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Wnt Signaling in Oncogenesis and Embryogenesis--a Look Outside the Nucleus
Mark Peifer1,*,
Paul Polakis2
1 Department of Biology and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599–3280, USA.
2 Genentech, 1 DNA Way, Mail Stop #40, S. San Francisco, CA 94080, USA.
Science 03 Mar 2000:
Mark Peifer
Paul Polakis
The Wnt cell-cell signaling pathway plays a critical and evolutionarily conserved role in directing cell fates during embryogenesis. In addition, inappropriate activation of the Wnt signal transduction pathway plays a role in a variety of human cancers. Many recent studies of Wnt signaling have provided mechanistic insight into these dual roles. Here we focus on two areas of rapid advance: (i) the machinery that regulates the stability of the key signal transducer, β-catenin, and (ii) the effect of Wnt signaling on cellular targets outside the nucleus, the actin and microtubule cytoskeletons.
Some of the most profound questions in biology were first asked by ordinary people confronting events in their everyday lives. The miracle of a newborn baby raises questions about how an egg assembles itself into an animal. The tragedy of a cancer diagnosis leads to questions about how normal cells go wrong. Although seemingly distinct, biological processes like embryogenesis and carcinogenesis both rely on cell communication via identical signaling pathways. For example, in the larva of the fruit fly Drosophila, cells determine their position within each body segment by communicating with one another. One of the key signaling molecules is the secreted Wnt family protein Wingless (Wg), which acts as a "Be posterior" signal [reviewed in (1)]. Fate decisions require changes in gene expression; thus cells must transmit information from the cell surface to the nucleus.
Wnt signaling also regulates cell proliferation in adult tissues. The epithelial cells lining the colon provides an excellent illustration of this [reviewed in (2)]. Colonic cells proliferate at a rate that perfectly balances the death of cells due to attrition. Cells are sent signals to proliferate when appropriate, and when sufficient cell numbers are attained, proliferation is halted. Colon cancer, like other cancers, results in part from mutations that cause cells to receive a continuous signal to proliferate. Mutations can lock a signal transduction component in an ON position or can inactivate a protein that normally keeps the pathway in an OFF position. For example, in most colon cancers, a negative regulator of Wnt signal transduction, APC [the tumor suppressor protein encoded by the adenomatous polyposis coli (APC) gene], is inactivated, and the Wnt pathway is aberrantly turned on.
In this review we will summarize the current model for Wnt signal transduction and then discuss two particularly active areas of investigation: the critical role of regulated protein destruction in determining where the Wnt pathway is ON or OFF and cytoskeletal targets of the pathway. There are many other aspects of Wnt signaling that we will not cover; for a more comprehensive review see (1). Detailed information about Wnt signaling can also be found on the Wnt gene homepage (http://www.stanford.edu/∼rnusse/wntwindow.html) and at the connections map at Science's STKE Web site (http://www.stke.org).
A Model for Wnt Signaling
Wnt signaling is regulated by the presence or absence of the intracellular protein β-catenin [Fig. 1; reviewed in (1)]. A large multiprotein machine that includes proteins of the APC and Axin families normally facilitates the addition of phosphate groups to β-catenin by glycogen synthase kinase-3β (GSK3β). Phosphorylated β-catenin binds to a protein called βTrCP, and is then modified by the covalent addition of a small protein called ubiquitin. Proteins tagged with ubiquitin are degraded by the proteosome, the cell's protein-recycling center. When Wnt signal is absent, the signal transduction pathway is OFF because β-catenin is rapidly destroyed. When cells are exposed to Wnt, it binds to cell surface receptors of the Frizzled family. Receptor activation antagonizes the APC-Axin "destruction complex" by an unknown mechanism that requires Dishevelled protein. This blocks β-catenin phosphorylation and its subsequent ubiquitination. β-Catenin is thus diverted from the proteosome, and it accumulates and enters the nucleus, where it finds a partner, a DNA binding protein of the TCF/LEF family. Together, they activate new gene expression programs. In the embryonic skin of a fruit fly, Wg signal turns on genes conferring "posterior identity," whereas in human colon cancer cells, inappropriate activation of the Wnt pathway drives cell proliferation by turning on genes encoding oncoproteins and cell-cycle regulators. As discussed below, Wnt signals also regulate the polarity and function of both the actin and microtubule cytoskeletons.
(left). The Wingless/Wnt signaling pathway. Schematic summary of the response of a cell to Wg signal. See text for details. The icon for β-catenin, an armadillo, is based on the name of its fruit fly homolog.
The β-Catenin Destruction Complex
The tumor suppressor protein APC is a critical component of the β-catenin destruction machinery. APC is inactivated in most colorectal cancers, resulting in activation of the Wnt pathway. The Wnt pathway also plays a role in other cancers: for example, viral activation of Wnt-1 causes mammary tumors in mice, and mutations that make β-catenin refractory to destruction are found in a variety of human tumors [reviewed in (2)]. Elucidation of the β-catenin destruction machinery may reveal how Wnt signaling regulates embryogenesis, and may uncover new oncogenes and tumor suppressors. Many proteins that work with APC in regulating β-catenin have been identified, including Axin, Dishevelled, GSK3β, β-TrCP, casein kinases 1ɛ and II, protein phosphatase-2A, and FRAT (frequently rearranged in advanced T cell lymphomas). These proteins all interact functionally or physically, but their specific roles in β-catenin regulation remain to be resolved.
A second key component of the destruction machinery is Axin [reviewed in (2)]. Mice lacking functional Axin have defects in development of the dorsal-ventral body axis similar to those elicited by Wnt overexpression in embryos of the frog, Xenopus laevis. In both Xenopus and mammalian cells, Axin and its paralog Conductin/Axil destabilize β-catenin, thus negatively regulating Wnt signaling (3–6). Axin and APC physically interact. The binding site in Axin is in its RGS (regulator of G protein signaling) domain. The RGS domain binds to three copies of a short protein motif in APC known as the SAMP repeat (4,6). The truncated APC proteins found in most colon cancers lack these SAMP repeats and thus cannot bind Axin. Likewise, APC protein fragments that destabilize β-catenin in cultured cells contain both β-catenin and Axin binding sites [reviewed in (2)]. It has been hypothesized that the truncation of APC observed in cancer may be driven by selection against Axin's ability to bind APC. Consistent with this idea, mice carrying truncating mutations in APC that remove the Axin binding sites develop colon tumors at a high frequency [reviewed in (2)], whereas mice carrying truncating APC mutations that retain an Axin binding site do not develop tumors (7).
Why is the physical interaction between APC and Axin so critical to β-catenin regulation? This issue is far from clear, as conflicting data have emerged from studies of different animal models. InXenopus, Axin mutants lacking the RGS domain not only fail to inhibit Wnt signaling but they act positively, promoting dorsal axis duplication. This is consistent with an obligate role of APC binding in Axin-mediated destruction of β-catenin [reviewed in (2)]. In contrast, mammalian Axin mutants lacking the RGS domain still destabilize β-catenin in cultured cells (4–6, 8, 9). Similarly,Drosophila Axin lacking the RGS domain inhibits Wg signaling when ectopically expressed (10). One interpretation is that APC stimulates or localizes Axin by binding to its RGS domain; overexpression of Axin mutants lacking this domain might overcome the requirement for APC. The differences observed in Xenopuscould relate to relative expression levels. Consistent with the hypothesis that APC family members localize the destruction complex, a mutation in Drosophila APC2 that disrupts its cortical localization also disrupts its function (11). Whereas most data suggest that APC negatively regulates Wnt signaling, experiments with Xenopus and the nematode Caenorhabditis elegans suggest that APC may also play an unexpected positive role in Wnt signaling [reviewed in (2)].
The APC-Axin complex is thought to modulate activity of the serine/threonine kinase GSK3β. Mutation of the GSK3β homolog inDrosophila and expression of dominant-negative GSK3β inXenopus activate Wnt signaling [reviewed in (1)], suggesting that Wnt signaling inhibits GSK3β (Fig. 1). One substrate of GSK3β is β-catenin. The GSK3β phosphorylation sites of β-catenin are mutated in tumors; these mutations stabilize β-catenin [reviewed in (2)]. Although β-catenin is a poor GSK3β substrate in vitro, β-catenin phosphorylation is significantly enhanced when a fragment of Axin containing β-catenin and GSK3β binding sites is included in the reaction (3, 4, 12). Thus, Axin may serve as a scaffold on which both GSK3β and β-catenin reside, facilitating their interaction (Fig. 2). Both APC [reviewed in (2)] and Axin (12) are also GSK3β substrates and their ability to bind β-catenin in vitro is enhanced by phosphorylation. Wnt signaling in mammalian cells leads to Axin dephosphorylation, as does incubation of cells with LiCl, a GSK3β inhibitor (12, 13). The mechanism by which Wnt signaling "turns off" GSK3β remains unclear. In fruit flies, at least, Dishevelled is critical for this. Recent demonstrations that Dishevelled is part of the "destruction complex" (8, 9, 14) and binds directly to Axin, may provide the first mechanistic clue. Other proteins such as FRAT, which binds GSK3β and Dishevelled and which antagonizes GSK3β function when ectopically expressed (14), may also play a role.
(right). Linear representation of Axin, APC, β-catenin, and Dishevelled (Dvl), with the approximate locations of the binding sites for various protein partners indicated with arrows. The length of each polypeptide in amino acids is shown to the right. Two numbering conventions have been adopted for Axin, as the initiation codon has not been identified. In Axin, the DIX domain is a domain shared with Dishevelled. In APC, HR is heptad repeat, ARM indicates seven Armadillo repeats (a motif first found in the fruit fly β-catenin homolog Armadillo), + indicates a region rich in basic amino acids, and Dlg/PSD is the binding site for the PDZ protein Discs large/PSD-95. The majority of APC-truncating mutations in colon cancer occur within the mutation cluster region (MCR), thus producing a truncated protein lacking Axin binding sites. In β-catenin, ST indicates serine and threonine residues that affect protein stability; ARM indicates 12 Armadillo repeats. LIT-1 is theC. elegans Nemo-like MAPK homolog; its binding has been demonstrated only with Wrm-1, a C. elegans relative of β-catenin. Dishevelled has three recognizable domains: a DIX domain shared with Axin, a PDZ domain, and a DEP (Dishevelled, EGL-10, pleckstrin) domain.
Phosphorylation by GSK3β is critical to β-catenin destruction. Other proteins are targeted for ubiquitination—and thus destruction—by phosphorylation. Sites for this phosphorylation have been mapped in some, such as the nuclear factor kappa B (NF-κB) regulator IκB. IκB and β-catenin share sequence similarity within the region required in both proteins for destruction. This is reminiscent of the situation in yeast, where the stability of certain proteins is regulated by E3 ubiquitin ligases that only recognize their substrates when phosphorylated. F-box proteins act as the recognition subunits. Mutation of theDrosophila F-box protein Slimb stabilizes the fruit fly β-catenin homolog (15), leading to speculation that Slimb and its human homolog β-TrCP form part of an F-box–containing ubiquitin ligase targeting phosphorylated β-catenin. This hypothesis has now been experimentally verified (16–18). β-Catenin binds β-TrCP only when phosphorylated by GSK3β, and oncogenic β-catenin mutants that lack GSK3β phosphorylation sites do not bind β-TrCP. Further, overexpression of a β-TrCP derivative lacking the F-box activates β-catenin–dependent gene expression. Studies with Xenopusembryos are also consistent with a role for β-TrCP as a negative regulator of β-catenin signaling (19). Thus β-TrCP is the penultimate stop on β-catenin's trip to the proteosome. These observations explain how oncogenic mutants of β-catenin avoid the proteosome and continue to signal.
Although this may seem complex enough, other protein players continue to be identified. Where there are kinases, there must be phosphatases. Wnt signal triggers Axin dephosphorylation, thus reducing its ability to bind β-catenin and trigger its destruction (12). The protein phosphatase PP2A dephosphorylates Axin in vitro, and the PP2A inhibitor okadaic acid blocks Wnt-mediated Axin dephosphorylation in mammalian cells, suggesting that PP2A may regulate β-catenin stability in response to Wnt signaling (12). PP2A's catalytic subunit binds directly to Axin; its B56 regulatory subunit interacts with APC (20, 21). Overexpression of the B56 subunit downregulates β-catenin protein levels and Wnt signaling (21). Finally, the Aβ regulatory subunit of PP2A is mutated in a subset of primary colon tumors (22). It remains problematic that PP2A's effect on Wnt-triggered Axin dephosphorylation is consistent with a positive role in regulating the Wnt pathway, whereas the downregulation of β-catenin by the B56 subunit is consistent with a negative role.
