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Hyperparathyroidism (HPT) is a common endocrine disorder with incompletely understood etiology, characterized by enlarged hyperactive parathyroid glands and increased serum concentrations of parathyroid hormone and ionized calcium. We have recently reported activation of the Wnt signaling pathway by accumulation of β-catenin in all analyzed parathyroid tumors from patients with primary HPT (pHPT) and in hyperplastic parathyroid glands from patients with uremia secondary to HPT (sHPT). Mechanisms that may account for this activation have not been identified, except for a few cases of β-catenin (CTNNB1) stabilizing mutation in pHPT tumors.
Methods and Findings
Reverse transcription PCR and Western blot analysis showed expression of an aberrantly spliced internally truncated WNT coreceptor low-density lipoprotein receptor–related protein 5 (LRP5) in 32 out of 37 pHPT tumors (86%) and 20 out of 20 sHPT tumors (100%). Stabilizing mutation of CTNNB1 and expression of the internally truncated LRP5 receptor was mutually exclusive. Expression of the truncated LRP5 receptor was required to maintain the nonphosphorylated active β-catenin level, transcription activity of β-catenin, MYC expression, parathyroid cell growth in vitro, and parathyroid tumor growth in a xenograft severe combined immunodeficiency (SCID) mouse model. WNT3 ligand and the internally truncated LRP5 receptor strongly activated transcription, and the internally truncated LRP5 receptor was insensitive to inhibition by DKK1.
Citation: Björklund P, Åkerström G, Westin G (2007) An LRP5 Receptor with Internal Deletion in Hyperparathyroid Tumors with Implications for Deregulated WNT/β-Catenin Signaling. PLoS Med 4(11): e328. https://doi.org/10.1371/journal.pmed.0040328
Academic Editor: Hans Clevers, Utrecht University, The Netherlands
Received: June 12, 2007; Accepted: October 3, 2007; Published: November 27, 2007
Copyright: © 2007 Björklund et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the Swedish Research Council, Swedish Cancer Society, and Lions Fund for Cancer Research. The funders had no role in study design, data collection and analysis, decision to publish or the preparation of the manuscript.
Competing interests: The authors declare the following competing interest: A patent is filed (USA) for "LRP5 and LRP6 Receptors in Cancer Treatment," number WO2005048913.
Abbreviations: APC, adenomatosis polyposis coli; ChIP, chromatin immunoprecipitation; HPT, hyperparathyroidism; LRP5, low-density lipoprotein receptor–related protein 5; pHPT, primary hyperparathyroidism; SCID, severe combined immunodeficiency; sHPT, secondary hyperparathyroidism
Editors' Summary
The parathyroid glands—four rice-sized glands in the neck—maintain a normal calcium balance in the body, to maintain strong bones and essential cellular functions. The glands release parathyroid hormone as a response to a decrease in blood calcium level. By stimulating calcium release from bone and its absorption in the gut, parathyroid hormone restores the blood calcium level. However, 100,000 new individuals in the US develop hyperparathyroidism (HPT) annually, characterized by enlarged, overactive parathyroid glands and high blood levels of calcium. Primary HPT (pHPT) is usually caused by a benign tumor (a non-life-threatening growth) in one of the parathyroid glands. Secondary HPT (sHPT) occurs in response to calcium regulatory disturbances, linked to vitamin D deficiency, and more or less invariably develops in patients with uremic kidney disease.
HPT is usually treated by surgical removal of the enlarged parathyroid glands, which is done with great efficiency. However, ideally, doctors would like to know what drives the overgrowth of the parathyroid glands to be able to develop drugs for treatment or disease prophylaxis. Researchers recently reported that the cells in enlarged parathyroid glands from patients with HPT contain high amounts of β-catenin. This protein is part of the Wnt signaling pathway, which has been found to be disrupted in many tumor entities in other organs. In the absence of Wnt proteins, a group of proteins called the β-catenin destruction complex marks β-catenin so that it is rapidly destroyed. When Wnt proteins bind to a cell-surface receptor called Frizzled and a coreceptor called LRP5, the destruction complex is inhibited and β-catenin accumulates. This accumulation induces the production of other proteins (in particular, c-Myc) that stimulate cell growth and division. The accumulation of β-catenin in the enlarged parathyroid glands of patients with HPT could, therefore, significantly contribute to the overgrowth of their glands—but what causes β-catenin accumulation? In this study, the researchers have investigated this question to try to identify a target for drugs to treat HPT.
The researchers looked for genetic changes (mutations) in β-catenin that stabilize the protein and measured the expression of LRP5 in abnormal parathyroid gland tissue from 37 patients with pHPT and 20 with uremia and sHPT. All the samples contained high levels of β-catenin, but only four contained a β-catenin–stabilizing mutation. All the sHPT samples and 32 pHPT samples (but none of the samples containing the β-catenin stabilizing mutation) expressed a mutated LRP5, with the central region deleted. To investigate the functional consequences of this internally deleted LRP5 protein, the researchers used a technique called RNA interference to block its expression in a human parathyroid tumor cell line. They found that expression of the mutated, short LRP5 is required for accumulation of β-catenin, expression of c-Myc, and continued growth of the cell line in test tubes and in animals.
The accumulation of β-catenin in all the enlarged parathyroid glands examined so far strongly implicates abnormal Wnt/β-catenin signaling in the development of pHPT and sHPT. These new findings identify which part of the signaling pathway is altered. The expression data and functional data together suggest that an internally deleted LRP5 coreceptor is often responsible for the accumulation of β-catenin. The functional data also show that expression of shortened LRP5 is necessary for the abnormal growth of parathyroid tumor cells. Exactly how the internally deleted coreceptor activates β-catenin signaling in parathyroid gland cells, or why a shorter-than-normal LRP5 is made, are not yet known. However, because these findings indicate that internally deleted LRP5 has a fundamental role in activating Wnt signaling in HPT, drugs that inactivate this aberrant protein but leave the normal protein unscathed might provide a nonsurgical treatment for this common hormone disorder.
Please access these Web sites via the online version of this summary at http://dx.doi.org/10.1371/journal.pmed.0040328.
- edlinePlus has encyclopedia pages on hyperparathyroidism, primary hyperparathyroidism, and secondary hyperparathyroidim (in English and Spanish)
- Information is available for patients from the US National Institute of Diabetes and Digestive and Kidney Diseases on hyperparathyroidism, which includes links to organizations that help people with hyperparathyroidism
- Wikipedia maintains pages on Wnt signaling pathway and on β-catenin (note that Wikipedia is a free online encyclopedia that anyone can edit; available in several languages)
Primary hyperparathyroidism (pHPT) is characterized by hypersecretion of parathyroid hormone and generally also hypercalcemia, due to one or several parathyroid tumors (adenoma). Secondary hyperparathyroidism (sHPT) develops in patients with uremia because of phosphate retention, hypocalcemia, and reduced 1,25-dihydroxyvitamin D3 levels, causing parathyroid hyperplasia and eventually development of parathyroid tumors and hypercalcemia [1–4]. Parathyroidectomy is the only considered therapy for most patients. We recently reported aberrant β-catenin (CTNNB1) accumulation in all analyzed parathyroid tumors from patients with pHPT and in hyperplastic parathyroid glands from patients with uremia secondary to HPT [5]. MYC, a direct target of the Wnt/β-catenin signaling pathway in colorectal cancer cells and established as the critical mediator of the early stages of intestinal neoplasia [6,7], was found to be overexpressed at the protein level in 79% of parathyroid tumors [5]. Maintained activity of endogenous β-catenin was found to be necessary for the expression of MYC and cyclin D1 (CCND1), as well as growth and survival of a unique human parathyroid tumor cell line [8]. Overexpression of cyclin D1 has been reported in 20%–40% of pHPT tumors [2], and overexpression of cyclin D1 in the parathyroid glands of transgenic mice caused development of pHPT [9]. In a small fraction of parathyroid adenomas, overexpression is due to activation of the CCND1 gene by pericentromeric inversions of Chromosome 11, involving the parathyroid hormone (PTH) promoter [10]. Augmented cyclin D1 expression in some parathyroid adenomas could also be a consequence of aberrant β-catenin accumulation [5], although it remains to be determined whether CCND1 constitutes a β-catenin target [11] in parathyroid cells. We also reported CTNNB1 stabilizing mutations in a few cases (3 out of 20) of pHPT tumors, while no mutation was found in uremic secondary HPT tumors, and inactivating truncations of adenomatosis polyposis coli (APC) were not seen [5]. Mutation or deregulated expression of other Wnt-signaling components leading to β-catenin accumulation was therefore anticipated.
Dysregulated Wnt signaling with accumulation of β-catenin in the cytoplasm/nucleus plays an important role in a variety of human cancers. The stability of β-catenin is regulated by Wnt ligands through a "destruction complex" consisting of APC/Axin/GSK-3β/Ck1/Dvl and other factors. In the absence of Wnt ligand, free cytoplasmic β-catenin is rapidly degraded by the proteasome after phosphorylation of its amino terminus at residues serine 33, serine 37, threonine 41, and serine 45 [12–15]. Wnt ligands bind to cell-surface Frizzled receptors and LRP5/6 coreceptors and result in changes in phosphorylation of several intracellular signaling components with the subsequent accumulation of nonphosphorylated β-catenin [16–19]. According to a current model, the destruction complex is inactivated through recruitment of Axin to the intracellular domain of LRP5 [20]. β-catenin binds the LEF/TCF family of transcription factors to positively or negatively regulate transcription of target genes. Many mutant proteins of the Wnt signaling pathway, such as β-catenin, APC, Axin, and beta-transducin repeat-containing protein (β-Trcp), are associated with specific forms of cancer. For instance, aberrant accumulation of β-catenin through stabilizing mutations in CTNNB1 or inactivating mutations in APC is strongly implicated in the cause of approximately 10% and 80% of colorectal cancers, respectively [13,14]. A mutant of LRP5 lacking the extracellular domain was demonstrated to be constitutively active in vitro [20]. In this study, we aimed at investigating the potential role of LRP5 in parathyroid tumorigenesis.
Tissue Specimens
Parathyroid adenomas (n = 37) and hyperplastic glands (n = 20) from patients with pHPT and sHPT, respectively, were acquired from patients diagnosed and operated on in the clinical routine. Each patient contributed with one tumor. All 57 tumors displayed aberrant accumulation of β-catenin (unpublished data), of which 14 parathyroid adenomas and all 20 hyperplastic parathyroid glands were described previously [5]. Normal parathyroid tissue (n = 6) was obtained from glands inadvertently removed in conjunction with thyroid surgery where autotransplantation was not required or as normal parathyroid gland biopsies in patients subjected to parathyroidectomy. All tissues were intraoperatively snap-frozen, and cryosections were used in the analyses. Written informed consent and approval of local ethics committee was obtained.
