MAP2c, MAP4 (Timm et al., 2003), and doublecortin (Schaar et al., 2004) since these MAPs also contain the 12E8-specific KXGS motifs in their microtubule-binding domains. However, 12E8 has strong affinity for tau, in which the microtubule-binding domain contains several KXGS motifs, the target sequence for 12E8 (Seubert et al., 1995; Yoshida and Goedert, 2002; Timm et al., 2003; Schaar et al., 2004). Rabbit polyclonal antibodies are ADF (1439), ADF (D8815; Sigma-Aldrich), Ser3-phosphorylated ADF/cofilin (Shaw et al., 2004), cofilin (C8736; Sigma-Aldrich), actin (A2066; Sigma-Aldrich), and total tau (A0024; Dako). Phalloidin 488 (Invitrogen) was used to visualize F-actin (Invitrogen). Anti-mouse and anti-rabbit secondary antibodies included Alexa Fluor (488, 555, and 647)-conjugated (Invitrogen) for immunofluorescence and horseradish peroxidase-conjugated (GE Healthcare) for immunoblotting. Lyophilized amyloid peptides were from Bachem.
Cell culture and treatments.
Primary chick neurons were prepared from freshly dissected chicken embryos [embryonic day 7 (E7)], as previously described, and cultured for 7 d in vitro (div) on polylysine-coated 30 mm culture dishes or glass coverslips (Goldsbury et al., 2008). Primary human neurons were prepared and cultured for 14–20 d as previously described (Guillemin et al., 2007). Primary (E18) rat hippocampal neurons were cultured for 5 d as previously described (Minamide et al., 2000). For ATP depletion studies, cells were treated with 1–2 μm antimycin (AM) (a mitochondrial complex III inhibitor; Sigma-Aldrich), 3 μm carbonyl cyanide 3-chlorophenylhydrazone (CCCP) (a mitochondrial uncoupling agent; Sigma-Aldrich), or 100 μm hydrogen peroxide (Sigma-Aldrich) in PBS containing 0.5 mm CaCl2 and 1 mm MgSO4. For ADF/cofilin phosphorylation studies, cells were treated for 15 min with 1 μm AM. Actin drugs were used at 1 μg/ml jasplakinolide (Jasp) (Calbiochem; 420127) or latrunculin B (Lat B) (Calbiochem; 428020). Cells left without a medium change or treated with PBS or DMSO acted as controls. Stock solutions of lyophilized Aβ peptides were solubilized to 2 mm in DMSO, aliquoted, and stored at −20°C. Immediately before application to cells, the stock solutions were diluted to 100 μm in PBS and agitated at 1200 rpm on a laboratory shaker for 30 min to generate mixtures of polymorphic oligomeric and fibrillar aggregates as previously described (Goldsbury et al., 2000). The peptide assemblages were then applied to cells in 24-well plates at final concentrations of 1 or 2 μm. Control wells were treated with the same volume of DMSO in PBS. After treatments, cells were immediately fixed (cells on coverslips) or prepared for Western blot analysis (plated cells).
Luminescent measurements.
After treatments (in triplicate), primary chick neurons were harvested and assessed for total ATP (ATPLite; PerkinElmer) using a Victor III Multilabel plate reader. Protein measurements were determined using the identical samples and the bicinchoninic acid assay (Sigma-Aldrich). Total ATP was normalized against total protein to account for any difference in cell density. Statistical analysis used Prism software (version 3.0; GraphPad Software).
After treatments, cells were lysed and prepared for SDS-PAGE. For equal gel loading, protein concentrations were determined by the Lowry assay (Bio-Rad). Proteins transferred to nitrocellulose membranes were detected with ECL Western Blotting Detection System (GE Healthcare) on a ChemiDoc XRS (Bio-Rad). For analysis, band densities were measured using ImageJ (version 1.38x; National Institutes of Health freeware; http://rsb.info.nih.gov/ij), and background intensity was subtracted and normalized to individual β-actin or tubulin loading controls. An average and SEM for each treatment condition was determined, and results are presented as a percentage of the mean control band intensities.
Plasmids and transfection.
Plasmid-mediated expression of wild-type human cofilin has been previously described (Davis et al., 2009). Human cofilin cDNA in a pET vector (a gift from Alan Weeds, Medical Research Council Laboratory of Molecular Biology, Cambridge, UK) was modified by PCR with a 5′-PCR primer containing an EcoRI site and a 3′-primer that removed the stop codon and introduced an XmaI site at the 3′-end. EcoRI and XmaI were then used to cut the PCR product and the cDNA was ligated into pEGFP-N1 (Clontech) in-frame with the green fluorescent protein (GFP) to encode cofilin-GFP. A plasmid vector for expressing small interfering RNAs (siRNAs) for chick ADF was made by inserting DNA oligonucleotides (Macromolecular Resources) into a plasmid expression vector (pSuper) (Brummelkamp et al., 2002) containing the H1 polymerase III promoter. The oligonucleotide product from the pol III promoter is a double-stranded hairpin RNA (antisense-linker-sense) that is processed into a functional siRNA in the cell. The modified inserts including the H1 pol III promoter from the pSuper vector were excised and ligated into the pAdTrack vector (He et al., 1998). The siRNA sequence contained within the hairpin used for chick ADF is 5′-GTGGAAGAAGGCAAAGAGATT-3′. A plasmid made identically but that makes a hairpin RNA to silence human Pak2 (5′-GTCTCTGGGTATCATGGCTAT-3′) was used as a transfection control in the chick cells. The ability of the small hairpin RNA (shRNA)-expressing plasmid to effectively knock down ADF in chick cells was tested in cultures of chick skin fibroblasts (B. M. Marsick, K. C. Flynn, J. R. Bamburg, and P. C. Letourneau, unpublished observations). For plasmid-mediated expression, neuronal cultures at 3 d were transfected using Lipofectamine 2000 (Invitrogen). Four days after transfection, neurons were treated with AM (1 or 2 μm for 10 or 20 min).
Cells were fixed with 4% paraformaldehyde at 37°C for 30 min, permeabilized with 0.05% Triton X-100 or 100% ice-cold methanol, blocked in 5% goat serum, and stained for immunofluorescence. To optimize visualization of ADF/cofilin–actin rods, 0.1% EM-grade glutaraldehyde was added to the fixative. Epifluorescence images were obtained on a Zeiss Axioplan 2 microscope, captured with a CCD camera driven by AxioVision software. Single-labeled cells/sections were used to check for bleed-through in all double-label immunofluorescence studies. All captured images were converted to tagged image files for subsequent presentation. For fluorescence resonance energy transfer (FRET) analysis, cells (plated on glass-bottomed MatTek dishes) were imaged in PBS using a Zeiss LSM 510 and C-Apochromat 40× water-immersion objective and the argon (488 nm) (for GFP donor) and HeNe1 (543) (for Alexa Fluor 555 acceptor) laser lines. For photobleaching of acceptor, regions of interest in the transfected cells were bleached using the 543 nm laser line at 100% power. FRET efficiency was calculated from the increase in the fluorescence intensity of the donor after the acceptor was selectively photobleached (Gervásio et al., 2008). The donor fluorescence was measured from at least five bleached and five unbleached rods per cell.
Analysis and statistics.
For quantification of cells containing rod structures, cells with rods were counted for each treatment condition from randomly selected fields on each coverslip, and cells containing rod structures were then expressed as a percentage of the total cell population or mean number of rods per cell. Treatments and measurements were repeated in triplicate using independently prepared cell cultures. For determination of relative fluorescence intensities for phalloidin or ADF, immunofluorescent intensity was measured using ImageJ. Cells with abnormal nuclei, as indicated by 4′,6′-diamidino-2-phenylindole (DAPI) staining, were excluded from the data. After adjustment to background, mean and SEM intensities were calculated for each condition. Significance was measured using Student's t test.
Human tissue and rat hippocampal slices.
Free-floating sections (45 μm) of formalin-fixed superior frontal cortex and basal forebrain from normal adult and confirmed Alzheimer's disease patients obtained from the New South Wales Brain Bank were immunostained as previously described (Cullen et al., 2005). Sections were incubated overnight at 4°C in primary 12E8 (see Fig. 1a,b) or AT8 antibody (see supplemental Fig. S1a,b, available at www.jneurosci.org as supplemental material). Bound antibody was visualized using Alexa Fluor-conjugated secondary antibody (Invitrogen) or using ABC-peroxidase (Vector Laboratories) and DAB (Sigma-Aldrich).
Organotypic rat hippocampal slices were grown for 14 d on membranes as described previously (Davis et al., 2009). Slices were treated with 2 μm AM in PBS for 1 h, fixed in 4% formaldehyde for 1 h, permeabilized 90 s in 0.05% Triton X-100 in PBS, and immunostained overnight with 1439 (cofilin) (2 μg/ml) and mouse 12E8 (4 μg/ml) antibodies. Secondary antibody incubations were for 2 h.
Neuritic pMAP accumulation in striated rods, resembling structures in postmortem Alzheimer brain, is induced in primary neurons by energy depletion