More recently, another serine/threonine kinase, casein kinase 1ɛ (CK1ɛ), entered the fray. CK1ɛ can stabilize β-catenin and thus activate Wnt-target genes, and dominant negative forms of CK1ɛ inhibit Wnt signaling (23, 24). CK1ɛ has been placed downstream of Dishevelled and upstream of GSK3β, although biochemical analysis suggests that it binds directly to Dishevelled's PDZ region (Fig. 2). Dishevelled is phosphorylated when coexpressed with CK1ɛ, implicating it as a possible target. Thus, CK1ɛ has the hallmarks of a Wnt signaling molecule and may represent a missing link between the Wnt receptor and β-catenin stabilization. Axin also binds a third serine/threonine kinase, the MAP kinase kinase MEKK1 [Fig. 2 (25)].
The APC-Axin Complex—Some Assembly Required
When considering the numerous interactions reported to occur within the APC-Axin complex (Fig. 2), one is confronted with many potential models for complex assembly and mechanisms of action. APC binds directly to itself, β-catenin [reviewed in (2)], Axin (4, 6, 18, 26), and the PP2A B56 subunit (21), whereas Axin binds directly to itself (5, 8, 20), APC, β-catenin, GSK3β (3, 4, 6, 27), the PP2A catalytic subunit (20), and Dishevelled (8,9, 14). In turn, Dishevelled binds to itself (8), Axin, CK1ɛ (23, 24), and FRAT (14). Moreover, β-catenin can bind β-TrCP while associated with the APC-Axin complex (16). Given this complexity, it is clear that linear models for signaling are not appropriate.
How might things work? The binding of APC to the Axin-GSK3β complex could promote APC phosphorylation and thus increase its affinity for β-catenin. This might allow proper presentation of β-catenin to GSK3β, resulting in its efficient phosphorylation and subsequent binding to β-TrCP. Upon activation by Wnt, CK1ɛ might phosphorylate Dishevelled, resulting in the displacement of FRAT from Dishevelled, thereby positioning FRAT to inhibit the GSK3β that is associated with Axin. These models are consistent with the literature, but so are many others. There are also unsolved mysteries: for example, what is the link between the destruction machinery and Wnt receptors? Perhaps we are still missing a G protein. After all, Frizzled proteins distantly resemble G protein–coupled receptors, Axin harbors an RGS domain, and Dishevelled contains a DEP (Dishevelled, EGL-10, pleckstrin) domain, which is also present in certain RGS proteins. However, researchers looking for a G protein connection to Wnt-1 signaling have so far come up empty-handed, though Wnt-5A may utilize one. A related mystery is the intracellular localization of the destruction complex. Is it assembled adjacent to the Frizzled receptor? Finally, thus far only APC and β-catenin have been found mutated in primary human tumors. Mutations in other destruction complex components may also be uncovered in cancer.
All Roads Do Not Lead to the Nucleus
Extracellular signals clearly affect gene expression, and they also induce changes in cell shape, in cell-cell interactions, and in cell migration. Many of these effects are mediated by the cytoskeleton. Wnt signals affect both the actin and microtubule cytoskeletons. The effect on the actin cytoskeleton occurs in epithelial cells, which possess two sorts of cell polarity. Apical-basal polarity distinguishes the top and bottom surfaces of the cell sheet; planar polarity allows cells to determine directions in the plane of the sheet, providing compass coordinates as on a map. Wnt signaling regulates planar polarity (Fig. 3A) [reviewed in (28)]. For example, the hexagonal cells of a fruit fly wing each secrete a single hair, an actin-filled plasma membrane projection. All hairs extend distally from the distal cell vertex. Certain mutations disrupt planar polarity, so that cells polymerize actin at random locations, thereby randomizing the position and direction of wing hairs. Two planar polarity genes encode the most upstream known components of the Wg pathway: Frizzled, a Wg receptor, and Dishevelled, which acts just downstream.
Wingless/Wnt signaling directly regulates the cytoskeleton. (A) In tissue polarity, Wnt signaling directs polymerization of the actin cytoskeleton at particular positions on the cell cortex with respect to the animal's body axes. (B) Flamingo protein is enriched on particular sides of the cell and thus may mark the side of the cell where actin polymerization should be directed. (C) Model in which Dishevelled acts as a branchpoint, with different signal transduction pathways activated to modulate cell fate choices or to regulate planar polarity. (D) Wnt signaling directs the positioning of the cell's mitotic spindle in both early C. elegans embryos (this example) and Drosophila sensory cells. (E) dAPC2 localizes to a crescent in larval neuroblasts undergoing an asymmetric cell division. This crescent localizes with one pole of the spindle.
An alternative signal pathway appears to mediate the effects of Wnts on planar polarity (Fig. 3C) [reviewed in (29)]. Genetic epistasis experiments suggest that the small guanosine triphosphatase Rho, and a kinase cascade including Misshapen (a Ste20 homolog), JNKK, and JNK (the JNK pathway) act downstream of Frizzled in planar polarity. Furthermore, Dishevelled can activate JNK (30,31), suggesting that there is a branchpoint at Dishevelled, with one branch leading to the canonical Wg pathway and regulating cell fate, and the other leading to the JNK pathway and regulating planar polarity. The discovery that Axin overexpression stimulates JNK signaling (25) suggests that Dishevelled may work with Axin in this process. Loss-of-function mutations in JNKK and JNK do not affect planar polarity directly (30), suggesting that other MAPK proteins play a redundant role in the process. Candidate genes have been identified in C. elegans: one encodes a MAPKKK related to vertebrate TAK1, and the other encodes a divergent MAPK family member related to Drosophila Nemo (32–35). These kinases are required for Wnt signaling but are postulated to act in a pathway parallel to the traditional Wnt pathway. TAK1 and Nemo thus could be part of an alternative kinase pathway that operates in planar polarity.
In the simplest model, Frizzled receptors could activate the canonical Wnt pathway in some tissues and the JNK pathway in others. This could occur if different Frizzled family receptors used specific signal transduction pathways. However, Drosophila Frizzled, originally implicated in planar polarity, acts upstream of Armadillo, the β-catenin homolog, in the embryonic epidermis and central nervous system (36–40), implying that individual receptors can act through either pathway. One remaining question is which Wnt, if any, is involved in planar polarity.
Other fruit fly proteins required for planar polarity are not obviously connected to Wg signaling. Most encode novel proteins with recognizable motifs but without homologs of known function: for example, Strabismus and Fuzzy are transmembrane proteins [reviewed in (28)], and Prickle has multiple Lim domains (41). These proteins may be the effector machinery activated by Wnt signaling to alter actin polymerization. The discovery of Flamingo (also known as Starry Night) (42–44) may reveal new insight. Flamingo's extracellular domain resembles that of the cadherins, a family of cell adhesion molecules, and Flamingo can mediate cell-cell adhesion in culture. COOH-terminal to Flamingo's cadherin repeats is an apparent G protein–coupled receptor domain, distantly resembling Frizzled family receptors. Perhaps most intriguing is Flamingo's subcellular localization. Before the initiation of planar polarity, it is found uniformly around the cell cortex. However, as planar polarity is being established, Flamingo becomes preferentially localized to the faces of the cell where actin polymerization will occur (Fig. 3B). It thus may mark those cell surfaces, distinguishing them from the others. Both genetic and cell biological tests suggest that Flamingo acts downstream of Frizzled. It could couple Wnt signal to cytoskeletal rearrangements, marking a site on the cell cortex where actin should polymerize.
Another possible effector that may couple Wnt signaling to actin rearrangement is the APC relative Drosophila APC2 (dAPC2). dAPC2 colocalizes with actin and is found in cell-cell adherens junctions of polarized cells (11, 45). dAPC2 colocalizes with actin during assembly of the embryonic denticle (11), an actin-based structure similar to the developing wing hair. Thus dAPC2 is properly positioned to influence actin polymerization. However, a role for it or other APC proteins in planar polarity remains to be tested.
Wnt signaling also affects positioning of the mitotic spindle. Certain cells divide with a fixed orientation relative to the body axes of the animal, requiring that the mitotic spindle be properly oriented. In both C. elegans and Drosophila this process can be directed by Wnt signaling. During division of an early embryonic cell in C. elegans called EMS (Fig. 3D) [reviewed in (46)], the mitotic spindle is oriented by contact with a neighboring cell, P2, such that one pole is directed toward the signaling cell. The polarizing signal is thought to be a Wnt molecule acting via a Frizzled receptor. Wnt signaling directly affects the cytoskeleton (47): inhibition of transcription does not block spindle positioning, indicating that this effect does not require activation of Wnt target genes. In C. elegans, Frizzled and GSK3β homologs are essential for spindle orientation (47), whereas in fruit flies Frizzled and Dishevelled are essential (48). In contrast, double-stranded RNA inhibition of the C. elegans β-catenin relative Wrm-1 or of the distant APC relative APR-1 does not block spindle orientation, nor do mutations in the Wnt mom-2 or the TCF relative pop-1. One interpretation of these data is that there is a branch in the pathway at GSK3β, with one pathway affecting gene regulation and the other influencing spindle orientation. However, differences in the genetic circuitry and divergence in the sequences of Wrm-1 and Apr-1 relative to their homologs suggest that the nematode pathway may be mechanistically different than that in mammals or fruit flies, so this may not apply to other animals. Regardless, because Dishevelled, GSK3β, and APC can all reside in the same protein complex (see above), output from this complex may affect signaling to both the nucleus and to the cytoskeleton.
The mechanism by which Wnt regulates spindle positioning remains to be determined. One possibility is that APC plays a role: mammalian APC binds and bundles microtubules in vitro, and in migrating cultured cells it localizes to spots at the plasma membrane where bundles of microtubules terminate [reviewed in (49)]. Further, in dividing stem cells of the Drosophila brain, dAPC2 localizes asymmetrically within the cell to a crescent adjacent to one spindle pole (11) (Fig. 3E). The functional significance of this localization remains to be tested. The cadherin-relative Flamingo is also required for spindle orientation in neural precursors (43), suggesting that it mediates the effects of Wnt signaling on both the actin and the microtubule cytoskeletons.
In summary, investigation of the Wnt signaling pathway has revealed insights into both embryogenesis and oncogenesis. Many challenges remain. We now have an outline of how this critical pathway operates, but fleshing out the precise mechanisms involved will occupy us for years to come.
↵* To whom correspondence should be addressed. E-mail: peifer{at}unc.edu
A. Wodarz,
R. Nusse
, Annu. Rev. Cell Dev. Biol. 14, 59 (1998).
P. Polakis
, Curr. Opin. Genet. Dev. 9, 15 (1999).
S. Ikeda
, EMBO J. 17, 1371 (1998).
M. J. Hart, R. de los Santos,
I. N. Albert,
B. Rubinfeld,
, Curr. Biol. 8, 573 (1998).
C. Sakanaka,
L. T. Williams
, J. Biol. Chem. 274, 14090 (1999).
J. Behrens
R. Smits et al., Genes Dev. 13, 1309 (1999).
S. Kishida et al., Mol. Cell. Biol. 19, 4414 (1999).
M. J. Smalley et al., EMBO J. 18, 2823 (1999).
K. Willert,
C. Y. Logan,
A. Arora,
M. Fish,
, Development 126, 4165 (1999).
B. M. McCartney et al., J. Cell Biol. 146, 1303 (1999).
S. Shibamoto,
, Genes Dev. 13, 1768 (1999).
H. Yamamoto et al., J. Biol. Chem. 274, 10681 (1999).
L. Li
J. Jiang,
G. Struhl
M. Hart et al., Curr. Biol. 9, 207 (1999).
M. Kitagawa et al., EMBO J. 18, 2401 (1999).
C. Liu et al., Proc. Natl. Acad. Sci. U.S.A. 96, 6273 (1999).
Y. Marikawa,
R. P. Elinson
, Mech. Dev. 77, 75 (1998).
W. Hsu,
L. Zeng,
F. Costantini
J. M. Seeling et al., Science 283, 2089 (1999).
S. S. Wang
J. M. Peters,
R. M. McKay,
J. P. McKay,
J. M. Graff
P. Leong,
L. Xu,
S. D. Harrison,
, Proc. Natl. Acad. Sci. U.S.A. 96, 12548 (1999).
Y. Zhang,
S. Y. Neo,
X. Wang,
J. Han,
S. C. Lin
S. Kishida et al., J. Biol. Chem. 273, 10823 (1998).
J. B. Weiss,
J. M. Shulman,
N. Perrimon,
J. D. Axelrod
, Trends Genet. 14, 452 (1998).
M. Boutros,
M. Mlodzik
N. Paricio,
D. I. Strutt,
, Cell 94, 109 (1998).
L. Li et al., J. Biol. Chem. 274, 129 (1999).
M. D. Meneghini et al., Nature 399, 793 (1999).
T. Ishitani
C. E. Rocheleau
T. H. Shin et al., Mol. Cell 4, 275 (1999).
K. M. Bhat
, Cell 95, 1027 (1998).
J. R. Kennerdell,
R. W. Carthew
H. Muller,
R. Samanta,
E. Wieschaus
, Development 126, 577 (1999).
P. Bhanot
C. Chen,
D. Gubb et al., Genes Dev. 13, 2315 (1999).
T. Usui
B. Lu,
T. Usui,
T. Uemura,
L. Jan,
Y. N. Jan
, Curr. Biol. 9, 1247 (1999).
J. Chae
X. Yu,
L. Waltzer,
M. Bienz
, Nature Cell Biol. 1, 144 (1999).
B. Bowerman,
C. A. Shelton
, Curr. Opin. Genet. Dev. 9, 390 (1999).
A. Schlesinger,
C. A. Shelton,
J. N. Maloof,
M. Meneghini,
B. Bowerman
M. Gho,
F. Schweisguth
A. I. Barth,
I. S. Nathke,
W. J. Nelson
, Curr. Opin. Cell Biol. 9, 683 (1997).
Work in the Peifer lab is funded by the NIH, the Army Breast Cancer Research Program, and the Human Frontiers Science Program. We regret that editorial policy precluded us from providing a more comprehensive list of primary references.
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By Mark Peifer, Paul Polakis
Science 03 Mar 2000 : 1606-1609


