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Is Warner's pulled app future of TV content purchases?
What might have been a feature that fans snapped up, wound up just being a preview. Warner Bros. accidentally released an app in the U.S. that was only intended for international markets, according to CNET.
The app allowed for streaming of Warner Brothers shows the day after they air, including "Big Bang Theory," "Gossip Girl," and "The Vampire Diaries." The app was yanked from U.S. iTunes stories right after it was made available, according to the report. It was intended solely for Belgium, the Netherlands and Luxembourg, an underserved market in terms of having capacity to stream TV programs.
Per World TV PC, the episodes stream over a wi-fi connection, but can be downloaded. Users will also have real-time news access on Twitter and Facebook.
But the social media tool may be the future model of television viewing. There is an increasing demand for an a la carte viewing system that cable operators are reluctant to provide. A recent refusal by the U.S. Supreme Court to hear a case on cable providers bundling channel packages demonstrates that cable is not going there anytime soon. So it may be up to content creators and consumers to create ways to circumvent the system.
According to CNET, the app was free to download and each show was individually purchased. It looks as though there won't be a set price for content, it will be based on its quality and popularity. Standard definition was $3, whereas HD was $4. Entire current seasons were also available, with "The Big Bang Theory" and "The Vampire Diaries" being priced at $38 for standard and $44 for HD. "Gossip Girl" was available for $25 standard and $31 HD.

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Osteoprotegerin (OPG) has been reported to prevent bone resorption by inhibiting the formation, function, and survival of osteoclasts in a variety of animal models. However, the effects of OPG on bone metabolism in avian species have not been described. The objective of this study was to investigate the effects of chicken OPG (chOPG) expressed in chicken embryo fibroblasts (CEFs) on chicken osteoclast function in vitro.
The chOPG sequence containing the open reading frame (ORF) was amplified from chicken embryo frontal bone and inserted into the pcDNA3.1 (+) vector. PcDNA3.1 (+)/chOPG was transiently transfected into CEFs by lipofectamine 2000. Transcription of OPG mRNA and expression of chOPG recombinant protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and indirect immunofluorescence. The level of chOPG recombinant protein was detected by enzyme-linked immunosorbent assay. The suspension of osteoclasts was separated from chicken embryos and divided into three groups (control group, pcDNA3.1 (+) group and pcDNA3.1 (+)/chOPG group). The percentage of osteoclast apoptosis was detected by flow cytometry. The tartrate-resistant acid phosphatase (TRAP) secreted by osteoclasts was measured by the diazol method. The resorbing activity of osteoclasts was evaluated by the area of lacunae on bone flaps and the concentration of calcium in the supernatant liquid of osteoclasts.
48 h after transfection, the exogenous OPG gene transcription was detected by RT-PCR. After 72 h, the CEFs transfected from pcDNA3.1 (+)/chOPG displayed green fluorescence and the concentration of chOPG protein was 15.78 ± 0.22 ng/mL. After chicken osteoclasts were cultured for 5 d in a medium containing supernatant from transfected CEFs, the percentage of osteoclast apoptosis was increased significantly, the concentration of TRAP, the area of lacunae on bone flaps and calcium concentration were decreased significantly in the pcDNA3.1(+)/OPG group compared to the control group and the pcDNA3.1 (+) group.
Constructed pcDNA3.1 (+)/chOPG transfected into CEFs expressed bioactive OPG protein that was able to inhibit osteoclast function.
Osteoporosis in laying hens is a condition that involves a progressive loss of bone resulting in bone fragility and increased risk of fracture. Surveys of laying flocks in Europe have indicated that about 30% of the birds experience one or more bone fractures due to osteoporosis during their lifetime. The high fracture rates show that osteoporosis not only leads to production losses, but also to severe welfare problems in hens [1].
In laying hens, the main types of bone providing structural integrity are cortical and trabecular bone. In addition to these, medullary bone, an extremely labile source for calcium that develops in specific bones of female birds at the onset of sexual maturity, provides a labile source of calcium for shell formation. Bones undergo a constant process of remodelling, which at the cellular level involves a coordinated regulation of osteoblasts and osteoclasts. As hens mature sexually, bone formation of osteoblasts switches from structural bone to medullary bone [2]. In the absence of structural bone formation, continued osteoclastic resorption of structural bone will result in a depletion of structural bone, ultimately leading to osteoporosis.
The differentiation and function of osteoclasts are regulated by soluble cytokines from osteoblasts, such as osteoprotegerin (OPG) and the receptor activator of nuclear factor ligand (RANKL; also called OPG ligand) [3]. OPG is a soluble decoy receptor that inhibits osteoclast formation, function, and survival by preventing the binding of RANKL to the receptor activator of nuclear factor κB (RANK), a membrane-bound protein that is found on chondrocytes, dendritic cells, osteoclast precursors, and mature osteoclasts [4]. Many cytokines and effectors are known to influence the osteoclastic bone resorption via the OPG/RANK/RANKL trio of proteins [5,6]. Changes of expression levels of OPG/RANK/RANKL would be expected to cause bone disorders such as postmenopausal osteoporosis, glucocorticoid-induced osteoporosis, and sporadic Paget's disease in man [7].
Although the importance of OPG in the osteoclastogenesis has been established in mammalian models, it is not yet clear how OPG regulates the function of osteoclasts in avian species. To elucidate the function of OPG in laying hens in vivo, we amplified the open reading frame (ORF) of the chicken OPG (chOPG) sequence, constructed the pcDNA3.1 (+)/chOPG plasmid and transiently transfected it into chicken embryo fibroblasts (CEFs). We tested whether pcDNA3.1 (+)/chOPG expressed OPG protein at a level able to inhibit the biological activity of osteoclasts in vitro.
