text:
- Research
- Open Access
Systematic screening of polyphosphate (poly P) levels in yeast mutant cells reveals strong interdependence with primary metabolism
© Freimoser et al.; licensee BioMed Central Ltd. 2006
- Received: 2 August 2006
- Accepted: 15 November 2006
- Published: 15 November 2006
Inorganic polyphosphate (poly P) occurs universally in all organisms from bacteria to man. It functions, for example, as a phosphate and energy store, and is involved in the activation and regulation of proteins. Despite its ubiquitous occurrence and important functions, it is unclear how poly P is synthesized or how poly P metabolism is regulated in higher eukaryotes. This work describes a systematic analysis of poly P levels in yeast knockout strains mutated in almost every non-essential gene.
After three consecutive screens, 255 genes (almost 4% of the yeast genome) were found to be involved in the maintenance of normal poly P content. Many of these genes encoded proteins functioning in the cytoplasm, the vacuole or in transport and transcription. Besides reduced poly P content, many strains also exhibited reduced total phosphate content, showed altered ATP and glycogen levels and were disturbed in the secretion of acid phosphatase.
Cellular energy and phosphate homeostasis is suggested to result from the equilibrium between poly P, ATP and free phosphate within the cell. Poly P serves as a buffer for both ATP and free phosphate levels and is, therefore, the least essential and consequently most variable component in this network. However, strains with reduced poly P levels are not only affected in their ATP and phosphate content, but also in other components that depend on ATP or free phosphate content, such as glycogen or secreted phosphatase activity.
- Additional Data File
- Total Phosphate
- Glycogen Level
- Vacuolar Protein
- Molecular Function Term
Inorganic polyphosphate (poly P) is a linear polymer that consists of phosphoanhydride linked phosphate residues and occurs ubiquitously in all organisms and living cells [1]. The functions of poly P range from its role as a phosphate store and buffer [2–4] to the activation of enzymes [5, 6] and regulation of chromatin condensation, gene expression and translation [1, 7, 8]. Poly P is also involved in bacterial pathogenicity [9, 10], survival during stationary phase in bacteria and yeast [9, 11, 12], or the adaptation to alkaline and osmotic stress [13–15]. In the slime mold Dictyostelium discoideum, poly P regulates development and predation behavior [16], and in humans blood coagulation is accelerated and fibrinolysis is delayed by poly P [17]. At the cellular level, poly P has been detected in the cytosol, the nucleus, mitochondria, as well as the endoplasmic reticulum [7, 18, 19]. Poly P is particularly prominent in the acidocalcisomes of trypanosomatids [20], and in Saccharomyces cerevisiae almost the entire poly P pool resides in the vacuole [21, 22].
Despite its universal occurrence and its broad functions, very little is known about poly P metabolism and its regulation. In S. cerevisiae (and all higher eukaryotes) it is, for example, still not known how poly P is synthesized, despite the fact that more than 20% of this organism's dry weight can be composed of poly P [1]. To gain a broad view of poly P metabolism in yeast and to identify pathways involved in the regulation of poly P levels, we extracted and quantified poly P in the knockout strains of almost all non-essential yeast genes.
Altogether, 4,765 strains from the YKO collection [23], consisting of knockouts of non-essential yeast genes, were initially screened. Strains that either hypo- or hyper-accumulated poly P were subjected to three consecutive rounds of screening. After the third round, 255 strains from the YKO collection had altered poly P levels in all three experiments (a complete list of all data is available online as Additional data file 1). Almost all of these 255 strains had reduced poly P levels and only occasionally, at specific time-points during growth, poly P hyper-accumulation relative to the wild-type strain was observed.
All cellular compartments are involved in maintenance of poly P levels
Vacuolar proteins are most important in determining poly P levels
All 255 genes were ranked according to their impact on poly P content. The mutant cells most strongly affected in their poly P content were highly enriched for knockouts of genes that encode vacuolar proteins (6 out of the 20 vacuolar proteins among the 10 most strongly affected mutants). Vacuolar function depends on the acidification of the vacuolar lumen, which is mediated by the vacuolar H+-ATPase (V-ATPase) complex. In this poly P screen, 9 out of the 14 V-ATPase subunits were required for normal poly P content (Vma5 (14), Vma8 (10), Vma10 (13), Vma13 (34), Vph1 (1), Cup5 (5), Tfp3 (15), Ppa1 (20), Vma6 (27); ranks given in parentheses). In addition, regulators (Rav1 (43), Vps34 (30), Fab1 (150) and Vac14 (213)) and assembly factors (Vma22 (16), Vph2 (23)) of the V-ATPase were also discovered as being important for normal poly P levels.
Next to the VPH1 knockout, the VTC4 and VTC1 deletion strains were the second and third most affected mutants, and the knockout of VTC2 also hypo-accumulated poly P (rank 219). Together with Vtc3, these are the four subunits of the vacuolar transporter chaperon (Vtc) complex, which was previously shown to be required for maintenance of poly P levels [3]. Other proteins involved in the hypo-accumulation of poly P are involved in membrane docking and fusion at the Golgi-to-endosome and the endosome-to-vacuole steps (Vps33, ninth most affected mutant), or represent an alternative pathway from the Golgi to the vacuole (effected by the AP-3 complex that consists of Apm3, Apl5, Apl6 and Aps3; ranks 21, 59, 57 and 55, respectively). Vam3, a vacuolar t-SNARE protein that traffics to the vacuole via the AP-3 complex [24], was also important for maintenance of poly P levels (rank 8).
In other strongly affected strains, genes encoding either of the two phosphofructokinase subunits (PFK1 and PFK2, ranks 28 and 17) or the pyruvate kinase PYK2 (rank 7) were knocked out. In three additional strains, other components of glycolysis were identified (GCR2, GPM3, YOR283W; 63, 201 and 181, respectively).
Poly P profiles of the identified mutant strains
Many strains with altered poly P levels are also affected in their total phosphate content, ATP levels, acid phosphatase activity and glycogen accumulation
Comparison of the 255 non-essential ORFs found to be important for poly P content with the results from published large scale analyses revealed that 56 strains were also affected in glycogen accumulation [25], but that only 4 strains with reduced poly P levels had previously been found in a screen for altered PHO5 (acid phosphatase (rAPase)) regulation, which serves as an indicator of the activation state of the PHO pathway [26, 27]. To verify the data from the screen, and to test if a poly P phenotype is accompanied by other complex phenotypes, we measured cell density, poly P levels, total phosphate content, ATP concentrations, glycogen levels and rAPase activity in the 30 most affected knockout strains.
Cell density was slightly reduced (to roughly half the OD600) in most strains, but only at the 2 h and 5 h time-points (Figure 3). Poly P content and total phosphate levels were strongly reduced in all 30 non-essential mutants except in the knockout of TFP3 (only slightly reduced poly P and total phosphate levels) and the Δecm14, Δmrp51,Δapm3 and Δsir3 strains had almost normal content of total phosphate (Figure 3). The ATP levels exhibited the highest variability at the 0 h time-point, with a cluster of knockouts having an increased content (knockouts of ERG6, VTC4, VTC1, PYK2, VPH1, CUP5 and ECM14). Strongly reduced ATP levels were only measured in the knockouts of the dubious open reading frame (ORF) YPR099c and the uncharacterized ORF YOL019w (Figure 3). Almost all of the 30 knockouts hyper-accumulated glycogen, either at the 2 h or the 0 h and 24 h time-points. An exception was again the mutation in the dubious ORF YPR099c, which had undetectable glycogen levels at the 0 h and 2 h time-points (which could be explained by the effect on MRPL51, which is encoded on the opposite strand). Most of the 30 knockout mutants also exhibited strongly increased rAPase activity at the 0 h and 24 h time-points. Exceptions were only the knockouts of ECM14, VTC4, VTC1 and VAM3 (Figure 3).
Previous to the screen described here, only very few yeast mutant cells had been analyzed with respect to their poly P content and, to our knowledge, this is the first time that a specific metabolite, that is, poly P, was isolated and quantified in an almost complete mutant cell collection. After three consecutive screens, 255 knockout mutant strains with altered poly P levels were identified. The genes affected in these mutant cells encode proteins from all intracellular compartments and components functioning in many processes of primary metabolism. This broad analysis thus defined novel biological functions for about 250 yeast genes and allowed, for the first time, a global view of the pathways and processes affecting poly P metabolism in yeast. But not only were many genes required for the maintenance of a normal poly P content, strains that hypo- or hyper-accumulated poly P had often a reduced total phosphate content, altered ATP and glycogen levels and an up-regulated rAPase secretion. From this analysis we conclude that poly P content is an extremely sensitive parameter that is highly intertwined with primary metabolism.
However, the number of knockout mutant cells with reduced poly P levels was probably even underestimated: Several of the few mutants previously identified to hypo-accumulate poly P were not found in this screen. For example, the knockouts of VMA4, VTC3, ARG82 or KCS1 were not discovered, as well as all knockouts of members of the PHO pathway (PHO3, PHO4 or, for example, PHO84), which have been shown to convey hypo-accumulation of poly P [3, 26, 28]. This is an indication of the stringency of the screening conditions and selection criteria and suggests that the true number of non-essential genes involved in poly P metabolism is even higher than the 255 genes reported here. The reliability of the high-throughput screening procedure was also confirmed by the fact that all 30 strains from the YKO collection that were individually tested indeed showed strongly reduced poly P levels.
Although poly P has been observed in and functions in different organelles [7, 18, 19, 29], 90% to 99% of all poly P is localized in the vacuole [1, 21, 22]. Consequently, we measure almost exclusively vacuolar poly P and the proteins from other organelles that were found in this screen must affect vacuolar poly P levels. Poly P storage is thus a central function of the yeast vacuole and, thus, mutants affected in vacuolar functions or morphology are likely to be impaired in their capability to store polyP. Important vacuolar functions include the maturation and activation of different proteins and physiological functions in the storage of metabolites and in cell homeostasis [30–32] that all depend on the V-ATPase. This large multimeric, partly membrane-embedded complex is conserved from yeast to man and is also relevant in human diseases such as osteopetrosis and distal renal tubular acidosis [33, 34]. In this screen, nine subunits of the V-ATPase and several of its regulators were identified. In addition, V-ATPase activity is also glucose regulated; in the presence of glucose, the V-ATPase is functional while the absence of glucose causes reversible dissociation of the V0 and V1 subunits and thus V-ATPase is inactivated [35]. This phenomenon could explain the decline of poly P levels as soon as the growth medium is depleted for glucose [36]. However, based on these data it is impossible to conclude whether V-ATPase activity regulates poly P levels directly or indirectly. As poly P levels in a VMA4 knockout strain could be slightly restored by growing the cells in acid buffered growth medium, Ogawa et al. [3] concluded that V-ATPase itself was not essential for poly P metabolism. Instead, these authors suggested that the Vtc complex (consisting of Vtc1, Vtc2, Vtc3 and Vtc4), which regulates vacuolar membrane fusion, morphology and function [37, 38], is directly involved in the synthesis of poly P in yeast [3]. The fact that mutants affected in various stages of retrograde and forward vesicle trafficking, as well as in autophagy, indicate that an intact secretory pathway is required for normal poly P content and supports the hypothesis that the Vtc complex influences poly P levels indirectly [38]. This secondary effect on poly P metabolism can be caused by: the mislocalization or misregulation of vacuolar proteins that are important for poly P synthesis and storage; or the impairment of poly P synthesis and transport along the secretory pathway. Thus, many pathways could indirectly affect poly P content via their impact on the vacuole or secretory pathway and, thereby, link seemingly unrelated pathways to poly P metabolism. The reduced poly P content in mutants of any one of the four subunits of the AP-3 complex, which mediates an alternative pathway from the Golgi to the vacuole [39] and is also medically relevant for one type of Hermansky-Pudlak syndrome [40], is another such case of indirect effects; mutations in proteins that are targeted to the vacuole via the AP-3 complex or that contain a putative di-leucine signal for AP-3 targeting hypo-accumulated poly P (data not shown).
Intervention at any point within this phosphate-energy-network inevitably causes many side effects that require readjustment of the allocation of resources between the different phosphate and energy pools. In this context, 'interventions' could represent genetic changes but also externally induced physiological changes as they occur, for example, during the progression through different growth phases. Both types of changes were found to affect poly P levels strongly, but also other characteristics were altered. Many mutants that failed to accumulate poly P seemed to invest more in the accumulation of storage carbohydrates (glycogen) and could no longer buffer the expression of phosphate regulated genes (as indicated by elevated rAPase activity). In contrast, total phosphate content closely followed poly P levels, but even in this case the correlation coefficient between all relative poly P and all total phosphate values (log2-transformed) was only 0.65. Even ATP levels per se were sometimes significantly altered, but compared to the other characteristics that were measured, these changes were smaller, confirming, at least partly, the general notion that, in viable and growing cells, ATP levels remain constant [43].
PolyP metabolism and primary metabolism are strongly interdependent: On one hand, poly P levels depend necessarily on the integrity of primary metabolism and reflect the physiological state of a cell. On the other hand, poly P itself influences cell metabolism through its importance for energy and phosphate homeostasis. This first genome-wide analysis of poly P content thus also implies poly P as an indirect link between different cellular pathways such as phosphate, energy and carbohydrate metabolism or transcription and translation activities.
Materials and methods
Strains and culturing conditions
The haploid yeast knock-out strains (YKO [23], based on the strain BY4741: MAT a his3Δ1 leu2Δ0 met15Δ0 ura3Δ0) were grown in 96-well deep well plates at 30°C in YPD medium (1% w/v yeast extract, 2% w/v peptone, 2% w/v glucose, with optional addition of 200 mg/l G418). For the screen, fresh YPD medium was inoculated with 40 μl of the precultured (3d) stationary cells and cells were harvested after 4 h (first and second round of screen) or after 2 h, 4 h, 8 h and 24 h (third round of screen). Growth and cell number was monitored by measuring the light scattering at 600 nm (OD600) in a BioTek PowerWave™ XS microplate spectrophotometer (BioTEK Instruments Inc., Vermont, USA). For detailed poly P measurements in selected strains, the culture was inoculated at a cell density of 1 OD600/ml (approximately 107 cells/ml), and at each time-point 1 OD600 equivalent of cells was collected for poly P quantification.
Poly P purification and quantification
Poly P extraction, purification and quantification were performed as described previously [36]. In short, cells were pelleted in deep well plates by centrifugation (approximately 640 g, 15 minutes, 4°C), the supernatant was discarded, 50 μl 1 M sulphuric acid were added and the suspension was neutralized with 50 μl 2 M NaOH and 100 μl Tris-malate buffer (1 M, pH 7.5, 6% neutral red solution (0.1% neutral red in 70% ethanol)). Cell fragments were pelleted by centrifugation (approximately 640 g, 15 minutes, 4°C) and 100 μl of the supernatant were removed. Poly P was purified, enzymatically digested and quantified using a colorimetric assay [36].