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ANZ Sports Scene: Kiwi paper plane thrower set to take flight at world champs
18 May, 2019 7:49am Quick Read
Blues star stands in support of Israel Folau
Netball: Noeline Taurua opens up on Maria Folau as World Cup selection looms
Maria Folau still looks set for Silver Ferns World Cup selection. Photo / Getty
Silver Ferns head coach Noeline Taurua said she would have no issue selecting Maria Folau in New Zealand's World Cup squad, despite her husband's ongoing saga with Rugby Australia.
Maria's husband, Israel Folau, had his four-year A$4 million contract terminated by Rugby Australia on Friday following the fallout to his anti-gay Instagram posts.
While Maria has not shared the same views as Israel publicly, the Silver Ferns veteran has supported him in the past through a post which read: "Stand with God no matter what ... Don't be afraid to stand up for the truth, even if that means you will be standing alone".
But the termination of Israel's contract has so far not bothered Taurua, who told Stuff Maria was still in the mix.
"I have no issue with her," Taurua said.
Silver Ferns coach Noeline Taurua. Photo / Getty
"I have no cause for concern at the moment. As long as she does what she needs to do out on court and is prepared as much as possible like anybody else, she's in the same [selection] boat.
"She's toiling away behind the scenes over here [in Australia]. She's learning a lot I know and being immersed in the Australian way will only be positive for her game."
Maria, who is currently playing for the Adelaide Thunderbirds in Australia's Super Netball competition, is all but set to be named in the Silver Ferns squad on Thursday as one of the game's best shooters.
Her experience with the British media, however, could pose problems with no restrictions around social media set to be placed on the Ferns during the World Cup - a timely reminder about using platforms wisely according to Taurua.
"Like everything we've got to be clear what our strategy is moving forward in regards to communication and regards to social media," she told Stuff.
'I'm very close to the end': Folau's massive hint
Kopua gets winning ending in front of home crowd
Maria and Israel Folau. Photo / Getty
"That's the given, I suppose. We're still in the throes of selection and we will work to make sure we have the plans sorted as to what that is going into the worlds or over in England."
Earlier this week, Maria revealed how she was nearing retirement, suggesting that July's World Cup could be her final outing in the black dress.
"Let's just say I'm very close to the end," she said, in an interview with former Australian goal shoot Catherine Cox.
"It's only been in the last couple of years where I've realised netball isn't everything.
"Netball is a huge part of my life, but there's other things in my life that have come into place that really need to take forward stead."