Cloning of the ORF of chOPG
Total RNA was extracted from chicken embryonic frontal bone (Animal Husbandry Industry Co., Nanjing, China) with TRIzol® Reagent (Invitrogen, Inc. Carlsbad, CA, USA) according to the manufacturer's instruction. RNA purity was determined by 260 nm and 280 nm absorbance ratios and integrity was checked by 1% agarose/formaldehyde gel electrophoresis. A Biometra DNA Thermal Cycler was used for reverse transcription polymerase chain reaction (RT-PCR). RT-PCR was performed in the presence of DTT, oligo(dT)18, dNTP, RNase inhibitor, first-strand buffer and Moloney murine leukaemia virus reverse transcriptase (TakaRa Bio Inc. Japan). The final mixture was reacted at 42°C for 60 min and at 70°C for 15 min to denature the enzyme. On the basis of the published nucleotide sequence of chOPG (DQ098013), one pair of PCR primers (Invitrogen) were designed. Primers P1 and P2 were used to amplify the ORF of chOPG sequence. Nhe| and Xho| (TakaRa Bio Inc.) restriction sites were inserted into primers P1 and P2, respectively:
GCTAGCATGAACAAGTTCCTGTGC-3' (sense strand, positions 10-27 of cDNA sequence);
CTCGAGTTAGACACATCTTACTTT-3' (antisense strand, positions 1,201-1,218 of cDNA sequence).
PCRmix (TakaRa Bio Inc.), primers P1 and P2, and cDNA were mixed and amplified for 30 cycles under the following conditions: denaturation for 30 s at 94°C, annealing for 45 s at 47°C, and extension for 50 s at 72°C. The products were subsequently sequenced (Invitrogen) after 1% agarose electrophoresis, recovery and purification.
pcDNA3.1 (+)/chOPG construction
The eucaryote expression vector pcDNA3.1 (+) (Invitrogen) and OPG products were digested with Nhe| and Xho|. After purification, two fragments were ligated with T4DNA ligase (TakaRa Bio Inc.) at 16°C (overnight). The ligation product was subsequently transformed into DH5α competent cells (Nanjing Agricultural University, Nanjing, China)). The transformed cells were plated on Luria-Bertani agar (Invitrogen-Gibco, Grand Island, NY, USA) containing ampicillin (Invitrogen-Gibco). The positive clones were identified by PCR (PCRmix, pcDNA3.1 (+) consensus primer P3 (5'-CTGGCTAACTAGAGAACCCAC-3'), P4 (5'-TAGAAGGCACAGTCGAGG-3')). DNA of positive clones were mixed and amplified for 30 cycles under the following conditions: denaturation for 30 s at 94°C, annealing for 45 s at 49°C, and extension for 50 s at 72°C and double restriction digestion, followed by agarose gel analysis. Then pcDNA3.1 (+)/chOPG and pcDNA3.1 (+) were prepared with non-endotoxin plamid extraction kit (Sigma Chemicals. St. Louis, MO, USA).
Cell culture and DNA transfection
CEFs were prepared from two 10-days old chicken embryos (Animal Husbandry Industry Co) and were grown according to standard procedures, cultured in Dulbecco's modified Eagle's medium (DMEM) (Invitrogen-Gibco) supplemented with 5% fetal bovine serum (Invitrogen-Gibco). The number of cells was adjusted to 2 × 105 cells/ml and incubated in 24-well tissue culture plates (Bo Quan Sci&Tech. Co. Ltd. Nanjing, China) containing cover glass at 37°C in a humid atmosphere of 5% CO2 for 24 h. Prior to each test, CEFs were washed three times with phosphate buffered solution (PBS), transfected with 1 μg/well pcDNA3.1 (+)/OPG plasmid and pcDNA3.1 (+) vector using 3 μl/well lipofectamine 2000 (Invitrogen), respectively, followed by incubation at 37°C in 5% CO2 for 48 h and 72 h. The culture medium was renewed every 2nd day.
RT-PCR analysis of chOPG mRNA
The cells (both floating and adherent cells) were harvested 48 h post transfection. The total RNA was extracted with TRIzol® Reagent according to the manufacturer's instruction. RNA samples were then treated with DNase I (1 U/μg) (TakaRa Bio Inc.) before the RT step to avoid the interference with contaminating genomic DNA. P5 (5'-ATGAACAAGTTCCTGTGC-3') and P6 (5'-TTAGACACATCTTACTTT-3') were subjected to PCR using upstream and downstream primers.
Immunocytochemical analysis of chOPG product in CEFs
CEFs (2 × 105) cultured for 72 h on glass coverslips (6 mm × 6 mm) were replated into 24-well plates. Glass coverslips were washed with 0.01 M PBS and fixed in 4% formaldehyde for 45 min. Detergent extraction with 3% Triton X-100 was performed for 10 min. Coverslips were saturated with PBS containing 5% bovine serum albumin (Wuhan Boster Biotechnology Company, China) for 1 h at room temperature with gentle rocking, processed with rabbit anti-chOPG polyclonal antibody (Nanjing Agricultural University) for 1 h at 37°C and followed by FITC-goat-anti-rabbit IgG (Wuhan Boster Biotechnology Company) for 1 h at 37°C and then stained by DAPI staining solution (Wuhan Boster Biotechnology Company). Coverslips were washed by PBS for 30 min prior to each treatment. Finally, coverslips were mounted on slides and fluorescence signals were analyzed by a Fluoview microscopy (Olympus, Japan).
ELISA analysis of chOPG product in supernatant
The concentration of the chOPG product in the supernatant was determined using an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer's instructions. The concentration was determined for three wells of each sample by measuring the optical density (OD) at 450 nm wavelength by an ELISA reader (Immuno Mini NJ-2300, InterMed, Japan).
Effect of chOPG on osteoclast bioactivity
Tibias and humeri were isolated from 15 18-days old chicken embryos. Osteoclast cultures were prepared as previously described [8]. Briefly, a cell suspension was seeded at a concentration of 2 × 105 cells per well in 24-well dishes containing either glass coverslips or bovine bone slices (4 mm × 4 mm × 50 μm) (Nanjing Agricultural University). Non-adherent cells were washed off after 2 h. The adherent cells were grown for another 2 d and then cultured in DMEM containing OPG supernatant. The medium was changed every 48 h. Glass coverslips, bovine bone slices and supernatant were harvested after 5 d. The percentage of osteoclast apoptosis was detected by flow cytometry. TRAP secreted to the supernatant by osteoclasts was measured at a OD of 530 nm by the diazol method using TRAP test kit (Bo Quan Sci&Tech. Co. Ltd. Nanjing, China). The resorption lacunae on the bone slice was visualized by toluidine blue staining after removal of osteoclasts using 50 mM NH4OH and brief sonication [9]. The concentration of calcium in the supernatant was determined by atomic absorption spectrometry (wavelength 422.7 nm, electric current 3.0 mA, spectrum width 0.4 nm) after 5 times dilution.