Screening procedure and data analysis
For all poly P measurements the poly P raw data were normalized to the cell density (OD600) measured for each corresponding culture. The poly P content was then expressed relative to either the median of all cultures in one 96-well plate (first screen) or the poly P content in the wild-type strain BY4741 (second and third screens). All strains with a relative poly P content 1.5-fold higher or a relative poly P content 0.67-fold lower were selected for the second and the third round of screening. In the third screen, data were collected at four different time points (2 h, 4 h, 8 h, 24 h). Only strains that fulfilled at least one of the following criteria were selected for subsequent data analysis: relative poly P content throughout the 24 h time-course varied more than two-fold; at least one time-point differed more than two-fold from the wild-type poly P content. Strains that did not grow were excluded. In some strains, the affected gene was reintroduced and expressed from a plasmid, which restored poly P levels (data not shown).
The poly P data of all selected strains and all screens and time-points were combined and clustered hierarchically (with the Pearson uncentered or Pearson correlation distance measure, respectively, and complete or average linkage in Genesis [44]). The selected strains were also ranked according to how strongly poly P levels were affected (calculated as the sum of the squares of the log2-transformed poly P ratios at the four time-points of the third screen).
The genes that were deleted in the strains with altered poly P content were categorized and studied by different computational tools and in comparison with available data sets. All genes were categorized according to the GO terminology by using the annotation given in the go_slim_mapping.tab file available from the Saccharomyces Genome Database [45]. Significantly overrepresented GOSlim terms (compared to their representation in the complete yeast genome) were determined by using Cytoscape and the BiNGO plugin (settings: hypergeometric test statistic, false discovery rate correction for multiple tests, P ≤ 0.05 confidence limit) [46, 47]. All data manipulations were performed in Microsoft Excel and FileMaker.
Measurement of total phosphate, ATP, glycogen and acid phosphatase activity
Detailed measurements of several metabolites and activities were performed in selected strains at various time-points (0 h, 2 h, 5 h, 24 h). Individual cultures for each time-point were inoculated to an OD600 = 1 (approximately 107 cells/ml). At each time point 1OD600 equivalent of cells was harvested and used for subsequent measurements. All data were obtained from duplicate measurements.
For measurement of total phosphate, cells were pelleted, resuspended in 200 μl of 1 MH2SO4 and heated in a boiling water bath for 20 minutes. Released phosphate was quantified with molybdate and malachite green as described [36]. For ATP quantification the same neutralized cell extract as for poly P quantification was prepared, and ATP was quantified by a method adapted from Hyswert et al. [48]. The neutralized sample (20 μl) were added to 80 μl Tris buffer (20 mM, pH8, 2 mM EDTA) and 4 μl phosphoenol pyruvate were added (2.5 mM, pH8, 0.125 M MgSO4, 0.312 M K2SO4). For the quantification of ATP, 5 μl of the sample were added to 45 μl luciferase buffer (10 mM Tris-H2SO4, pH 7.4, 3.5 mM MgSO4). After the addition of 50 μl luciferase solution (Roche ATP Bioluminescence Assay kit CLS II, Roche Diagnostics GmbH, Mannheim, Germany) relative light units emitted were measured in a luminometer (Lumat LB 9507, Berthold Technologies GmBH and Co. KG, Bad Wildbad, Germany).
To quantify glycogen the cell pellet was first frozen and glycogen was extracted as described [49]. The extracted glycogen was then digested by adding 10 U alpha-amylase and 12.6 U amyloglucosidase in a sodium acetate buffer (pH 4.8, 220 mM) at 55°C for 16 h. Released glucose was quantified with the D-Glucose HK kit (Megazyme International Ireland Ltd., Bray, Ireland).
Acid phosphatase activity was assayed according to Huang and O'Shea [27] in 50 μl cell suspension by adding 200 μl p-nitrophenyl-phosphate (20 mM). After 15 minutes at room temperature, 200 μl of 10% ice-cold trichloracetic acid and 400 μl sodium carbonate solution (2 M) were added and the OD420 was measured. All results were expressed relative to the respective values of the wild-type cells (BY4741).
Additional data files
The following additional data are available with the online version of this paper. Additional data file 1 contains all poly P data of the three screens.
We thank S Eicke, M Stadler, S Ernst, C Neupert and S Clerc for help at different stages of this research. M Aebi is acknowledged for helpful and motivating discussions, SZeeman and his group for providing a spectrophotometer and help with glycogen quantification, and W Gruissem and J Fütterer for making a luminometer available. M Peter and M Sohrmann are given thanks for providing yeast strains. This work was supported by the Swiss National Science Foundation (grant 3100A0-112083/1) and ETH Zurich.
Authors' Affiliations
- Kornberg A, Rao NN, Ault-Riché D: Inorganic polyphosphate: a molecule of many functions. Annu Rev Biochem. 1999, 68: 89-125. 10.1146/annurev.biochem.68.1.89.PubMedView ArticleGoogle Scholar
- Neef DW, Kladde MP: Polyphosphate loss promotes SNF/SWI- and Gcn5-dependent mitotic induction of PHO5. Mol Cell Biol. 2003, 23: 3788-3797. 10.1128/MCB.23.11.3788-3797.2003.PubMedPubMed CentralView ArticleGoogle Scholar
- Ogawa N, DeRisi J, Brown PO: New components of a system for phosphate accumulation and polyphosphate metabolism in Saccharomyces cerevisiae revealed by genomic expression analysis. Mol Biol Cell. 2000, 11: 4309-4321.PubMedPubMed CentralView ArticleGoogle Scholar
- Thomas MR, O'Shea EK: An intracellular phosphate buffer filters transient fluctuations in extracellular phosphate levels. Proc Natl Acad Sci USA. 2005, 102: 9565-9570. 10.1073/pnas.0501122102.PubMedPubMed CentralView ArticleGoogle Scholar
- Kuroda A, Nomura K, Ohtomo R, Kato J, Ikeda T, Takiguchi N, Ohtake H, Kornberg A: Role of inorganic polyphosphate in promoting ribosomal, protein degradation by the Lon protease in E. coli. Science. 2001, 293: 705-708. 10.1126/science.1061315.PubMedView ArticleGoogle Scholar
- Wang L, Fraley CD, Faridi J, Kornberg A, Roth RA: Inorganic polyphosphate stimulates mammalian TOR, a kinase involved in the proliferation of mammary cancer cells. Proc Natl Acad Sci USA. 2003, 100: 11249-11254. 10.1073/pnas.1534805100.PubMedPubMed CentralView ArticleGoogle Scholar
- Kulaev IS, Vagabov VM, Kulakovskaya TV: The Biochemistry of Inorganic Polyphopshates. 2004, Chichester, West Sussex: John Wiley and Sons, Ltd, 2View ArticleGoogle Scholar
- McInerney P, Mizutani T, Shiba T: Inorganic polyphosphate interacts with ribosomes and promotes translation fidelity in vitro and in vivo. Mol Microbiol. 2006, 60: 438-447. 10.1111/j.1365-2958.2006.05103.x.PubMedView ArticleGoogle Scholar
- Kim KS, Rao NN, Fraley CD, Kornberg A: Inorganic polyphosphate is essential for long-term survival and virulence factors in Shigella and Salmonella spp. Proc Natl Acad Sci USA. 2002, 99: 7675-7680. 10.1073/pnas.112210499.PubMedPubMed CentralView ArticleGoogle Scholar
- Rashid MH, Rumbaugh K, Passador L, Davies DG, Hamood AN, Iglewski BH, Kornberg A: Polyphosphate kinase is essential for biofilm development, quorum sensing, and virulence of Pseudomonas aeruginosa. Proc Natl Acad Sci USA. 2000, 97: 9636-9641. 10.1073/pnas.170283397.PubMedPubMed CentralView ArticleGoogle Scholar
- Crooke E, Akiyama M, Rao NN, Kornberg A: Genetically altered levels of inorganic polyphosphate in Escherichia coli. J Biol Chem. 1994, 269: 6290-6295.PubMedGoogle Scholar
- Sethuraman A, Rao NN, Kornberg A: The endopolyphosphatase gene: essential in Saccharomyces cerevisiae. Proc Natl Acad Sci USA. 2001, 98: 8542-8547. 10.1073/pnas.151269398.PubMedPubMed CentralView ArticleGoogle Scholar
- Pick U, Weiss M: Polyphosphate hydrolysis within acidic vacuoles in response to amine-induced alkaline stress in the halotolerant alga Dunaliella salina. Plant Physiol. 1991, 97: 1234-1240.PubMedPubMed CentralView ArticleGoogle Scholar
- Rohloff P, Montalvetti A, Docampo R: Acidocalcisomes and the contractile vacuole complex are involved in osmoregulation in Trypanosoma cruzi. J Biol Chem. 2004, 279: 52270-52281. 10.1074/jbc.M410372200.PubMedView ArticleGoogle Scholar
- Weiss M, Bental M, Pick U: Hydrolysis of polyphosphates and permeability changes in response to osmotic shocks in cells of the halotolerant alga Dunaliella. Plant Physiol. 1991, 97: 1241-1248.PubMedPubMed CentralView ArticleGoogle Scholar
- Zhang H, Gomez-Garcia MR, Brown MR, Kornberg A: Inorganic polyphosphate in Dictyostelium discoideum : influence on development, sporulation, and predation. Proc Natl Acad Sci USA. 2005, 102: 2731-2735. 10.1073/pnas.0500023102.PubMedPubMed CentralView ArticleGoogle Scholar
- Smith SA, Mutch NJ, Baskar D, Rohloff P, Docampo R, Morrissey JH: Polyphosphate modulates blood coagulation and fibrinolysis. Proc Natl Acad Sci USA. 2006, 103: 903-908. 10.1073/pnas.0507195103.PubMedPubMed CentralView ArticleGoogle Scholar
- Beauvoit B, Rigoulet M, Guerin B, Canioni P: Polyphosphates as a source of high energy phosphates in yeast mitochondria: a 31 P NMR study. FEBS Lett. 1989, 252: 17-21. 10.1016/0014-5793(89)80882-8.View ArticleGoogle Scholar
- Harold FM, Miller A: Intracellular localization of inorganic polyphsophate in Neurospora crassa. Biochim Biophys Acta. 1961, 50: 261-270. 10.1016/0006-3002(61)90324-9.PubMedView ArticleGoogle Scholar
- Docampo R, de Souza W, Miranda K, Rohloff P, Moreno SN: Acidocalcisomes - conserved from bacteria to man. Nat Rev Microbiol. 2005, 3: 251-261. 10.1038/nrmicro1097.PubMedView ArticleGoogle Scholar
- Indge KJ: Polyphosphates of the yeast cell vacuole. J Gen Microbiol. 1968, 51: 447-455.PubMedView ArticleGoogle Scholar
- Urech K, Durr M, Boller T, Wiemken A, Schwencke J: Localization of polyphosphate in vacuoles of Saccharomyces cerevisiae. Arch Microbiol. 1978, 116: 275-278. 10.1007/BF00417851.PubMedView ArticleGoogle Scholar
- Winzeler EA, Shoemaker DD, Astromoff A, Liang H, Anderson K, Andre B, Bangham R, Benito R, Boeke JD, Bussey H, et al: Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis. Science. 1999, 285: 901-906. 10.1126/science.285.5429.901.PubMedView ArticleGoogle Scholar
- Darsow T, Burd CG, Emr SD: Acidic di-leucine motif essential for AP-3-dependent sorting and restriction of the functional specificity of the Vam3p vacuolar t-SNARE. J Cell Biol. 1998, 142: 913-922. 10.1083/jcb.142.4.913.PubMedPubMed CentralView ArticleGoogle Scholar
- Wilson WA, Wang Z, Roach PJ: Systematic identification of the genes affecting glycogen storage in the yeast Saccharomyces cerevisiae : implication of the vacuole as a determinant of glycogen level. Mol Cell Proteomics. 2002, 1: 232-242. 10.1074/mcp.M100024-MCP200.PubMedView ArticleGoogle Scholar
- Auesukaree C, Tochio H, Shirakawa M, Kaneko Y, Harashima S: Plc1p, Arg82p, and Kcs1p, enzymes involved in inositol pyrophosphate synthesis, are essential for phosphate regulation and polyphosphate accumulation in Saccharomyces cerevisiae. J Biol Chem. 2005, 280: 25127-25133. 10.1074/jbc.M414579200.PubMedView ArticleGoogle Scholar
- Huang S, O'Shea EK: A systematic high-throughput screen of a yeast deletion collection for mutants defective in PHO5 regulation. Genetics. 2005, 169: 1859-1871. 10.1534/genetics.104.038695.PubMedPubMed CentralView ArticleGoogle Scholar
- Auesukaree C, Homma T, Tochio H, Shirakawa M, Kaneko Y, Harashima S: Intracellular phosphate serves as a signal for the regulation of the PHO pathway in Saccharomyces cerevisiae. J Biol Chem. 2004, 279: 17289-17294. 10.1074/jbc.M312202200.PubMedView ArticleGoogle Scholar
- Lichko LP, Kulakovskaya TV, Kulaev IS: Inorganic polyphosphate and exopolyphosphatase in the nuclei of Saccharomyces cerevisiae: dependence on the growth phase and inactivation of the PPX1 and PPN1 genes. Yeast. 2006, 23: 735-740. 10.1002/yea.1391.PubMedView ArticleGoogle Scholar
- Kane PM: The where, when, and how of organelle acidification by the yeast vacuolar H +-ATPase. Microbiol Mol Biol Rev. 2006, 70: 177-191. 10.1128/MMBR.70.1.177-191.2006.PubMedPubMed CentralView ArticleGoogle Scholar
- Klionsky DJ, Herman PK, Emr SD: The fungal vacuole: composition, function, and biogenesis. Microbiol Rev. 1990, 54: 266-292.PubMedPubMed CentralGoogle Scholar
- Eide DJ, Clark S, Nair TM, Gehl M, Gribskov M, Guerinot ML, Harper JF: Characterization of the yeast ionome: a genome-wide analysis of nutrient mineral and trace element homeostasis in Saccharomyces cerevisiae. Genome Biol. 2005, 6: R77-10.1186/gb-2005-6-9-r77.PubMedPubMed CentralView ArticleGoogle Scholar
- Sobacchi C, Frattini A, Orchard P, Porras O, Tezcan I, Andolina M, Babul-Hirji R, Baric I, Canham N, Chitayat D, et al: The mutational spectrum of human malignant autosomal recessive osteopetrosis. Hum Mol Genet. 2001, 10: 1767-1773. 10.1093/hmg/10.17.1767.PubMedView ArticleGoogle Scholar
- Stover EH, Borthwick KJ, Bavalia C, Eady N, Fritz DM, Rungroj N, Giersch AB, Morton CC, Axon PR, Akil I, et al: Novel ATP6V1B1 and ATP6V0A4 mutations in autosomal recessive distal renal tubular acidosis with new evidence for hearing loss. J Med Genet. 2002, 39: 796-803. 10.1136/jmg.39.11.796.PubMedPubMed CentralView ArticleGoogle Scholar
- Parra KJ, Kane PM: Reversible association between the V1 and V0 domains of yeast vacuolar H+-ATPase is an unconventional glucose-induced effect. Mol Cell Biol. 1998, 18: 7064-7074.PubMedPubMed CentralView ArticleGoogle Scholar
- Werner TP, Amrhein N, Freimoser FM: Novel method for the quantification of inorganic polyphosphate (iPoP) in Saccharomyces cerevisiae shows dependence of iPoP content on the growth phase. Arch Microbiol. 2005, 184: 129-136. 10.1007/s00203-005-0031-2.PubMedView ArticleGoogle Scholar
- Cohen A, Perzov N, Nelson H, Nelson N: A novel family of yeast chaperons involved in the distribution of V-ATPase and other membrane proteins. J Biol Chem. 1999, 274: 26885-26893. 10.1074/jbc.274.38.26885.PubMedView ArticleGoogle Scholar
- Müller O, Neumann H, Bayer MJ, Mayer A: Role of the Vtc proteins in V-ATPase stability and membrane trafficking. J Cell Sci. 2003, 116: 1107-1115. 10.1242/jcs.00328.PubMedView ArticleGoogle Scholar
- Odorizzi G, Cowles CR, Emr SD: The AP-3 complex: a coat of many colours. Trends Cell Biol. 1998, 8: 282-288. 10.1016/S0962-8924(98)01295-1.PubMedView ArticleGoogle Scholar
- Huizing M, Boissy RE, Gahl WA: Hermansky-Pudlak syndrome: vesicle formation from yeast to man. Pigment Cell Res. 2002, 15: 405-419. 10.1034/j.1600-0749.2002.02074.x.PubMedView ArticleGoogle Scholar
- Rudra D, Warner JR: What better measure than ribosome synthesis?. Genes Dev. 2004, 18: 2431-2436. 10.1101/gad.1256704.PubMedView ArticleGoogle Scholar
- Warner JR: The economics of ribosome biosynthesis in yeast. Trends Biochem Sci. 1999, 24: 437-440. 10.1016/S0968-0004(99)01460-7.PubMedView ArticleGoogle Scholar
- Ditzelmüller G, Wohrer W, Kubicek CP, Rohr M: Nucleotide pools of growing, synchronized and stressed cultures of Saccharomyces cerevisiae. Arch Microbiol. 1983, 135: 63-67. 10.1007/BF00419484.PubMedView ArticleGoogle Scholar
- Sturn A, Quackenbush J, Trajanoski Z: Genesis: Cluster analysis of microarray data. Bioinformatics. 2002, 18: 207-208. 10.1093/bioinformatics/18.1.207.PubMedView ArticleGoogle Scholar
- Saccharomyces Genome Database. [ftp://genome-ftp.stanford.edu/pub/yeast/data_download/literature_curation/]
- Maere S, Heymans K, Kuiper M: BiNGO: a Cytoscape plugin to assess overrepresentation of Gene Ontology categories in biological networks. Bioinformatics. 2005, 21: 3448-3449. 10.1093/bioinformatics/bti551.PubMedView ArticleGoogle Scholar
- Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, Amin N, Schwikowski B, Ideker T: Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003, 13: 2498-2504. 10.1101/gr.1239303.PubMedPubMed CentralView ArticleGoogle Scholar
- Hyswert DW, Morrison NM: Studies on ATP, ADP, and AMP concentrations in yeast and beer. J Am Soc Brew Chem. 1977, 35: 160-167.Google Scholar
- Wang Z, Wilson WA, Fujino MA, Roach PJ: Antagonistic controls of autophagy and glycogen accumulation by Snf1p, the yeast homolog of AMP-activated protein kinase, and the cyclin-dependent kinase Pho85p. Mol Cell Biol. 2001, 21: 5742-5752. 10.1128/MCB.21.17.5742-5752.2001.PubMedPubMed CentralView ArticleGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