We used the monoclonal phosphorylation-dependent 12E8 antibody raised against the pMAP tau, an established early marker for neuropil threads in AD (Augustinack et al., 2002), to determine whether pMAP accumulates in primary neuronal models derived from human, rat, or chick after mitochondrial inhibition. Probing AD brain sections with the 12E8 antibody (Fig. 1a,b) or AT8 antibody (supplemental Fig. S1a,b, available at www.jneurosci.org as supplemental material) showed numerous striated rod-like inclusions within neuropil threads and cytoplasmic accumulations of pMAP, indistinguishable from previous observations in human AD (Augustinack et al., 2002). Additionally, linear striations of rod accumulations labeled with cofilin antibodies were also observed in AD brains (supplemental Fig. S1c, available at www.jneurosci.org as supplemental material).
Treatment of primary human CNS neurons cultured for 7 d in vitro, with the mitochondrial complex III inhibitor AM elicited a rapid accumulation of 12E8-labeled protein into rod inclusions (Fig. 1c, arrows) comparable with those seen in the AD neurons (Fig. 1b, arrows). The rods sequestered the fraction of MAP serine-phosphorylated in the microtubule-binding domain KXGS motifs specific for the 12E8 antibody, rather than total MAP/tau because a polyclonal antibody against total tau yielded a more uniform labeling along neurites of both human and chick neurons (Fig. 1c,d). During AM treatment, primary chick CNS neurons derived from embryonic tectum also generated pMAP-positive rod-like structures (Fig. 1d,e) morphologically identical with those from human neurons. Thus, the readily accessible chick neurons are a useful model system for studying the mechanism by which pMAP accumulates into rods. The conserved 12E8 epitopes in the pMAP sequence reside within the microtubule binding domain in human tau (Ser262/Ser356 residues) and chicken tau (Ser253/Ser378) (supplemental Fig. S2a,b, available at www.jneurosci.org as supplemental material) (Yoshida and Goedert, 2002). Primary chick tectal neurons express five isoforms of tau that are highly homologous to human tau isoforms and exhibit conservation of phosphorylation-specific epitopes recognized by antibodies against human tau (supplemental Fig. S2c, available at www.jneurosci.org as supplemental material) (Yoshida and Goedert, 2002).
pMAP-containing striations induced by ATP depletion colocalize with ADF/cofilin–actin rods
Actin dynamics are highly dependent on ATP availability (Bernstein and Bamburg, 2003), and ATP depletion is associated with an increase in cellular F-actin and an increase in ADF/cofilin activity (Minamide et al., 2000). It follows that acute inhibition of mitochondrial function used here to generate pMAP-immunostained rods, may result in changes to the actin cytoskeleton. Consistent with this, AM-treated chick neurons exhibited a 1.9-fold increase in overall phalloidin staining in cell bodies indicative of increased F-actin content (supplemental Fig. S3, available at www.jneurosci.org as supplemental material). The pMAP-positive rods, however, did not overlap with phalloidin labeling (supplemental Fig. S3b, available at www.jneurosci.org as supplemental material).
In ATP-depleted rat hippocampal neurons, the rapid dephosphorylation of cofilin leads to the development of cofilin–actin rods in neurites (Minamide et al., 2000). These rods contain actin, as evidenced by both antibody immunostaining and ultrastructure, but do not stain with phalloidin, indicating they are probably saturated with ADF/cofilin, which stabilizes the "twisted" form of the filament and eliminates the phalloidin binding site (McGough et al., 1997). We therefore asked whether rods induced by ATP depletion contain both pMAP and ADF/cofilin. Double labeling revealed that ADF/cofilin–actin rods in part colocalize with pMAP in chick (Fig. 2a–c, asterisks), human (Fig. 2d, asterisk), and rat hippocampal neurons (Fig. 2e, asterisk). Colocalization of cofilin and pMAP in rod structures in ATP-depleted organotypic rat hippocampal slice cultures was also revealed (Fig. 3). Moreover, staining for β(III)-tubulin did not accumulate at rods, further suggesting the specificity of pMAP and ADF/cofilin in formation of neuritic rods (supplemental Fig. S4b,c, available at www.jneurosci.org as supplemental material). However, ADF/cofilin-stained rods that do not stain for pMAP (Fig. 2c,e, arrowheads) and pMAP-stained rods that do not stain for ADF (Fig. 2b, arrow) were both observed. Because optimal immunostaining for pMAP requires Triton X-100 permeabilization (avoiding methanol) and optimal immunostaining for ADF/cofilin requires methanol (avoiding Triton), structures that stain for one and not the other may be suboptimally permeabilized or immunostained. However, we cannot exclude that the detected inclusions may represent a heterogeneous population of rod structures. Surprisingly, in hippocampal neurons derived from rat, there was no change in the diffuse and uniform distribution of pMAP staining after ATP depletion, fixation in 4% paraformaldehyde and 0.1% glutaraldehyde, and brief (90 s) 0.05% Triton X-100 permeabilization, which looked identical with the staining of pMAP in untreated human, rat, and chick neurons (Fig. 1f). However, rat neurons subjected to AM-induced ATP depletion, followed by fixation in 4% formaldehyde and permeabilization as above, did exhibit rod-like 12E8 immunostaining (Fig. 2e), suggesting that glutaraldehyde fixation of pMAP when it is in a rod-like structure masks its epitope from 12E8 binding. Together, these results link ADF/cofilin–actin and pMAP inclusions within the same rod structures in neurites, which can be induced in cell culture by the common mechanism of acute ATP depletion through mitochondrial dysfunction.
To establish the activation state of ADF/cofilin during pMAP accumulation, we used an antibody specific for phosphorylated (inactive) ADF/cofilin (Minamide et al., 2000; Maloney et al., 2005). ADF/cofilin actin-binding activity is negatively regulated by phosphorylation of the N-terminal Ser3 (Huang et al., 2008; Kim et al., 2009). On ATP depletion with AM, ADF/cofilin was rapidly dephosphorylated (activated) in chick tectal neurons, as early as 2 min after treatment (Fig. 4a,b), consistent with studies in primary rat hippocampal neurons (Minamide et al., 2000). Peroxide (H2O2)-treated tectal neurons also showed nearly complete ADF/cofilin activation after 30 min (Fig. 4a,b). The time course for the redistribution of pMAP immunostaining into the rod-like inclusions (Fig. 4c) correlated with the rapid dephosphorylation of ADF/cofilin and the formation of ADF/cofilin–actin rods.
Based on the above results, we expected that the extent of pMAP incorporation into rods would be proportional to the level of intracellular ATP. To explore this relationship, we measured ATP levels in tectal neurons before and after treatment with AM, CCCP (a mitochondrial uncoupling agent), or H2O2 (Fig. 4d). Consistent with the notion that activation of ADF/cofilin induced by intracellular ATP depletion correlates with pMAP accumulation, there was an inverse relationship between the percentage of cells in the culture that contained pMAP-stained rods (Fig. 4c) and the level of ATP detected in the cell treatment groups (Fig. 4d). Cell viability tests revealed 95% cell survival in 10 min AM-treated chick neuronal cultures (compared with control cultures) and 77% survival in 30 min treated cultures (supplemental Fig. S4d, available at www.jneurosci.org as supplemental material). Likewise, abundant pMAP-positive rod formation was recapitulated in primary human neurons, after 30 min AM treatment (mean ± SD, 24 ± 2 rods per field for AM-treated compared with 8 ± 1 rods per field in controls; p < 0.001). Together, these results suggest ATP depletion is an important correlate for redistribution of pMAP into rods.
Activated cofilin sequesters and is closely associated with pMAP in rods
Since ADF/cofilin–actin rods costained for pMAP and rod formation depends on activation of ADF/cofilin, we asked whether transfected cofilin-GFP could sequester pMAP into rods. Previous work has shown that expression of cofilin-GFP leads to increased pools of activated cofilin and cofilin–actin rod formation in neurites of rat hippocampal neurons (Minamide et al., 2000; Bernstein et al., 2006). We investigated the effects of ATP depletion on the reorganization of pMAP in relation to cofilin-GFP rods by inhibiting mitochondria. ATP depletion of neuronal cultures led to rapid cofilin-GFP rod formation in transfected cells and strong pMAP colabeling (Fig. 5a,b; supplemental Fig. S5a, available at www.jneurosci.org as supplemental material). Conversely, cofilin-GFP rods did not sequester β(III)-tubulin label when all other conditions were identical (Fig. 5c). When non-neuronal cells in culture formed cofilin-GFP rods, these were negative for both pMAP and β(III)-tubulin (Fig. 5d), neither of which is expressed in non-neuronal cells. Together, these results suggest that activated cofilin first accumulates into rods and subsequently sequesters pMAP into these cytoskeletal inclusions.