Statistical analysis
All values were expressed as means ± the standard deviation (SD). Differences between mean values of normally distributed data were assessed by the one-way ANOVA test and Student's t-test. Statistical difference was accepted at P < 0.05.
Cloning of the ORF of chOPG and construction pcDNA3.1 (+)/chOPG
The size of the specific gene fragment amplified was, as expected, about 1.2 kbp (Figure 1A). The positive clones were identified by PCR amplification and the double restriction digestion with Nhe| and Xho| (Figure 1B and Figure 1C). Analysis of the PCR products by agarose gel electrophoresis showed that both constructs contained a DNA insert of the correct size and in the correct orientation. The result of sequencing showed that it had 100% homology with that reported in GenBank (DQ098013) indicating that the OPG gene has an extensive hereditary conservation and that no mutations were present in this region of the vector.
Figure 1. Gel electrophoresis of chOPG. 1A: Gel electrophoresis of reverse transcription polymerase chain reaction (RT-PCR) product. Total RNA extracted from chicken embryo frontal bone was analyzed using RT-PCR with specific primers. About 1.2 kbp gene of chicken osteoprotegerin (chOPG) was amplified (lane 1); DL2000 marker (lane 2); 1B: Gel electrophoresis of pcDNA3.1 (+)/chOPG PCR product. chOPG fragment was inserted into the eucaryon expression vector pcDNA3.1 (+) between Nhe| and Xho|. Negative plasmid (lane 1) and positive plasmid (lane 2) were chosen using PCR; marker (lane 3); 1C: Gel electrophoresis of pcDNA3.1 (+)/chOPG double restriction enzyme assay. Positive plasmid (lane 3) was identified by Nhe|and Xho|double restriction digestion and showed pcDNA3.1 (+) and OPG (lane 2); marker (lane 1); 1D: Gel electrophoresis of RT-PCR analysis showing the expression of chOPG gene at 48 h. Amplification of chOPG using cDNA from lane 1 (control group) and lane 2 (pcDNA3.1 (+) transfected CEFs group) showing negative result. Amplification of chOPG using cDNA from lane 4 (pcDNA3.1 (+)/chOPG transfected chicken embryo fibroblasts group) showing about 1200 bp gene of chOPG. Lane 3: marker
Expression of chOPG in CEFs transfected with pcDNA3.1 (+)/chOPG
RT-PCR analysis indicated that CEFs in the group with pcDNA3.1(+)/chOPG transfection expressed OPG mRNA, but there was no expression of OPG mRNA in the control group and pcDNA3.1(+) group (Figure 1D).
Immunofluorescence studies showed that chOPG protein was distributed in the cytoplasma and CEFs showing green fluorescence were observed in the pcDNA3.1 (+)/chOPG group, but were not present in the other groups (Figure 2A).
Figure 2. The expression of chOPG protein and effect on osteoclast morphology, apoptosis and resorption. 2A: Immunofluorescence assay for a possible chicken osteoprotegerin (chOPG) protein. Chicken embryo fibroblasts (CEFs) were grown on coverslips, fixed, and examined by indirect immunofluorescence. Cells were incubated with rabbit anti-chOPG serum. The secondary antibody was fluorescein-conjugated goat anti-rabbit immunoglobulin G (green). The nuclei of the corresponding cells were visualized by DAPI staining (blue). Fluorescence signals were analyzed by Fluoview microscopy (×200). Negative results are shown on card l (control group) and card 2 (pcDNA3.1 (+) transfected CEFs group), positive green fluorescence for CEFs are shown on card 3 (pcDNA3.1 (+)/chOPG transfected CEFs group). 2B: The morphology of osteoclasts was observed by inverted phase contrast microscope (×200). The adherent osteoclasts were cultured in Dulbecco's modified Eagle's medium (DMEM) containing supernatant of control group (l), pcDNA3.1 (+) transfected CEF group (2) and pcDNA3.1 (+)/chOPG transfected CEF group (3) for 5 d. 2C: Effect of the supernatant of three groups on the apoptosis of osteoclasts by flow cytometry. 2D: Toluidine blue staining of bone slices showing resorption lacunae (×200). The adherent osteoclasts were cultured in DMEM containing supernatant for 5 d in three groups.
In the culture supernatant of the pcDNA3.1 (+)/chOPG group transfected from CEFs, the concentration of chOPG was 15.78 ± 0.22 ng/ml, whereas chOPG was not be demonstrated in media from the control group or the pcDNA3.1 (+) group.
Effect of product from transfected CEFs on chicken osteoclast bioactivity in vitro
The morphology of osteoclasts after culturing for 5 d is shown in Figure 2B. Osteoclasts grew well in the control and pcDNA3.1 (+) transfected CEF groups, whereas major nuclei disappeared, many vacuoles and lipid droplet appeared in the cytoplasm and many non-adherent and dead osteoclasts were observed in the culture solution of the pcDNA3.1 (+)/chOPG transfected CEF group. The percentage of osteoclast apoptosis in the control, pcDNA3.1 (+) and pcDNA3.1 (+)/chOPG groups was 10.32%±1.50%, 12.61%±0.95%, 20.59%±2.83%, respectively (Figure 2C). TRAP enzyme activity in the pcDNA3.1 (+)/chOPG group was significantly decreased compared to the control group (P < 0.01) (Figure 3A). An individual resorption event was seen as a dark border of toluidine blue stain surrounding an excavation. The data were recorded for each resorption event separately (Figure 2D). The quantity and area of lacunae reflected bone resorption by osteoclasts (Table 1). DMEM culture solution did not contain Ca2+ until after culturing thus suggesting osteoclast activity (Figure 3B).