text:
From Publishers Weekly
Collin Deverell is one of two twin sons, heir to a fortune that his father, an oil tycoon, expects him to take part in. While his brother Justin readily involves himself in the family business, Collin has little wish to tie himself down to an executive life, preferring a carefree life of adventure and his love of fencing. With the sudden death of their parents on a business trip, Collin takes the chance to live life on his own terms, turning over shares in the family company and all responsibility to his brother in exchange for the family mansion, the art collection, and his mother's jewelry. He lives abroad for some years, rarely settling down, living his carefree life, seducing whatever woman crosses his path. When he returns home, he finds that the valuable paintings and jewels have all gone missing, sold off by his devious brother. Collin vows to take back what's rightfully his, even if it means breaking the law and going after some very dangerous people to do it.
Ashley Gordon is an artist from the Napa Valley in California who establishes a career for herself in San Francisco. After becoming a success in the art world and on the social circuit, she falls in love with Brandon Hollister. They're happy together, and Brandon wants to marry her, though she's puzzled by his complete estrangement from his parents. When we meet them, it's not hard to understand: Bradley and Claudia Hollister are downright nasty to the core. Ashley and Brandon marry, have a son, Robert and are happy together, until Brandon is killed in a plane crash. In the wake of her grief, Ashley is hit again when her in-laws launch a custody battle for their grandson, using bribery, lies, and their connections to take him away from Ashley. Ashley is, understandably, devastated.
It's into this mix that Ashley and Collin meet. Collin's been busy recovering what was stolen from him by becoming a thief himself, learning the trade from a master who saves his life. What started out for him as a mission to take back what's his becomes something more, as he discovers his father's company has been mismanaged by his brother, and is falling into the hands of a criminal syndicate who are readily dismantling it. The syndicate are made up of the same people who have possession of his property, and what began as thefts to recover property gradually shifts, as Collin realizes he does, in fact, have a responsibility to save the company his father built. And since Bradley Hollister is a member of the syndicate, Collin decides to enlist his former daughter-in-law as a partner to bring down the syndicate, save his family company, and restore Ashley's son to her custody.
It's a wise decision to keep the two from really meeting until mid way through the book. We, the reader, get to see both characters develop fully on their own, so we care about them and what happens to them (Ashley's loss of her husband and her son are particularly painful, which is one of the reasons the book works so well). When Collin and Ashley start working together, we see a growing connection between them, an emotional intimacy that comes across as very real. This is a testament to how human the two characters feel. They have depth, quirks, and flaws. As Ashley learns the tricks of the trade, of sleight of hand and the use of disguise, she and Collin find themselves drawn closer and closer. The bond and growing love between them comes across to the reader as the real thing. We come to root for them to achieve all they're after, and it's because of how well both of them have been written.
In every heist story, to root for the person pulling off the heist, it requires that the target be unsympathetic. Certainly having the target be a criminal syndicate is a very good way of having the reader dislike the target. And the primary targets, Bradley and Claudia Hollister, are more then worthy of our dislike. Both of them, particularly Claudia, are cruel and malicious. It's not hard to understand why their son broke ties with them, and as readers, we want to see them brought down, broken, and defeated.
Justin Deverell is another interesting character in the book. Early on it felt like he'd be the primary antagonist of the book, but as things go on, it's made clear that he's the dupe, the tool for the syndicate to dismantle the family company after they're done using it. I enjoyed the premise Norma used that Collin and Justin aren't the kind of twins we're used to in fiction... they have nothing in common but blood, barely speak for years, and ultimately are so far apart that it's doubtful they'll ever bridge that gap. There's no closer then blood mental connection sort of bond between these two twins, and it's a refreshing change.
There is a wild card sort of character I thought I'd make mention of. Anton DeVries, an insurance investigator, lurks in the background of the story. He first comes into the picture after Collin discovers the theft of his possessions. Through the rest of the book, he suspects Collin, looks for proof, and takes part in a pivotal moment towards the climax. He's an interesting character, something of a bloodhound, or a Javert to Collin's Valjean. DeVries is a good adversary, conflicted by catching a man who he knows to be morally right.
The attention to detail throughout the book is spot on, and perhaps never as much as during the various thefts that take place in the book. From training sequences in which both Collin and Ashley learn how to become thieves to the heists themselves, each act feels intricate, and brings a lot of variety to the table. An escape from a time lock safe and a judicious use of a mirror stand out particularly for me during the theft sequences. And the attention to detail also reflects itself in the early sequences featuring fencing and the artistic process.
Angels At Midnight is a beautifully written book that you'll enjoy reading. The plot and pacing of the novel keeps the reader on the edge. The details drawn out in the book about technique, places, and situations give it a very real world sensibility. And the characters really make the novel. Collin and Ashley are a winning couple that we can't help but sympathize with, to root for, and to cheer.
And who knows? Perhaps Robert has siblings... and all of them have grown up to take after Ashley and Collin's habit of breaking into high security vaults....
Final Hours fooled me: Despite the title, it isn't really about the giant asteroid that's about to wipe out human civilization. On the contrary, if there was ever a story that's all about the journey, it's this one.
Jamie Randall has to make a decision in the hours leading up to the apocalypse: Retreat to a secretly built bunker, where he might survive to continue his loveless marriage, or seek out the woman he's loved for the last fourteen years and die with her? We soon know his decision – the story is about why he made it, and as we wait to know his fate the story flashes back to the events that led him there.
It turns out Jamie is – let's face it – a jerk, although as we learn more about his history we get to know why. He married his wife to get ahead, to get revenge over those who once had power over him. The events that keep him in the marriage are believable, if tragic.
He's rescued in every way when Kate appears, quite literally saving his life. The rest of the book is a love story, as Jamie woos Kate but is stymied again and again in his attempts to make her more than "the other woman".
The truth is, Jamie probably doesn't deserve either of the women at first, and by the time he starts trying to do the right thing he's dug himself into a hole deeper than the one the asteroid's going to make. Kate is practically a saint, while Jamie's wife is trapped just as much as he is, and I kept rooting for a way for them to all get away happy.
That says something about the story – that we want to know how it all comes out, even though we already know it from the very beginning.
Forced to choose between his own happiness and the happiness of those he cares about, Jamie spends most of the book torn between the woman he loves and the woman he needs. His wife, the mother of his sons, was able to give Jamie everything he thought he wanted out of life, but when a free-spirited photographer named Kate saves his life, he begins to realize that maybe his priorities were wrong all along, and it's time to start living the way he now knows he needs to.
Despite some bad choices all of the characters make, they really are what makes the story golden. Everyone makes bad choices, and these characters are all willing to face up to their mistakes, which makes them all the more admirable. They're doing what they think is right in the current situation, and that's really what sets them apart. The story really makes you think about life and love, and what it really means to be alive. And most importantly, when everything is stripped away, what truly is important enough for us to spend our final hours doing?