To further evaluate the interaction between cofilin and pMAP in rods, we exploited FRET between the GFP (donor) and Alexa 555 (acceptor) fluorophors. FRET between colocalized cofilin-GFP and pMAP/Alexa 555 was measured using acceptor photobleaching (Fig. 5b). The cofilin-GFP fluorescence signal in rods increased significantly after bleaching the Alexa 555 fluorophor (Fig. 5b, right, arrows), with a measured FRET efficiency of 41 ± 3% (mean ± SEM; n = 52; p < 0.0001) as measured by the percentage increase in donor fluorescence after acceptor bleaching. In contrast, cofilin-GFP fluorescence did not significantly increase in adjacent rods in the same cells in which the Alexa 555 fluorophor was not bleached (Fig. 5b, right, asterisks). Furthermore, a much reduced FRET signal was detected in neurons between cofilin-GFP and Alexa 555 immunostained pMAP in adjacent non-rod-forming regions of neurites and cell bodies (supplemental Fig. S5b, available at www.jneurosci.org as supplemental material). Control experiments demonstrated no FRET between cofilin-GFP and control antibody-labeled epitopes—β(III)-tubulin/Alexa 555 or Src/Alexa 555—even in regions in which the fluorescence appeared colocalized (supplemental Fig. S5c,d, available at www.jneurosci.org as supplemental material). Cells expressing free GFP and treated with AM to generated pMAP-positive rods labeled with 12E8/Alexa 555 also never produced FRET (supplemental Fig. S5d, available at www.jneurosci.org as supplemental material). These results demonstrate that cofilin-GFP and the secondary antibody labeling pMAP are colocalized <10 nm apart within the cytoskeletal rod complex, indicating a close proximity of pMAP and cofilin in these inclusions.
Silencing ADF prevents pMAP accumulation in rods
Since induction of ADF/cofilin rods leads to the reorganization and sequestering of pMAP, we asked whether silencing ADF or cofilin would prevent sequestering of pMAP under the same conditions. During chick brain development, ADF comprises ∼75% of the total ADF/cofilin from embryonic day 14 onward (Devineni et al., 1999). We therefore chose to knock down the total cellular pool of ADF specifically, using a plasmid for expression of a shRNA that yields an siRNA when expressed and processed. The shRNA was transfected into chick neurons and coexpressed from the same plasmid as GFP for visualization of transfected cells. Four days posttransfection, cells were immunolabeled for ADF (Fig. 6a) and quantified, revealing 75% knockdown of ADF (Fig. 6e). Treating transfected cultures with AM (1 μm for 10 min) and staining for pMAP revealed that significant silencing of ADF inhibited accumulation of pMAP into rod structures, whereas surrounding untransfected cells contained an abundance of rods (Fig. 6d, asterisks). In control chick neuronal cultures transfected for 4 d with shRNA specific for human PAK2, immunolabeling revealed no decrease in total cellular ADF (Fig. 6b,e). pMAP accumulated in rod-like inclusions both in surrounding nontransfected neurons and in shRNA PAK2 transfected neurons (Fig. 6c, asterisks and arrowhead). Together, these results suggest that the presence of ADF/cofilin is necessary for sequestration of pMAP into rod-like structures.
Latrunculin B enhances and jasplakinolide represses pMAP staining in rods
Since activation of ADF/cofilin coincided with pMAP recruitment into ADF/cofilin–actin rods, we asked whether other manipulations of F-actin assembly could influence pMAP sequestration to rods. To address this, we used pharmacological manipulation (enhancing or suppressing) of F-actin pools and stained for pMAP. Latrunculins sequester monomeric G-actin to form a nonpolymerizable 1:1 complex, thereby inhibiting F-actin reassembly and thus promoting overall F-actin depolymerization (Coué et al., 1987). Latrunculins, however, compete weakly with ADF/cofilin for actin binding (Bernstein et al., 2006) and can actually induce ADF/cofilin–actin rod formation (Pendleton et al., 2003). The decline in the phalloidin-stainable F-actin pool after Lat B treatment is indeed evident (Fig. 7a), although it should be noted that phalloidin cannot stain ADF/cofilin saturated F-actin. In contrast, Jasp binds and stabilizes F-actin resulting in a net decrease in the G-actin pool (Bubb et al., 1994). Since Jasp competes for the phalloidin-binding site on F-actin, stabilized F-actin cannot be visualized with fluorescent phalloidin in the presence of Jasp (Fig. 7a). However, ADF/cofilin cannot bind to phalloidin-stabilized F-actin (Minamide et al., 2000) and thus also would not likely bind to the Jasp-stabilized actin filaments.
Antibody labeling of Lat B-treated neurons revealed formation of both pMAP- and ADF-positive rods, which frequently colocalized in double-labeling experiments (Fig. 7b). In contrast, neurons treated with Jasp (or cotreated with Jasp and AM) exhibited a total absence of pMAP and ADF rods (Fig. 7a). We then compared the rate and abundance of pMAP-stained rods after Lat B compared with AM treatments and found the chronology of their formation indistinguishable between conditions (Fig. 7c). That Lat B treatment only moderately reduced ATP in neurons (Fig. 7d) yet still induced ADF/cofilin–actin and pMAP rod formation comparable with mitochondrial inhibitors suggests that ATP depletion is not a direct cause of rod formation, but an upstream event. These data collectively suggest that subunit release from F-actin is required for the generation of pMAP-positive rods and that in neurons this can be induced by ADF/cofilin activation. It also appears that ADF/cofilin binding to F-actin is essential for the formation of cytoskeletal rods. Dissociated actin subunits saturated with activated ADF/cofilin thereby form cytoskeletal rod inclusions that appear to sequester and accumulate pMAP.
Aβ peptides enhance pMAP accumulation at ADF/cofilin rods
In AD, the relationships between pMAP/tau pathologies, cofilin–actin pathologies, and amyloid pathologies arising from Aβ peptide oligomerization and/or aggregation are poorly understood. Synthetic Aβ1-42 has been shown to induce cofilin–actin rod pathology in up to 20% of neurons in dissociated rodent hippocampal cultures (Maloney et al., 2005), with the majority of these neurons located in the dentate gyrus (Davis et al., 2009). We therefore asked whether Aβ could influence the sequestering of pMAP to ADF/cofilin–actin rods. Primary chick neurons were exposed to 1 or 2 μm Aβ1-40 or Aβ1-42. Before applying to cells, peptide solutions were agitated on a shaker for 30 min to generate mixtures of oligomeric and fibrillar structures (Goldsbury et al., 2000). The solutions were then diluted into cell culture medium and added to cells at a final concentration of 1 or 2 μm. After exposure to the peptides for 20 h, the cells were fixed and immunostained for pMAP. Less rods were generated in cultures treated with Aβ compared with those treated with the mitochondrial inhibitors, consistent with previous studies of ADF/cofilin–actin rod generation in hippocampal neurons treated with Aβ peptides (Maloney et al., 2005). However, when mean numbers of pMAP-positive rods per cell were quantified on vehicle control and Aβ-treated coverslips, a significant increase in the number of rods was revealed in Aβ-treated cells (Fig. 8a). No difference was observed between cells treated with 1 and 2 μm peptide or between cells treated with Aβ1-40 and Aβ1-42. The presence of F-actin-rich active growth cones in Aβ-treated neurons containing pMAP rods in the neurite shafts indicates the continuing viability of the neurons, despite the formation of pMAP-positive rods (Fig. 8b, white arrows). We confirmed that rods in Aβ-treated neurons contained both pMAP and ADF (Fig. 8c, arrows). In conclusion, these results show that synthetic Aβ aggregates can induce the recruitment of pMAP to ADF/cofilin–actin rods in a subset of neurons in these primary cultures.
Neuropil threads of the AD brain are composed of linear arrays of cytoskeletal inclusions containing the pMAP tau (Velasco et al., 1998; Augustinack et al., 2002). These structures are positive for Ser262/356 tau labeled with the 12E8 antibody before extensive staining by antibodies marking hyperphosphorylation of tau that is observed in late-stage disease (Augustinack et al., 2002). Corresponding to this, formation of neuropil threads is associated with the early clinical stages of dementia (Giannakopoulos et al., 2007). The structure of neuropil threads implicates a disrupted cytoskeletal network that spans the width of the neurite likely contributing to dementia by blocking cargo trafficking to synapses that underlie memory formation and cognition, causing subsequent retraction of distal neurites (Velasco et al., 1998) (for review, see Terry, 1998; Bamburg and Bloom, 2009). Our data open the possibility that abnormal activation of cofilin, an actin-binding protein, in the AD brain and generation of cofilin–actin cytoskeletal rods is a triggering factor for the sequestration and accumulation of Ser262/356 phosphorylated tau in some neuropil threads. This process may well represent an early pathogenic event in AD neurodegeneration.