Figure 3. The change of TRAP enzyme activity and concentration of Ca2+ in three groups. 3A: Effect of culture supernatant from chicken embryo fibroblasts transfected on osteoclastic TRAP enzyme activity (Mean ± SD; n = 8). ** indicates P < 0.01 compared with the control group. 3B: The concentration of Ca2+ in the supernatant containing bovine bone slices (Mean ± SD; n = 8). ** indicates P < 0.01 compared with the control group.
Table 1. Effect of culture supernatant of chicken embryo fibroblasts on the quantity and area of osteoclast resorption lacunae in three groups.
Bone is an exceedingly complex tissue with multisystemic regulation. Skeletal metabolism depends on the dynamic balance of bone formation by osteoblasts and bone resorption by osteoclasts. The discovery of the OPG/RANKL/RANK system in the mid 1990s has led to major advances in our understanding of how bone modeling and remodeling are regulated [10]. Current research has focused on OPG in humans and mice, while reports on avian OPG are lacking. In our laboratory, chOPG mRNA was extracted from chicken embryo frontal bone. The OPG coding region was successfully amplified and sequence analysis indicated that OPG is highly conserved evolutionary. The sequence reported here had a 68.76%, 68.60% and 68.29% homonology to human, rat and mouse OPG, respectively. The sequence similarity suggests a similar function across species.
Bone is particularly intriguing in laying hens because of the huge demands for calcium for eggshell formation and the occurrence of medullary bone. On the surface of the medullary bone, osteoclasts undergo cyclical functional modifications during the egg-laying cycle [11].
In this study, chOPG induced osteoclast apoptosis after in vitro incubation for 5 d. This result was similar to that reported by Lacey et al. [12], who demonstrated that OPG inhibited bone resorption and induced osteoclast apoptosis though inhibition of F-actin ring formation of mature osteoclasts or altered interaction between stroma cell and osteoclasts.
The results suggest that the secretion of TRAP by osteoclasts was significantly decreased; further demonstrating that recombinant chOPG could inhibit the activity of osteoclasts in vitro. Chamber et al. [13] and Boyde et al. [14] provided evidence for a direct association between the quantity, area and depth of absorption and the capability of osteoclasts to resorption bone. The present study showed that chOPG inhibited osteoclast bone resorption and consequently the concentration of Ca2+ in the supernatant was significantly reduced. However, the mechanisms by which OPG exerts its biological activity as well as the nature of its molecular interactions with osteoclasts are not well defined. Hakeda et al. [15] reported the first evidence of a direct biological activity of OPG on isolated osteoclasts via a 140 kDa OPG-binding protein. The exact nature of osteoclastic OPG receptors was not further characterized. Direct biological activities of OPG on osteoclasts were recently showed by Wittrant et al. [16] demonstrating OPG enhanced proMMP-9 activity along with several other parameters (TRAP, TIMP, cathepsin K) in purified osteoclasts. Theoleyre et al. [17] showed that OPG stimulates proMMP-9 activity of osteoclasts by the ras/MAPK pathway involving p38 and ERK1/2 phosphorylations. Moreover, OPG-induced MAPK pathway depends on RANKL. In general, OPG is not only a soluble decoy receptor for RANKL as described in the literature but may be also considered as a direct effector of osteoclast functions.
ChOPG is capable of inhibiting bone resorption as well as promoting osteoclast apoptosis. The study also indicates that pcDNA3.1 (+)/chOPG may be a target for regulating bone metabolism in chicken bone metabolic diseases such as osteoporosis.
CEFs: chicken embryo fibroblasts; chOPG: chicken OPG; DMEM: Dulbecco's modified Eagle's medium; OD: optical density; OPG: osteoprotegerin; ORF: open reading frame; PBS: phosphate buffered solution; RANKL: receptor activator of nuclear factor κB ligand; RANK: receptor activator of nuclear factor κB; RT-PCR: reverse transcription polymerase chain reaction; TRAP: tartrate-resistant acid phosphatase.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
LH and JH conceived of the study, and participated in its design and coordination and helped to draft the manuscript. ZZ participated in the data collection. JY cultured the chicken embryo osteoclasts. LH performed the other experiments. All authors have been involved in drafting the manuscript and have read and approved the final manuscript.
This work was supported by the National Natural Science Foundation of China (30972234, 30671546) and the key program of Education Ministry of China (200803070021).
Beck MM, Hansen KK: Role of estrogen in avian osteoporosis.
Poult Sci 2004, 83:200-206. PubMed Abstract | Publisher Full Text
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Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, Elliott R, Colombero A, Elliott G, Scully S, Hsu H, Sullivan J, Hawkins N, Davy E, Capparelli C, Eli A, Qian YX, Kaufman S, Sarosi I, Shalhoub V, Senaldi G, Guo J, Delaney J, Boyle WJ: Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation.
Cell 1998, 93:165-176. PubMed Abstract | Publisher Full Text
Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Luthy R, Nguyen HQ, Wooden S, Bennett L, Boone T, Shimamoto G, DeRose M, Elliott R, Colombero A, Tan HL, Trail G, Sullivan J, Davy E, Bucay N, Renshaw-Gegg L, Hughes TM, Hill D, Pattison W, Campbell P, Sander S, Van G, Tarpley J, Derby P, Lee R, Boyle WJ: Osteoprotegerin: a novel secreted protein involved in the regulation of bone density.
Cell 1997, 89:309-319. PubMed Abstract | Publisher Full Text
Kwan Tat S, Padrines M, Théoleyre S, Heymann D, Fortun Y: IL-6, RANKL, TNF-alpha/IL-1: interrelations in bone resorption pathophysiology.
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Anderson DM, Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, Roux ER, Teepe MC, DuBose RF, Cosman D, Galibert L: A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function.
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Hofbauer LC, Schoppet M: Clinical implications of the Osteoprotegerin/RANKL/RANK system for bone and vascular diseases.
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Yao J, Zhang J, Hou JF: Effects of ipriflavone on caged layer bone metabolism in vitro and in vivo.
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Wang Y, Hou JF, Zhou ZL: Chicken receptor activator of nuclear factor-κB ligand induces formation of chicken osteoclasts from bone marrow cells and also directly activates mature osteoclasts.