subset: commoncrawl


text:
Nothing is certain except change itself.
John Steinbeck once wrote that it is the nature of humans to complain about change. Change has a considerable psychological impact on the human mind. To the fearful it is threatening because it means that things may get worse. To the hopeful it is encouraging because things may get better. To the confident it is inspiring because the challenge exists to make things better. We all have big changes in our lives that are more or less a second chance.
Chloe' wasn't what I was looking for that day last August. I wasn't even sure I was doing the right thing – looking for a puppy so soon after Katie's passing. But there she was – bounding down the dry, dusty gravel hill on a farm outside of Kalamazoo. Her bright blue eyes full of piss and vinegar. Her round body barely keeping up with her chubby legs as clouds of dust rose up around her. She sprang onto my leg and into my heart in one full swoop – I never knew what hit me!
I had just gotten out of the hospital and was looking at a long, up-hill climb to regain my health and my life. The last thing I needed – everyone close to me advised – was a puppy. But I wanted – needed – my life to change. And it certainly did. Chloe' landed in the middle of my struggle for normalcy like a heat-seeking missile.
Chloe' is instinctive and dominant, in the way of all well-bred border collies – and explosive. You don't simply coexist with her – as you can with most dogs – you have to react to her. Like all border collies, she needs work – and like it or not she will invent her own work if not given something productive to do. She somehow knows what annoys me the most and commences to have at it if left alone too long. Eating my zucchini plants is a current favorite "job".
I have been around dogs enough to know that the problem with dogs is almost always the people who own them. Chloe' and I are both impulsive, impatient, distractible, and restless. We are also both redheads of Scottish descent. Need I say more?
Recognizing early on that I needed help with Chloe' – a far greater challenge than I had ever imagined – I started reading books. Books on training, books on border collie history, books on trick learning, books on recognizing "calming signals". Our coffee table is piled high with dog books.
And then there are the classes. Puppy Head-Start was my very first introduction to the world of the American Kennel Association and its members. Dog People – but a totally different bred of dog people. These are not "pet dog" people. These are serious dog people. I have been told by more than one trainer that I don't always take my "work" as a handler serious enough (Just when did I transition from being a dog lover to being a dog handler????).
At 11 months Chloe' is well on her way to becoming a champion Agility Dog. At 52 years I am wise enough to know that I may be the only thing that stands in her way. But I am also confident because the challenge exists to make things better for both of us. I have had other big changes in my life that were more or less second chances – this time change just happened to arrive on four legs.
Dogs may not be our whole lives. But they make our lives whole.
- Roger Caras


text:
Always a fan of the arts, I, like all of the other starving artists out there, am just trying to cut out a place in this world. I devised the FEATURED ARTIST concept to diversify the scope of the website and give credit to up-and-coming graphic artists and photographers who work as tirelessly on their crafts as I do on mine.
Every four-to-five months, a new artist will be featured and interviewed as part of the series. To inquire about becoming a FEATURED ARTIST, see the CONTACT page.
I had the privilege of interacting with Ontario-based artist Paula Sherry on Twitter after being taken by her work.
The following is a biography for Featured Artist Paula Sherry via her website PetalsbyPaula.com:
Petals by Paula grew out of a passion and love for nature, paint and clay. How these three elements combine and ignite the imagination is different yet each one is sensually alive in its own right. Flowers are seasonal by nature so a pairing of artistic works grew alongside this endeavour. It is a labour of love and getting all the pieces to work together has not been an easy task but a definite goal.
To have an imagination, to not know limitations and hard work always win out.
Cut flowers are available and sold at local markets as season permits. The erection of a greenhouse will provide a lengthened season and hard to find nursery plants will be grown here.
This is the nature part of Petals by Paula.
I paint using acrylics and acrylic mediums. I try to breathe life into imagination showing scenes of nature in impressionistic and abstract form. All paintings are ready to hang if no frame is your preference, all are varnished for UV protection and packaged with great care.
Please visit Petals Art Shoppe,this is imagination in visual form becoming reality on canvas.
My hand built ceramics feature my Soul Fusion figurines all one of a kind sculptures depicting our soulconnection to one another. Also available are my handbuilt Petals signature bowls and sculptural handbuilt functional ware.
All ceramics are fired to Cone 6 and are stoneware. Most are dishwasher safe and food friendly. Each piece is unique and has its own characteristics and character that can be recreated but never imitated. Custom orders are available upon request.
Please visit the Mudroom to see imagination come to life in three dimensional form with the use of clay.
See Paula Sherry on the following:
Her website Petals by Paula