Neurodegenerative stimuli including oxidative stress, mitochondrial dysfunction, excitotoxic glutamate, ischemia, and soluble forms of Aβ lead to activation of cofilin and the related protein ADF and the generation of ADF/cofilin–actin rods in primary neuronal cell culture (for review, see Bamburg and Bloom, 2009). These rods resemble the cofilin aggregates that are widely distributed in the AD brain (Minamide et al., 2000). At a low ratio to actin and in an active (dephosphorylated) state, ADF/cofilin maximally enhance subunit turnover of F-actin, dynamically remodeling the actin cytoskeleton and thus playing an integral role in regulating actin-dependent synaptic stabilization (for review, see Bamburg and Bloom, 2009). ADF/cofilin bind cooperatively to F-actin, and, at higher ratios to actin, they can saturate regions of filaments and stabilize the pieces of filaments that remain after severing (Andrianantoandro and Pollard, 2006; Chan et al., 2009). Because ADF/cofilin bind to a minor, slightly twisted conformation of F-actin, they stabilize this "twisted" form, which prevents binding of the commonly used F-actin stain, phalloidin (McGough et al., 1997). In the brain, mitochondrial dysfunction and energy depletion associated with AD (Smith et al., 2005; Wang et al., 2009) could feasibly serve as a pathway for F-actin remodeling in neurons, since up to 50% of neuronal energy is dedicated to actin dynamics (Bernstein and Bamburg, 2003). We propose mitochondrial dysfunction is one potential pathway upstream of the assembly of cytoskeletal rods because direct electron transport chain inhibitors elicit cofilin activation, concomitant with rod assembly in primary neurons and organotypic rat hippocampal brain slices. This pathway results in a precipitous drop in ATP, and the release of the cofilin phosphatase chronophin from an inhibitory complex with Hsp90, resulting in cofilin–actin rod formation (Huang et al., 2008). However, cofilin–actin rods can also be induced while maintaining higher levels of cellular ATP, such as with peroxide (Fig. 4), which activates the cofilin phosphatase slingshot by oxidizing and removing inhibition by 14-3-3 and leads to almost complete cofilin dephosphorylation and cofilin–actin rod formation (Kim et al., 2009). The common thread is an increased pool of active ADF/cofilin with respect to the amount of F-actin such that ADF/cofilin-saturated pieces of F-actin are available and coalesce into rods.
What is the role of pMAP at the cofilin–actin rods? Rods isolated from neurons and non-neuronal cell lines contain ADF/cofilin:actin in a 1:1 complex and indeed can be formed in vitro from these purified proteins (L. S. Minamide, S. Maiti, J. A. Boyle, R. C. Davis, J. A. Coppinger, Y. Bao, T. Y. Huang, J. Yates, G. M. Bokoch, and J. R. Bamburg, unpublished results). Thus, as is also shown here, pMAP is not essential for cofilin–actin rod formation. Nevertheless, pMAP association with most cofilin–actin rods occurs very early in the rod formation process. Rods isolated from cortical neurons and those formed from endogenous proteins from a non-neuronal cell line both have similar stabilities to alterations in pH, ionic strength, reducing agents, Ca2+/EGTA, ATP, and detergents. Therefore the presence of pMAP on the neuronal rods does not appear to confer in them any special property related to their stability. The question then arises—do the rods confer any special property on the associated pMAP? The tandem arrays of the rapidly formed cofilin–actin rods are similar in size and distribution to the neuritic striations of the neuropil threads in AD. The ultrastructure of most striated neuropil threads from AD brain clearly shows these to consist of the ∼20-nm-diameter paired helical filaments (PHFs) (Velasco et al., 1998), not of the cofilin–actin bundles containing ∼10-nm-diameter filaments that we observe in organotypic hippocampal slices treated with Aβ1-42 peptide oligomers (Davis et al., 2009). Thus, we suggest that the cofilin–actin rods serve as a template for recruitment and binding of pMAP, and that this association facilitates additional phosphorylation of pMAP and its self-assembly leading to the eventual replacement of the cofilin–actin rods with PHF bundles. Additional long-term studies are needed to test this hypothesis.
Supporting the concept that pMAP/tau accumulation and toxicity is linked to interactions with the actin cytoskeleton is work in Drosophila models that showed that toxicity of overexpressed hyperphosphorylated tau could be modified by genetic ablation or coexpression of actin-associated proteins (Fulga et al., 2007). Interestingly, tau phosphorylation at the 12E8 antibody epitope was shown to be essential for the initiation of tau toxicity and its downstream hyperphosphorylation in Drosophila (Nishimura et al., 2004). Mutated tau nonphosphorylatable at the 12E8 epitope did not confer toxicity in Drosophila and exhibited reduced downstream phosphorylation at other AD-relevant phospho-tau epitopes (Nishimura et al., 2004). These findings are consistent with the premise that the sequestration of 12E8-specific pMAP to cofilin–actin rods demonstrated here could represent an essential early event in a neurodegenerative pathway. The recent finding that tau reduction can ameliorate toxicity induced by APP (amyloid precursor protein) overexpression or exposure of cells to Aβ oligomers is also consistent with the key involvement of pMAP/tau in response to toxic events (King et al., 2006; Roberson et al., 2007). Potential deleterious effects of cytoskeletal rods on neurons are multifold. First, rods that assemble in axons and dendrites of cultured neurons form transport blockades inhibiting the free movement of vesicles and organelles leading to synaptic dysfunction (Jang et al., 2005; Maloney et al., 2005, 2008). Second, the accumulation of actin and cofilin in rod structures could directly impinge on the availability of cofilin for its function in modulating actin dynamics at the synapse (Hotulainen et al., 2009). Likewise, the sequestration of pMAP/tau to the rod matrix could adversely affect the role of tau in stabilizing and regulating the axonal MT network of tracks for fast axonal transport.
The generation of cytoskeletal rods in response to stressors may be an initial compensatory response to slow down energy-consuming actin dynamics and thereby protect neurons under conditions in which ATP supply is compromised (Bernstein et al., 2006). However, repeated or prolonged stress would likely be detrimental. Increasing evidence suggests declining mitochondrial function and reduced energy metabolism to be early events in the AD brain, preceding severe pathological changes (Smith et al., 2005; Wang et al., 2009). Aging is the major risk factor for sporadic AD, and on the one hand, electron transport chain activity declines with age, whereas on the other, oxidative stress increases, potentially impacting on the availability of ATP in cells (Lin and Beal, 2006). Moreover, the reduction of glucose uptake in AD brain cells and concomitant decline in glycolysis would also negatively impact ATP levels. Precedents of local mitochondrial dysfunction could be multifold and varied in the aging brain, for example, low brain perfusion attributable to vascular insufficiency, stroke, and Aβ peptide-mediated perturbation of mitochondria (Cullen et al., 2005; Du et al., 2008; Cho et al., 2009). Mechanisms linking this metabolic dysfunction with the formation of neuropathological lesions in the AD brain need to be established. Interestingly, it was demonstrated that environmental toxin inhibitors of mitochondrial respiratory chain complexes induce somatic inclusions of phosphorylated tau in neuronal cell culture and neurodegeneration in tau-related disease (Escobar-Khondiker et al., 2007).
Advanced neuropathology involving pMAP and cofilin accumulation and aggregation may be a result of increased vulnerability to normal environmental stress in brains expressing mutant tau (i.e., in hereditary frontotemporal dementias) or from elevated oxidative/mitochondrial stress in brains expressing wild-type endogenous tau (in sporadic AD). In a healthy brain, a fine balance between these stressors and compensatory defense mechanisms must normally occur, given that development of tau pathology and AD are not an inevitable part of aging. Recent evidence showing that neuronal cell bodies attached to dystrophic neurites in AD-related transgenic mice are still viable and otherwise healthy (Adalbert et al., 2009) holds promise for future work to develop methods to disrupt the formation of the cofilin–actin rods and/or the interaction between cofilin and pMAP for establishing new therapeutic strategies to combat this disease early in its development.
This work was supported by The Sir Zelman Cowen Universities Fund, The Judith Jane Mason and Harold Stannett Williams Memorial Foundation, The Rebecca Cooper Foundation, Sydney Medical School (C.G.), Alzheimer Drug Delivery Foundation Grant 281201 (J.R.B.), and National Institutes of Health–National Institute of Neurological Diseases and Stroke Grants NS43115 and NS40371 (J.R.B.). We thank Prof. Peter Seubert (Elan Pharmaceuticals, South San Francisco, CA) for providing the 12E8 antibody.
- Correspondence should be addressed to either of the following: James R. Bamburg, Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523, james.bamburg{at}colostate.edu; or Claire Goldsbury, The Brain & Mind Research Institute, School of Medical Sciences, University of Sydney, 100 Mallet Street, Camperdown, NSW 2006, Australia, E-mail: cgoldsbury{at}usyd.edu.au