Detection of Normal and Internally Truncated LRP5 Transcripts by PCR and DNA Sequencing
Total RNA was extracted with TriZol Reagent (Gibco BRL, Life Technologies) according to the manufacturer's instructions and the RNA was subsequently treated with RQ1 DNase I (Promega) and proteinase K. Alternatively, DNA-free RNA was prepared using the Nucleospin RNA II kit (Macherey-Nagel). Successful DNase treatments were established by PCR analysis of all RNA preparations. Reverse transcription of total DNA-free RNA was performed with random hexamer primers using the First-Strand cDNA Synthesis kit (Amersham Pharmacia Biotech) according to the manufacturer's instructions. cDNA was amplified by primary or nested PCR using mRNA-specific primers spanning positions 1992–2932 of LRP5 (GenBank accession number AF064548; http://www.ncbi.nlm.nih.gov/Genbank). A total of 1%–2% of the primary PCR product was used for nested PCR. Primers used were the following: forward primer, 5′-CTTCACCAGCAGAGCCGCCATCCACAG-3′; nested forward, 5′-GGATCTCCCTCGAGACCAATAACAACG-3′; and reverse, 5′-CCGGGATCATCC GACTGATG-3′. The PCR amplifications were performed with cDNA, 25 pmol of each primer, 0.2 mM dNTPs, 1× PCR buffer, 1.5 mM MgCl2, and 0.25 U Platinum Taq DNA polymerase (Invitrogen). The PCR conditions were: denaturation at 95 °C for 60 s, followed by 40 cycles of denaturation for 20 s, annealing at 58 °C for 20 s and extension at 72 °C for 90 s, and a final extension at 72 °C for 7 min. An annealing temperature of 61 °C and 40 cycles were used for nested amplification. Two LRP5wt and six randomly chosen truncated LRP5 cDNA fragments from parathyroid tumors, as well as from sHPT-1 cells, were cloned into pCRII-TOPO (Invitrogen), and sequenced on ABI 373A using the ABI Prism Dye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems). All fragments encoded an open reading frame, and the seven truncated fragments contained the same in-frame deletion (Δ666–809). Normal tissue cDNA or RNA were purchased from BD Biosciences Clontech and Ambion.
Cell Growth Determination and Flow Cytometry
Cells (2 × 105) were distributed onto 35-mm dishes in DMEM/10% fetal bovine serum and subsequently harvested at the indicated time points. The number of viable cells were determined by using the NucleoCounter (ChemoMetec). Cells were collected at 84 h after siRNA transfection and incubated with FITC-labeled annexin V to assess phosphatidylserine externalization as a marker for apoptosis.
Propidium iodide was added to distinguish tumor cells that had lost membrane integrity. Cells were analyzed by flow cytometry on a Becton Dickinson FACS Calibur flow cytometer (BD Biosciences).
RT-PCR Analyses
cDNA was prepared as described above. The following mRNA-specific PCR primers and labeled probes (5′FAM-sequence–3′TAMRA) were used for quantitative real-time RT-PCR analysis. For LRP5wt: forward: 5′-CCTGAAGACCATCAGCCGCG-3′; reverse, 5′-CCCGCTCCTGACCCAGCATG-3′; and probe, 5′-TCCCACCAAGGGCTACATCTACTG-3′. For LRP5tot: forward, 5′-ATCGACTGTATCCCCGGGGC-3′; reverse, 5′-CACCACGCGCTGGCACACAA-3′; and probe, 5′-CGGACTGTGACGCCATCTGCCTGC-3′. For MYC: forward, 5′-AAGACTCCAGCGCCTTCTCTCCGT-3′; reverse, 5′-TGGGCTGTGAGGAGGTTTGCTGTG-3′; and probe, 5′-AGCGACTCTGAGGAGGAACAAGAA-3′. For 28S rRNA, the Ribosomal RNA Control Reagents (VIC probe) was used (Applied Biosystems). PCR reactions were performed on ABI PRISM 7700 Sequence Detection System or MyiQ Single-Color Real-Time PCR Detection System (Bio-Rad) using the TaqMan PCR core Reagent Kit (Applied Biosystems). Each cDNA sample was analyzed in triplicate. Standard curves for the expressed genes were established by amplifying a purified PCR fragment covering the sites for probes and primers. Messenger RNA-specific PCR primers for WNT1 were as described [21]. Primers for WNT3 were as follows: forward, 5′-GGCTGTGACTCGCATCATAA-3′; reverse, 5′-CAGCAGGTCTTCACCTCACA-3′.
Transfection Experiments, Western Blotting, and Immunoprecipitation
sHPT-1 parathyroid tumor cells [8] were transfected with siRNA at least in triplicates at 2 × 105 cells/35-mm dish with jetSI-ENDO according to the manufacturer's recommendations (Poly-Plus-Transfection SAS). The following siRNAs were used: control nonsilencing siRNA (Qiagen Operon), siLRP5Δ666–809; 5′-TAACAACGACCUCACCAUUdTdT-3′ and 5′-AAUGGUGAGGUCGUUGUUAdTdT-3′ (synthesized by Thermo Electron, Ulm, Germany), siLRP5wt; 5′-CAACCACAUCUACUGGACAdTdT-3′ and 5′-UUCCAGUAGAUGUGGUUGdTdT-3′ (Thermo Electron), siLRP5tot; stealth 5′-CCUGCAUGGACUGAGGAACGUCAAA-3′ and stealth 5′-UUUGACGUUCCUCAGUCCAUGCAGG-3′, control siLRP5tot; stealth 5′-CCUGGUAGUCAAGGAUGCACCGAAA-3′ and stealth 5′-UUUCGGUGCAUCCUUGACUACCAGG-3′ (Invitrogen). A very high transfection efficiency (virtually all cells) was obtained for sHPT-1 cells by this protocol [8]. HeLa cells were transfected in the same manner, with a transfection efficiency of approximately 90% (data not shown). Conditioned medium was produced in HEK293T cells transiently transfected for 24 h by pCIN4/Wnt1 (kind gift from Dr. R. Kemler), pLNC Wnt-3HA (kind gift from Dr. J. Kitajewski), PON-Wnt-3a (kind gift from Dr. B. Williams), or pCS2/Dkk1 (kind gift from Dr. S. Y. Sokol) using Fugene 6 (Roche Diagnostics). sHPT-1 cells were transfected with the FOPFLASH or TOPFLASH TCF [22] luciferase reporter (Upstate) and the CMV-LacZ reference plasmid [23] for 24 h using jetPEI (Poly-Plus-Transfection SAS). In some experiments, TOPFLASH was transfected; transfection of siRNA to the same culture was done after 24 h and cells were harvested 96 h later. Luciferase and β-galactosidase activities were determined luminometrically, and luciferase activity was normalized for differences in β-galactosidase activity. Expression plasmid LRP5Δ666–809 was constructed by replacing the XhoI/KpnI fragment of pcDNA3.1/LRP5 and pcDNA3.1/V5-His/LRP5 (expressing tagged LRP5) with a XhoI/KpnI-digested PCR fragment harboring the deletion Δ666–809. Western blotting analyses were done on extracts prepared in Cytobuster Protein Extract Reagent (Novagen) supplemented with Complete protease inhibitor cocktail (Roche Diagnostics) or on a cytosolic protein extract [24]. Anti-V5–HRP antibody (Invitrogen), anti-active–β-catenin [25] mouse monoclonal antibody (Upstate; catalog number 05–665), anti-LRP5 goat polyclonal antibody (Santa Cruz Biotechnology; catalog number sc-21390), anti-actin goat polyclonal antibody, and anti–β-tubulin rabbit polyclonal antibody (Santa Cruz Biotechnology) were used. For tumor specimens, protein extracts for Western blotting were prepared from 10 consecutive frozen tissue sections (6 μm) in Cytobuster Protein Extract Reagent (Novagen) supplemented with Complete protease inhibitor cocktail (Roche Diagnostics). For immunoprecipitations, cells were resuspended in 300 μl buffer (50 mM Tris [pH 8.0], 150 mM Nacl, 0.5% NP-40, 50 mM NaF, and 1 mM EDTA, supplemented with Complete protease inhibitor cocktail), kept on ice for 20 min, and centrifuged for 20 s at 14,000 rpm. After addition of 20 μl anti-LRP5 goat polyclonal antibody (Santa Cruz Biotechnology; catalog number sc-21390), the lysate was incubated overnight at 4 °C with gentle agitation. A total of 50 μl Protein G PLUS–Agarose (Santa Cruz Biotechnology; catalog number sc-2002) was then added to the lysate and further incubated for 6 h. After centrifugation, the sample was boiled in 40 μl Laemmli sample buffer for 10 min and 10–20 μl was subjected to Western blotting analysis.
ChIP Assay
Chromatin immunoprecipitation (ChIP) of transfected cells was performed using a protocol from Upstate, but with immunoprecipitation conditions as described [26]. The anti-active–β-catenin mouse monoclonal antibody [25] was used (Upstate; catalog number 05–665) and MYC promoter DNA, containing TCF-4 binding site 2 [22], was PCR amplified in the linear range by the following primers: forward, 5′-ACGTGGCAATGCGTTGCTGGG-3′; and reverse, 5′-ACACAGAGAACGCACTGCGCG-3′.
Mouse Xenograft Model
Female Fox Chase severe combined immunodeficiency (SCID) mice (2- to 3-wk-old) were used (Taconic). The mice were anesthetized with isoflurane (Forene; Abbott ) during the manipulations. One flank or both flanks of each animal were injected subcutaneously (total 200 μl) with sHPT-1 cells together (1:1) with BD Matrigel Matrix (BD Biosciences Clontech) after transfection of 106 cells for 24 h. The animals were monitored every day and humanely killed after 8–9 wk. The animal experiments were approved by the Uppsala University board of animal experimentation and were performed according to the United Kingdom Coordinating Committee on Cancer Research guidelines for the welfare of animals in experimental neoplasia [27].
An Internally Truncated LRP5 Receptor Is Expressed in Parathyroid Tumors
A shorter LRP5 transcript (LRP5Δ666–809) was detected in 32 out of 37 pHPT tumors (86%) and 20 out of 20 sHPT tumors (100%) by using RT-PCR primers located in exons 9 and 13 of LRP5 (Figure 1A). Of the analyzed parathyroid tumors (n = 57) which all displayed aberrant cytoplasmic/nuclear accumulation of β-catenin ([5] and unpublished data), four pHPT tumors without the LRP5Δ666–809 transcript harbored the stabilizing mutation of the β-catenin exon 3 (S37A), and one pHPT tumor had neither. Three out of the four tumors with β-catenin mutation were reported previously, and the one tumor without β-catenin mutation or the LRP5Δ666–809 transcript displayed wild-type APC expression [5]. Six normal parathyroid tissues and a panel of 17 normal tissues expressed only the normal LRP5 (LRP5wt) transcript (Figure 1B). Accordingly, a smaller LRP5 protein was detected by immunoprecipitation followed by Western blot analysis of the parathyroid tumors expressing the LRP5Δ666–809 transcript and in the human parathyroid tumor cell line sHPT-1 (Figure 1C), where CTNNB1 is not mutated in exon 3 [8]. The expression level of the smaller form of LRP5 varied in relation to LRP5wt among the analyzed tumors. In the sHPT-1 cell line, it was expressed at a higher relative level (see also Figure 2C). Only the LRP5wt receptor was detected in HeLa cells (Figure 1C), which did not express the truncated LRP5Δ666–809 transcript (unpublished data). No apparent relationships between expression of LRP5Δ666–809 or CTNNB1 stabilizing mutation and clinical characteristics, such as glandular weight or serum levels of parathyroid hormone or calcium, were found.
(A) RT-PCR analysis of RNA from pHPT tumors (n = 20) and sHPT tumors (n = 20) using primers located in exons 9 and 13 of LRP5. The wild-type or truncated LRP5 mRNAs were detected by primary PCR for most of the tumors or with nested PCR using an additional overlapping forward primer. The truncation comprised the last 93 bp of exon 9, all 227 bp of exon 10, and the first 106 bp of exon 11.
(B) Nested RT-PCR of normal parathyroid tissue (n = 6), and parathyroid tumor cell line sHPT-1 [8] as marker (top panel). Nested RT-PCR analysis of 17 normal tissues, and the parathyroid tumor pHPT21 as marker (lower panel).
(C) Immunoprecipitation followed by Western blot analysis of LRP5. Transiently expressed (HEK293T cells) LRP5 and LRP5Δ666–809 shown as size markers. sHPT-1, human parathyroid tumor cell line [8]; pHPT, pHPT tumor. The lower panel shows additional pHPT tumor samples analyzed on a 5% SDS–polyacrylamide gel, where the proteins separate more clearly.