redpajama_set_name: RedPajamaCommonCrawl


In our PD lesson about mindset, we focus more on what it's called 'Growth Mindset".
What is growth mindset all about?
Growth mindset is the belief that qualities can change and that we can develop our intelligence and abilities. There are two types of mindsets. Fixed mindset and growth mindset.
Here is the chart showing the different between these two mindsets.
Intelligence isn't fixed but something that can grow and can be developed.
Challenge your brain can increase your intelligence and make it stronger.
Students with growth mindset focus on learning over just looking smart. They see effort has the key to success and thrive in the face of a challenge.
Growth mindset leads to a desire to learn, embrace challenges, persist in the face of setbacks and learn from criticism.
They find lessons and inspiration in the success of others. As a result, they reach even higher.
Here is another link to a very great YouTube animation that I really like about How to help every child fulfill their potentials.
The research shows that we should praise students for their effort, strategies, focus, persistence - not their intelligence.
Praising intelligence encourages a fixed mindset while praising effort, strategies, focus and persistence encourages a growth mindset.
As educators, it is our task to create a growth mindset classroom. In the safety of these classrooms, students can begin to leave behind their fixed mindset and try out the idea that they can develops their abilities.
"The best thing we can do is to teach children to love 'challenges, be intrigued by mistakes, enjoy effort and keep on learning."
The best way to help students develop a growth mindset is by teaching them about the amazing properties of the brain and neuroplasticity. Aaini, how would you explain the concept of neuroplasticity to a grade 5 student?


Bhastrika is the Sanskrit word for bellows which is why this technique is also referred to as the 'bellows breath'. It is a Pranayama breathing technique where the breath is forcibly drawn in and out as if using a blacksmith's bellows. Hence the name. Bhastrika pranayama is considered one of the most important breath exercises in yoga and pranayama.
In other Pranayama breathing techniques, inhalation sets the pace, the pattern, and the rhythm for exhalation. In bhastrika pranayama, exhalation sets the force and the pace. In bhastrika, both inhalation as well as exhalation are vigorous and forceful.
When practiced regularly, bhastrika improves digestion, immunity, and energizes the body and the mind.
There are four stages to this practice. In this article, we'll explore the technique and steps involved in all four stages of bhastrika pranayama.
In this stage, the nostrils are kept open throughout.
Sit in a comfortable position. You can choose to sit in padmasana or sukhasana, if you're comfortable in these asanas.
Sit quietly for a while, taking in slow, deep breaths. Keep your eyes open.
Lower your head towards the trunk and lift up the inner frame of the chest towards the descending chin. Rest the chin in the notch just above the breastbone (this is the chin-lock aka jalandhara bandha). If you find this difficult, just keep the head down as low as you can without causing any strain.
Keep the arms down and rest the back of your wrists on the knees.
Close your eyes and direct the gaze inwards. Breathe deeply.
On your next breath, exhale quietly as far as possible. Exhale whatever breath is in the lungs.
Now, take a short, strong breath and expel it with a quick strong blast. Repeat this and you'll find the second in-breath quicker and more forceful than the first one.
One quick inhalation and exhalation, taken together, completes one blast of bhastrika.
Do four to eight such blasts at a stretch to complete one cycle, ending with an out-breath.
Now take a few slow, deep breaths just as in ujjayi pranayama, for ten seconds. Then exhale slowly and completely. This rests the lungs and the diaphragm and prepares them for fresh blasts of bhastrika. This ends one round of bhastrika.
Repeat three to four rounds of bhastrika blasts interspersed with ujjayi breath (step #10). Then take a deep breath and lie in savasana (corpse pose).
This completes the stage 1 of bhastrika pranayama.
In this stage, both nostrils are kept partially closed throughout.
Sit in a comfortable position, following step 1-7 of Bhastrika Stage 1. Exhale whatever breath is in the lungs.
Bring the right hand to the nostrils.
Partially close both nostrils with the tips of the thumb, ring, and little fingers. Make sure that both sides of each nostril are even.
Now perform bhastrika blasts as described in points 7 to 11 of stage 1 above.
Repeat five to six times, take a few deep breaths, then lie down in savasana.
This completes bhastrika stage 2.
In this stage, bhastrika is done through alternate nostrils, with interspersion of ujjayi breaths.
Now block the left nostril completely and the right one partially.
Inhale and exhale through the right nostril vigorouslly, doing four to eight blasts at a stretch. Make sure that no breath escapes from the left nostril. End with a blast of out-breath.
Next, block the right nostril, partially block the left. Inhale and exhale through the left nostril vigorously, doing four to eight blasts at a stretch. See that no breath escapes from the right nostril. End with a blast of out-breath.
These two together complete one round of bhastrika, stage 3. If you're a beginner, take a few deep breaths after each round to rest the lungs.
Do three to five rounds of bhastrika, stage 3, take a few deep breaths and lie down in savasana.
This completes stage 3 of bhastrika.
In this stage, the in-blasts and out-blasts are done through alternate nostrils. In other words, if the in-breath is done from the right nostril, then the out-breath should be done from the left, and vice versa.
Block the left nostril, half-open the right and take a quick, but strong in-breath through it. Quickly close the right nostril, half-open the left and breathe out quickly and vigorously through it. Do four to five blasts in quick succession. This forms the first half-round.
Now, do the other half-round, repeating the same procedure as above, but breathing in from the left and out through the right. Do four to five blasts to complete the second half-round.
Do three to five rounds of bhastrika, stage 4, take a few deep breaths and rest in savasana.
This completes bhastrika stage 4.
It invigorates the liver, spleen, pancreas, and abdominal muscles.
It energizes the body and the mind.
It benefits the nervous system.
Drains the sinuses and stops the nose running.
Creates a feeling of exhilaration.
Hope you found this guide on bhastrika pranayama useful. How has been your experience with bhastrika? Do you practice it on a regular basis? Please share your thoughts on the practice in the comments section.