subset: commoncrawl


text:
In the Crochet for Charity group on Ravelry I came across a post mentioning the Snuggles Project. I had never heard of this before but fell in love with the concept. I know I can't adopt all the abused or neglected animals in the world nor am I rich enough to fund all the animal shelters in the world, but snuggles? I can make some snuggles.
Basically, people create snuggles (little blanket pads) in any size from a small kitten size to a large mastiff size, that are then donated to participating animal shelters around the area. It's all over the United States and if your favorite one isn't listed, there's a FAQ sheet on how to get them involved.
-Great for beginner knitters/crocheters as you're creating simple rectangles and can make them any size.
-You can use up stash yarn very quickly.
-It is very inexpensive, as cheap acrylic, washable yarns are a good thing here (yea, red heart super saver prices); even recycled yarns from old afghans, sweaters or terrible unfinished projects can be used.
-Some little animal gets something soft and their own to make their stay a bit more comfortable and secure.
So the pic above is of my first Snuggle. It's a little lopsided (I should not have tried to crochet the border at the same time as the main body with the eyelash yarn…too easy for me to miss stitches) but I think a little kitty or puppy would be quite happy laying on it. It was made with one strand of red heart super saver yarn and one strand of eyelash yarn crocheted together in a hdc stitch (half double crochet).
Oh! And I found a Snuggles Project group on Ravelry too!


text:
The man heading the Obama administration's response to the oil spill in the Gulf of Mexico says a containment cap installed on the BP rig is helping to limit the leak.
Speaking at a Washington briefing, Adm. Chad Allen of the Coast Guard said that more than 14,800 barrels of oil have been kept out of Gulf waters in the last 24 hours. Allen told reporters the amount of oil kept from spilling into the Gulf "has climbed steadily" from the first day the containment cap was installed.
He also said officials will be meeting with BP to assess how well it is handling claims for relief from people hurt by the spill. Allen said the aim is "to see if we need to provide any oversight." He had said Monday BP was struggling to handle claims.


text:
They may not be as deep as last year's squad, but the Lovell Bulldogs wrestling team still has some formidable firepower, which they showed off at the Powell Roundtable Invitational Friday and Saturday in Powell.
The wrestlers were divided into varsity and junior varsity divisions and competed in round robin wrestling Friday, which allowed officials to seed them into a bracketed tournament Saturday, coach Daniel Robertson said.
Lovell produced two champions and one additional placer at the large tournament as seniors Jacob Beck and Dino Collins captured individual titles and Jacob Asay placed fourth.
Beck won it all at 170 pounds, going 3-0 on the final day. He started with a pin in 1:33 over Skyler Landerman of Cody, pinned Tyler Patterson of Powell in 1:30 and beat Brecken Biggs of Casper Natrona 5-1 for the championship.
"We discovered some things we can do on the mat to neutralize an opponent," Robertson said. "We were testing out a new ride that will pay dividends, a cross-body ride. At this point teams didn't have an answer.
"Jacob wrestled really smart. He's working hard to fine tune some things that got him into trouble in the past."
Beck beat the number two (Patterson) and number three (Landerman) wrestlers in Class 3A and the number two wrestler in Class 4A (Biggs) on Saturday.
Collins cruises
Collins dominated the competition at 182 pounds, pinning most of his opponents over the weekend. On Saturday he pinned Jackson of Columbus, Mont., in 37 seconds, beat Thomas Myler of Casper Natrona (third ranked in 4A) 12-2 and defeated Sterling Baker of Dubois 6-1 for the title.
"We wrestled him (Baker) seven times last year," Robertson said. "This was the eighth time in two years. Everywhere they go, they meet. They are the two best 182-pounders in the state."
Asay beat Smith of Rawlins 7-0 to start Saturday action, then fell in 30 seconds to eventual champion Yates of Colstrip, a defending Montana state champion. In the third/fourth-place match he fell to Stringer of Powell, 5-3.
"I hope this is Jacob's coming-out party," Robertson said. "He's gotten out of a double tie that heavyweights seem to be fond of. It's completely neutral and there's nothing you can do from there. This puts him in a better position to defend himself, and now he's attacking, too. It's so important to take the initiative and score points, take advantage of opportunities."
Lovell's other senior, Nathan Grant, had to withdraw after Friday's action with a back injury, Robertson said.
"We didn't wrestle incredibly well," he said. "We finished eighth in varsity and were open at 106, 113, 126, 138 – and Nathan was hurt. We're still learning stuff we need to work on, especially the JV kids. We have a lot of JV kids with up side. They just need experience. They're now finding out what wrestling is about. Drilling with your friends is a lot different than wrestling someone who doesn't like you."
Host Powell won the tournament with 341 points, followed by Worland with 291, Natrona with 210 and Casper Kelly Walsh with 207 points. Lovell was the top 2A team with 103 points.
Wrangler Duals
The Bulldogs will be off to Shoshoni this Saturday for the Wrangler Duals – a series of dual matches against mostly Class 2A teams. Extra wrestlers will have the opportunity to fill holes in other teams' lineups, Robertson said.
Action begins at 9 a.m.
By David Peck


text:
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I have a website already setup, but now it needs the content changed. We have to update a number of pages, maybe about 20. I also have 27 pictures that need changing. Slider Images need changing. The most tricky change is that the site hasn't been udpated since May last year, and it we need to get about 70 new pieces of content from the old
...website at realmlabs.net. The site is running an old version of Joomla and virtue mart e-commerce plugin and both need to be updated. It currently has 800 users that have signed up over time. Occasionally, they log into the website so the protected content and users need to remain and cannot be "broken" because of an update. Here is what needs to be
*** I am seeking an honourable Joomla expert. The total work time for this task should take you NO LONGER than one hour to do the entire work and we are paying $30 for this job --> this equates to a rate of $30 an hour (maximum 1 hour). If you need longer than an hour, then your work is either too slow or you are not experienced enough so do NOT bid
...issues with getting into site as it is so outdated. I just need updated to 3.0 This is a simple site, so nothing complicated. Admin access has been difficult due being version 1.5 Once we get it on a 3.0 platform I can do the major content upgrades that are needed but I would like to minimize downtime as it is a money making site as well as keep up on
...building a website using a Joomla template and a music player that was another 3rd party component I bought. It's a subscription based site and the template has that installed, but the problem I have is that the music player also has a Login function that I need to bridge with the Joomla template. So that when they login to my site it will automatically


text: This video is best watched at YouTube in Full Screen HD so you can see all the details: http://www.youtube.com/watch?v=bN9ThVOiWZU Now, there is no method here in this post. I've just been seeing A LOT of disinformation regarding SB thrown around and felt like making this video & posting it. I'll have a full method WITH video instructions (I'll literally walk you through it) posted in the next couple of days. It will be freely posted in the BlackHat SEO section. Of those 50k or so indexed links you see in the post, all but roughly 5 to 6k are from using SB. So, I guess the title of the thread should have been entitled "over 44k indexed links insdie of a month" but 50 had a better ring to it. Besides, what's 6k links anyway? To answer any potential questions before they are asked: No, I didn't do anything at all to get those links indexed faster than normal. That is entirely natural backlink indexing....alright, I gave it a LITTLE push (I'll post that trick in a second in another thread). But seriously, it's a little trick...nothing serious. See here: http://www.blackhatworld.com/blackh...get-your-backlinks-indexed-faster-google.html It didn't hurt my site. In fact, it helped it significantly. The site is 6+ months old (more than 6 months old, less than a year old). I can't think of anything else...I'm sure you guys will.