text:
Danger of Death for Anglers
17 April 2007
Anglers, fishery owners and landowners are being reminded about the dangers of overhead power lines by the electricity network industry national representative body. The Energy Networks Association is re-launching its campaign to hammer home the risks that there can be to life and limb.
Every year cases are reported of angling equipment contacting overhead electric power lines. Some of these have resulted in horrific burn injuries, some in death. The main cause of these incidents is a lack of awareness on behalf of the angler.
Findings from inspections suggest that there are considerably more minor incidents or 'near miss' incidents that are occurring that should but never get reported.
Although angling is generally regarded as a safe sport, there have been a number of instances where anglers have suffered serious and fatal injuries due to contacting overhead electric power lines whilst fishing. The increasing length of carbon fibre fishing poles has often been a factor in these unfortunate incidents.
Examples of the type of incidents that have been reported are:
- Whilst returning from fishing across fields to parked car, fishing rods came into contact with an overhead electric power line - sustained major injuries. Boy aged 13 made contact with 11,000 volt wood pole overhead electric power line with carbon fibre fishing rod. Suffered burns. Angler made contact with 33,000 volt overhead electric power line with 13 metre long carbon fibre pole from peg on side of lake - fatal injuries sustained. Fishing line became entangled with overhead electric power line - line cut clear from rod - no injuries sustained.
- In recent years the number of incidents has been reduced, mainly as a result of campaigns supported by the electricity industry, angling groups, landowners and the tackle trade. But every year throughout the country, fishing line is found hanging from overhead electric power lines with no report of incident or injury.
- At the higher voltages, electricity can jump short distances through the air. This means it may not be necessary to touch an overhead electric power line with a fishing rod or pole to suffer an electric shock and severe burns that could lead to death.
Energy Networks Association head of safety, health and environment Peter Coyle said, "Electricity is a vital form of energy in today's high technology world. Distribution of this energy requires an extensive network of overhead electric power lines which cross all parts of the UK. Electricity is also carried along overhead lines for specific uses such as transport.
"Overhead electric power lines are often difficult to see, particularly at night and against a dark or very bright background. They are normally bare (not insulated) and can be mistaken for telephone wires, with disastrous consequences.
"Currently the higher voltage lines normally have yellow 'Danger of Death' warning notices fitted to the poles or towers but lower voltage lines may not be marked. At higher voltages electricity may mjump short distances through the air. This means it is not necessary to touch an overhead electric power line to suffer an electric shock and burns which could result in death.
"Fishing rods and poles made from carbon fibre and similar materials will conduct electricity. Many other materials will allow electricity to flow along them when they are wet.
"Detailed advice is available at the Energy Networks Association website, at www.energynetworks.org and, for landowners and fishery managers, from local electricity companies."


text: Wounded trees, steel mesh almost completely disappeared into the trees. Man nailed the mesh against the tree, the tree grew around the mesh. Clay is supposed to be healing, it is said that clay sticks to the organism, and literally sucks out what doesn't belong in it. Inflammations would cure in hours and wounds would close. I don't know, but I take clay, roll it out, stagger with the slab in my arms to the tree, and I wrap the clay and myself around the tree. When I take the slab of again, I take it back in to the studio, turn it inside out and reconnect the ends: a ring with the inverse imprint of the tree's scars is carefully made. The rings are fired, some in a field kiln at a lower temperature, most in a regular gas kiln. They have different diameters, various thicknesses, are lower and higher. Those variations result in each ring having its own tone now they are turned in stone fossils of a curing process that continues to take place inside the tree. Together the fossils can be played like an instrument, its sound vaguely reminding of an Indonesian Gamelan. Thanks to Anne Ausloos, Rob Muylaert en Curdin Tones


text:
THQ Ships Rugrats Go Wild for GBA and PC
As fans eagerly await the next installment in the Rugrats feature film franchise, gamers can get the first taste of the mayhem in store. THQ Inc. has released Rugrats Go Wild for Game Boy Advance and PC. The interactive adventure is now available at retail outlets throughout North America.
Based on Paramount Pictures and Nickelodeon Movies' upcoming film Rugrats Go Wild, produced by Klasky Csupo, the video game features the Rugrats and The Wild Thornberrys shipwrecked on a deserted island. Gamers are able to play as both Rugrats and Thornberrys characters as they travel through an expansive island wilderness with jungles, caves, beaches and swamps.
Rugrats Go Wild also features seven replayable action games that allow players to beat their best score on three difficulty levels.
According to THQ Sr. VP of worldwide marketing Peter Dille, THQ's Nickelodeon video game sales have exceed $300 million at retail to date.
The feature film Rugrats Go Wild is scheduled to hit theaters on June 13.


text:
We believe that the key to nurturing a student's love for performing arts is to provide them with opportunities to perform on stage. Caper offers exciting and varied performance opportunities aimed at fostering our students' love of dance. These events become special occasions and treasured memories for the whole family.
Caper students are a regular attraction at many community events and a wide cross section of Caper students are given the opportunity to perform at these shows. Each year we stage a show at Darling Harbour for all our school-age jazz students. We also have our Annual Concert that features all our students.
Caper kids are also regulars at the Royal Easter Show, Crestwood Fair and the Orange Blossom Festival. However lots of fun performances are scheduled each year including charity events, sports games and community concerts. All performance opportunities are optional.
Whilst participation in shows is spread out to include as many students as possible, Caper also has dedicated performance groups consisting of students who are selected to represent the school at competitions and high profile events.
Caper has a proud history of success at competitions. Students compete in every age category across multiple genres and attend some of the most renowned eisteddfods on the Eastern seaboard. Caper also has a team of soloists who have established themselves as prize getters in prestigious competitions.


text:
We conclude this year's column at Amnesia, Space, DC-10 and Sankeys.
Amnesia closing
It seemed as though every party on the island extended for a week this year, and almost all the major clubs stayed open a week longer. But did all the extra partying wear everyone out? Not really. The closing parties were still wild. At one point Marco Carola was forced to make a one-handed catch of an errant, silver cowboy hat headed directly for his face at Amnesia. And for the first time in several years, we weren't squeezed in like sardines; something no one can argue is a bad thing.
Though he's best known for playing at Space, this year Steve Lawler chose the Amnesia terrace instead. It couldn't have worked better. Lawler's slightly darker, deeper style in The Basement at Sankeys is great, but I was excited to see what he would do in the big room. Upbeat, and jacking, he kept the energy up from start to finish with tracks like Skylark's remix of "Westworld," one he's been rinsing all summer long. After Lawler, crowd favorite Marco Carola was on, and he slammed home dubbed-out bombs like Chris Wood & Meat's "Triple S" and an edited version of Stefano Noferini's "Koro Koro," only letting the saxophone riff come in and out at his command.
Possibly having learned a lesson from Music On closing with "You Spin Me Round," towards the end of the night he dropped in INXS's "Need You Tonight," bringing the room to a complete fever pitch. I've only seen Amnesia tear itself apart that hard one or two other times this season. After he finally wrapped up at around 6 PM, several in the audience began to chant "Music On!" over and over, reminding us why he was picked to finish the season off.
Photos: Amnesia closing
Space closing
Unlike Amnesia, when we first arrived at Space, the inside seemed like a slow night in June, so we headed to the car park where the action was taking place. And though not as full as the opening, the chance to see Carl Cox do his thing in an outdoor, festival-like arena in early October is something special. Dubfire and Carlo Lio were already at work, performing a massive techno set, with giant Matrix-like visuals flashing behind them. The two kept things dark, dirty and bass-heavy early on, picking up the energy mid-set with tracks like Agent!, Vito and Danito's "Dopeman," and heading back to deep and twisted territory later with tracks like Ovi M's "Point of No Return."
Cox was up next, and predictably brought the car park to its knees, with wild screams echoing out of all corners of the place. Cox kept the dark techno vibes rolling along with tracks like Hollen's "Crossroads" as he urged the crowd on with his trademark "oh yes, oh yes!" I couldn't help but get a bit emotional every time he said that, realizing I may not hear his voice again for some time. Later he headed into house territory with the piano-laden, Salsa-tinged groove "Transformation" of Tim Deluxe, then traveled back into techno towards the end of his set with Paco Osuna's "Transheet." Needless to say, Cox was on form.
Once the car park closed, things picked up in the club. Over in the terrace, Joris Voorn and Nic Fanciulli kept things bouncy and fun for most of the night, throwing in classics like Lil Louis' "French Kiss." In the discoteca, Richie Hawtin played a rare closing set at Space after a fantastic season at the club with ENTER. He weaved his way through more minimal grooves than I've seen most of the season, with rolling bass and tripped out synths, a la fASTER's "Fatima," Fabio Florido's "Dipendence," and BaAus' "Bombolone."
Photos: Space closing
DC-10 closing
One party that wasn't lacking in attendance or energy was DC-10, which saw the garden totally rammed by early evening. Though the club has a reputation for being the most overcrowded opening and closing party spot on the island, this year things were just about perfect in the terrace and main room, though the garden got to be a bit much from time to time. Early on, however, Soul Clap kept things relaxed and grooving out in the garden with tracks like "Lost In Music" by Sister Sledge as clubbers laughed, bobbed and got in the mood. Over in the terrace, those who were already in the mood held up inflatable planes, while others wore neon clown wigs and red clown noses as Andrew Grant and Sossa played tracks like Markus Homm's "Take Off" and Lomez's "Real People" to get things going. One girl even brought a homemade sign that read "God Save This Club," which Andrew took from her to hang on the DJ booth, where it stayed almost the entire night.
Later on The Martinez Brothers got the terrace absolutely jacking, playing tunes like their own twisted house bomb "H 2 Da Izzo." Over in the garden, Seth Troxler kept it melodic and groovy, fitting the warm festival vibe out there perfectly. Closing the night out in the terrace was Maceo Plex, smashing the place with what I've seen as somewhat of a DC-10 anthem this year, Barnt's "Geffen." And just before him, Derrick May absolutely rocked the main room with the disco treat "I Feel Love" by Donna Summer. Crosstown Rebels boss Damian Lazarus finished off the night with tunes like the forthcoming Claude VonStroke remix of Infinity Ink's "Infinity" to a still-packed house as Claude himself celebrated the night behind him, clearly enjoying the end of a great season.
Photos: DC-10 closing
Sankeys closing
Just hours later (or what felt like it), we headed over to the first part of the Sankeys closing party. This year they threw down the gauntlet with 24, a 24-hour fiesta split into two 12-hour parties with 24 DJs in total. The big news was that Danny Tenaglia made a special appearance, and though the place wasn't quite as packed as I've seen it earlier on in the season, the crowd was ready and up for it. Tenaglia kept things upbeat and pumping throughout. The roof terrace, meanwhile, gave you the chance to plop down on a couch and watch the iconic raver film Human Traffic on a projector screen. (Shouts of "Nice one bruva!" echoed from around the roof.) Part two took a while to get going, and though it didn't quite reach part one levels, the ability to watch DJ Sneak and Cajmere in such an intimate setting was an experience unto itself. Despite most of the clubbing tourists having gone home for the winter, it rounded out the season nicely.
Photos: Sankeys closing


text:
My cricut can never cut enough hearts!!
All my side of the family lives out of state, so I made this for my mom recently. Yes, another frame from Ben Franklin ( 2 for $14.00)...If ya know of any other great places to get frames I'm all ears!!.... It's another simple gift and the hearts could be the butterflies I did on a previous project or any little figure ya like! You could put all the hearts in and say, "I love you" .... use your imagination and beautiful things will happen!!!