February 16, 2016 – [Los Angeles] CineConcerts and Warner Bros. Consumer Products are announcing the world premiere of The Harry Potter Film Concert Series, a new concert tour celebrating the Harry Potter films. The tour will feature Harry Potter and the Sorcerer's Stone and will kick off this summer, travelling to destinations across North America. The world premiere will take place on June 23rd, 2016 at The Mann Center with the Philadelphia Orchestra performing. Tickets go on sale Saturday, February 20th at 10AM via MannCenter.org, Ticketmaster.com, 800-745-3000, or the Mann's Center Box Office. Soon thereafter the tour will appear with such orchestras as the San Diego Symphony, Nashville Symphony, Milwaukee Symphony Orchestra, Grand Rapids Pops and Symphony Silicon Valley.
The tour will feature a live symphony orchestra performing, to picture, every note from Harry Potter and the Sorcerer's Stone. Audiences will be able to relive the magic of the film in high-definition on a 40-foot screen while hearing the orchestra perform John Williams' unforgettable score.
This exciting concert tour series begins with Harry Potter and The Sorcerer's Stone with a score composed by award-winning film composer and Oscar winner, John Williams.
CineConcerts is one of the leading producers of live music experiences performed with visual media. Founded by producer/conductor Justin Freer and producer/writer Brady Beaubien, CineConcerts has engaged millions of people worldwide in concert presentations that redefine the evolution of live experience. Recent and current live concert experiences include Gladiator, The Godfather, It's a Wonderful Life, DreamWorks Animation In Concert, Star Trek: The Ultimate Voyage 50th Anniversary Concert Tour, andBreakfast at Tiffany's. Justin Freer has quickly become one of the most sought-after conductors of film music with a long list of full symphonic live to projection projects. He has appeared with some of the world's leading orchestras including the Chicago Symphony Orchestra, London Philharmonic Orchestra, New York Philharmonic, Philadelphia Orchestra, Philharmonia Orchestra, San Francisco Symphony and Sydney Symphony Orchestra. From full-length movie screenings with live orchestra to music-interactive sporting event experiences to original 3D-environment holiday programming, CineConcerts is at the forefront of live entertainment.


text:
The Rat Pack meets the Rat Patrol
Sheriff's CJ-3B in Ocean's 11
The 1960 heist film Ocean's 11 , starring Frank Sinatra and the "Rat Pack," became popular again as a result of the success of the 2001 remake. The new version has the edge in acting and suspense, but the original has some unique features including one of the most primo CJ-3Bs I've seen on the big screen. It's a vehicle of the Clark County Sheriff's Jeep Posse. Not exactly the Rat Patrol though; instead of jumping the dunes, it's only seen parked outside the Dunes hotel.
As Sammy Davis Jr. slips through a police roadblock the morning after the big heist, driving a garbage truck, the CJ-3B sits in the background ready for action. The '59 Ford (foreground) and '59 Olds cruisers look better prepared for any high-speed chase that might develop, but the Jeep looks better equipped in case somebody runs out of gas and needs a push....
The Jeep is also glimpsed at the roadblock during the night (below) while power to the casino is blacked out, and a Caddy passes in the foreground. In this shot you can see the "Sheriff's Jeep Posse" lettering on the hood, and what appears to be a loudspeaker mounted at the top of the grille.
The Jeep Posse
Lt. Dennis Larsen of the Metropolitan Police Museum Association of Nevada describes the Jeep Posse: "There was a Jeep Posse in Clark County from the early 40's till the early 80's. It was composed of volunteers from around the community and they performed Search and Rescue missions in conjunction with the Clark County Sheriff's Department Search and Rescue unit."
"The Sheriff's Department did not have enough personnel to perform all the rescues around the 8,000 square miles in Clark County so they would use the volunteers to help out. These people became very skilled in their rescue techniques and were recognized throughout the country. They were all deputized as Clark County Sheriff's Deputies and carried credentials identifying them as members of the Department. Their vehicles were painted Black & White with red lights and sirens. It was not uncommon for them to respond to a rescue with lights and siren."
Thanks to Mark Browder on Flickr for these early-sixties photos of the Jeep Posse. The CJ-3B in the photo above appears to be the same Jeep that appeared in the movie, minus its hardtop.
Dennis Larsen also provides the recent history of the Posse: "In the 1980's the Clark County Sheriff's Department was no longer in existence and the members of the Jeep Posse were under the control of the Las Vegas Metropolitan Police Department's Search and Rescue Section. Due to liability concerns they were disbanded and the department created a volunteer Search & Rescue unit. They are still involved in all search and rescues that take place in Clark County and even our neighboring states, but they are no longer commissioned police officers, and do not drive marked personal vehicles."
It's interesting to see the smaller but more glamorous Las Vegas of 1960 in Ocean's Eleven , very different from the sprawling family resort city with many huge hotels, which it is today. The Vegas of that era had only a few casinos -- and it had CJ-3Bs in the Jeep Posse. "How cool baby, as Frank (Ol' Blue Eyes) would say!" comments Jim Veen, who spotted the Jeep in this film.
Michael Coy at the Internet Movie Database describes the Ocean's 11 story: "Sketchy and virtually irrelevant, the plot concerns a bunch of paratroopers-turned-playboys who, craving money and thrills, mount simultaneous burglaries of five of Las Vegas' big casinos. The slackness of the plotting is part of the film's relaxed, tongue-in-cheek style as Sinatra and Dean Martin indulge themselves in what is almost a home movie."
The 2001 remake stars Brad Pitt and Don Cheadle, with George Clooney as Danny Ocean. A reviewer at Crazy for Cinema says, "Though I'm usually a sucker for the classics, I think Clooney and company out-cooled the Chairman and his cronies. I expected the 1960 pic to have class and style. Unfortunately, it doesn't have much of either... which makes the thin plot and phoned-in performances even harder to watch. It's Frank's mystique and power that got this film made, giving the boys something to do to fill their downtime while they performed in Vegas."
So don't rush out to rent the film except for nostalgia purposes. Elsewhere on the web, you can read more about the original Ocean's 11 .
Thanks to Jim Veen, Dennis Larsen, and to 1960 Clark County Sheriff "Butch" Leypoldt. -- Derek Redmond
See real Los Angeles Sheriff's Jeeps in 1957, in Anytime, Anywhere on CJ3B.info.
Return to The CJ-3B in the Movies.
Visit CJ3B.info on Facebook.
CJ3B Home | Contents | History | Photos | Toys | Links | 3A and 3B Community
Last updated 28 June 2014 by Derek Redmond redmond@cj3b.info
https://cj3b.info/Movie/Oceans11.html


text:
Burke Scholar Wins Statewide Logo Contest for Shred360
By The Charleston Chronicle | May 29, 2018 | 0
Hesham Hikal
Micah Brown helped start the Digital Arts program at Burke High School (BHS) in the 2016-2017 school year. If his students continue to win contests, the Digital Arts program is going to become a popular option for scholars there. Earlier this month, Shred360, a paper shredding company based in Columbia, announced Burke senior Hesham Hikal as this year's logo design contest winner.
Over 1,000 entries were submitted for this year's contest, which is open to every student in the state of South Carolina. Hikal's design will be placed on the back of one of Shred360's trucks by the end of June, and will remain there until June 2019. He earned $500 for the school as well.
"It feels really good to be selected as the winner out of all those entries," said Hikal. "We have a great thing going thanks to Mr. Brown, and I hope this award shows what we are all capable of in this class, and at Burke."
Shred360 opened its doors 11 years ago, and has used this state-wide contest to determine their company's logo each year of operation.
"It's an honor winning two logo competitions consecutively since the inception of the Digital Arts program here at BHS," Brown stated. "I look forward to the coming years and connecting with the community to enhance learning in this way."
Last year, a student from Brown's class won a contest sponsored by the local branch of the insurance brokerage and consulting firm, NFP. The business was looking for a logo to place on souvenir bags for a health expo in which they were participating.
Brown also pointed out another one of his students, Mia Washington, finished in the top eight in this year's Shred360 competition.
For more information about Hikal's accomplishment, or the Digital Arts program at Burke, contact Executive Principal Cheryl Swinton at (843) 579-4815.
Posted in Spotlights, Tri-County News (Berkeley, Charleston, Dorchester)
Claflin University Becomes a Regional Hub for Coding and Creativity
Dominion Energy Commits $35 Million to Initiative Supporting Historically Black Colleges and Universities, Minority Student Scholarships
African American High School Music Group Releases Groundbreaking Animated Video Addressing Current Social Issues
SBA Celebrates the Launch of New South Carolina Women's Business Centers
HBCU Buzz Presents: 2020 HBCU Top 30 Under 30


I do alot of caterings and this I swear I can never have enough. Always asked for this recipie and perfect when made with fresh fruit!
I try to use all fresh fruits but as we know sometimes its not feasable depending on what time of year this is made. So drain mandarine oranges, cut up apples and strawberries, clean grapes, toss in a bowl I usually cut the banana and stir in just before serving. Add whipped cream and serve you can add in nuts marshmellows anything but I found these fruits to be the best. Even add in some cream cheese with whip cream enjoy !!!


Show up to a party wearing a shirt that announces how you truly feel: Drink First, Pork Later. The bacon graphic in between those two phrases shows your true love for the ultimate meat candy. This apparel says it all. Get your drink on. And then... well... eat your bacon later. Uh... yeah. That's exactly what we mean. *Wink, wink.


text:
Q: Puppet vcsrepo git checkout branch I am using puppet to clone a repo on an agent node. My site.pp contains
node foobar{
   vcsrepo{"home/user1/gitrepo1":
     provider=>git,
     source=>"https://github.com/foobar/foo.git",
     revision => "remotes/origin/bar",
   }
}

On the agent when I do git branch after the catalog has been applied, it shows
*(no branch)
master

When I do git branch -a it shows
master
remotes/origin/HEAD -> origin/master
remotes/origin/bar

I want to be able to checkout the remote "bar" branch. Only specifying the revision=>bar gives the following error. The documentation is not too clear on this either.
err: /Stage[main]//Node[foobar]/Vcsrepo[/home/user1/gitrepo1]: Could not evaluate: Execution of '/usr/bin/git rev-parse bar' returned 128: fatal: ambiguous argument 'bar': unknown revision or path not in the working tree.
Use '--' to separate paths from revisions
bar

EDIT: There was a typo in what I had initially posted. I had in fact specified revision=>bar in my site.pp
Thank you.

A: The branch can be specified with 'revision' (can be a commit SHA, tag or branch name):
vcsrepo { "/path/to/repo":
    ensure => present,
    provider => git,
    source => 'git://example.com/repo.git',
    revision => '0c466b8a5a45f6cd7de82c08df2fb4ce1e920a31'
}

vcsrepo { "/path/to/repo":
    ensure => present,
    provider => git,
    source => 'git://example.com/repo.git',
    revision => '1.1.2rc1'
}

vcsrepo { "/path/to/repo":
    ensure => present,
    provider => git,
    source => 'git://example.com/repo.git',
    revision => 'development'
}

https://github.com/puppetlabs/puppetlabs-vcsrepo/blob/master/README.GIT.markdown

redpajama_set_name: RedPajamaStackExchange


text:
Sanford students delving into mystery of child's grave
Teacher Paul Auger sets them to work – and learn – after a child's remains are found in a long-abandoned cemetery.
By Gillian GrahamStaff Writer
Paul Auger explains some of the coffin artifacts to seniors, from left, Shealyn Kane, Vanessa Hodge and Sydney Littlefield in his classroom at Sanford High School. Sanford students are helping to unravel the mystery surrounding the remains of a child found May 4. Staff photos by Brianna Soukup
SANFORD — In the early 1930s, more than six dozen coffins were exhumed from the overgrown Woodlawn Cemetery, carried a mile away and re-interred at the town's new municipal cemetery.
The grave of one child was left behind, an apparent oversight that would not be discovered for more than 80 years.
Related Headlines
Human remains found at Sanford construction site belong to a child
For decades, children attended classes at Sanford's Emerson School and ran on a playground that was created on the same grounds once occupied by the original town cemetery. Eventually, the school closed and last month was torn down to make way for a Cumberland Farms gas station and convenience store.
On a Thursday afternoon in early May, a construction crew digging near the new store foundation unearthed the remains of the forgotten child and the remnants of a Victorian-era coffin. The discovery halted the project and touched off a delicate process of excavating the grave, documenting the bones and cataloging each artifact found with the remnants of the coffin.
Students researching braces from the coffin have determined that the hardware is nickel-plated. They're still researching a pair of rare, Victorian-era coffin keys that also turned up during excavation of the former Emerson School playground.
Now, local high school students born more than a century after the child was buried at Woodlawn Cemetery are helping city officials unravel the mystery of the child's identity. After examining the remains and studying the artifacts, they are pursuing DNA tests to see if the child was the great-aunt of a Maine woman who reached out to city officials after hearing the child's remains had been found.
"I don't think anyone wanted to see whoever this person is just re-interred in Oakdale Cemetery in some anonymous plot when there is technology to find out who it is," said Paul Auger, the local historian and high school teacher who exhumed the remains. "What a tragedy it would be if we had the opportunity and didn't take advantage of it."
On June 6, City Manager Steven Buck will ask the Sanford City Council to use about $4,800 from the proceeds of the Emerson School sale to pay for the DNA tests.
"I'm very positive we're going to be able to identify who this is," Buck said. "The great part of the story is we know where the family is and she could be re-interred in the family plot at Oakdale Cemetery. That's our driver."
A GRAVE IS UNEARTHED
The land's history as Sanford's first municipal cemetery was well-known to local historians and city officials, but finding remains there still came as a shock. Auger had asked the construction company building the Cumberland Farms to keep an eye out for anything unusual, but didn't really expect them to find anything.
Buck was in a meeting on May 4 when he got a call on his cellphone from Pete Smith, the public works foreman. A few blocks down Main Street from City Hall, a construction crew had just unearthed the grave.
"This was a surprise because we had records showing everybody was moved," Buck said.
Paul Auger stands in the pit at a Main Street construction site a day after a child's remains were unearthed there May 4. He enlisted students to help clean the bones, study the coffin remnants and pursue clues to identification of the child. He estimates that the grave is from sometime between 1880 and 1906.
When the city sold the property to Cumberland Farms last year, the purchaser had raised concerns about the land's previous use as a cemetery. But the city had records showing 77 bodies were exhumed and re-interred at Oakdale Cemetery by 1933, when Emerson School purchased the lot to build a playground. Everyone was satisfied with that and the city sold the property for $800,000.
The Emerson School, which closed in 2013 and needed extensive repairs, was demolished in April. Construction workers had just finished the new store's foundation – inches from the remains, it turns out – and shifted their focus to excavating for a water line when they found the grave. The PM Construction foreman was watching every scoop of earth coming out of the trench when a piece of skull poked out of the soil. He halted work and called police.
"Within a few minutes, the entire police department and most of the fire department were surrounding the hole," Auger said. "No one had ever seen this happen before."
Sanford police contacted the state Medical Examiner's Office and provided information showing the grave was found in a known cemetery. The ME's office told the city the remains had to be hand excavated, the local historical society notified and the remains re-interred. Auger, a member of the city's Historical Committee, was called in to help.
Auger, who was a police officer for a decade before becoming a teacher 19 years ago, remembers only one other time that an old grave was found during construction in Sanford. Back around 1980, a crew digging an elevator shaft at the Town Hall annex found a femur. Construction was stopped while the Historical Committee determined the area had once been a cemetery. The leg bone was reburied in an anonymous plot at Oakdale Cemetery.
'GRAMPY'S LITTLE SISTER'
To prepare for the exhumation, Auger found three coworkers willing to cover his classes at Sanford High School the next day, went to the hardware store for supplies to make a sifter and arranged to have a tent put over the grave because rain was in the forecast. By 7 the next morning, he was climbing into the pit.
For hours, Auger, Buck and Smith sifted through the dirt a handful at a time, pulling out shards of glass, pieces of metal and bones caked in dirt and encased in root hairs. A large root from a nearby oak tree had grown around the coffin, perfectly outlining three sides of the grave. Auger measured everything: the length and width of the grave, the placement of the bones, the size of the coffin braces, hinges and keys. They found ribs and finger bones and pieces of jawbone with a half-dozen teeth still intact.
Their work was halted for several hours in the middle of the day as Buck and the city attorney dealt with last-minute questions from the Medical Examiner's Office and the Department of Health and Human Services, which has purview over cemeteries and cemetery relocations. The city clerk issued a permit to exhume and move the remains, and work resumed.
Late that afternoon, as the small group tried to finish their work before it started raining, Buck's cellphone rang. It was his assistant back at City Hall with an urgent message to call a Maine woman who saw news of the discovery and had a story to share.
"She said she was always told in her family that when the cemetery was moved in 1931, they never found Grampy's little sister," Buck said.
The woman, whose name city officials aren't releasing until the remains are identified, lives in southern Maine and is an avid researcher of her family genealogy, Auger said. She provided many details about her family, including names going back generations. Auger was able to research the family and discovered the mother of the child had five living children at the time of the 1900 census, but only four of her children are shown on other records.
"We don't even know the name of this child," he said.
Auger said the child was likely born between 1890 and 1895. The woman who contacted local officials also provided information about a second possible match, a different girl in the extended family who died in Sanford in 1902.
STUDENTS STEP IN
With a few clues about the possible identity of the child, Auger turned to a group of students at the high school to assist in the investigation. He sees it as a unique learning experience and way to connect students with history.
One student went with Auger to a local jewelry store to determine the metal hardware used for the coffin's handle braces was nickel-plated. They're still researching a pair of rare, Victorian-era coffin keys.
Paul Auger holds a nail and a bracket from a grave discovered at a construction site on Main Street.
Two honors students, both headed to nursing school in the fall, cleaned and sorted the bones with help from Auger's two children.
"We wanted to learn as much as we could from the remains," Auger said.
Seniors Kristen O'Connell and Sydney Littlefield had never held real human bones before the afternoon they arrived at Carll Heald & Black Funeral Home in Springvale, where the remains are being stored until they are re-interred. During their anatomy classes at Sanford High, they used plastic bones and photos to learn about the 206 bones in the human body.
Using soft-bristled toothbrushes, chopsticks and water, they spent several hours cleaning dirt and roots from the bones.
"We had to try to make them recognizable," said O'Connell, 18.
They used their anatomy textbook to help identify each bone before laying them out on an examination table to form the skeleton. The pelvis was missing, making it impossible for Littlefield and O'Connell to determine the gender of the child. They were encouraged to see the teeth that could possibly provide DNA to confirm the child's identity.
"It's interesting to think you could help someone find a missing relative and help put someone to rest," said Littlefield, 17.
Officials have not allowed the bones to be photographed because of privacy concerns for the family.
Auger turned to science teacher Beth Marras and her Advanced Placement biology text to make a DNA tree to show which living family members would have the same mitochondrial DNA, which is inherited through the mother. The woman who contacted the city cannot provide the sample because the family connection would be through her grandfather, but a first cousin twice removed from the child could, the students determined after reviewing a family tree provided by Auger. Students will perform the buccal swab to collect cells from the inside of the cheek of the cousin, who has agreed to provide a DNA sample.
George Pouravelis, who teaches anatomy and physiology at the high school, said it has been amazing to watch students be part of a once-in-a-lifetime experience that has generated a lot of interest in the community.
"This is an exciting statement about this community to do the right thing here," he said. "If it was someone in my family, it would be nice to know that someone cared enough to do this the right way."
Gillian Graham can be contacted at 791-6315 or at:
Twitter: grahamgillian
sanford maine
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By Telephone – Please have the student's ID number, appropriate credit or debit card information, and the amount to be applied to the account.
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Payments will be due on the 5th of each month.
Choose Payment Method - Automatic deduction from a bank account (ACH), or credit/debit card. You must owe at least $300 for the Fall term to set up a payment plan.
Know Your Fees - $35 set up fee, $30 return payment fee if your financial institution denies the payment.
How to Set Up a Payment Plan - Go to OwlNet > All About Me > My Bill-Payment Plan > Payment Plan-Click Here. On the next page select the Fall 2019 term from the drop down box, then click the E-Cashier link at the bottom of the page.
NOTE: All down payments and set up fees are processed immediately!
For help setting up a payment plan or to make changes to your plan after it is setup call Nelnet at (800) 609-8056.
Financial aid covers tuition, fees and books.
Any remaining funds will be refunded using the student's select a refund preference thru www.refundselection.com.
Students can add funds to their HCC ID cards to purchase items in the College Store, in the Globe Cafe, and from some of the vending machines. To add funds, visit the Cashier's Office with appropriate payment method and ID card or call 443-412-2208 to add funds by credit card over the phone. Please note that unspent funds on campus ID cards will not be refunded.
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When submitting your registration, you assume responsibility for tuition, fees, and charges. Additionally, if your charges become delinquent and are sent to collections, you will be responsible for the collection costs. Tuition and fees are non-refundable after the refund deadline. In order to avoid an outstanding financial obligation to the college, you must officially drop your class within the stated refund period. Non-attendance does not constitute a refund or removal of the debt. Should a student have a financial hold on their account, and they submit full payment through the HCC portal, they must contact the Cashiers Office to remove their hold; web payments do not automatically release holds on student accounts. To check refund dates for credit, see below.
Important note: Refunds are processed approximately four weeks after a class is dropped. Withdraw dates are posted on the Academic Calendar.
We are located on the ground floor of the Student Center, behind the staircase.