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I Believe in Being Passionate About Something
In today's fast paced world, it's rare that someone finds something they can be truly passionate about. Everyone seems to be too busy to figure it out, but I have been lucky enough to find something that moves me in a way nothing else can. Music has been a part of my life since I was just six years old. It was at that age that I asked my parents if I could take what would become the first of countless piano lessons. And the rest, as they say, is history. I'm twenty-one years old now, and I am still as enamored by the sound of the instrument as I was the first time I played Mary Had a Little Lamb start to finish.
Since that first lesson I have moved on to perform in recitals, entertain at parties, play in bands, and gain an absolute respect for music that I hold very dear to my heart. But of all the things I have accomplished, the most important to my appreciation of music, and the most worthwhile, has been teaching the piano. When I was sixteen I had my first opportunity to do so when my teacher, the owner of my music school, asked if I had any interest in taking a few students. Over the next three years I taught students ranging in age from seven to thirty-seven, and the experience had a profound effect on me. It was hearing that sigh of relief when a young student finally learned those difficult few bars, or seeing the big smile on their face when they looked over at me after playing a song perfectly that made me remember why I had stuck with it for so many years.
As I have gotten a little older, music has become many different things to me. It has become an escape from the mundane, a stress reliever, and a form of personal expression. And slowly, it has grown to be my greatest passion. I find that if there's a piano around somewhere, it's impossible for me to be bored. My most satisfying moments are those spent sitting at that bench playing. It doesn't matter if it's Bach, Billy Joel, or just something I made up on the spot; If I'm playing, I'm happy. Years ago, my great aunt embroidered a framed picture for me that said "Music is a fair and glorious gift of God." That frame is still mounted above my piano, and it is a constant reminder of how fortunate I am to have found something to be passionate about.
If you enjoyed this essay, please consider making a tax-deductible contribution to This I Believe, Inc.


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Born in New York City, Samantha Heydt's interest in photography stems from her experiences overseas and the influence of her father who is also an artist. In 2004, she commenced her studies at La Sorbonne in Paris, after which she transferred to the Universidad de Buenos Aires in Argentina where she studied dance and fine art. In 2005, she began her degree in communications at The New School, spending several years abroad in Amsterdam and later in Florence. As a result, she speaks several languages. Currently, she finishing up her second degree at Parsons the New School for Design in photography. She has had work exhibited at the State Hermitage Museum in St. Petersburg, as well as several galleries in Italy and New York City.
Areas of Expertise
Photoshop, InDesign, Bridge, Medium Format, Large Format, Raw, 32 MM, Digital Archiving, Studio Lighting, Mac/PC proficient, Microsoft Office (Word, Excel, Powerpoint), IWork: (Pages, Numbers, Keynote). IMovie, IPhoto, IWeb, Directors Cut, Font Lab, Alien Skin, Automator, Fetch, TextEdit, Wordpress, Typography, Bookbinding, Printmaking, Collage.
Fluent in French, Conversant in Dutch & Italian, Comprehensive Spanish
Professional Affiliations
W Magazine
Visionaire Magazine
The State Hermitage Museum (exhibited in two group shows 2010/2011)
Milk Gallery (exhibited in MGU 2011)


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Introduction to mod_perl \(part 6\): Even More Perl Basics
April 08, 2003 - This article is a third one in a series talking about the essential Perl basics, that you should know before starting to program for mod_perl.
Please discuss this article in this thread. You can read the article here .


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Come see our students' artwork on display at the Princeton Public Library on Witherspoon Street in downtown Princeton (on the 3rd floor – children). Art pieces on display were made by our Preschool to Kindergarten students (3 to 5 years old).
Our school will be featured until November 27th, so please stop by to see their work. We are very proud of our students!


text:
(What's on this website.)
E-Z Inquiry Form
A word from the poet.
I've been writing it since I was 18. It's a condensed form of expression employing elevated diction, striking imagery, sound effects and allusions to the intellectual and emotional library all humans share. The best language to write it in is Chinese, where it's totally free of syntactical rigidities. No verb declensions or connective elements. Only the "hot" words, the words with color, and impact. I try to write like this, doing the best I can in English.
It's ten years since I gathered together Poems For My Kids. On this page I've collected a lot of what I've done since then, starting with the latest stuff. Dislike poetry or no experience with it? Look at my "How to Read Poetry," for important help. Comment or share your stuff? Thanks.
P.S. See me on YouTube!
RECENT POETRYMary of the Circle Brooklyn NY
Green Pavilion New York NY
HAWAII Honolulu HI
GUNTER GRASS Havre de Grace MD
KAFKA Prague CZ
SAMUEL BUTLER Renssalear NY
WALMART Secaucus NJ
FEEDING BIRDS IN WINTER Greenpoint Bklyn
GNATS Sugar Camp
JOSE ITURBI AND OTHER GHOSTS 7th & 56th
DRONE 1829 Stone House
MACEDONIA 7th & 23rd
JOHN BERRYMAN IN THE VILLAGE Sheridan Square
The Bangkok Lady Visits Champlain SUNY Plattsburgh
CAMP The Cabin
WAINWRIGHT MOUNTAIN Plattsburgh NY
RAIN (For Francesca) Plattsburgh NY
FRAGMENTS AND EXPERIMENTSBoy on a Bike
'05 Eye of the Storm
Boys Clubhouse
Stone and Sand
CLICK AND GO! (More on this page.) San Felipe del Rio, Jan 18, 2009 HEB, Feb 18, 2009 Tornado, Feb 21, 2009 Souvenirs, March 5, 2009 (From A Gentleman's Travel Notes.) (Email Exchanges.) "The location is gorgeous . . . yet your poetry had a strong negative current throughout every page." "Are you working on those memoirs yet?"
"The location is gorgeous . . . yet your poetry had a strong negative current throughout every page." (An Email Exchange.)
Subject: Re: Your Fourpeaks visit 12/18/01 to Ridge Camp
Date: Fri, 11 Jan 2002
From: firstname.lastname@example.com
To: firstname.lastname@example.com
Hi Martin-
Dave, Ellie, Adam, and I thoroughly enjoyed our visit at Ridge Camp last month, and we wouldn't have changed anything! Even the
weather was perfect! We hope to return next year, ideally in the fall, but that will depend on crops, etc. I believe our only
disappointment was that you don't seem to appreciate everything around you. The location is gorgeous, your two kids were very
appealing, yet your poetry had a strong negative current throughout every page. We would have purchased several volumes, as we
enjoyed the imagery and your style, but you never seemed to consider people or events positively and we found it depressing. My
family enjoys people, animals, and the outdoors immensely, but we view most in a positive light. Perhaps you would appreciate your
guests,your family, and your surroundings more if you could be less cynical. Life is good! Thanks for a great experience. We hope
to be back soon. Forgive me for being blunt, but I think that's what you wanted. -Sue Th***
Subject: Re: Your Fourpeaks visit 12/18/01 to Ridge Camp
Date: Fri, 11 Jan 2002
From: Your Adirondack Guide email@example.com
Organization: Fourpeaks--Adirondack Camps & Guest Barns
To: email@example.com
Thanks for your email.
My bad disposition is because I have a failed relationship with my wife.
But that's getting fixed. Pretty soon I may be viewing things as positively
(well, maybe not all that positively!) as you.
Please come back.
Did you take any photos? Share them if you have. See
http://4peaks.com/fgstbndx.htm I'll make a webpage of your visit.
Best wishes, Martin
"Are you working on those memoirs yet?" (An Email Exchange.)
Connie Francis, Among my Souvenirs
There's nothing left for me
Of days that used to be
They're just a memory
Among my souvenirs
Some letters tied with blue
A photograph or two
I see a rose from you
Among my souvenirs
A few more tokens rest
Within my treasure chest
And, though they do their best
To give me consolation,
I count them all apart
And, as the teardrops start,
I find a broken heart
Among my souvenirs
I count them all apart
And, as the teardrops start,
I find a broken heart
Among my souvenirs
"I would like to interest you in re-starting a relationship with this." (An Email Exchange.)


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Bear Grizzly Recurve 58in
Brand: Bear
Fred Bear Grizzly. Fred Bear shot with this bow. Why settle for any thing else
Bear Grizzly Recurve 58inPrice: $586.95
$533.59 excl. GST**
** Your price if you do not reside in Australia
Bear Grizzly Recurve 58in Description
Hand crafted legendary performance. Often rated a top buyer's choice for quality and bowyer craftsmanship, the traditional line up from Bear Archery continues to satisfy those recurve enthusiasts looking for a time tested quality hunter. These are the same handcrafted bows that Fred Bear designed and tested all over the world. Exotic hardwoods have been used to create stunning bows that are both superior in the field and almost certainly to become tomorrow's collectibles.
Grizzly Hunter Series recurves are finished in satin gloss to protect the bow and provide a shield to minimise dampness and abrasion. A dependable Dacron string makes the 58-inch Grizzly a tough, economical "working man's bow." The classic Grizzly design has not changed in 50 years. Includes cut-past-centre shelf covered in Bear Hair.
Fred Bear Grizzly Recurve 58" Technical Specifications
- AMO Length 58"
- Draw Weights RH 30lb, 35lb, 40lb, 45lb, 50lb, 55lb, 60lb. LH 35lb, 40lb, 45lb, 50lb, 55lb. Please specify draw weight
- Available in right hand and left hand. Please specify RH or LH
- Riser has rich wood tonings
- Limbs are of hardwood laminates with black fibreglass overlays
- Soft Bear hair rest
- Leather side plate
- Dacron Flemish Twist String, a Bowstringer and easy to follow instructions on how to string the bow
Also available in a complete bow package of
- Arrow rest
- Bow quiver, side quiver, back quiver or hunting side quiver
- Whisker string silencers or puff silencers
- Martin Deerskin Leather Glove, Vista Deerskin Glove or ASAT Glove
- ASAT Leafy or True Timber camo suits, headnets and camo paint
- Bowhunters gear belt or Rangefinder case
- Hunting or ventilated armguard
- Easton Carbon or Aluminium hunting arrows with two, three or four blade broadheads to suit
- String Nok Set
- Bjorn 60cm Target Face
- Bowstring Wax
- Fred Bear Padded Recurve Soft Bow Case or Martin Recurve Bow Case
- 3D Animal target, ShotBlocker block target or DeadStop bag target
- Bow will be set up and ready for you to shoot.
- Please specify which accessories you would like included in your package and we will be only too pleased to provide you with a quote. If you need further advice or assistance, please enquire or contact us during business hours.


text:
Official School Video
Bodwell High School is a Canadian private co-educational boarding school, offering grade 8-12 instruction, and university preparation. Founded in 1991, Bodwell High School is situated in a picturesque corner of Vancouver, British Columbia. The school is certified and inspected by the British Columbia Ministry of Education and is a recognized leader among Canadian boarding schools. Most Bodwell graduates (97%) successfully enter universities, and many of them receive scholarships.
While striving to fulfill the main purpose of education – attainment of academic knowledge, Bodwell also focuses a lot of attention on students' physical, aesthetic and cultural development so as to mold students into well-rounded, balanced, young adults.
East Wing Opening Ceremony - Sept. 7, 2017
Find out what makes Bodwell one of the best boarding schools in Canada in your own language:
"Thank you for taking care of my daughter, Irina who spent four wonderful terms at the school. Words can't express my enormous gratitude to you as a parent. My daughter attended classes with pleasure – thanks to the teachers. She had fun and it was interesting for her to spend weekends and vacations at the school. She lived in a clean room, ate delicious breakfasts and dinners – thanks to the cooks and the technical staff. She has made a lot of friends from different countries – thank you to everyone at the school."
Bodwell's university guidance & career team has developed strong relationships with many post-secondary institutions. Through one-on-one counselling, workshops and presentations, they support students in selecting and applying to universities and colleges across Canada, the U.S.A. and around the world.
In 2016, 163 graduates received a total of 337 offers of admission from 92 universities and colleges in 12 countries.
of graduates are accepted to college or university right after graduating
[visualizer id="18094"]
Bodwell High School's curriculum is carefully verified, meaningful, diverse, well-balanced and, most importantly, promotes harmonious personal, intellectual, and physical development. Bodwell constantly ensures improvement and adherence of its curriculum to the standards of the British Columbia Ministry of Education.
Committed to innovative education, Bodwell provides all students with laptops optimized for academic research, which includes essential software like Microsoft Office 365 and Adobe Creative Cloud. Our dedicated IT team provides comprehensive support, as well as scheduling and monitoring high-speed Internet for appropriate content.
Bodwell teachers encourage their students to take initiative, and help them to develop leadership skills and innovative approaches to problem solving. The school also has experienced counselors, who are ready to listen and help.
Trimester Schedule – The school year is divided into three terms: fall (September-December), winter (January-April) and summer (April-July). New students can start school at the beginning of any term. The trimester system allows students to finish the school program faster and thus complete 3 grades in 2 years, if desired.
All students are required to participate in extracurricular sports and other activities. The school organizes a large number of cultural and recreational events which include outings to theatres, sports competitions, excursions to different cities in Canada, etc.
All students attend three hours of enrichment classes every Saturday morning, on top of 30 hours of academic classes Monday to Friday. Students enrol in a Saturday course each term in one of the following areas: -English Enrichment -Global Citizenship -Career Exposure -Outdoor Education & Sports -New Media and Performing Arts. Students are guided to select and register for Saturday courses each term, which are included in the report card.
Students live in onsite dormitories. (designed for more than 500 people). The rooms (2-4 person capacity) have spectacular views of the mountains and the bay. Girls and boys live in separate sections of the dormitory under constant supervision of youth advisors.
Our boarders gain skills for life in four areas: self-care, interpersonal skills, personal planning and leadership.
All new students start in boarding. Upon completion of the Boarding Program, students may apply to move to a Bodwell homestay.
Homestay program – Another accommodation option following time in boarding is staying in a homestay with carefully selected Canadian families, where the student is given a full room and board in a private room.
Our 6 L's – Becoming well-rounded | The essence of a successful life | At Bodwell, we strongly believe that there are several powerful aims of education all working together to guide the development of young people towards a balanced character. This concept shapes everything that we do at Bodwell, represented by "6 L's." Each L is an essential part of the Bodwell experience in both the academic and boarding programs.
The "House System" – On arrival students will be placed in one of the six "houses" where these future world citizens learn about the values needed to live together, values common to many societies around the globe. They learn about fairness and social justice, non-violence and peaceful resolution to conflict, intercultural understanding and acceptance, compassion and service to others, and respect for the dignity of the individual.
Learn to Excel through English
Liberate the Academic Mind
Live an Active and Healthy Lifestyle
Lead for Positive Change
Look Inward and Forward
Love the Earth and One Another
Boarding Program Benefits
|Wake up
|7:30 - 8:10
|8:15 - 11:30
|11:30 - 12:10
|12:15 - 15:30
|15:30 - 16:50
|Extracurricular Clubs & Teams
|17:00 - 18:00
|Free Time
|18:00 - 19:00
|19:00 - 21:15
|Study & Life Skills
|21:15 - 22:15
|Free Time
Boarding Life - Student Perspective
The City of North Vancouver is a safe and friendly community with a rich cultural heritage and numerous places of natural beauty. It is surrounded by spectacular mountains in the north and picturesque beaches in the south. The campus area of 70000 square meters overlooks the sea and is close to city parks.
Modern facilities with an indoor swimming pool, 2 gymnasiums, 2 lecture theatres, 2 cafeterias, science labs, a library, and a music room, Bodwell also features the latest technology with all classrooms equipped with either SMARTBoards or high definition TV screens.
• Type of school: Private co-educational boarding school for students grades 8-12
• Founded: 1991
• Age of students: 13-18
• Number of students: Around 600 from more than 40 countries
• Average class size: 18 (max. 24)
• Number of boarding spaces: 500
• 24-hour student support led by a team of over 100 professionals
• Effective and accelerated trimester system allows the completion of a three-year program in two. Students have the opportunity to start school in September, January or April
• Programs offered: Certificate of Secondary Education (Dogwood Diploma) issued by the Ministry of Education of British Columbia, academic preparation, secondary school curriculum, and university preparation.
• The majority of graduates enrol in prestigious universities in Canada, USA and other countries.
• Individual approach to each student, friendly atmosphere, and spacious classrooms.
• Accommodation in onsite dormitory or full room and board with a Canadian family.
• Enhanced sports and cultural activities program with an indoor swimming pool, 2 gyms, 2 lecture theatres, science labs, a library and a music room.
• Tuition (all-inclusive) for 2 terms are $38,600 CAD in boarding.
Contact us!
For Admission details, prices, and other information about the school, please contact Bodwell Admissions, or one of our education representatives in or near your city.


text:
Louisa Blue
Grab a pen and paper before reading Jeanie Doyle Singler's third mystery novel, Louisa Blue, because it will be helpful to take notes on the action, as this investigation into an unsolved murder grows more complex with each turning page. The list of suspects is extensive, and the potential motives are many, including jealousy, greed, and religion.
Singler opens Louisa Blue with an unusual missing persons report. The wealthy Spencer Townshend has been missing for over a year, after disappearing during a ski trip in Whistler, British Columbia. His wife, Louisa, still wonders: Did he take off intentionally, or did something happen to him? Louisa flees Spencer's astoundingly dysfunctional family and moves to Tacoma, where she meets pastor Dark Ansgreth, whose own wife has recently died in a mysterious car accident. Meanwhile, Spencer's illegitimate cousin Alexander LaGuardia receives an odd directive from his father's will: If you want to receive your inheritance, find out what happened to Spencer.
Pulling all of these threads together is Singler's most difficult task, one she performs ably, if somewhat slowly, over three-hundred-plus pages. Her explanation of events is clear, as Alexander retraces Spencer's steps and interviews friends, relatives, and business associates. She uses dialogue to deliver new information, as Alexander talks to Louisa, Louisa talks to Dark, and Dark talks to his friend Davy, a detective.
As the mystery deepens, so do the relationships among the truth seekers. Louisa is enigmatic, vacillating between passionate, emotional declarations and ice-cold, logical evaluations. This volatility, along with Louisa's fairy-tale beauty—long, curly hair and emerald-green eyes—draws men to her side. Alexander and Dark are not immune to her charms, and the potential love triangle adds tension to the search for answers.
Singler is comfortable with details, and perhaps a bit too enthusiastic about the particulars of each scene. For instance, Alexander doesn't just walk in the front door of a new house, but rather surveys the layout of the house and the contents of each room. Run-on sentences describe the floral patterns in the sofa fabric, along with the carpet color and the type of curtains in the windows. The surplus of domestic details distracts from the action and slackens the pace.
Singler alternately creates and extinguishes tension as the list of likely victims grows. On one hand, the reader follows Alexander and Dark closely, hoping along with them for a breakthrough with each new clue. On the other hand, however, the reader loses sight of the goal when Singler shifts into flowery prose. The gritty details of car chases and police barricades lose their power against the "apprehensive mist" of the moonlight and the "apricot sun" on the horizon.
Resolution comes late in the game, and a reader might turn to her notes in the last pages, curious to see if all the loose ends have been tied up. Mysteries both murderous and romantic are indeed wrapped up with satisfying logic, while leaving ample room for future adventures.
Reviewed by
Sheila M. Trask
Disclosure: This article is not an endorsement, but a review. The author of this book provided free copies of the book and paid a small fee to have their book reviewed by a professional reviewer. Foreword Reviews and Clarion Reviews make no guarantee that the author will receive a positive review. Foreword Magazine, Inc. is disclosing this in accordance with the Federal Trade Commission's 16 CFR, Part 255.


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The young and unpredictable military leader of North Korea is placed in the crosshairs of an assassin's scope at the Great Wall of China. The killing attempt points to the CIA and President Harris O'Neil. A revenge plot unfolds aimed at Secret Service vulnerabilities; the president's teenage daughter, Bibi, the victim.
A brief and flirtatious encounter with 17-year-old Baltimore superstar athlete Dante Brown the night Bibi is abducted offers Homeland Security a surprising clue. But is it enough to unravel the mysterious kidnapping, the identity of the abductors, and the reason behind the revenge?
Author novelist Kent Politsch sends his readers on a new adventure with federal bureaucrat Jack Fitzgerald and his team of teenage basketball players.


text: The best Pennsylvania adult video on demand with hundreds of videos to choose from and no signup ever necessary. Introducing Free Adult Store, featuring live webcasts by thousands of girls around the world on the largest adult entertainment website. Click a city within Pennsylvania or browse below and start watching the best adult entertainment.


text:
- Product Description
- Specification
- Shipping Options
- Reviews
Jaco 3 Light Semi Flush Antique Brass Ceiling Light
Exclusive to Pagazzi, the Jaco 3 Light Semi Flush Antique Brass Ceiling Light is a gorgeous ceiling light with semi flush fitting. The light features an antique brass finish and white glass shades upon a twisted frame.
- PAGAZZI Exclusive.
- Antique Brass Finish.
- Frosted White Glass Shades.
It measures a height of 155mm, a diameter of 370mm and requires 3 x Golfball SES 40w bulbs.
Additional Information
|Weight (kg)
|Height (mm)
|Width (mm)
|Diameter (mm)
|Number of Bulbs
Orders of £50 and over qualify for free next day UK Mainland delivery* with DPD.
We also offer free delivery to BT postcodes and Highlands and Islands addresses on orders of £100 and over.
|Orders of £50 and over placed before 3pm
(UK Mainland*)
|FREE Next Working Day Delivery
|Orders of £100 and over placed before 3pm
(BT Postcodes, Highlands & Islands*)
|FREE Two Day Delivery
|Orders under £50 placed before 3pm
(UK Mainland*)
|£5.95 - Next Day Service
|Orders under £100 placed before 3pm
(BT Postcodes, Highlands**)
|£9.95 - Two Day Service
|Orders under £100 placed before 3pm
|£14.95 - Two Day Service
|Saturday or Sunday Delivery - Orders placed before 3pm on Friday
(UK Mainland*)
|Belgium, France, Germany, Ireland, Luxembourg, Netherlands
Standard (2-3 Working Days)
See our full delivery details page for detailed information.
For further details please contact us


text:
|City Archives
New Orleans Public Library
Photographs Arranged by Street Names
Piety Street
Click on the thumbnail to see a larger image. Use your browser's back button to return to this page.
Captions taken from the back of photographs
|Piety Street from Miro Street to Florida Ave "before"
Paving Lien-6th Phase
February, 1961
[Photography by J.R. St. Julien]
|Return to Photographs Arranged By Street Names


text:
$17 million STEM focused expansion opens at Colorado high school
Caitlin Hendee
Digital Producer / Social Engagement Manager- Denver Business Journal
A $17 million expansion meant to help alternative high school students focus on building a business or science, technology, engineering or math (STEM) focused career opened Thursday.
The 98,000 square-foot space at the Englewood School District's Colorado's Finest High School of Choice was designed by Slaterpaull / Hord Coplan Macht, a Denver architectural firm, and built by Centennial-based Haselden Construction LLC. Read more.


text:
Thompson, William Boyce
Thompson, William Boyce, 1869–1930, American financier, b. Virginia City, Mont. He operated silver and copper mines in Montana and Arizona before moving to New York City. He was (1914–19) a director of the Federal Reserve bank of New York and was twice (1916, 1920) a delegate to the Republican national convention. In World War I, he helped finance and accompanied (1917–18) a Red Cross mission to Russia. He contributed money to the government of Aleksandr Kerensky and, after the Bolsheviks seized power, advocated U.S. recognition of the Soviet government. In 1919, he founded the Boyce Thompson Institute for Plant Research in Yonkers, N.Y.
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