(D) A schematic structure of LRP5 is shown. The truncated mRNA contains an in-frame deletion of LRP5 between amino acids 666 and 809 (Δ666–809), encompassing the third YWTD β-propeller domain. The truncation (Δ666–809) is flanked by imperfect direct repeat sequences (horizontal arrows) with putative cryptic donor (Ac-GTG) and acceptor (AC-cT) RNA splice sites in exons 9 and 11, respectively (arrows). The Δ666–809 is between nucleotide positions 2039 and 2466 of the LRP5 mRNA (GenBank accession no. AF064548).
(A) Locations of siRNAs and probes for quantitative real-time PCR. tot mRNA PCR probe determines both LRP5wt and LRP5Δ666–809 transcripts.
(B) Expression of LRP5wt and LRP5tot mRNA assessed by quantitative real-time PCR. (C) Immunoprecipitation and Western blot analysis of LRP5.
The tumor-associated shorter transcript LRP5Δ666–809 contained an in-frame deletion of 142 amino acids (Δ666–809), encompassing the third YWTD β-propeller domain [28,29] between the second and third epidermal growth factor repeats of LRP5 (Figure 1D). The deletion (Δ666–809) was flanked by imperfect direct repeat sequences with putative cryptic donor and acceptor RNA splice sites located in exons 9 and 11, respectively (Figure 1D). The 5′ putative cryptic splice site (Ac-GTG) related to the consensus sequence of major-class introns and the 3′ cryptic splice (AC-cT) related more to the minor-class [30]. The genomic sequences of exon/intron junctions and branch points in question as well as exons 10 and 11 did not reveal any abnormalities in several tumors. Also, no deletion was noted by Southern blotting of genomic DNA (unpublished data). The LRP5 internal truncation is most likely the result of cryptic splice site usage.
The Internally Truncated LRP5 Receptor Contributes to Active β-Catenin Signaling
To investigate the functional consequences of LRP5Δ666–809 expression, if any, specific and control siRNAs were efficiently transfected [8] to the recently established unique human parathyroid tumor cell line sHPT-1. Three different siRNAs against LRP5 mRNA were used (Figure 2A): one against the LRP5wt transcript (siLRP5wt), one against the internally truncated transcript (siLRP5Δ666–809), and one recognizing both the LRP5wt and LRP5Δ666–809 transcripts (siRNAtot). siLRP5wt was directed to exon 10, which was excluded from the truncated transcript (Figure 1A); siLRP5Δ666–809 was directed against the unique sequence created at the aberrant exon junction; and siRNAtot was directed to exon 13, which was present in both LRP5 transcripts (Figures 1D and 2A). A nonsilencing siRNA (control siRNA) and a mutant siRNAtot (control siRNAtot) were also used. Specificity and silencing potential of the siRNAs were ascertained both at the mRNA and protein level (Figure 2B and 2C).
Transfection of siLRP5Δ666–809 reduced the nonphosphorylated active β-catenin level markedly as well as decreased the growth of sHPT-1 cells. siLRP5wt showed a tendency toward a lowered β-catenin level, but no significant effect on cell growth was observed (Figure 3A and 3B). In agreement with these results, transfection of siRNA specific for both LRP5 transcripts (siLRP5tot) resulted in similar diminished β-catenin expression and cell growth as for siLRP5Δ666–809 (Figure 3A and 3B). The growth of HeLa cells, which did not express the internally truncated LRP5 receptor (Figure 1C), was not affected by siRNA transfections (Figure 3B). As shown previously for siRNA to CTNNB1 [8], sHPT-1 cells transfected with siLRP5Δ666–809 but not with control siRNA showed accumulation of dead cells as well as a smaller population of apoptotic cells 84 h after transfection (Figure 3C). Furthermore, the endogenous β-catenin activity in sHPT-1 cells (11-fold), as measured by FOPFLASH/TOPFLASH TCF/β-catenin luciferase reporter transfection, was dependent on maintained expression of LRP5Δ666–809 and not of LRP5wt (Figure 4A). Dependence on maintained expression of LRP5Δ666–809 was also observed for expression of the endogenous β-catenin target gene MYC [22], which is commonly overexpressed in parathyroid tumors [5], as the MYC mRNA level was significantly reduced in sHPT-1 parathyroid cells transfected with siLRP5Δ666–809 and siLRP5tot, but not with siLRP5wt, control siRNA, and control siLRP5tot (Figure 4A). Expression of MYC in sHPT-1 cells has been shown previously to depend on β-catenin [8]. Thus, maintained expression of the internally truncated LRP5 receptor in sHPT-1 cells appeared necessary for accumulation of transcriptionally active β-catenin, continued cell growth, and MYC expression. An effect of the internally truncated LRP5 receptor on β-catenin signaling was further substantiated by experiments in HEK293T cells. Transient expression (Figure 4B) of LRP5Δ666–809 and not of LRP5wt in these cells resulted in increased level (5-fold) of nonphosphorylated active β-catenin (Figure 4C), a 5-fold increase of endogenous MYC mRNA expression (Figure 4D), and enhanced association (4-fold) of β-catenin to the MYC promoter, as revealed by ChIP (Figure 4E).
(A) Western blot analysis of active β-catenin [25], 60 h after transfection. The β-catenin–actin signal ratio is shown.
(B) Effects on sHPT-1 cell growth. HeLa cells were used as control for toxic effects. *p < 0.05.
(C) Flow cytometry analysis of sHPT-1 cells at 84 h after transfection after staining with annexin V–FITC and propidium iodide. Accumulation of dead cells in the upper left quadrant; population of late apoptotic cells (upper right quadrant) and early apoptotic cells (lower right quadrant).
(A) Transient cotransfections of FOPFLASH or TOPFLASH TCF/β-catenin reporters, the CMV-LacZ reference plasmid, and indicated siRNAs to sHPT-1 cells (left panel). FOPFLASH contains mutated binding sites for TCF factors, while TOPFLASH does not [22]. Luciferase activities were normalized to β-galactosidase activities. Effects on endogenous MYC expression in sHPT-1 cells (right panel). *p < 0.05.
(B) Western blot analysis of V5-tagged LRP5 and LRP5Δ666–809 transiently transfected to HEK293T cells. LRP5 and LRP5Δ666–809 were expressed at similar levels.
(C) Western blot analysis of cytosolic nonphosphorylated active β-catenin in HEK293T cells, 24 h after transfection.
(D) Endogenous MYC expression in transfected HEK293T cells. *p < 0.05.
(E) ChIP of the MYC promoter in transfected HEK293T cells. An anti-active–β-catenin monoclonal antibody was used [25].
WNT3 Ligand and the Internally Truncated LRP5 Receptor Strongly Activate Transcription with Impaired DKK1 Inhibition
The effects of WNT ligand stimulation of the LRP5wt and LRP5Δ666–809 receptors were determined by transient cotransfection of the TOPFLASH TCF/β-catenin luciferase reporter and the LRP5 receptor expression vectors to sHPT-1 cells, followed by incubation in WNT1, WNT3, or WNT3A conditioned medium. Transfected LRP5Δ666–809 displayed activation of β-catenin driven transcription in sHPT-1 cells (9-fold), and transcription was most prominently stimulated (80-fold) in the presence of WNT3 ligand, where it was 4-fold higher than for LRP5wt (Figure 5A). In addition, only WNT3 stimulated the endogenous β-catenin activity (9-fold). WNT3A ligand stimulated transcription only in the presence of LRP5wt (10-fold), and WNT1 showed no effects. WNT3 but not WNT1 was expressed by RT-PCR in parathyroid tumors (Figure 5B).
(A) sHPT-1 cells cotransfected with TOPFLASH, LRP5wt, or LRP5Δ666–809 expression vectors and CMV-LacZ reference plasmid, followed by incubation in WNT1, WNT3, or WNT3A conditioned medium (CM). CM was from HEK293T cells transfected transiently with expression vectors for the various WNT ligands. The 11-fold (Figure 4A) endogenous β-catenin activity is set to 1 (unstimulated, empty vector).
(B) Representative RT-PCR analysis of RNA from three pHPT and three sHPT tumors using primers for WNT3. No expression of WNT1 was detected by the conditions used [21].
(C) Cotransfection of TOPFLASH and CMV-LacZ reference plasmid to sHPT-1 cells. Incubation in WNT3 and DKK1 CM.
(D) Cotransfection of TOPFLASH, CMV-LacZ reference plasmid, and LRP5wt or LRP5Δ666–809 expression vectors to HEK293T cells followed by incubation in WNT1, WNT3, or DKK1 CM. HEK293T cells do not express the internally truncated LRP5 receptor.
Several amino acid residues in the deleted part of LRP5 (Δ666–809) were shown to be required for inhibition of β-catenin activity by the WNT antagonist DKK1 [31]. Therefore, we examined the effect of DKK1, which is expressed in parathyroid tumors (unpublished data) on WNT3-stimulated β-catenin activity in sHPT-1 cells. The presence of DKK1 in the culture medium resulted in inhibition of WNT3 activation in HEK293T control cells as expected (Figure 5D), but not in sHPT-1 cells (Figure 5C). In agreement with these results, WNT3-induced TOPFLASH transcription was inhibited by DKK1 in HEK293T cells cotransfected with LRP5wt and not with LRP5Δ666–809 (Figure 5D). Also, the activity of LRP5Δ666–809 in unstimulated cells (7-fold) was insensible to DKK1. In contrast to sHPT-1 cells (Figure 5A), HEK293T cells responded to WNT1-conditioned medium (7-fold) and was sensitive to DKK1 inhibition as expected. However, the small increase of LRP5Δ666–809 activity in the presence of WNT1 (1.7-fold) was insensible to DKK1 (Figure 5D). The impaired DKK1-mediated antagonism of β-catenin activity may contribute to overall signaling of the internally truncated LRP5 receptor.
Inhibition of Tumor Growth in SCID Mice
The in vivo growth properties of sHPT-1 cells were evaluated in a xenograft SCID mouse model. Significant tumor growth inhibition was observed in transplants of cells when pretransfected with siLRP5Δ666–809 or siLRP5tot, but not with siLRP5wt or control siRNA (Figure 6A and 6B). Thus, the internally truncated LRP5 receptor appeared to be required for tumor growth in this animal model. Combination of the very high siRNA transfection efficiency of sHPT-1 cells [8] and generally prolonged gene silencing in slowly dividing cells [32] are likely prerequisites for the sustained tumor growth inhibition observed 8–9 wk after transplantation.
(A) Injection of sHPT-1 cells pretransfected for 24 h with siLRP5Δ666–809 or control siRNA. Arrows indicate representative parathyroid tumor growth at site of transplantation.
(B) Parathyroid tumors from SCID mice (n = 35) injected with sHPT-1 cells pretransfected for 24 h with the indicated siRNAs were excised and weighed. *p < 0.05. The animals were monitored every day and killed after 8–9 wk.
β-catenin accumulation has been observed by us in all so far analyzed parathyroid tumors of primary origin and hyperplastic parathyroid glands of HPT secondary to uremia ([5] and this work), strongly suggesting dysregulated WNT/β-catenin signaling as a common pathogenic pathway for these different conditions. We reported CTNNB1-stabilizing mutations in a few cases (3 out of 20) of pHPT tumors, while no mutation was found in uremic secondary HPT tumors, and inactivating truncations of APC were not seen [5]. Recently, parathyroid adenomas from 97 patients with pHPT who had undergone parathyroidectomy in the United States were analyzed regarding CTNNB1-stabilizing mutations in exon 3 [33]. In agreement with one previous smaller study of Japanese patients [34], no mutations were identified. Taken together, these results indicated a low CTNNB1 mutation frequency in pHPT tumors [35]. We are currently analyzing a large number of additional pHPT tumors from Swedish patients. A few additional tumors with CTNNB1-stabilizing mutations has been found so far, suggesting an overall mutation frequency of approximately 6% (Björklund et al., unpublished data). This frequency is slightly lower than that observed for colorectal cancers, in which approximately 10% of the tumors harbored missense mutations or interstitial deletions of exon 3 [13,14].