text:
Selecting the Best Battery for Embedded-System Applications article tells that non-rechargeable (primary) batteries find wide use in mainstream embedded-system applications. This category of batteries includes alkaline devices, which manufacturers typically fabricate using manganese dioxide and zinc powder with a caustic alkali of potassium hydroxide as an electrolyte. The nominal voltage of an alkaline cell is 1.5V, with a discharge voltage of 0.9V.
The capacity of an alkaline (regular) AA battery is typically approximated at anywhere between 700 and 3,000 mAH. "Heavy duty" batteries have much less powerful than alkaline batteries: for example Dynamic Energy Ultra AA say 258 mAh and Grunding Plus AAA 325 mAh.
FAQ – Batteries tells that the Mallory charts give the AA size alkaline cell a capacity of about 2800 mAH, but that's at room temperature, under light loading, with end of life at about 1.0 or 0.9 volt. But if you draw more current (over 500 mA) the alkaline really sucks, and dies after less than 900 mAH. Under similar conditions the NiMH are around 1500 mAH. Battery Life article give out tips how to get maximum battery life.
AA battery capacities from Wikipedia
|IEC name
|ANSI/NEDA name
|Capacity under 500mA constant drain
|400-1000 mAh
|1800-2600 mAh
|2700-3400 mAh
|600– 1000 mAh
|2200– 2900 mAh
|1500-1800 mAh
|Nominal voltage
If you use lots of batteries you might wonder what is the most economical way to buy batteries. Are Expensive Batteries Worth the Extra Cost? That's a good question that Are Expensive Batteries Worth the Extra Cost? article tries to answer. The writes tries to find out if cheap batteries from "dollar" stores are economically feasible or not. Here is a plot of the voltage vs. time for three different AA batteries: Energizer (Alkaline), Duracell (Alkaline), and DG (the one from the Dollar Store "heavy duty").
You will see clearly that these cheaper batteries are not nearly as good as the more expensive ones. What about the energy per dollar? Really, you are paying for energy – right? Very interesting. So it seems that all three of these batteries have about the same money-energy density.
But there is a bit more to batteries than just the energy stored in it. Your battery selection also depends on what you are using it for. Because the voltage on cheap "heavy duty" battery drops quite quickly, it drops very quickly to value that is too low for electronics devices to work properly (what is the limit depends on device): Even through battery will still have energy in it, but when it won't run the device correctly.
There are applications where cheap batteries can make sense. According to Are Expensive Batteries Worth the Extra Cost? article comments cheap batteries is the only way to go for kids' toys when kids constantly leave toys running when not playing with them. Much better to let cheap batteries go dark that way than expensive batteries.
One Are Expensive Batteries Worth the Extra Cost? article comment points out that there is not one Duracell and one Energizer battery; each have a range of battery types/technologies at widely varying prices. Article comment also points out that the measured data is only valid when batteries are used continuously and then discarded when usable power falls beneath a set threshold. Usually the normal use is slightly different (dry batteries can have a certain level of re-activating the charge when in rest).
How about rechargeable batteries? A typical rechargeable NiMH is 1.2V 2.5Ah, so it would be a very close race in capacity with alkaline batteries and better capacity than "heavy duty". The biggest problem with normal rechargeable batteries (NiCd and NiMH) is self-discharge, which means that after few months a flashlight tossed to drawer will be empty without any use. There are low discharge rechargeable batteries (for example Eneloop) that solve this self-discharge issue quite well.
Rechargeable battery vs regular battery article tells that most modern-day devices can compensate for minor differences in voltage (a common one is 1.2 and 1.5 V), but since there are so many "chemically different" regular and rechargeable batteries out there today. The simple solution: know how much voltage your device needs, then pick out the appropriate battery.
Rechargeable battery vs regular battery article says that when it comes to rechargeable batteries, though, you should keep in mind that the numbers indicated by the manufacturer are those seen under optimal conditions. Naturally, this varies depending upon the actual conditions of use. Factors like temperature and current-used have a large impact in determining just how long it will last. This applies to both rechargeable and to non-rechargeable batteries. Selecting the right battery article gives some more details on different battery types.
Select the right battery fuel-gauge for smart phones and tablets article tells that a highly accurate battery fuel-gauge is needed in many electronics devices because it can can increase customer satisfaction, reduce battery costs, and extract the maximum run-time. Also the electronics needs to be designed so that it consumes as little power as possible. How Long Will My Battery Last? Standardized benchmarking methodology compares microcontroller power consumption in battery-powered applications informs you how to compare power consumption of different microcontrollers.
For many uses I can agree on the conventional wisdom used to be that most alkaline batteries have fairly similar energy levels. Alkaline Battery Shoot Out data seems to support that that the differences between most alkaline batteries in capacity are quite small (much smaller than price difference on shop). Based in that usually the best deal is finding the cheapest alkaline you could find. It is a good idea to avoid cheap "heavy duty" batteries that have low capacity and many seem to leak out too easily. I have tried to find cheap as possible "decent quality" alkaline batteries.
If you are shopping for rechargeable batteries for applications where the device is not constantly used, look for low-self-discharge rechargeable batteries. Even though their listed capacity is usually somewhat lower and price higher than other rechargeable batteries, in many cases they are worth of that. If you use rechargeable batteries all the time, then go for high capacity batteries that can be charged quickly using a smart charger.


text:
I picked up the book with a bit of apprehension, though. We know the girl and her boyfriend murdered her part-time lover. Open and shut case, despite a bit of initial bumbling by the Mumbai Police. How do you build a 229-page book around it? Baghel approaches it quite cleverly though. After the initial description of the crime and the victim going 'missing', she changes track and goes into the background of the key players as well as the key situations/settings.
Ekta Kapoor and her chaotic production company occupies a large part of the book. While Maria Susairaj and Emile Jerome's lives in Mysore are the back-story of the crime and directly linked to it, Ekta Kapoor was supposed to be just a passing background. Instead, Baghel portrays Balaji Telefilms as a magnet for hundreds (if not thousands) of small-town youngsters who land up in Mumbai every single day in the hope of stardom. It is against this high-pressure Holy Grail that the high-stakes games of casting couches, false promises and eventual heartbreaks are played out.
Death In Mumbai builds this background deftly but strongly for us to realise that why it is almost 'natural' to kill off a casting assistant because he took advantage of a girl after promising her a role. It also explains the ambition of small towns really well though it tends to drag a bit after a point. The shortcoming of the book is the belabouring of the small-town back stories to establish the motivations of the victim and the two accomplices. We know the murder has happened. We know who the murderer is. We want to get on with the investigation and unravel the 'mystery'.
But despite that, the story remains gripping. The side characters, who remain on the periphery, contribute only nuggets to narrative (and the police investigation) and make us realise how difficult it must be to trace witnesses and build a case (outside Criminal Minds, that is). Despite the Mumbai Police's initial reluctance and bumbling, they are clearly the heroes (identified by name) who solve the seemingly simple case with relentless groundwork and interrogation.
You could argue that here is a story that is nearly five years old and we all know the outcome. Why a book on it?
Think. A woman and her boyfriend killed her lover in her flat. And before chopping his body in small pieces, they had sex in the blood-splattered room. Then, they disposed off the body and feigned complete ignorance for several weeks before relentless police investigation exposed them. If this story doesn't get a book, what will?


text:
When I saw these pics of my friend Scott and his fiancé Sarah, I just had to post them. The vintage 60s feel of these photographs is just too beautiful not to. I especially love how, despite the 60s vibe, they still have a modern touch to them. The suit is classic and handsome. But let's face it, it's all about the bride! Her dress is vintage Algo-ettes and is absolutely beautiful. The bodice is a type of embroidery finished with a full skirt. Classic 60s! Together with the hair, the pearls and the shoes adorned with a bow, the look is just so pretty. Congratulations you two!
[Photo credit via VadimDaniel.com]


text:
The Brassy Broadcast podcast is an exploration of ways we create, engage, and communicate.
Oct 1, 2017
Meet Jen's parents. Everyone is a little slap happy and silly
after a full day of traveling from West Yellowtone down to Ogden,
Beth gets control of the mic in this episode. Will she let it
go, or hang on to it forever?
Recent Facebook Posts
The Brassy Broadcast is an exploration of ways we create, engage, and communicate more effectively using technology, social media, and non-traditional broadcast platforms.
Having trouble logging in?


text: Sustainability Programs Sustainability Makes Business Sense. For over 30 years, Centurion Medical Products has been at the forefront of developing programs and products that help healthcare providers optimize the profitability of their green initiatives. That's made us the performance leader in supporting environmentally focused business strategies. We help our customers discover they don't have to choose between profit and sustainability. They can control costs with sustainable healthcare. Centurion Medical Products' On-Going "Green" Initiatives: Instrument Recycling & Rebate Program - Centurion offers free collection and pick up of used medical instruments, eliminating instrument disposal costs, reducing landfill and raw material usage, and issuing cash rebates for returned instruments. To date six million pounds of instruments have been reprocessed. In the past three years this program has paid out over $3 million in cash rebates to Centurion customers. Original Soiled Linen Collection System - Centurion pioneered the soiled linen and liner recycling program and to date has recycled over 50 million pounds of plastics bags used for soiled linens.