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Boyc BF, Xing LP: Functions of RANKL/RANK/OPG in bone modeling and remodeling.
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Miller SC: Osteoclast cell-surface specializations and nuclear kinetics during egg-laying in Japanese quail.
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Lacey DL, Tan HL, Lu J: Osteoprotegerin ligand modulates murine osteoclastsurvival in vitro and vivo.
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Chamber TJ, Thomson BM, Fuller K: Resorption of bone by isolated rabbit osteoclasts.
J. Cell Sci 1984, 66:383-399. PubMed Abstract | Publisher Full Text
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Br Dent J 1984, 156:216-230. PubMed Abstract | Publisher Full Text
Hakeda Y, Kobayashi Y, Yamaguchi K, Yasuda H, Tsuda E, Higashio K, Miyata T, Kumegawa M: Osteoclastogenesis inhibitory factor (OCIF) directly inhibits bone-resorbing activity of isolated mature osteoclasts.
Biochem Biophys Res Commun 1998, 251:796-801. PubMed Abstract | Publisher Full Text
Wittrant Y, Couillaud S, Theoleyre S, Dunstan C, Heymann D, Redini F: Osteoprotegerin differentially regulates protease expression in osteoclast cultures.
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Theoleyre S, Wittrant Y, Couillaud S, Vusio P, Berreur M, Dunstan C, Blanchard F, Redini F, Heymann D: Cellular activity and signaling induced by osteoprotegerin in osteoclasts: involvement of receptor activator of nuclear factor κB ligand and MAPK.
Biochimica Biophysica Acta 2004, 1644:1-7. Publisher Full Text


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Corn Stuffed Pork Chops
|Pork loin chops
|4 , cut 1 inch thick
|Chopped onions
|1⁄3 Cup (5.33 tbs)
|Chopped green pepper
|3 Tablespoon
|3 Tablespoon
|Whole wheat bread slices/Rye bread
|1 , toasted and cubed
|Frozen whole kernel corn
|1⁄3 Cup (5.33 tbs) (Loose Pack)
|1 Tablespoon, diced
|Ground cumin
|1⁄4 Teaspoon
|1⁄8 Teaspoon
|1⁄8 Teaspoon
|Apple juice concentrate
|1 Tablespoon
Trim fat from pork chops.
Make a pocket in each chop by cutting horizontally into the chop from the fat side almost to the bone.
For stuffing, in a saucepan stir together the onion, green pepper, and water.
Bring to boiling; reduce heat.
Cover and simmer about 4 minutes or till the vegetables are tender.
Do not drain the vegetable mixture.
In a small mixing bowl stir together the toasted bread cubes, corn, pimiento, cumin, salt, and pepper.
Add the cooked vegetable mixture; toss lightly.
Spoon about 1/4 cup of the stuffing into each pocket.
If necessary, securely fasten the opening with wooden toothpicks.
Place the chops on a rack in a roasting pan.
Bake, uncovered, in a 350° oven for 35 to 40 minutes or till the juices run clear.
If desired, brush the chops with apple juice concentrate the last 5 minutes of baking.
Before serving, remove toothpicks


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Red light ruling means stop
July 1, 2011 in Letters, Opinion on Page A15 OK, who blew it? The red light photo program was ruled invalid by a local judge. Either the City Council ignored existing law or they were given incompetent advice. Every … 5
Personally foul
July 1, 2011 in Letters, Opinion on Page A15 To the intoxicated dad from the 9-year-old boys team at Hoopfest. 1
Diploma sticker shock
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Time to move on
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Articles tagged belén rueda
27 Apr 2008 // 11:00 PM
One would think the whole 'spooky child' setup would have run its course, but this film finds a few new wrinkles to explore.READ more
9 Jan 2008 // 10:00 PM
Beginning and ending with the superlative filmmaking of Jia Zhang-ke, traversing the nooks and crannies of the globe, PopMatters presents the 20 best international and indie films of 2007.READ more
31 Dec 2007 // 3:25 AM
At once mother and child, victim and antagonist, space and inhabitant, Laura (Belén Rueda) is remarkable, but also traumatized.READ more
//Mixed media
Colette Carr "Static" (video) (Premiere) // Music
House of Blondes "First of July" (audio) (premiere) // Music
Digisaurus 'No More Room For Love' (EP stream) // Music
Eszter Balint 'Airless Midnight' (album stream) (premiere) // Music
//Pm Picks
PopMatters is on a short summer publishing break. We resume Monday, July 6th.
// Announcements
"PopMatters is on a short summer publishing break. We resume Monday, July 6th.READ the article


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Five Favorite Fast Meals (continued)
Those are my favorite time-saver meals. Now take advantage of these time-saver tips and you'll find yourself with new time on your hands.
- Plan meals. Yes, I know you hear about meal planning all the time, but that's because it's definitely the key to successful healthy eating. For example, let's focus on dinner. If you sit down for less than 20 minutes you can plan dinners and make a grocery list for the week. The health benefits of getting more of the nutrients you need will be worth those 20 minutes.
- Make use of leftovers. If you bake a chicken use the leftover chicken to make chicken soup or chicken salad. Don't throw away leftovers! Get creative. If you make pork tenderloin for dinner, transform it into pork tacos the next night.
- Eat more one-pot meals. One-pot meals can be convienient and healthy time savers because you can load up on high-fiber veggies, throw everything in the pot and walk away.
- Try crock-pot cooking which is another of my favorite time savers. Add your ingredients in the morning and come home to a cooked meal and a great-smelling house. dLife's Recipe Finder has lots of great crock pot (a.k.a. slow cooker) recipes.
- Do batch cooking and prep ahead of time. Batch cooking involves cooking a few meals at once and freezing them for a later date. There are various websites available for batch cooking tips and recipes. Prepping ahead of time can be great for chopped veggies such as onions, peppers, and other veggies that can be added to various dishes during the week.
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Recipe Shuffle
Black Bean Quesadillas Almond Spice Pound Cake Corn Tortilla Pepperoni Pizza (Gluten Free) Salmon with Citrus Cabbage Cornmeal Coated Fried Snapper Barbecue Pork on Buns Carrot and Cabbage Slaw Seven-Layer Beef Skillet Chicken and Mushroom Pizza Easy Mexicali Pork Chops
When last we left this saga, my doctor had me staying out from work for an additional week, but with the head swelling down, I was bound to be back on the bicycle as soon as possible (you know what they say about getting back on the horse that threw you...). I had planned to take my sister on a short (under a mile) ride — both for the company and to get us both on the bicycle. She turned chicken and I ended up going the "flat" route to the greengrocer — a little...


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Aliases for OR5H7P Gene
Subcategory (RNA class) for OR5H7P Gene
Aliases for OR5H7P Gene
External Ids for OR5H7P Gene
- Entrez Gene: 79291
- Ensembl: ENSG00000187900
ORGUL Members for OR5H7P Gene
Summaries for OR5H7P Gene
Olfactory receptors interact with odorant molecules in the nose, to initiate a neuronal response that triggers the perception of a smell. The olfactory receptor proteins are members of a large family of G-protein-coupled receptors (GPCR) arising from single coding-exon genes. Olfactory receptors share a 7-transmembrane domain structure with many neurotransmitter and hormone receptors and are responsible for the recognition and G protein-mediated transduction of odorant signals. The olfactory receptor gene family is the largest in the genome. The nomenclature assigned to the olfactory receptor genes and proteins for this organism is independent of other organisms. [provided by RefSeq, Jul 2008]
GeneCards Summary for OR5H7P Gene
OR5H7P (Olfactory Receptor, Family 5, Subfamily H, Member 7 Pseudogene) is a Pseudogene, and belongs to lncRNA RNA class.


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Comment: Re:Current level of Japanese debt? (Score 2) 189 189
Comment: Supercomputers already do drug design (Score 1) 57 57
Comment: Why is the hardware so complex/expensive? (Score 2) 217 217
From what I read the dongle is merely the interface from the camera (USB) to the smartphone (USB). That should be trivial. (For my setup a USB OTG cable + adapter to mini USB is sufficient, there are tons of apps to control cameras).
The article states that they had to use a beefier micro controller etc., but I wonder: why not do all the processing on the smart phone? These days our phones have so much processing power AND sensors, there should be no need to do any kind of non-trivial logic outside, especially when you're just trying to launch your first product.
Comment: C++14 != C++98 (Score 3, Interesting) 407 407
Comment: We're no longer at the origin (Score 1) 181 181
Comment: Clickbait Caption, but Valid Arguments (Score 3, Insightful) 181 181
Of course general purpose CPUs exist, simply because we call them that way. But it is also true that each design has it's own strengths, and "dark silicon" is another driver for special purpose hardware. Efficiency is another. Andrew Chien has published some interesting research on this subject. In his 10x10 approach he suggests to use 10 different types of domain-specific compute units (e.g. for n-body, graphics, tree-walking...), each of which is 10x more efficient than "general purpose CPUs" in its domain (YMMV). Those compute units bundled together, make up one core of the 10x10 design. Multiple cores can be connected via a NoC.
Let's see how software will cope with this development...
ps: can special purpose hardware exist if general purpose hardware doesn't?
Comment: Is the joke here... (Score 1) 292 292
Comment: Re:Hai! (Score 1) 111 111
One reason might be that railways are more efficient in densely populated areas. There express trains can even compete with airplanes. Yesterday we went from Tokyo to Osaka. Flight time would have been ~1h, plus 1h checkin and transfer to/from the airport (~45min. each). The Nozomi Shinkansen took us there in 2:30, and both stations were directly at the center of the cities.
Most of Japan's population is situated in coastal regions, so just a hand full of routes can service all major cities. Imagine how many connections you'd need in the US...
Comment: Need many compilers? Try Gentoo... (Score 1) 739 739
Comment: Yesno? (Score 1) 89 89
It's not that specialized. It's just plenty of DSPs strapped together on a torus.
Actually Anton uses ASICS, their cores are specially geared at MD codes. This goes way beyond just "strapping together DSPs". They have IIRC ~70 hardware engineers on site. (Source: I've been to DE Shaw Research last year).
Unlike what wikipedia claims, you could probably achieve comparable performance using a more classical and general-purpose supercomputer setup with GPU or Xeon Phi accelerators, provided the network topology is well tuned to address this sort of communication scheme
No, you can't, and here is why: Anton is built for strong scaling of smallish, long running simulations. If you ran the same simulations on a "x86 + accelerator" system (think ORNL's Titan) then you'd observe two effects:
- The GPU itself might idle a lot as each timestep only involves few computations, leaving many shaders idle or waiting for the DRAM.
- Anton's network is insanely efficient for this use case. IIRC it's got a mechanism equivalent to Active Messages, so when data arrives, the CPU can immediately forward it to the computation which is waiting for it. That leads to a very low latency compared to a mainstream "InfiniBand + GPU" setup.
(most recent supercomputers don't use tori)
Let's take a look at the current Top 500:
- #1 Tianhe-2: Fat Tree
- #2 Titan: 3D Torus
- #3 Sequoia: 5D Torus
- #4 K Computer: 6D Torus
- #5 Mira: 5D Torus
- #6 Piz Daint: 3D Torus
- #7 Stampede: Fat Tree
- #8 JUQUEEN: 5D Torus
- #9 Vulcan: 5D Torus
- #10 nn: 3D Torus
So, torus networks are the predominant topology for current supercomputers.
Comment: Missing: Project Pluto (Score 3, Interesting) 133 133
Comment: /Very/ different hardware (Score 2) 89 89
Computational drug design and bitcoin miners have in common that both run best on custom hardware. The crux is, that both require very different types of hardware. As an example, please refer to Anton, designed by DE Shaw Research exactly for molecular dynamics (MD) codes.
Bitcoin mining is classified as a so called embarrassingly parallel algorithm, while MD is a tightly coupled problem. Hence an efficient parallelization for MD codes is much harder to speed up: communication gets in the way, and communication is essentially always bound by the speed of light.
ps: fun fact: bitcoin mining and MD can be carried out (at least somewhat) efficiently on GPUs.


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Originally written on BlackSportsOnline | Last updated 3/9/13
Leodis McKelvim is a solid cornerback but has struggled to command a starting spot with the Buffalo Bills. However, he has proven to be a dangerous kickoff and punt returner, ; leading the NFL last season in in punt return yards average while scoring a combined four touchdowns. The Bills decided that he was worth keeping signing him to a contract extension. Cornerback Leodis McKelvin is staying in Buffalo after the Bills reached an agreement to re-sign their 2008 first-round draft pick on Saturday. A person familiar with the negotiations said McKelvin agreed to a four-year contract worth a little over $20 million. The person spoke on condition of anonymity because the Bills did not reveal the contract terms in announcing the agreement with McKelvin. Used mostly as a third cornerback last year, McKelvin had one interception in 13 games, including four starts. He missed the final three games because of a groin injury. Overall, he has six interceptions in 64 games, with only 33 starts.
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VIEWS: 0 PAGES: 2 CATEGORY: Entrepreneurship & Business Planning POSTED ON: 2/12/2009 Public Domain
SOUTH FAYETTE ELEMENTARY PTA PROPOSAL TEMPLATE The following is a guide to be used when making a proposal to the South Fayette Elementary PTA. Cover Sheet Name Address and phone number Date State what your proposal is for Proposal Sheet Proposal What is the reason/need for this proposal? Who (which grade levels or student population) will benefit, keeping in mind that the majority of the student body, parent/guardian or teacher population should be positively impacted? Objectives/Benefits/Criteria Why should the SFEPTA pursue this proposal? (Which objects or goals of the National, State or SFEPTA will be furthered by the implementation of this proposal? Where will this proposal be implemented, i.e., school or township property, classroom, etc.? When is the targeted implementation (date)? How long will implementation take to complete (estimate of lapsed time from approval to implementation)? Cost Cost of implementation (include comparisons from at least 2 vendors, as appropriate)? On-going maintenance or upkeep (future estimated expenses) Replacement or replenishment costs, if applicable. Conformance to PTA Standard Guidelines for Spending Additional information (if possible) Attach examples or samples, photographs, brochures, etc. Summary of research regarding proposal (other school district's success, Administration or School Board approval, other organizations, as appropriate). How many people will be needed to implement (include number of PTA volunteers, Teachers/Administration/School board, township or community constituents)? Who will need to maintain? Confirm agreement to do so with appropriate party or include request for SFEPTA to maintain in the proposal document. For proposals relating to school property, please include information regarding easement, site preparation, liability and potential vandalism issues. Proposals are required to be verbally submitted by the author at a SFEPTA meeting along with a written proposal. Then the membership may vote on the request. As much of the above information should be used when making your verbal and written proposal. Missing information may put your proposal in jeopardy of being denied. Any questions may be directed to the Leadership Team prior to submission of your proposal. PTA Standard Guidelines for Spending SFEPTA is a nonprofit 501(c) 3 organization; it must adhere to the guidelines set by the IRS for such groups. Violation of the IRS guidelines may result in the loss of your unit/council nonprofit status. Educational ● Supplements the educational instruction of students such as field trips, reading programs, science fair, PTA publications, etc. Also may provide officer training and parenting workshops. ● Anything involving the "education" of children or parents Charitable ● Pennsylvania Life Membership, Past President's Pin, Student Programs ● Not a gift to a charity but instead services or programs otherwise unobtainable for students and members such as assemblies. Operational ● Postage, minute books, supplies, committee expenses, membership flyers, etc. ● Anything having to do with administration of programs and projects
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Select Local Merchants
Camp GladiatorRead MoreCamp Gladiator founder Ally Davidson?s childhood obsession with the outdoors and sports led her to pursue a degree in sports science. After her 2008 win on the TV show American Gladiators, Ally decided to share her love of fitness with the world at large. Camp Gladiator reflects Ally's passion for fitness in its mission, which espouses their desire "to positively impact the physical fitness and ultimately the lives of as many people as possible." Davidson also leads boot camps in 10 U.S. cities, with each hosting multiple training locales. The upbeat staff of certified personal trainers lead patrons of all ages and fitness levels through four-week outdoor boot camps, earning Camp Gladiator Best Boot Camp Dallas 2014 from Dallas Observer, Best Boot Camp from Rare magazine in 2010, Best Outdoor Boot Camp 2012 from Austin Fit magazine, and placing them #643 on Inc.com's 2014 list of the 5000 fastest-growing private companies in America.302 Dartmouth AvenueOldsmar, FL US
DUMMY ADDRESSRead MoreFit Body Boot Camp's instructors all over the world share a common goal: to help clients hit their fitness and weight-loss goals with regimented training programs. Trainers lead group workouts that combine cardio with resistance exercises, strength training, and light alligator wrestling. They design the workouts to be as time-effective as possible and vary the routines constantly to avoid workout plateaus that can leave clients feeling uninspired. Trainers empower their students with healthy eating guidelines to keep motivation high outside of class. During one-on-one sessions, certified personal trainers plan out tailored fitness programs, which clients can take with them to Fit Body Boot Camp's gyms to continue shaping up on their own.640 Douglas Road EastOldsmar, FL US
My Way Fitness 24/7Read MoreSteve Orlando founded My Way Fitness 24/7 to help his local community to tone muscles and improve overall health through personal workouts, classes, and nutritional guidance. His team of trainers strolls through the 4,000-square-foot weight room and cardio area, granting new members their undivided attention during complimentary personal-training sessions that maximize fitness results without the use of dark magic. Inside newly completed classrooms with tall, plum-colored walls, certified instructors help patrons to sleeken silhouettes and clear busy minds with fun Zumba choreography or a variety of body-stretching poses during yoga. Nutritional counselors teach patrons to decipher their body's dietary demands and implement safe and effective eating habits. Steve allows the doors to hang open 24 hours a day, and the facility's IHRSA membership affords members access to any other IHRSA gym⎯a benefit when trying to maintain fitness routines during vacations or long-distance fast-food runs.4058 Tampa RdOldsmar, FL US
Vibe Fit ClubRead MoreThe team of passionate and certified personal trainers at Vibe Fit Club works with each of their clients to craft a personalized fitness and nutrition plan to help them achieve total wellness. Their philosophy is grounded in an inside-out approach that addresses the underlying causes of fat retention, whether it is due to nutrition, lifestyle behaviors, or a combination of both, such as licking butter sculptures. To help patrons toward their goals, the team combines nutritional programs with high-energy group fitness classes that range from Pilates and core workouts to cycling and personal training classes. Touting the philosophy that exercise can be fun and empowering, the lovely lady instructors lead classes that sculpt arms by using body weight as resistance, as well as develop flexibility by performing vertical splits. The intimate studio also acts as home for pole-dancing parties complete with a DJ and champagne.13529 Prestige PlaceTampa, FL US
Gracie Tampa WestRead MoreAt Gracie Tampa, instructors mainly emphasize clients' safety, working to ensure that students can execute technically-sound chokeholds without actually inflicting harm. As far as disciplines are concerned, the MMA studio focuses on Muay Thai kickboxing and Brazilian Jiu-Jitsu, with a flexible curriculum that can accommodate competitive fighters and fitness buffs alike. The trainings take place in a state-of-the-art facility, complete with a full-size ring, heavy bags and speed bags, and a fight cage ideal for holding your opponent captive and avoiding a match altogether. Available in group class and open mat formats, these trainings are designed to build functional muscle as well as intangibles, such as discipline and perseverance.141 Stevens AvenueOldsmar, FL US
John Soderman FitnessRead MoreProfessional boxer. Walking inspiration. Mustache grower. John Soderman has worn many hats in his career, all of which play a role in his current capacity as a personal trainer. A certified strength and conditioning specialist, Soderman boasts more than 20 years in the training game, and he has worked with everyone from high-level athletes to devoted laypeople.225 Pine Avenue NorthOldsmar, FL US


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NEW YORK— The U.S. government is seeking to stop Electrolux from buying General Electric's appliance division, saying the combined company would dominate sales of ovens and other cooking-related kitchen appliances, especially to customers like home builders, property managers, hotels, and governments. General Electric Co. and Electrolux said they will...» Read More
Nike and Adidas cash in on Chinese consumers' desire to be seen in foreign brands.
Americans often check their receipts to make sure they've bought everything they need, and probably to see if what they paid this time is any different from the last trip. The government does the same with the Consumer Price Index. Here are the details.
The rising labor costs for companies that supply Chinese goods to the West may result in higher consumer prices. The NYT reports.
Coach, the U.S. accessories brand, is planning to shift up to half of its manufacturing out of China to escape rising labour costs at the same time as it moves aggressively to expand its sales in the country. The FT reports.


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Supreme Court
Can I be the first one to make a Law & Order reference?
It's really reassuring to know that, if there's a 911 call that I've been stabbed in the middle of the night in DC's most prominent urban neighborhood, just on the edge of the downtown business district, it will take the cops "only" 13 minutes to reach me. Sheesh.
I had the privilege of graduating Xaverian High School with Rob. My condolences to his wife and family. Rob was a person of unlimited talent and impeccable character. I held him in the highest esteem, and cannot believe this tragedy. I am utterly at a loss.
I wouldn't call this neighborhood DC's "most prominent urban neighborhood.". The house is in an "up and coming" area. Definitelty not an excuse for our lackluster emergency response services - just a clarification.
This was a senseless act of rage, obviously done out of jealousy by one (or more) of the residents of the home, and another (others) there perhaps covering up for him. My wish is that law enforcement officials take their time and make this an air tight case without errors.
The homes in that neighborhood cost a $1 million. The area has
Re: Mark's comment. Before making off-handed comments you'd do well to be able to substantiate your accusations. If you know that to be fact, then you'd better do your civic duty and get off your couch and alert the police...shouldn't you?!
Can anyone said 'jealous rage'? What was a young married man doing staying at the home of a gay couple? Hmmm...
I had the honor and privilege of working under the supervision of Rob at Covington for 1 and 1/2 years. I am still unable to get over the shock that Rob is no longer with us. I wish I was able to take part in this investigation. I wish I had more answers. I pray that he did not suffer. But there are so many unanswered questions. Was he asleep when he was attacked? Were there any marks on him to show that he fought his attacker? Did the police confirm with his wife that he called to say he was staying over at the house? Did they go over his phone records? It does not seem like Rob to say that he was too tired to drive home. At Covington, we worked till 3 and he'd be in the office by 8. Did he have a change of clothes? It would make more sense to say that he was drunk but tired, that does not add up. I pray that the police will do their best to apprehend his murderer. The story that he was attacked by an intruder does not add up. Did anyone hear anything? How did they discover him? If everyone was already asleep why did they go to his room? What alerted them. All these questions. But I just need answers so my soul can rest. I'll never forget Rob. He was always there for me. I'll remember his smile most of all. I just cannot believe this. Can someone answer any of these questions for me please.
It's hard to find the words to describe how good, kind and ethical Robert was; it must be easy, for people who did not know him, to imagine that all the praise is only so much hyperbole brought on by his untimely death. I knew him from law school -- he was a mentor, a big brother and a friend. He is sorely missed. I cannot imagine what his family is going through and my thoughts go out to them. I hope that those who took him from this world, which so benefited from his presence and which so needed him, are swiftly brought to justice.
The Law Blog covers the legal arena's hot cases, emerging trends and big personalities. It's brought to you by lead writer Jacob Gershman with contributions from across The Wall Street Journal's staff. Jacob comes here after more than half a decade covering the bare-knuckle politics of New York State. His inside-the-room reporting left him steeped in legal and regulatory issues that continue to grab headlines.
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Here is what you need to know about Friday's gay-marriage ruling.
If blockbusters Supreme Court battles ended the way sports championships do, U.S. Solicitor General Donald Verrilli would be soaked in champagne and showered with Gatorade. And Jones Day partner Michael Carvin, the lawyer for the losing plaintiffs, would glumly file back into a hushed locker room.
Who are the Americans accused of trying to join ISIS? A new study released this morning by Fordham Law School's Center on National Security breaks down the numbers.
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