The problem is that there is no heat source. I tried dye-ing soup with it, but it would not dye the liquid, only the fat floating on top. However, after submerging it in clean water 20 times, the item is still bleeding a bright blue hue. And here's the thing: We only mention services and products that we think are truly worth your attention, whether they're free, paid, or otherwise. How to Make Concentrated Liquid Homemade Food Coloring Another option for making a more concentrated dye is to reduce a deeply colored liquid, such as beet juice or the juice from purple sweet potatoes, into a syrup.
The classical dye known as Phoenician Red was also derived from murex snails. Retrieved 8 November 2017 — via Google Books. During the colonial period the production of cochineal in Spanish, grana fina grew rapidly. Deep brown-black from oregano Greek widows traditionally wear black, and not from purchasing a new wardrobe! The yellow constituent disappears upon drying the root, and dissipates while boiling. The ammonia should be extremely dark at this point.
You can use it to dye clothes, yarn and basic fabrics. Chamomile tea can help you do just that, and is especially effective for those who already have blonde hair. Estimates are that it took 8,500 shellfish to produce one gram of the dye, hence the fact this dye was worth more than its weight in gold. To make a concentrate, place about 1 cup of freshly squeezed juice over very low heat. I visited your other websites and found some very interesting things! This technique of dyeing is unique, in that the final color is controlled by the choice of the diazoic and coupling components. This expensive dye was also mentioned in the bible, in Acts, where Lydia is a seller of purple. So if you dye your fabric, the hot water dying process would be more or less the same process as felting.
Madder, weld and other dye plants have been used for thousands of years. Before washing, weigh your fabric or yarn. Oak galls produce a strong, black dye. The dyers of , a great cloth town in the , produced the cloth associated with by dyeing wool with woad and then overdyeing it yellow with weld or dyer's greenweed , also known as dyer's broom. This form is often colorless, in which case it is referred to as a , and has an affinity for the textile fibre. You might also consider wild foraging if you really want to get back to nature, or starting a garden with some of these dye ingredients.
By a self-educated English naturalist; with the author's charming botanical paintings. Hair or fur is another excellent protein fibre. Natural Dyeing Method 2 — Solar Dyeing Solar Dyeing is a really cool method for dying at home! It was fun to experiment with different shades. The acid will unlock the ability of the brown dye to color the cloth. However this is perfect for dyeing wool, as very little felting will take place.
How do you like to use homemade natural food coloring? You need a mordant to actually get colorfast colors. At home spread them on a cookie sheet and toast for about 10 minutes in the oven, till they dry up. Wear rubber gloves to handle the fabric that has been dyed, the dye can stain your hands. . New to this, thanks for all of the great info.
I use natural colorants in my soaps too. Then the kids and I transferred each dye bath into a mason jar with a lid. Woollens were frequently dyed in the fleece with woad and then piece-dyed in kermes, producing a wide range colors from blacks and grays through browns, , purples, and. Throughout history, people have dyed their textiles using common, locally available materials, but scarce dyestuffs that produced brilliant and permanent colors such as the natural invertebrate dyes, and crimson , became highly prized luxury items in the ancient and medieval world. I rarely mordant wool or protein fibers like silk. The primary commercial species in Asia was true indigo.
This is achieved by treating a fiber with both diazoic and coupling. Just a side note, the chamomile dye bath smelled heavenly. Dye Bath: Place wet fabric in dye bath. Henna comes from a tree grown in hot, dry areas with leaves that contain a natural red-orange pigment. Get triple nourishment any natural textured diva could surely use from avocado, olive, and shea oils.
Rinse the material and squeeze out excess. The Madder which was previously sold as root pieces and Natural Indigo which used to be in chunk form are now both available in ground powder form. You then mix that with some egg yolks for a classic tempera paint method. Two level tablespoons equal one ounce of alum. And, like all the other colors, for whatever reason, the color stayed on an elastic lace that bordered some of the fabric scraps.
If a light is shone through it, it will look ruby red, but this is deceiving; the majority of the dye color is brown. Since I cooked the berries microwave with the seeds still attached I am carefully cleaning all of my prep equipment. Rinse the fabric a little bit in the sink, then give it a bath in white vinegar. Juniper, , ashes provide brown and yellow dyes for , as do the hulls of. To create a green dye bath, the entire artichoke is used.