Several lines of experimental evidence presented here support a fundamental role of the internally truncated LRP5 receptor in regulating β-catenin–driven transcription in parathyroid tumor cells. The truncated LRP5 receptor was frequently expressed both in parathyroid tumors of primary origin (86%) and in hyperplastic parathyroid glands of HPT secondary to uremia (100%). Of all 57 analyzed parathyroid tumors with accumulated β-catenin, 52 expressed the internally truncated LRP5 receptor without CTNNB1-stabilizing mutations, four tumors had CTNNB1-stabilizing mutations but no truncated LRP5 receptor, and 1 tumor displayed neither aberration. Thus, it seems that mutation of CTNNB1 and expression of LRP5Δ666–809 is mutually exclusive.
All reported LRP5 missense mutations, which cause a high-bone-mass phenotype, are located before the first epidermal growth factor–like repeat in the amino terminal part of the receptor, not including the third YWTD β-propeller domain, which is absent in the truncated LRP5 receptor. Several mutations of LRP5 known to cause osteoporosis-pseudoglioma syndrome is located within amino acids 666–809, highlighting a functional role for this part of the protein [36]. Osteoporosis-pseudoglioma mutations seem to cause varying degree of reduced Wnt or Norrin signaling by unknown mechanisms, as determined by the TOPFLASH reporter assay [37]. Clearly, the deletion of 142 amino acids in LRP5Δ666–809 and a missense mutation in the same part of the receptor have opposite effects. A missense mutation may for example interfere with binding of cell-type–specific factor(s), while a larger deletion may result in structural changes of the receptor with a different impact on cofactor interactions and signal outcome. The mechanism(s) by which the internally truncated LRP5 receptor activates β-catenin signaling in tumor cells remains to be elucidated, but may involve impaired inhibition by DKK1. The WNT3 ligand activated transient transcription more strongly in the presence of LRP5Δ666–809 than LRP5wt, possibly suggesting favored interaction of WNT3 with the internally truncated receptor. We emphasize that the conditioned medium of WNT1, WNT3, WNT3A, and DKK1 used throughout this study might contain uncharacterized signaling molecules induced by the WNTs or DKK1.
RNA splicing defects in cancer are common, but seem rarely to be caused by somatic mutations in splice sites and regulatory elements. Alternative splicing may potentially present as a diagnostic marker [38,39]. Interestingly, aberrant splicing between direct repeat sequences with potential cryptic splice sites and without mutations in selected parts of the gene has been described also for the MDM2 oncogene in breast cancer [40]. We found no mutations or polymorphisms in the sequenced exons/branch points/intron–exon junctions of the LRP5 gene in the analyzed tumors that potentially could explain the aberrant splicing. Mutations in other areas of the LRP5 gene affecting splicing specificity, accuracy, or efficiency, or perhaps more likely interference of trans-acting factors of the splicing machinery, are other possibilities. Alternative splicing may be affected in a splicing factor concentration-dependent manner [41], and this may also apply to aberrant splicing in tumors. Identification of splicing oncogenes or splicing tumor suppressor genes is being pursued.
The results support a fundamental role of the aberrantly spliced internally truncated LRP5 receptor in activating WNT/β-catenin signaling in parathyroid tumor cells. LRP5 receptors could be attractive targets for the development of antitumor drugs that specifically inactivate the truncated receptor and leave the normal protein unaffected.
We are grateful to B. Bondeson and P. Lillhager for skillful and extensive technical assistance. We thank Drs. R. Kemler, J. Kitajewski, S. Sokol, and B. Williams for providing expression plasmids.
Author Contributions
GW designed the study. PB performed experiments and analyzed data. PB, GÅ, and GW interpreted the data. GÅ reviewed clinical data. PB, GÅ, and GW contributed to writing the paper.
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text:
Michigan St Preview: Now with more charts!
Michigan St Preview: Now with more charts!
- Read more about Michigan St Preview: Now with more charts!
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All numbers included in this preview are using my PAN metric, Points Above Normal. PAN is essentially how many points above an average FBS team was a team/unit/player worth. For reference, an average FBS is approximately equal to Illinois or a top team from the MAC.
All games against FCS teams are excluded for all teams, as well as any plays in the second half where one team leads by more than 2 touchdowns or any end of half run out the clock situations.
At this point adjustments for strength of opponent are directional but still highly uncertain. They will be now be used in all situations except otherwise noted.
Rush Offense vs Michigan St
Michigan Off: +8 PAN, 2nd nationally, 1st Big Ten
Michigan St Def: -0 PAN allowed, 54th, 6th
[Chart note: positive numbers mean good performances for both offense and defense.]
Despite Michigan St holding Wisconsin to 24 points last week, they fared worse against Wisconsin's running game than the average team has in 2010.
Michigan has been between good and ridiculous in every game this year. Michigan St's rush defense is definitely an upgrade over the Hoosiers from last week, but it's more of an "allow touchdowns in reasonably length drives" as opposed to a "regularly keep us out of the end zone" type matchup.
Robinson will obviously be the catalyst for Michigan. His rushing PAN is +9, over 3.5 points better than anyone else in the nation. Vincent Smith and Michael Shaw have both hovered right around zero, with Smith grading out higher due to a combination of the long run against Indiana and Shaw's best performance coming against an FCS opponent.
Michigan will face a better rush defense than they have seen in the last month or so but Michigan St hasn't shown anything yet to indicate they are good enough to slow Michigan's run game down more than anyone else has. Look for at least a full touchdown worth of advantage from the Michigan rushing attack.
Pass Offense vs Michigan St
Michigan Off: +7, 4th, 2nd
Michigan St Def: –1 allowed, 46th, 4th
After struggling for the first month of the season, Michigan St's pass defense was the difference in shutting down Wisconsin last week. Michigan's pass game has continued to be very potent and last week at Indiana was the best game yet. Because Michigan's passing success is built so much off the success of the running game, it's not as clear as to how a good pass defense will be able to defend the Wolverine passing game.
Robinson has been +6 PAN on the season and is the 20th ranked passer in my ratings, which can reward for volume, which Michigan has very little of in the passing game. Robinson will be aided by a group of receivers who have been much more productive than last year. Hemingway and Roundtree are both averaging a solid +6 PAN per game and Odoms is at +3. Stonum is potentially a threat but has only been worth +2 against FBS competition.
Michigan St stepped up last week and did a great job limiting Wisconsin through the air. Although both Michigan and Wisconsin's passing games are set up by strong ground games, the spread and shred is very different from old school Big Ten rushing. If Michigan St can replicate last week's success this matchup could be a draw instead of the +6 for Michigan the numbers indicate.
Rush Defense vs Michigan St
Michigan Def: +1 allowed, 72nd, 7th
Michigan St Off: +3, 28th, 5th
This chart, with the exception the MSU-WMU game is a chart of averageness. Michigan hasn't been gouged in the running game but they haven't been closing the door on anyone either. Michigan St's two-headed running attack came out big in Week 1 but has been relatively quiet since.
Michigan St's +3 PAN is essentially split between Le'Veon Bell and Edwin Baker with both players contributing equally to the success. Bell had the big day against Western with a +7 showing but was shut down against Wisconsin by going –4 on the day. Baker has been much more consistent with all four games going between +1 and +3.
If you take out the beatdown Michigan St's backs administered in Week 1, the Spartan ground game looks much more tameable. Michigan can not sleep on this matchup but is in better shape than I thought. Michigan should have a chance to reasonably contain the Spartan backs, shifting the challenge to the…
Pass Defense vs Michigan St
Michigan Def: –1 allowed, 42nd, 3rd
Michigan St Off: +2 allowed, 32nd, 6th
As I have stated previously, the high rating for Michigan's pass defense against Indiana will not hold. I do agree with Brian that Indiana will probably still have the best pass offense in the Big Ten this year, but when you are compared to Akron and Western Kentucky in your performance, you should always come up looking good, even if you allow nearly 500 yards through the air.
Michigan's pass defense is very difficult to assess right now. Indiana doesn't have good comps to measure against. The BG performance mostly looks bad because of one fluke play where BG got away with a massive hold. ND and UConn both look like respectable performances in comparison with how other teams have defended them.
Michigan St has really stepped up their passing game in the last two FBS games, with a pair of +7's against legitimate opponents. After two sub-zero PANs for Kirk Cousins to start the season, he has been +11 and +12 in his last two games (the difference between the team and his score is that sacks count against the team but not the player).
The receiving has been pretty balanced with three Spartans checking in at +4 on the season. BJ Cunningham, Keshawn Martin and Mark Dell have carried most of the load this year. All three have a game-rated +9 or higher on the season.
If Kirk Cousins can keep up his recent success this where it starts to get scary for Michigan. The Spartan passing attack will not be as good we saw last week in Indiana but if Michigan does improve in the secondary, the results could be nearly as bad, especially if Michigan St can keep Michigan looking in the backfield on play action.
Special Teams vs Michigan St
Keshawn Martin. That's the two big green bars on the graph. He has three big returns on the year, one going the distance against Wisconsin last week. Let's not kick to him, although he has lost a fumble on a return this season. Michigan's blue bars have gotten closer to zero, mostly because we have chosen to forgo special teams altogether. The Spartan kicking situation is a polar opposite to Michigan. Michigan's kickers have cost the team 5 points vs an average kicker while Michigan St's kicker has been 5 points better than the average kicker. If this game is decided by special teams it is very unlikely to be a Wolverine win.
Predictions Almost Certain to Cost You Money if Taken Seriously
At a neutral site, this matchup is pretty much a tossup. Luckily its in Ann Arbor. If Michigan can keep Keshawn Martin from breaking a long return and is at least even in turnovers I would feel really good about the chances.
Michigan 35 Michigan St 32
Elsewhere in the Big Ten:
Northwestern 27 Purdue 20 – Northwestern continues to be college football's worst undefeated team
Ohio St 40 Indiana 21 – Chappell has a bunch of yards but also throws a couple picks while Ohio St runs all over Indiana
Penn St 13 Illinois 10 – Basically a repeat of every game Penn St has played this year
Wisconsin 31 Minnesota 24 – Going with my numbers. I think they are underrating Wisconsin but I still think they are overrated overall.
I guess I'll pick the Bama game again since Game Day dissed Ann Arbor for them this week.
Alabama 24 South Carolina 20 – A virtual mirror of the Alabama/Arkansas game.

subset: commoncrawl


text:
Names: Twitchgrass, Quickgrass, Doggrass.
Habitat: Grows in many parts of the world, and all too frequently ingardens.
Collection: The rhizome should be unearthed in spring or early autumn. Wash it, carefully dry in sun or shade.
Part Used: The rhizome.
Indications: Couchgrass may be used in urinary infections such as cystitis, urethritis and prostatitis. Its demulcent properties soothe irritation and inflammation. It is of value in the treatment of enlarged prostate glands. It may also be used in kidney stones and gravel. As a tonic diuretic, Couchgrass has been used with other herbs inthe treatment of rheumatism.
Ellingwood says that "its action is solely upon the urinary apparatus. It exercises a soothing, diuretic influence, greatly increasing the flow of the watery portion of the urine without to the same extent influencing the actual renal secretion. It is bland, mild, unirritating, and is used whenever urine, having a high specific gravity, causes irritation of the kidneys or bladder, more especially of their mucous surfaces." He recommends it for the following conditions: pyelitis, catarrhal and purulent cystitis, gonorrhoea, lithaemia, dysuria, tenesmus, prostatitis, strangury and haematuria.
Combinations: For urinary tract infections it may be used with Buchu, Bearberry or Yarrow. It can be combined with Hydrangea for prostrate problems.
Preparations & Dosage: Decoction: put 2 teaspoonfuls of the cutrhizome in a cup of water, bring to boiling and let simmer for l0 minutes. This should be drunk three times a day.
Tincture: take 2-4 ml of the tincture three times a day.


text:
To facilitate a platform for discussion and idea exchanges related to the library's role in the new student's first year experience in higher education. This could include, but is not limited to, discussions related to student transition and preparedness issues, transfer student populations, information literacy programs, outreach, and cross campus collaboration. The group could be of interest to any librarians working with this population at a variety of schools such as community colleges, four- year colleges, research universities, and even possibly those working in a K-12 setting.
Displaying active committee roster as of 12/14/2018. Last retrieved on 12/14/2018. Members can log in to view full contact information for committee members.
Discussion Group composition include group leaders and membership based on interest.
Subcommittees & Task Forces
Discussion Group composition include group leaders and membership based on interest.
The First Year Experience Discussion Group was established at the 2012 Fall Executive Committee Meeting.
Get Involved
Join a Discussion Group
If you would like to join the First Year Experience Discussion Group or have any questions, suggestions, or ideas for discussion topics, please contact the convener(s).
Staff Liaison
Mariel Colbert
Program Coordinator
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Work: 312-280-5285
E-mail: email@example.com


text: Markham, Ontario is one of the most culturally diverse and fast growing cities in the Greater Toronto Region with over 353,000 people. A multiple award winning municipality celebrating its 220th anniversary, Markham is recognized as Canada's High-Tech Capital anchored by 1000 high-tech and life sciences companies that generate 37,000 jobs. Of the 10,400 companies in Markham, there are more than 400 Canadian head offices located here - including industry leaders such as IBM, Aviva Insurance, AMD, Allstate, The Miller Group, Parsons, Huawei Technologies, MMM Group, Lenovo, GE Energy, Bank of China, Toshiba, J+J, Hyundai and Honda. Defined by its rich heritage, well-planned communities, high quality infrastructure, a talented workforce, and a pro-business environment, Markham's business community includes outstanding companies and organizations providing globally competitive banking, financing, engineering, design, technical and commercialization services.


text:
The OWB browser really is a blessing for the Amiga and MorphOS platforms, because it's a massive step forward from the CSS-less browsers that came before it. There's still a lot of work to do when it comes to rendering and especially rendering speed, but overall it's pretty impressive to see the progress the developers are making. Interface-wise, it's all very minimalist, but it's native. It lacks a few key features though, such as tabbed browsing.
Note that the below screenshots were made on AmigaOS 4.1, and not on MorphOS.
In any case, this new release for MorphOS runs on both version 2.x and 1.3 of MorphOS, but it only works on the later with a recent MUI snapshot (enthusiasts will know what that means).
Update: This is a screenshot of the actual latest version of OWB on AmigaOS4; as commenters pointed out, I wasn't using the latest version. This new one does have tabbed browsing and an improved interface.


text:
"...The primary purpose of the Eagle court of honor is to honor the new Eagle Scout. He should be the center of attention, and the spotlight should never stray far from him.
But the ceremony should fulfill other purposes as well. Besides presenting the Scout with a badge, it should also present him with a challenge: to use his skills and experience to be a leader in his troop and in his community. This challenge, or charge, helps him answer the question "Now What?" much as a commencement speech does for high school graduates.
While the new Eagle is the focus of attention, a few other people, specifically his parents and Boy Scout leaders, should briefly share the spotlight. The honoree may be the one who climbed the trail to Eagle, but his Scout leaders and parents pushed him at least part of the way."
The Eagle Court of Honor Book, The Complete Guide to Scouting's Greatest Moments.Mark Ray
This book is available at all Scout Shops or can be purchased online through www.scoutstuff.org.
Eagle Courts of Honor are usually held separately from troop Courts of Honor, since the focus of the Eagle Court of honor is on the Eagle Scout. The senior troop leadership, troop adult leadership, and/or the parents of the Eagle Scout may organize the ceremony.
But you can give them ideas to make the ceremony special!


text:
Parthenon Hostel
This hostel is the only hostel in downtown and walking distance to the train, Greyhound Bus, Amtrak, Michigan Avenue and the Loop. In fact, the airport shuttle train is just $3 to our block.
We listened to your reviews and just completed brand new renovations for the entire property. Enjoy modern and clean rooms and so much more!
We offer free breakfast, free Wi-Fi and Internet, free maps of Chicago, discounts to museums and attractions and more!
We're open 24 hours, with 24hr luggage storage and no curfew.
Our huge common room has plasma TV and there's a large kitchen as well.
The rooms are all new and we have private rooms to suit up to eight people, with special group rates available. Our bathrooms are granite. All rooms include free linen.
We've got discounts on bike rental!
Located just a few blocks from the Amtrak, on the bus line to the airport and our downtown location make us the perfect hostel to stay at when in Chicago.
Parthenon Guest House' is a winning choice for your lodging needs. Nestled in the shadow of the Sears Tower it is centrally located to many of Chicago's finest attractions. Just blocks from the Loop and close to restaurants, banks and shops all within walking distance. Michigan Ave/Magnificent Mile shopping, Chicago Planetarium, Art Institute of Chicago and Navy Pier are located a short distance away.
We're close to Union Station and Greyhound Bus and one block from the CTA El train. The CTA El is a $2 fare from both O'Hare International Airport and Midway Airport. Arriving by Amtrak or Greyhound? We are only a few blocks away and an easy walk!
Parking no longer available.
There's no age restriction and no membership fee, unlike H.I. locations.
Availability & RatesTo view availability and prices please select dates and click 'Update Dates'
Hostel Facilities & Policies
- 24 Hour Reception
- Age Restriction
- Bar
- Free Breakfast
- Cable TV
- Common Room
- Credit Cards Accepted
- Free City Maps
- Free WiFi
- Fridge/Freezer
- Hair Dryers
- Hot Showers
- Linen Included
- Security Lockers
- No Curfew
- Non Smoking
- Restaurant
- Taxes not included
- Vending Machines
Cancellation PolicyThis property has a 2 day cancellation policy. Failure to cancel within this time will result in a cancellation charge equal to the first night of your stay.
Must-Read Hostel InfoThis property has specific Terms & Conditions.
We have a 48-hour cancellation policy. Late cancellations and no-shows shall be charged the equivalent of the first night's accommodation. Cancellations must be made via email.
The local city tax of 17.4% is not included here and will be added to your balance, due at check in.
All guests paying their balance by credit card will be charged an additional 3% merchant fee. Cash accepted.
Local Chicago Residents are not permitted at the hostel.
Minors under the age of 18 are not allowed in the dorms. Minors can stay at the hostel in a private room with an adult.
****Please note***** In the months of January and February the front desk is closed from 12 am to 7 am. If you will be arriving during these times, please send us an email, letting us know so we can accommodate you.
DirectionsShow MapHide Map
From O'Hare airport:
Take the CTA blue line ($2.25) and exit at the Halsted Street Station. We're one block from the station.
From Midway airport:
Take the Orange Line to the Clark/Lake stop. Go all the way downstairs to the subway and take Forest Park blue line to UIC/Halsted stop. We are one block north.
When you arrive we'll provide you with a transit map full of bus and train routes on our block!
From Amtrak:
Walk west on Jackson 3 blocks and then turn left on Halsted.
We're right on the street at 310 S. Halsted.
From Greyhound:
Walk 1 block west on Harrison Street and then take a right on S. Halsted and we are 1 block up on
310 S Halsted St
Chicago, IL 60661
Located Near
We are located in Greektown, near the blue line CTA Halsted exit.
You can use the metro to access The Loop, Sears Tower, Michigan Avenue, Magnificent Mile, shopping, restaurants, pubs and bars and music.
All are accessible via the blue line.


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CHARLESTON, S.C. (April. 21, 2005) - The Southern Conference announced today the 2005 Men's and Women's Tennis All-Conference teams as well as the Players of the Year, Coach of the Year, Freshman of the Year and the Sportsmanship Awards. A total of 11 ETSU student-athletes were recognized as voted on by the league's head tennis coaches.
East Tennessee State picked up the men's player and coach of the year. Sophomore Oscar Posada (Valencia, Venezuela) earned the Player of the Year honor with a perfect 10-0 record at No. 1 singles in conference matches, 14-2 overall. He was the 2004 Freshman of the Year and is currently ranked 49th in the ITA national rankings. The Bucs head coach Yaser Zaatini was tabbed the Coach the Year by his peers after leading his team to its third-straight regular season title and eighth since joining the SoCon in 1978.
All six members of ETSU's line-up received accolades as All-Conference Selections. Posada and fellow sophomore Felix Insaurralde (Asunci�n, Paraguay) were named as First Team All-SoCon singles selections, while the pair of Posada and team captain Fernando Fiuri (Villa Maria, Argentina) was named as a doubles selection. Second Team All-SoCon selections included at singles Fiuri, Lisandro Picardo (Mar del Plata, Argentina) and Luis Sebastian Serrano (Cartagena, Colombia). At doubles the teams of Insaurralde and Picardo along with the brother duo of L. S. Serrano and Mo�ses Serrano (Cartagena, Colombia) were named to the second team.
On the women's side, two-time SoCon Women's Tennis Player of the Week Sandie Knight (Adamstown, Australia) earned a SoCon First Team All-SoCon singles team selection, while the pair of Sylwia Ziolkowska (Warsaw, Poland) and Masha Ilina (Kyiv, Ukraine) was tabbed as a doubles first team selection. Doubles partners Olya Batsula (Minsk, Belarus) and Gemma Halim (Jakarta, Indonesia) were named as an All-SoCon Second Team doubles selection.
The Southern Conference tennis championships are being held in Charleston, S.C. at the Earle Tennis Center (ETC) on the campus of The Citadel and at the City of Charleston/Farmfield Tennis Center (FTC). Both the ETSU men's and women's tennis teams will begin tournament action tomorrow at 10:00 a.m. The men will face No. 9 seed Appalachian State (ETC), while the women will face No. 5 UNC Greensboro (FTC).


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by Todd Norlander
Art: Al Weisner
"OI-VAY! The sound of distress carries to Mt. Israel and the Shin Rock trembles... OI-VAY!... and slowly turns into the Kosher Crusader... SHALOMAN!"
Not since the Superman of the '60s has there been a hero so sincere. (Note that when Siegal and Schulster, two young Jewish men from the Midwest, created Superman they gave him dark curly hair. No aryan hero here.)
For more than five years and 25 issues, Al Wiesner of Philadelphia, Pa. has chronicled the legend of Shaloman. Clad in skintight white and light blue, gold belt and matching yarmulke, Shaloman sports the Hebrew letter Shin where Superman has his S. Over the years readers have thrilled to adventures like "The Porkees of Planet X," "Kipa of Doom," "The Cybergs (SCI/FI)" and "The Man From L.A.T.K.E." In my opinion, no issue has yet matched Volume 3, number 5, The Matzoh Chronicles. This issue contains a trilogy of Passover stories: "The Great Missing Matzoh Mystery," "The Destruction of HMZ" and "The Affordable Afikomen."
The first story of the trilogy begins with the bald Dr. Traif, having been defeated again, raising his fists swearing, "Someday, Shaloman, I shall overcome you!" Years pass and on the eve of Passover Reb Nussy is baking matzoh "with a beedle-deedle bum-bum." We next meet Brad Krumm, who along with young runaways Hoffy and Komen, steals all the matzoh in the town of Kutzi Petrik. Crafty Krumm hides his booty in a huge cave lined with 24K gold, for you see, Shaloman cannot see through this precious metal! Upon discovering the theft Rabbi Yom Tov utters "OI-VAY." Enter Shaloman.
Meanwhile, a young boy, Velvel, and his matzoh-loving dog, Mutty, follow an odd trail of crumbs in the woods. Stumbling upon Krumm's hideout, Velvel learns of the villain's plan to ruin Passover by dropping the roof of the cave, thus making "five thousand pounds of farfel of all this! HA HA HA HA HA." Mutty escapes, alerting Shaloman just as Brad Krumm pulls the lever on a large dynamo destined to bring down the ceiling. The Kosher Crusader's great strength burns out the machinery. He then tackles the fleeing Krumm, who hits the ground with a loud "ZETZ!" Off comes the mastermind's bushy blond wig and mustache: "Well, well, Dr. Traif!" Passover is saved. Fartig.
Story two, "The Destruction of HMZ" takes place in the year 2057, when the pollution of our planet is rocketed to the sun, "prompted by the Jewish custom of burning unwanted food to cleanse the house before Passover." Yellow peril villain Ho Metz captures the rocket and brings it to his cave (which lacks 24K gold lining). Ho demands a ransom, or he will detonate the missile, polluting "over three quarters of New Seattle." As his demands are not met, the rocket begins to stink up Ho's cave. Increasingly anxious, Ho Metz guards the front of his lair while Shaloman sneaks through a hole in the back, grabbing the stolen HMZ. It blows up "in outer space" with a thundering "KER-ZETZ!"
The third and final part, "The Affordable Afikomen," is a fable not involving our hero. An afikomen is the dessert matzoh hid and ransomed by the youngest son after the Passover meal. Al Weisner posits a time when afikomen are collected, bronzed and stored in monogrammed, non stick carrying cases. In this Gogolian world afikomen are sold by used car salesmen, swapped like baseball cards and traded on the stock exchange. But then comes the crash..." Grossman Afikomen down ten points."
In a shift, "with sincere apologies to O'Henry" (sic) we get "The Gift of the Magi." Here, Al and his son have the same birthday and Dad wants to get his son a video game, "a sports one, a comic one or a travel one...what about the money for it?" Dad pawns his silver Afikomen holder, while Ben pawns his "gamemaster deluxe" to buy a now rare bronzed afikomen. D'oh! This story abruptly ends with an ad for what I consider the ultimate gift, "a quality cotton/poly Fruit of the Loom T-shirt with a five color picture of Shaloman" - just eight bucks!
Shaloman comics and T-Shirts can be purchased from Mark 1 Comics P.O. Box 5097 Philadelphia, PA 19111 or at your local enlightened comic shop.
© 2000 WFMU. All rights reserved.


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G9481, Revised July 2012
This deer conservation guide is one in a series developed jointly by MU Extension and the Missouri Department of Conservation.
Implementing a sound, quality deer management program depends on obtaining accurate survey data. Obtaining accurate census data requires the ability to collect reliable estimates of deer density on your property. Many hunters use infrared-triggered digital trail cameras to scout and pattern bucks on their property. This type of trail camera may be the best tool available to landowners for determining important demographic characteristics of the deer herd on their property. A camera survey can be used to monitor population demographic trends of deer densities, age structure, sex ratio and fawn recruitment. This guide outlines how to conduct a camera survey on your property.
The two time periods during which camera surveys are typically conducted are preseason and postseason. Because attractants or bait such as corn are used during camera surveys, preseason surveys should be conducted in August so as not to be in conflict with baiting regulations. Scheduling a postseason survey in Missouri is much more of a challenge. Postseason surveys should be done before the casting, or shedding, of antlers but not at a time that would violate baiting regulations. For these reasons, we recommend a preseason, late-August camera survey.
To lay out a camera survey, you will need a map or aerial photo of the property. Aerial photos are available through the following websites:
On the map or aerial photo, lay out a grid that divides the property into cells equal to the number of cameras you intend to use. Usually, one camera per 100 acres is sufficient. Locate each camera station near the center of a cell in an area with convenient access that is likely to be visited by deer.
Once you have identified the locations of the camera stations, bait each area with about 50 pounds of corn about one week before beginning the survey. Replenish corn as necessary to ensure bait is always present.
Set up the cameras when deer begin to visit the bait site on a regular basis.
Run the camera survey for 10 to 14 days. Check stations every four to five days to ensure bait remains and cameras are working properly and have sufficient battery life and memory card space. Replenish bait and replace camera batteries and memory cards as needed. Each camera will function differently, so rely on previous experience regarding battery life and memory card capacity.
Figure 1. Identifying individual bucks is easier with high-resolution photographs that are date- and time-stamped.
As of Sept. 30, 2012, grain, salt products, minerals and other consumable natural or manufactured products used to attract deer are prohibited in the chronic wasting disease (CWD) containment zone, which comprises Adair, Chariton, Linn, Macon, Sullivan and Randolph counties. Using such products to attract deer artificially concentrates deer within a small area, thus increasing the chance of CWD spreading from one deer to another or from the soil to deer. Because bait is required for camera surveys, population demographics on properties located in the CWD containment zone should be collected through another method, such as recording observational data (refer to MU Extension publication G9482, Estimating Deer Populations on Your Property: Observational Data).
When the survey period is complete, begin analyzing the photographs.
Managing thousands of photos can be a challenge. To make it easier, develop a system for organizing all the photos taken during the survey. One good way to manage the photos is to create a folder for each camera station and put all the photos from each station in the station's folder. Then, copy all the buck photos into a "buck" folder for further analysis.
Use the time and date stamp, as well as location, to help differentiate bucks. Yearling bucks are especially difficult to identify as many of them look exactly the same. Opening multiple pictures for viewing, or printing all buck pictures may allow for better differentiation of individuals. After all photographs have been analyzed, calculate population estimates as illustrated in Table 1.
Table 1. Example of calculating population estimates using a camera density of one camera per 100 acres for 10 nights on a 1,000-acre property.
Order publications online at http://extension.missouri.edu/explore/shop/ or call toll-free 800-292-0969.


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"Trajectory, Inc.'s Classics Illustrated has diversified and strengthened its already solid reputation as a thoughtful, intelligent publisher of canonical, traditional titles" said Dr. Katie Monnin, Professor of Literacy, University of North Florida.
Cambridge, MA (PRWEB) May 15, 2012
Trajectory, Inc. announced today that the iconic Classics Illustrated comic and graphic novel series are now available in the Apple iBookstore. The series consists of over 120 digital graphic novels presented in a comic book format with robust color illustrations that introduce great literature to readers of all ages. Trajectory is collaborating with Apple, Inc., the leading provider of smartphones and tablets to deliver the series on the iPad, iPhone, and iPod touch.
Comic book lovers worldwide get ready; the Classics Illustrated digital graphic novels published by Trajectory are now available for purchase in the iBookstore. The digital comic series features the world's greatest stories by the worlds greatest authors including: "The Last of the Mohicans," "War of the Worlds," "The Pearl Princess," "Treasure Island," "The Count of Monte Cristo," "Robin Hood," "Snow White", "Jane Eyre", "The Three Musketeers" and hundreds more.
"Trajectory, Inc.'s Classics Illustrated has diversified and strengthened its already solid reputation as a thoughtful, intelligent publisher of canonical, traditional titles" said Dr. Katie Monnin, Professor of Literacy, University of North Florida. "From the perspective of an educator I foresee limitless potential in bringing Classics Illustrated into the classroom. Probably well known to teachers, parents, and librarians the Classics Illustrated titles are sure to be engaging and exciting comic book stories for an entirely new generation of readers. An A+ idea and comic book execution eBook format, I highly recommend these titles make their way into your child or student's hands."
Novelist Anne Rice states, "I remember reading Jane Eyre in the Classics comics and how much I loved the details and seeing the madwoman in the attic in those little panels and seeing the whole novel play out. What always drew me were very detailed, representational drawings, rather than something abstract. I wanted to see a lot of richness and a lot of depth." Classics Illustrated graphic novels, she said, "Have a freedom that is all their own to create an entire universe in detail fairly inexpensively, compared to a film."
"Making the Classics available in digital form brings these brilliant works to where people live now, on their mobiles. The iPad and iPhone are great for interacting with one of the most beloved comics and graphic novel series of all time." said Jim Bryant, Trajectory CEO.
About Trajectory, Inc. –
Trajectory is a Cambridge, MA based technology and marketing oriented publishing company that collaborates with leading authors, artists, and traditional publishers. Trajectory has 130 eBooks and apps in distribution. Become a fan of Trajectory on Facebook.
Press Contacts: Maura K. Phelan Trajectory, Inc. (617) 564-1550
One Broadway – 17th floor, Cambridge, MA 02141 media(at)trajectory(dot)com


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The possibility of there being something wrong with your baby is a fear that plagues virtually every pregnant woman—and it's why so many of us choose to undergo some form of prenatal testing, either to be prepared to welcome a child with special needs or to make the tough decision to terminate the pregnancy. But two parents of a girl with Down syndrome say that they were misled into believing their baby was healthy and have successfully sued their doctors for $2.9 million, ABC News reports.
Plus: New Test Could Detect Down Syndrome From a Blood Sample
Ariel and Deborah Levy of Portland, Oregon had filed a "wrongful birth" lawsuit against Legacy Health System, claiming that their daughter was misdiagnosed prenatally and that they would have terminated the pregnancy had they known they would have a child with special needs. A jury has agreed that Legacy Health was negligent and the court documents cite five major mistakes that were performed by the hospital, including Deborah's doctor taking an incorrect tissue sample to the lab and a botched analysis.
Plus: A Short Life of Suffering vs. No Life at All
"These are parents who love this little girl very, very much," said the couple's lawyer. "Their mission since the beginning was to provide for her and that's what this is all about."
The couple, who have two older sons, will use the money from their lawsuit to pay for their daughter's care.
Plus: Prenatal Tests, Trimester by Trimester
The hospital issued a statement saying, "While Legacy Health has great respect for the judicial process, we are disappointed in today's verdict. The legal team from Legacy Health will be reviewing the record and considering available options. Given this, we believe that further comment at this point would not be appropriate."
Have you had prenatal testing that was handled incorrectly or came back with the wrong result? How did you handle it


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It comes out the same day as Crosby's DVD (10/07/08)
From deepdiscount:
All the action, all the highlights, and all the drama of the 2007-2008 Stanley Cup Championships are captured here. It's a must have for any fan of the sport since this production goes above and beyond any typical coverage of the event, including interviews, insights, and extras.
From Warner Brothers:
Come along for a magical ride with the 2008 Detroit Red Wings with this special collector's edition DVD set. This four-disc compilation includes the entire broadcast of all six games from the memorable Stanley Cup Final against the Pittsburgh Penguins. Relive all the sights and sounds from Detroit's 11th Stanley Cup championship -- their fourth title in 11 years -- and go inside the glass with one of hockey's greatest franchises. Experience all the nail- biting drama all over again, all the big goals and the show-stopping saves. . .and celebrate one more championship run with the heroes of Hockeytown.
I know it has the Red Wings on the cover, but it still has the Penguins playing in the finals inside, so I will be getting it for the nostalgia and Game 5.
It's shown in widescreen.
Now to give you the sources:
Warner Brothers


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Route 29 Corridor StudyCreating a transportation blueprint for the Route 29 corridor
November 2009
Corridor Study
$1.5 million
Lengths and Limits
Interstate 66 to North Carolina state line
Parsons Transportation Group
Albemarle, Amherst, Campbell, and Nelson
Culpeper, Lynchburg, and Northern Virginia
Marsha Fiol
The Route 29 Corridor Study takes a comprehensive look at the transportation needs for the entire Route 29 Corridor as it travels through Virginia, from the North Carolina state line to Interstate 66 in Prince William County.
The goal is to create a Route 29 corridor blueprint that includes a short-term action plan, intermediate recommendations and a vision plan that identifies long-range goals, policies and recommendations.
The Route 29 corridor is a vital link in Virginia's transportation network and ensuring its long-term viability is essential to the future development of the state's economic potential.
Along with identifying the numerous local needs along this regional corridor the study will also develop a vision for the future for how to best meet those needs.
The corridor study will consider a range of mobility options and needs along the corridor, including personal and commercial vehicles, rail and public transit as well as other modes of transportation.
The results of the study will be used to help establish local, regional and statewide goals for a long-range transportation blueprint that will form the basis for future projects along the Route 29 corridor.
Route 29 Corridor Study Report
The Commonwealth Transportation Board (CTB) considered a resolution on the Route 29 Corridor Study at its Dec. 17, 2009, meeting.
The CTB approved the consensus strategies and goals of the final report and directed the Virginia Department of Transportation (VDOT) and the Department of Rail and Public Transportation (DRPT) to use the corridor-wide strategies as a high-level guide for local and regional planning within the corridor.
Secretary of Transportation Sean T. Connaughton established a subcommittee of CTB members to direct additional work by VDOT and DRPT to develop a draft process for future corridor studies and to address the cited deficiencies in the Route 29 Corridor Study.
The corridor study process will ensure that the CTB will take the lead role in developing consensus among the affected localities, planning district commissions, metropolitan planning commissions and transit agencies along the corridor being studied.
The CTB resolution also directed VDOT and DRPT to provide recommendations for future action on the following specific issues in the Route 29 corridor:
- A prioritized list of intersections to be replaced by grade-separated intersections and interchanges
- A plan to improve mobility and accessibility north of Charlottesville, evaluating various alternatives and not limited to prior proposals
- A plan to improve mobility and accessibility in the Gainesville, Haymarket and Buckland region, evaluating various alternatives and not limited to prior proposals
- A plan to minimize the number of traffic-control signals in the corridor.
The recommendations are to be presented to the CTB by July 1, 2010. The CTB will authorize a corridor implementation plan once the preceding issues have been addressed and the final study is approved.
- Cover (297 KB)
- Table of contents (126 KB)
- Chapter 1 - Executive summary (291 KB)
- Chapter 2 – Study background and approach (841 KB)
- Chapter 3 – Vision for the Route 29 Corridor (508 KB)
- Chapter 4 – Getting to the vision (3.5 MB)
- Chapter 5 – Applying the vision (9.5 MB)
- Chapter 6 – Next steps and implementation (277 KB)
- Appendix A – Land use planning tools (197 KB)
- Appendix B – Transportation-efficient land use planning (578 KB)
- Appendix C – Land use planning for enhanced rail service (1 MB)
- Appendix D – Corridor-wide planning concepts (1 MB)
- Appendix E – Public involvement efforts (268 KB)
Study Addendum: The CTB subcommittee on Route 29 completed its work on the directives of the December 2009 CTB Resolution. This Addendum is the compilation of the work of the CTB Subcommittee which was accepted by the CTB on June15, 2011. Information on meetings of the CTB Subcommittee is available on the CTB's website, www.ctb.virginia.gov/meetings.asp
Local comprehensive plans and other documents for localities within the study area can be found at:
- Greene County multimodal study
- West Piedmont Planning District Commission
- Thomas Jefferson Planning District Commission
- Rappahannock/Rapidan Regional Commission
- Northern Virginia Regional Commission
- Region 2000 Partnership
- Danville Metropolitan Planning Organization
- Central Virginia Metropolitan Planning Organization
- National Capital Region Transportation Board
- Town of Amherst
- Places 29 Master Plan
- Rappahannock-Rapidan Regional Commission
- Ambriar Area Traffic Impact Overlay Plan (PDF, 2 MB)
Route 29 Citizen Information Meeting Materials (Sept. 22 - Oct. 6, 2009)
- Displays from citizen information meetings (ZIP, 13 MB)
- Route 29 Corridor Blueprint brochure, Fall 2009 (PDF, 2 MB)
- Public comment sheet, Fall 2009 (PDF, 42 KB)
- Map displays (ZIP, 18 MB)
- South Lynchburg map (PDF, 2.5 MB)
- Meeting presentation (PDF, 375 KB)


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The charter school that opened for class Thursday, Aug. 15, in North Memphis is unique for several reasons.
Former Memphis mayor and Memphis City Schools superintendent Willie Herenton on Thursday visited Thurgood Marshall High School – a school for teens in Juvenile Court custody – to talk about his upbringing and how students can pursue their own success story.
(Daily News/Andrew J. Breig)
A total of 130 children, all of them in the custody of Memphis-Shelby County Juvenile Court, are assigned to Thurgood Marshall High School of Career Development. Of that number, 86 had registered on the first day of classes at the school within a school at Northside High School.
The two schools are among seven that opened last week as charter schools run by the W.E.B. DuBois Charter Schools Consortium. The consortium's founder and CEO is former Memphis mayor and Memphis City Schools superintendent Willie Herenton.
Herenton saw a teenager in the school uniform of blue shirt and tie and khaki pants Thursday that he had talked with several weeks ago about the school's uniform.
"He said he's not wearing that," Herenton remembered and said that's when he remarked on the orange scrubs children in detention wear.
"I said, 'Did you tell that man that you weren't going to wear that orange uniform? You didn't have a choice did you?'" Herenton said. "When you come to Thurgood Marshall you are going to wear what we tell you to wear."
Sonny Hicks, a Memphis City Schools veteran who is principal of the high school, said the school is able to keep the children enrolled beyond the time that they are in Juvenile Court custody.
"At the point that they are assigned to us, they have to finish with us," he said. "It's not like they come and spend nine weeks or a month and go back to a home school. Once they are assigned they have to carry their programs through."
Juvenile Court officials as well as Herenton indicated last year they wanted a school that would continue to be the school for juvenile offenders after they left detention, citing the problems they often have when they return to their old schools.
There are some unique challenges although the school is held to the same state Common Core standards with the same achievement tests required and the same strategy of immediate intervention when students begin to fall behind.
The school year for the students begins with assessment batteries of tests "to get some kind of diagnostic indication of where they are," Hicks said. "We know that we have to be more of in a nurturing kind of mode and more about healing."
Hicks is realistic about the challenges but also said he sees them as "diamonds in the rough."
"I look at them as being no different than the kids that are in school that haven't come by way of the court system. There are some kids that are challenged and struggle there," he said. "Once we remove that mindset from them and have them to understand that this is the opportunity they need – as long as they take advantage of that, we can help them."
Northside is one of 13 schools the countywide school board will consider closing in the 2014-2015 school year. If Northside closes, interim superintendent Dorsey Hopson has said the W.E.B. DuBois consortium would continue to lease part of the building under its present contract terms.
Juvenile Court is also changing with last year's settlement with the U.S. Justice Department on due process issues the department's attorneys found in court procedures and racial disparities among those juveniles detained and those transferred for trial as adults.
As the Justice Department issued its findings last year, the court had already reduced the number of juveniles it held in detention and dropping that number even more has been one of the key terms in the settlement.
The court already had an arrangement with the old Memphis City Schools system to educate children in detention. But Juvenile Court Judge Curtis Person Jr. was receptive to the idea of a charter school that retained children who had been in detention after they were released.
"The juvenile justice system in the state of Tennessee is about rehabilitation," Person told the teens. "It's not about punishment."
Earlier Herenton had noted that the boy he talked with several weeks ago referred to Juvenile Court detention as being "in jail."
"In life we all make mistakes," Herenton added. "Prison shouldn't be your destination."
A few of the students looking elsewhere looked toward Shelby County Mayor Mark Luttrell when he mentioned that he had worked for 30 years in prison as a warden and director.
"The common denominator in prisons is a lack of education," he said. "Look around here at all of the people who believe in you. Don't take that lightly."
Copyright 1995 -
by The Daily News Publishing Co. Inc. - All Rights Reserved


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PORTLAND — Oregon's health exchange and insurance carriers are looking at extending deadlines to allow more people to get coverage starting Jan. 1, Cover Oregon's acting director said Thursday.
The news came two days after Bruce Goldberg and Gov. John Kitzhaber acknowledged that, despite earlier promises, thousands of people who applied for coverage by a Dec. 4 deadline would not be insured by January because the state wouldn't have enough time to process their applications.
The exchange's online portal has yet to enroll anyone, and the state is relying on paper applications and manual reviews to sign people up. Goldberg said he's working with regulators and insurers to push back the Sunday deadline for people to select a plan after the state has informed them of their options.
State officials estimate there are about 32,000 people whose applications have not had a first-level review, in which the state determines whether they're qualified for the publicly funded Oregon Health Plan or for tax credits to offset insurance premiums. After the first-level review, the state gives applicants who qualify for private insurance their options.
Goldberg told the Cover Oregon governing board that he's not sure what the new deadline would be, but he hopes there will be enough time to get through "the vast majority" of applications submitted in time for January coverage.
Goldberg also told the board that he's hired a private law firm to look into Cover Oregon's options to hold technology contractors accountable for problems delaying the launch of the online enrollment system by at least four months. He did not name specific contractors, but state officials have publicly and privately laid much of the blame on Oracle.
The company has a lucrative contract to provide hardware, software and consulting services for the insurance exchange and a related computer modernization project for the state human services agencies.
"It's only prudent that we get the best expertise," Goldberg said.
Emails obtained by The Oregonian show a growing sentiment among state and other officials that Oracle is at the root of the exchange's struggles and a strong effort to put political pressure on the company.
According to the emails, Oracle continued to tell Cover Oregon that it could deliver the website up until the weekend before the exchange should have gone live on Oct. 1. Subsequent deadlines also were missed.
Oracle spokeswoman Deborah Hellinger declined to comment on the missed deadlines, and Oracle has declined numerous interview requests by The Associated Press.
The emails also show that the governor, U.S. Sen. Jeff Merkley and U.S. Rep. Kurt Schrader called Oracle's top executives in October and November to demand that the company deliver the exchange it had promised.
John Cvetko, a quality assurance contractor hired by Cover Oregon, warned in a November email that implementing meaningful control over Oracle wouldn't be easy.
Cover Oregon "will need assistance riding herd over Oracle," he said.
Despite the private emails, top state officials declined to lay the blame on Oracle — or anyone else.
"To say that Oracle is the problem would not be accurate," Cover Oregon's chief information officer, Aaron Karjala, told The Associated Press. "There's a lot of challenges that we're looking to overcome, we have a lot of management in place, and we're closely partnered with Oracle."
All Cover Oregon advertising is scheduled to stop on Monday, Chief Communications Officer Amy Fauver said.
Associated Press writer Gosia Wozniacka contributed.
Copyright 2013 The Associated Press. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.
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