text:
It is known for its versatile collection of products. The various categories of products available are:
Amazon Offers: ELECTRONICS
Electronics have always been an important part of Amazon sales and this year also they are offering some great discounts on electronics.
|Amazon Offers
|Rs.2000/- off on Mobiles
|Click here
|Televisions: Up To 40% Off
|Click here
|Home Appliances at 20% off
|Click here
|Tablets at 35% off
|Click here
|Flat 22% Off on Washing Machine
|Click here
|HeadPhone at 30% Off
|Click here
|Camera at Flat 25% Off
|Click here
|Up To 30% Off on Refrigerators
|Click here
|SpeakersUp To 60% Off
|Click here
Samsung double door refrigerator
Product specifications:
- The Samsung double door refrigerator features a cooling technique that maintains the temperature throughout.
- This refrigerator has digital inverter technology that ensures reliable application
- The smart connect inverter function provides with backup in case of power down.
- The design is very user-friendly and accessible.
- The stabilizer-free operation provides protection in the case of power fluctuation.
- Energy efficiency – 2 star
Price, Offer and Discount: Its MRP is rupees 25,800 but is available for rupees 23,699 and a discount of 10% is available when payment is done with ICICI credit card in the mode of EMI.
Beats have always been a pioneer in the headphones department and it continues to show its quality of product with the Beats X. These are a set of Bluetooth earphones that provide you with the music experience which you just can't miss. With clear acoustics and power packed bass, these are a must have earphones. They come with 2 sets of extra ear tips and can be used to attend calls. Up to 8 hours of battery life. Amazon is giving RS. 2000 discount over these.
Amazon Offers: LIFESTYLE AND FASHION
Apparently, clothes are the one thing that is bought the most over the internet and Amazon gives you a perfect reason as to why buy it from their website or app. Get amazing deals like flat 70% off on men's clothing, 60% off on men's shoes, Up To 30% Off on Women's shoes and many more.
|Amazon Offers
|Flat 20% off on Men's clothing
|View Offer
|Up to Rs.1000/- off on Women's Clothing
|View Offer
|Minimum 30% off on kid's clothing
|View Offer
|Men's accessories at 50% off
|View Offer
|Flat 22% Off on Women's accessories
|View Offer
|Kid's accessories at 30% Off
|View Offer
|Men's Shoes at Flat 25% Off
|View Offer
|Kid's shoes 60% Off
|View Offer
It is easy to wear fashioned using premium cotton T-Shirt which we can wear the whole day. Save Rs 700/- and get 58% OFF on this premium cotton T-Shirt. You…
It is easy to wear night time and comfortable Raas Pret Women's Crepe Nightgown. Save Rs 951/- and get 45% OFF on this Rass Pret Women's Crepe Nightgown. Hurry up…
Arrow is a brand famous for its cloth quality and fit. Bring in a sense of luxury with 100% comfort and attractiveness with Aarrow's wide range of shirts at a discount of up to 50% off.
About the product: This melange blue shirt is made up of 100% standardized cotton, ensuring a comfortable wear with extensive ventilation to restrict sweating. This is a slim fit shirt to provide you with the best edge to edge fitting experience with a fish cut at the bottom.
Price: This is available at a discounted price of 999.
Marks and Spencer women's printed tee
Marks and Spencer is a brand whose name is enough to ensure the quality of the product. It is a brand with enough followers to make it under the list top 5 women brand.
About the product: This red mix colored tee is produced using a special type of cotton produced by Marks and Spencer which provides with optimal fitting, great comfort and does not affect the environment during its production. It is an extremely light and comfortable material and suitable for every body type.
Price: It is currently available at a discount of 40% and going out for 600.
Amazon Offers: LAPTOPS
Laptops are an essential item nowadays, for both students as well as office goers. You can search laptops on the basis of Brand, Processor, RAM or storage. Look for your dream laptop and get the best deal over it. And don't forget, if you have a laptop spare in your house, then don't just let it rest there, exchange it with a new one and get amazing discounts over it.
|Amazon Laptops
|Lenovo Ideapad 320E 80XH01FHIN 15.6-inch Laptop
|View Offer
|HP 15-BU004AU 2017 15.6-inch Laptop
|View Offer
|HP 15-BE002TX 15.6-inch Laptop
|View Offer
|Apple MacBook Air MQD32HNA
|View Offer
|Lenovo 110 -15ACL 15.6-inch Laptop
|View Offer
|Lenovo IdeaPad Thin and Light 320S
|View Offer
|HP Notebook 14-BU008TU LED Backlit 14-inch HD Laptop
|View Offer
|Dell Vostro 3468 14-inch Laptop
|View Offer
|HP 15-be016TU 15.6-inch Laptop
|View Offer
Check out the Laptop specifications & Amazon offers:
Apple is known for its fluidity and comfort for the user. MacBook is considered to be the most user-friendly laptop and its sleek design just attracts you towards it.
Product specifications:
- All new 2017 Apple MacBook Air
- 1.8GHz Intel Core i5 processor
- 8GB LPDDR3 RAM, 128GB hard drive
- 13.3-inch HD screen, Intel HD Graphics 6000
- MacOS Sierra operating system
- Weighing just above a kg, it's a very light laptop.
- 720p FaceTime HD camera
- 1-year manufacturer's warranty.
Amazon Offers: Get no-cost EMI on this laptop and also you can avail 26% off on the MRP.
HP makes it easy for the user to do more work with less effort. Attractive design with textured body and extensive performance capabilities, HP just gives you the best computing experience. HP ensures the quality of its product and provides you with the best segment features.
Product specification:
- 2GHz Intel Core i3-6006U 6th Gen processor.
- 4GB RAM and 1TB hard drive.
- 15.6-inch screen display.
- DOS operating system
- 4 hours battery life with media streaming, internet surfing and many more.
- Lightweight
- HD camera with WDR (Wide dynamic range) feature.
- 1-year warranty from the manufacturer.
Amazon Offers : No cost EMI available on this product. Discount of 15% can also be availed
Lenovo Ideapad is built to pass any test that you throw at it. With discrete design, powerful performance and sturdy structure, Lenovo Ideapad is just all you need. Now surf the internet, watch movies or play games, each and every experience would just be the best on the Lenovo Ideapad.
Product specifications:
- Watch movies, play games, browse the web and, most importantly, be more productive at work
- Surf the internet at 3x faster speed.
- Intel 6th gen i3 processor.
- 15.6-inch screen with integrated graphics
- 1TB Hard drive and 4 GB RAM.
- Complimentary 1-year warranty and 1-year accidental damage protection.
Amazon Offers: You can avail Cashback up to 15% by paying your bill Through ICICI bank.
This series of laptop is there to fulfill all your needs and to not bother you with any kind of bloatware. This is primarily a business-oriented laptop with top of the line security enhancements and enough memory to store all the data you need.
Product specification:
- 2.4GHz Intel Core i5 7th Gen processor
- 8GB RAM and 1 TB hard disk.
- Fingerprint reader
- 14-inch screen, Intel HD Graphics
- Anti-glare display
- TPM chip for bit locker security
- 1-year warranty from the manufacturer from date of purchase.
Amazon Offers:
- No cost EMI available
- Discount of 7%
- 1 year subscription of ADP
Amazon offers: BOOKS
Since childhood, we are advised that everyone should develop the reading habit and there is the better way to start than to start reading books. From fiction to children's books, every book you want is available on the Amazon site and really easy to grab. Amazon is offering great deals and discounts of up to 50% on books.
|Amazon Offers
|Flat 20% off on Fiction books
|View Offer
|Up to Rs.100/- off on Classic books
|View Offer
|Minimum 30% off on kid's Books
|View Offer
|Text Books at 50% off
|View Offer
|Amazon best reads
|View Offer
|Kid's accessories at 30% Off
|View Offer
|Exam central books at Flat 25% Off
|View Offer
|Up To 30% Off on Business books
|View Offer
|Romantics books at Up To 60% Off
|View Offer
About the book: This is the second volume of the Ramachandra series and you can expect the same type of matter from it. In this book, author Amish display the character of adopted women who later began the prime minister and then a goddess. It shows the struggles of how an orphan was adopted by a family and then her struggles to fight against the world for the right. You will also see a lot of the known characters from Ramayana like Lord Hanuman and Lord Ram.
Price: This book Is currently on a 50% price cut off and the best to buy at.
About the book: In the 1990's, a sitcom was released by the name of FRIENDS and it became the biggest soap opera hit this world has ever seen. The show continued for a whole decade and the number of its fans just kept on increasing. Even today, more than a decade after its finale, the fan following of this show is just immense. In this book, you can find various behind the scene stories and photographs. This book also contains the special interviews of each of the cast member.
Price: This book is available at a discount of 30% on Amazon.
Amazon Offers: MOBILES
Amazon has declared a discount of up to 40% on smartphones in the amazon everyday sale. An exchange discount of up to 10,000 is also available. Like last year this year smartphone sales are predicted to be major for iPhone followed by Redmi and Samsung. Best smartphones to consider this sale are:
|Amazon Offers
|Flat Rs.2000/- cashback on OnePlus
|View Offer
|Up to 25% off on Mi Phones
|View Offer
|Minimum 20% off on Samsung brands
|View Offer
|Motorola at 20% off
|View Offer
|Flat 19% off on Nokia
|View Offer
|Apple iPhones at flat 26% Off
|View Offer
|Lenovo mobiles at Flat 17% Off
|View Offer
|Up To 30% Off on Lava Mobiles
|View Offer
|Honor mobiles at Up To 10% Off
|View Offer
Originally priced at 14,999, Xiaomi mi max 2 is a power-packed smartphone with all the features of the high-end phone. Now it is being offered at a discounted price of 12,999.
Product description:
- 12MP primary camera.
- 6.44-inch display with corning gorilla glass protection
- 4GB RAM, 64GB internal memory expandable up to 128GB.
- 5300mAH battery
- Dual speakers to enhance the music and gaming experience.
- 1-year manufacturer warranty.
iPhones are considered to be a style statement and have the maximum followers in the smartphone market. Amazon is giving a super discount of 11,000 on iPhone 7.
Product description:
- 12MP primary camera and 7MP front-facing camera.
- 4.7-inch display.
- iOS 10 and 32GB internal memory.
- Non-removable Li-Ion 1960 mAh battery.
- 1-year manufacturer warranty.
Redmi 4
It is known as the best budget phone. There is no smartphone better than Redmi 4 in the less than 10,000 departments. With its stylish looks and powerful processor, it can take anything you'll throw at it.
Product description:
- 13MP primary camera and 5MPfront-facing camera.
- 5-inch capacitive touchscreen display.
- Qualcomm Snapdragon 435 octa-core processor,4GB RAM, 64GB internal memory expandable up to 128GB.
- 4100mAH battery.
- 1-year manufacturer warranty.
Asus Zenfone 3 is a complete package. It has got the amazing processor, 3GB of ram and a camera module that can make high-end mobile cameras insecure. It was launched at a price of 22,000 but is now available at a price of 10,999.
Product description:
- 16MP primary camera and 8MP front facing camera.
- 5.2-inch capacitive touchscreen display.
- Snapdragon 625, 3GB RAM, 32GB internal memory expandable up to 2TB.
- 2650mAH battery
- 1-year manufacturer warranty
The site along with some incredible sale on products also offers gift cards and discount coupons. Save this offer season more using Amazon Coupons.