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Through the Pain > Time Heals Nothing > Reviews
Time Heals Nothing
An embarrassment to DSBM - 20%
Slater922, June 11th, 2020
Depressive-suicidal black metal (or DSBM for short) is a more depressed version of black metal. The first DSBM song I've listened to was "Time Heals Nothing" from the German band Through the Pain. I thought the song was decent overall, and decided to check out their debut album. To say I was disappointed would be an understatement. I was mind blown by how awful it was. I try not to be too harsh on debut albums, but they could've done better.
One major problem of the album is the repetition. By that, I mean all the songs sound the same. They start off with a slow guitar riff with lots of distortion. As the song progress, the guitar increases in speed while Bael'vect screams his head off. Then, halfway through the song, they stop to have some sort of powerful moment before going back to what they were playing before. I am not kidding when I say all of the songs on this album sound exactly like that. The only different being that they play in slightly different tempos. It's like the band only played one song, then copy and pasted it three more times with only minor changes, then called it a day.
Another problem is Bael'vect's vocals. Black metal is known for having high-shrieking vocals, but here, it's just screams. What makes this problematic is that Bael'vect doesn't try to make something out of it, so it sounds unnecessary. One example of this is in the song "No Way Out". It sounds like all the other songs on this album, but the atmosphere on this song is more depressing. Bael'vect could've sung a poetic story about depression and suicide, but that would've took effort. Bael'vect's screams are forced and unnecessary, and should've been cut out in the final product.
If I had to pick one good song off the album, it would be "Time Heals Nothing". Once again, it follows the same tiring formula, but the way it sounds makes it feel like the band had actually put some effort into it. The guitar riffs offer a different sound with more distortion, giving it more of a black metal feel. Bael'vect's vocals also have some purpose, as they flow well through the progressing instruments. However, the break in the middle of the song is probably one of the most powerful moments on the entire album. The slow-paced guitar followed by Bael'vect's screams of agony bring an atmosphere of hopelessness and anger. I was dumbfounded by this, as by that point, I've basically gotten used to the repetitive formula. Overall, I would give this song a 90%. I would give it a higher score, but it uses the same formula again, though executed in a better way.
Every genre I've listened to has some bad albums, but they also include some good albums. However, after hearing this, I thought that all DSBM sounded like this. It wasn't until I listened to Xasthur and Thy Light that I got to hear what good DSBM sounds like. Here, I'm not sure what direction the band was trying to go for. It doesn't work as DSBM, it doesn't work as bringing awareness to suicide and depression, and even though it includes a couple of emo elements, it isn't even good at that either! I'm kinda glad that the band eventually broke up and moved on to better things, because while they had some interesting ideas, the execution was awful.
Time spent hearing this EP not soothed either - 40%
NausikaDalazBlindaz, June 9th, 2014
As befits a band with a name like Through The Pain, the style and themes are within the ambit of depressive BM and the actual music itself is a stony creature that you could certainly imagine bashing your head against without getting a response. Listeners are very likely to feel this way all the way through this album: it's not only time that heals nothing here. The steaming noise-guitar music grinds through its chosen path dragging tinny drumming and a set of shrieking vocals - always shrieking - in its wake.
The first track is a relentless plod for its first half until its sudden collapse just after the sixth minute. It slowly recovers with effort and with a soothing counter-melody that softens the edges of the ragged vocals and brittle instruments. Apart from this change, the song doesn't vary much throughout. The next track, appropriately named "No Way Out", continues in similar plodding manner save for a slight change in key and the vocalist continues to screech as though nothing's happened in the meantime. Yep, there's no way out of this lumbering repetitive grinding guitar drone and the wailing voices all right.
So we plough on ahead with "Absence of Will" where the opening riffs are a little more cheering than in previous songs but it turns out to be slower and more lethargic as well. The ragged screeching is a little less and there are more pauses in-between the screams. The bass rhythm is more noticeable and has a definite groove so at least there is something to hold on over the duration of the song. The repeating melody has something of the quality of a lullaby and can be pleasant to listen to if you ignore the wailing in the background.
After four tracks of pain - the vocalist's existential pain and your more viscerally felt pain at having lost 37 minutes of your life to hearing out this music - you realise you feel just the same as you did before you started listening to this music: you feel no more enlightened about the nature of suffering, anguish or the loneliness and alienation of modern life. None of the four songs develops much over its course; there seems no attempt on the musician's part to try to connect with you and atmosphere throughout the album is lacking. Sure, he's shouty but constant screaming in agony can have the effect of driving listeners away: it communicates no emotion and you feel the fellow could have just pre-recorded his parts and left his Moebius-strip tape to play over and over. Each track ends up being far too long and on each also, a good two or even three minutes could have been chopped off and the point of each piece would still have been made.
At least the album's title is appropriate ... time heals nothing, certainly not my disappointment that this recording could have been edited and worked over a bit more.
Has this all gone too far? - 35%
autothrall, November 9th, 2010
The major snafu with an album like Time Heals Nothing is that there is simply nothing to it. Anyone would be hard pressed to convince me that there was any noticeable effort involved in its creation. But to be fair, I realize that's kind of the point of such a piece. Through the Pain's material is fully intended to sound as barren and lifeless as possible, to create a paradox in which its own vacuum of melancholy sucks the listener into a droning, dull hypnosis. Unfortunately, most artists working in such an aesthetic have the chops and wherewithal to actually draw you into that lucid state, to offer some tantalizing surprise, for good or evil, hidden beneath the stone, some wriggling thing that compels the witness to hoist it from its muddied bank.
Here on this German band's debut, I'm simply not finding any such elusive trait. Essentially, you could take any trio of musicians on Earth with electric guitar, bass, drums and even a primordial notion of composition, have then spend 37 minutes repeating simplistic chord patterns, and produce something with nearly as much merit as Time Heals Nothing. It's that simple. Yet where some bands shine in simplicity (fuck, look at the Ramones), and even overcome these limitations with brilliant ideas, Through the Pain simply plod along at a null pace, about as exciting as watching rust peel from some long abandoned machine. In fact, aside from the fact that Baelvect utilizes the torturous strain of vocals reminiscent of Burzum or Weakling, I wound find it difficult to define this as black metal at all, but more like a droning alternative to the basic indie rock music you'll hear coming out of any handful of teenagers in a garage or basement in suburbia.
Time Heals Nothing consists of four compositions, all of which range from 7-11 minutes, so there would seem plenty of time to absorb the listener into the proceedings, but it fails to do this. Instead of sounding in pain in the opener "Dwell in Agony", Baelvect just sounds like he's fucking around, and he continues to do so in the clean guitar sequence at 6:30, which despite itself is the clear highlight. "No Way Out" is slightly more painful due to the desperation of the chords, but it's just too basic to compel. The later tracks, "Absence of Will" and "Time Heals Nothing" are the stronger of the four, but even these operate on a truly primitive foundation, with nothing below the surface, just steady melodic bass lines affixed to the repetitious chords. At some points deep in the album, you do feel like something is on the edge of happening, but the slight shifts in riffing continue to cry wolf until the album's end.
For all I know, Through the Pain might be a trio of virtuosos who were simply attempting to capture a particular mood when writing this, but I found it ultimately as empty as a stripped down Svarti Loghin, as mind numbingly vacant as the hideously overrated Alcest, sans the melodic course those bands weave toward a greater accessibility. I am not opposed to minimal aesthetics in metal music, and if you'll sift through these pages you'll find a great amount of my praise for black metal bands who explore that path while keeping the listener on edge, but I do draw a line when something becomes as vapid as this, and the raw production doesn't really help here, since there is no real atmosphere with which it can capitulate. Time Heals Nothing is not long enough to become frustrating, or a staggering bore, but its ineffectual nature fosters none of the satisfying, negative emotion so intrinsic to this sub-genre.
-autothrall
http://www.fromthedustreturned.com
Three Average Songs Eclipse One Monumental Song. - 60%
Perplexed_Sjel, June 11th, 2010
I'm not sure what I was expecting from Through The Pain. After the split with Trist, the iconic Czech Republic based one man band, I was on the fence as to whether or not I thought this band could succeed in this sub-genre. It seems that their first full-length, entitled 'Time Heals Nothing', has been well received by the masses, but I find it to be really ponderous and not at all charismatic. The material seems to take an influence from Trist, as well as Hypothermia, but without the gruelling, punishing and downright devastating affect those two iconic bands in the sub-genre draw out through their agonising music. Although Trist have seemingly gone down the shitter with their lead man, also named Trist, being somewhat psychotic (I do believe he actually does have a few mental issues) and Hypothermia becoming more and more secluded from the scene, there is room for someone to take their place while they look to be on hiatus. Through The Pain could have it all if they actually worked on being powerful instead of clichéd. There is only one true highlight to this album, otherwise it's very much a by-the-book affair with little in the way of innovation from the standard. It's not exactly well paced, seeing as the songs are quite long, but it is sufficiently melancholic and emotional, though it could have done with stronger riffs because, on songs like 'Absence of Will', the bass tends to eclipse the performance of the guitar.
The fact that they have Hypothermia's drummer, Richard, enlisted into the newly formed line-up, though only as a session member, the three-some could have shared the spoils of taking advantage of a real leading inspiration in today's present scene, but 'Time Heals Nothing' is a fairly formulaic display of negative emotions and really doesn't spark any new energy, or life into the sub-genre which has seen a recent improvement in technical ability and vision through the likes of Pensées Nocturnes and so forth. With bands now starting to introduce elements of classical, neoclassic and other various genres into this negative spectrum, Through The Pain are beginning to gather dust by reproducing what bands like Hypothermia have already covered. Whilst I'm not against the idea of taking after your role models, after all, that is how many numerous sub-genres and fresh sounds are created, there must be an extra added element to the music to make transform it from an overused, formulaic branch of music, to an extraordinary display of negative emotions, grace, subtleties and do forth.
Bands like Gris, for example, have worked tirelessly on producing a top-quality sound by introducing a wide range of elements into their music that don't normally get associated with depressive black metal. In fact, their music is so adventurous and unique that it really seems to transcend black metal altogether. If it were not for the agonising vocals of Icare, I would wonder whether or not it can be pigeon holed into a genre like depressive black metal. Through The Pain do some things differently, as shown well on 'Absence of Will', which actually feels far more upbeat than your normal song in this sub-genre. The bass on this particular song is very good. It gives the atmosphere a bouncy, vibrant quality despite the nature of the song attempting to be downright depressed. The song title, the rasped vocals and even the structure of the song should all point to a negative conclusion, but the material feels somewhat, and refreshingly so, jovial, hopeful and upbeat. Not something I would have expected from a sub-genre, or band like this who revolve around sullen emotions.
This song seems to stand alone however. None of the other songs conjure up such a happy atmosphere. The bass, on this song, is incredibly important. It gives a positive, bright outlook and a sunny feel to the material, as if we're gazing through a clean window onto a beautiful landscape with the sun shining down on it and on us, making life all the more glorious. This song doesn't really fit into the general mould of the album, but I love it! It's definitely the best song on the album by far. It shows an inventive side to Through The Pain which isn't always on offer during the rest of the songs. Unfortunately, this one break-away song is the second shortest of the four available to us and isn't as groundbreaking as it may seem. There have been upbeat black metal and songs for years, perhaps dating all the way back to Ulver's iconic 'Nattens Madrigal - Aatte Hymne til Ulven i Manden'. Once again, the song lengths also become an issue because, seeing as the songs are so repetitive from the drums, to the guitars, the songs could quite easily convey the same message in a much shorter space of time rather than eclipsing the eleven minute mark. This album is decent, in places, but very unsurprising in most others.


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