text:
Page Two
This simply blew my mind.
Thanks Skye, from the bottom of my heart
Another wonderful surprise !
Marvaline & Krystal, my fellow music lovers
I thank you for this beautiful award.
An awesome award
Thank you, Sherry
Thank You Joe
This is a beautiful award
Thank You Joanne888
Thanks Trickerchic, For The Above Two Awards !
Thanks Donna
Teri, It Is An Honor To Display Your Award..Thanks
Thank you so much, Marvaline
Tom, I am honored to have received your award ! Thank You.
I treasure your award, Marvaline ! Thank you.
Margie, It is a pleasure to display this award. Thanks
JazzyGma's Websites


text:
Canton Economic Development
What happens when economic development has real power behind it?
CMU is not only a faithful servant to the city of Canton, MS, but CMU also aids the global economy through many large Canton businesses including the world's largest Nissan plant. CMU serves over 10,500 customers with the four utilities: water, electricity, natural gas and waste water.
With that said, our goal is not to be the largest utilities company. Instead, CMU strives to provide its customers with reliable service in all four utilities at the most economical price. CMU provides Canton businesses with all of the resources they need to compete in any market, and we would love to have your business join our community. Canton economic development is a priority for CMU.
Our site-selection team is ready to connect your company to a vibrant, growing economic region that offers competitive tax rates, multi-modal transportation systems and skilled, affordable labor.
Contact us for more information at 601-407-3214.
(Visit CMU Economic Development Web Page)


text:
Every business owners want to succeed his business and generate better revenues and for the purpose, they need to market their business. The modern world demands from every owner to market his business online as technology is grown rapidly and every business owner will move towards SEO of the business i.e. Search Engine Optimization. TheRead More
You are here
Home > Posts tagged "internet services"


text:
36 York Street London, ON N6A 1A5 ()
Random reviewRead all reviews
Escape Canada on York St. in London is fabulous. We've done all but one of their rooms and can speak to the cleverness of the puzzles and the quality of the games. These guys know what they are doing. I highly recommend this Escape Room if you are trying to decide which one to choose in London. I especially love the new Pandemic room but Flipped is another personal fave as my husband and I did it without a single clue and escaped with less than a minute to spare!! very exciting :-)RowFamily


text:
Related Pages: Drain Cleaning in Pleasanton
When it comes to calling a plumber in Pleasanton or surrounding CA areas we understand it is usually a stressful time in your home or business. Choosing the right plumbing company should help to alleviate some of the stress. At Clog Gone we have served the home and business owners of the community for over 15 years. We are locally owned, family operated and all of our contractors are licensed for your safety.
Do you have a plumbing problem? We specialize in both residential & commercial services 24/7. We pride ourselves on our fast, friendly & affordable service. Call Clog Gone today and ask about our 26 month guarantee!
Services we provide include:
- Tankless water heaters
- Drain cleaning
- Sewer repair & replacement
- 24/7 emergency plumbing
- trenchless sewer line repair
- real estate inspections
- water backflow prevention
- custom maintenance plans
- Eco Friendly Plumbing
Why Clog Gone Plumbing Services?
Clog Gone is a leading Pleasanton plumbing contractor. Our company has been operating in the area for 15+ years, and since starting has been instrumental in aiding thousands of residential and commercial customers with their various needs. As the name suggests, drain cleaning and unclogging services are the bread and butter of our business, but not for a second does that mean we are not capable of handling a large-scale sewer line replacement project (we have been hired to do this numerous times), install a tankless water heater, or any other plumbing job for that matter.
Our residential and commercial customer base includes:
- Homeowners
- Bars
- Restaurants
- Municipal buildings
- Nursing homes
- Gas stations
- Grocery stores
- Shopping centers
- Education facilities
The job is not done until your are satisfied
Every one of our technicians has undergone extensive training and certification, and each is required to keep current with the latest industry technologies and pertinent industry-wide information.
Along with enhancing their on-site capabilities, our commitment to continual growth is what allows our firm to keep offering low rates, all without skimping on quality. If you were wondering what distinguishes the team at Clog Gone from the myriad of other plumbers in the Pleasanton region, this is it!
We never expected to become one of California's top-rated plumbing contractors, but now that we are, we credit this to our unrivaled commitment and dedication to our clients, as well as an undying passion for what we do and the communities we serve on a daily basis. Clog Gone is deeply honored to provide the very best residential and commercial drain cleaning and repair services, in addition to modern, trenchless sewage pipe replacement methods.
The difference is clear: we are not your everyday plumber!
So, the next time you are stuck with a plumbing problem in your home or office; who are you going to call? We sincerely hope you consider getting in touch with the professional crew at Clog Gone, not because it would be great to add another client to our consistently growing network, but simply for the fact that we know it would be an excellent investment on your part. With us, the supreme quality is just the start. Our Pleasanton plumbers always go above and beyond what is asked of them to ensure the customer is left more than satisfied!


text:
Wednesday, October 24, 2012
Monday, October 22, 2012
Tuesday, October 16, 2012
Group Picture of Olde Time Dolls
A little paint can make a big difference!
Monday, October 15, 2012
Best Friends
Friday, October 12, 2012
My New Doll Tags
Thursday, October 11, 2012
My Izannah Doll Has a New Bonnet!
Tuesday, October 9, 2012
Monday, October 8, 2012
This is Why I Make Dolls
Hello Judi!
I just want to let you know that I have enjoyed all of the dolls I recently purchased from you...Especially Bethany, Sadie and Laura. My sister LOVED her birthday doll - a very belated but worthwhile wait! Your dolls are indeed unique and beautiful. Every detail is artistically created and lovingly applied. I am definitely a life-long fan!
Laura Garcia
Thursday, October 4, 2012
Subscribe to: Posts (Atom)


text:
History And Designers Behind Vintage Ladies Compacts
14/02/2016 | Fashion Fair | No Comments
If you are a brunette with brown eyes and pale skin, you can boost your youthful organic beauty with a light makeup routine. To set my face, she used the Fashion Fair pressed powder, not sure what the shade was but once again she didn't have any in stock so I bought the 'Perfect finish illuminating powder' in the shade 'Earth Terre'. At Brooklyn's Fulton Street shop, she asked the sales staff to suggest yet another Fashion Fair colour — a close to-match to suffice until her shade was offered. We will cover makeup guidelines for eyebrows, foundation and concealer, eye makeup, blush colors, and lip colors.
Honestly I'm not a huge Mac fan I do have also significantly of their item and I definitely will be supporting Fashion Fair a lot more in the future. Fashion Fair was developed in 1973 by Johnson Publishing founder John Johnson and his wife, Eunice, to cater to women of color" at a time when other American beauty businesses have been significantly less interested in the buyers. I will try and get some Fashion Fair from now on. Is Milani black owned I like their goods also.
Eunice Johnson, who was from a socially prominent household in Selma, loved fashion and style, and she utilised it for black self-empowerment. Just around the corner from the planet-renowned Bondi beach sits the spacious, air-conditioned Westfield Bondi Junction mall. How to Score a Deal: Funan DigiLife Mall is considered a lot more reliable than Sim Lim Square, but is not constantly recognized for its sales. Final year, the Chicago History Museum went via racks and racks of Eunice Johnson's garments for a massive exhibit called Inspiring Beauty: Fifty Years of Ebony Fashion Fair.
Aiming to shed its image amongst young African-American girls as the brand their grandmothers and mothers put on, Fashion Fair Cosmetics is in the midst of a reset. The fashion world was left with a void in 2010 when the runways of the Ebony Fashion Fair had been shut down for great. We had a foundation leak after and fix it with flex seal the liquid rubber spray. If you want a smooth, even finish with 23 shades to choose from, this is a foundation to try. Females with fair skin frequently battle with blue or purple coloration below their eyes. The centre is located on Pitt St beneath the Sydney Tower and continues along to Pitt St Mall.
To anyone else who may study this comment then please note that you must not use a warm compress during the 1st 36 hours of a black eye as the blood vessels may possibly nonetheless be broken and this will just make the black eye worse! Eunice Johnson took more than as producer and director in 1963, and under her path, the traveling show took on new heights as she expanded her cachet and energy inside fashion circles. If you have any of these beauty concerns, very carefully and evenly apply concealer to the region just before you layer foundation on. This is for 1 new complete size 1oz Fashion Fair Correct Finish Liquid Makeup in the shade listed in the title and shown beneath.behind, compacts, designers, history, ladies, vintage


text:
And NO I am not taking down these next three posts before or after said events taking place, by your request or via abusing clergy and/or laypeople. Only the Office of the Archdiocese of Toronto, via its civilly legal, and or episcopal representatives, or the episcopacy themselves, can place an honest request that I will comply to, and I will comment publicly such a request was made in a future post. or modify/update the existing content.
Hello Everyone .... if you are still around.
I know I am pretty late to the game on these topics, but hey, this is a combination of things that have poked at my craw, what is happening in the Archdiocese of Toronto (and a little beyond) with regard to some Latin Mass stuff, and the Trads Behaving Badly that deter people like myself from wanting full participation, or even part-time participation in the Latin Mass. With a few good items on the docket, it's time I make an appearance again. Cue some sarcasm or wittiness.
4 topics to post, the 3rd and 4th together in the third post, with 3 posts over 3 days.
Topic 1: A Cardinal goes publicly Pro-Life .... and there are still Trads Behaving Badly out there poo-pooing him.
This past week, he created two posts where he smears my beloved ++Collins. Here's the first, and the follow up after with my Eminence fictitiously writing to Hitler. While the posts remaining online aren't highly vitriolic, you can tell "reading between the lines" that it is detraction of His Eminence and his efforts. Another smack in the face to my Eminence. How nice of him to harass people online from miles away on a computer, likely in his domicile, from an island in Canada.
1) Eric Alcock/Contra demonstrates once again why the heck most Catholics do NOT want to be a part of Traditional Catholicism, which, not always, though many a time is tied to the Latin Mass. Sadly, it's these people's online and in person example that make such a path undesirable and only strengthens TBBs' ``enemies`` and what the TBBs call the Novus Ordo/NewChurch" of heresy and false lies. We young people .... are on the Internet a lot you know, and what you show us on here determines how we respond to things and even, what guides us in our faith when so many of our spiritual leaders fail at what they are doing, or are hypocrites/lukewarm themselves.
Alcock is clearly in favour of the Latin Mass as seen in this post here. And so, here we have another example of the majority of these cases, as an online/in person Latin Mass TBB.
What would you desire all parties involve do as a Catholic?
Summary: Eric Alcock/Contra/dticion!: Another reason for Catholics and I, NOT to re-enter or even dabble in the Latin Mass and/or fully embrace the Latin Mass/Traditional Catholicism, even ``part time.`` This is even despite being not in my archdiocese.


text:
The BriteTap Feeder is a significant improvement over mason jar feeders.
- It's unique shield blocks chicks from standing on the feed tray and pooping into their food. The shield can be raised as chicks grow so they can eat comfortably but can't stand on the feed tray.
- It fills through a lid in the top so it's much easier to fill and wastes less feed than Mason jar feeders.
- Finally, it can be used long after other chick feeders have been put into storage. The shield also prevents rain and snow from getting into the feed tray so it can be used to give adult chickens scratch, meal worms, oyster shells, grit etc.
Quantities Limited. Available for pre-sale on our web site: