text:
Welcome to Brenda's Natural Foods, your source for natural groceries, supplements, locally grown foods, organic produce & plants, and more.
We serve our community by providing a place where health and local foods are available in abundance. We also assist those with special dietary needs and link them with local producers.
Our cafe is open to the public and provides a full service lunch and dinner menu. Dine in or take out.


text:
EAGAN — Playing just its second match of the season, the Stillwater volleyball team jumped right into the deep end while getting swept by Eagan 3-0 in Tuesday's nonconference showdown. The third-ranked Wildcats defeated Stillwater by scores of 25-14, 25-15 and 25-12.
"It was just a learning experience for our girls," Ponies coach Jamie Suapaia said. "We knew it was going to be a tough game, but we're still a little disappointed in our execution. I thought we were prepared, but I may have put too much pressure on them and some of that is my fault. We told them it's a long season and we can't base the season off this loss. This gives us some motivation, knowing where we're at."
Bri Horwath led Stillwater with 12 kills in 25 attacks with four errors, providing much of the offense on a night when less-than precise passing made it difficult for the Ponies to get into rhythm.
Maddie Brandon finished with nine digs to lead the way defensively and Susan Evans provided 23 set assists.
"Susan was running around quite a bit trying to set to our outsides and Bri, but we just weren't in system enough to be effective," Suapaia said. "Our serving and our passing is one thing I've been stressing throughout the entire season and those two skills are going to lead us there and those two shut down (on Tuesday)."
All three games played out similarly, with the Ponies hanging in there until Eagan went on extended runs midway through each game.
"Game 1 we were right in it, but down 11-8, and made a few mental mistakes passing and just shut down and couldn't get our feet underneath us again," Suapaia said. "They did a good job of finding our holes and taking advantage of single blocks and what not. We just let them get on too many runs instead of stopping them and getting after them ourselves. We let them stay in their game and they took us out of ours."
Middle hitter Steph Dietrich added four kills in eight attempts for the Ponies (1-1), who host Apple Valley in a nonconference match on Thursday before traveling to Apple Valley to compete in a tournament on Friday and Saturday.

subset: commoncrawl


text:
Quaker Valley youth wrestlers flourishing
In its second year of existence, the Quaker Valley youth wrestling team is finding unprecedented success each weekend at meets throughout the region. Although the boys are accustomed to coming home with hardware from Open meets, where each wrestler is competing for an individual trophy, this season has been highlighted by team wins in dual and team competitions.
Feb. 9 was a special day in program history, when Quaker Valley youth wrestling hosted its first wrestling meet in program history. Facing off against Burgettstown, Seneca Valley and Beaver, among the best competition in the region, the boys earned a resounding victory by besting the competition in 58 percent of the matches at the high school.
"It was an exciting day for Quaker Valley wrestling, and our boys really showed incredible toughness and passion for the sport," coach Jason Richey said. "We are thrilled to have such a strong wrestling foundation. This team is going to do great things as these boys grow and mature into scholastic wrestlers."
The youth team consists of boys ages 5-12. Ten QV students are taking advantage of a co-op, which allows seventh-12th graders to wrestle for Moon.


text:
Ask a Lawyer and Get Answers to Your Legal Questions
Thank you for your questions:
1. In Florida, the license plates continue to belong to the debtor, and the debtor has a right to possession of those plates. If the recovery agency (the repossessor) did not leave the plates for the debtor, the debtor can recover them from where the vehicle was taken. The debtors should receive notice within 5 days of where the vehicle is being held.
2. As a co-debtor, you are entitled to notice of the repossession and how it will be disposed of. If you don't receive that notice, that would be a defense to any action against you for a deficiency judgment.
In what time frame is the dealer required to notify me of the repossession and sale?


text:
" Manufacturer: Electrolux / Upright / Container "
bought this vacuum from a well known cash and carry hence the lower price , and even then what a complete waste of money !
it is advertised as a 2000w bag-less vacuum and I though great a powerful vacuum , a bit heavy but very powerful just what I need ... How wrong I was
Assembly wasnt too bad , and it worked ok for the first few days , then the belt broke , so changed it not thinking too much of this .
after a second week and yet another belt change plus several times having to clear the hose at the back from blockages , I still persevered however after the THIRD belt change and more dust being blown out of the front of the machine , I realized that there was a problem with the thing
Not only does it block at the back it also blocks at the front under the brush which means taking the whole thing apart , to clear the blockages
it seems although the power is there , there is some kind of design fault that keeps breaking both the belts and making it block as much over all a machine that after 6 weeks of use ended up in the bucket


text:
Quantitative utilization of L-cysteate (2-amino-3-sulphopropionate) as the sole source of carbon and energy for growth of the aerobic, marine bacterium Silicibacter pomeroyi DSS-3T was observed. The sulphonate moiety was recovered in the medium largely as sulphite, and the appropriate amount of the ammonium ion was also observed. Genes [suyAB (3-sulpholactate sulpho-lyase)] encoding the known desulphonation reaction in cysteate degradation were absent from the genome, but a homologue of a putative sulphate exporter gene (suyZ) was found, and its neighbour, annotated as a D-cysteine desulphhydrase, was postulated to encode pyridoxal 5′-phosphate-coupled L-cysteate sulpho-lyase (CuyA), a novel enzyme. Inducible CuyA was detected in cysteate-grown cells. The enzyme released equimolar pyruvate, sulphite and the ammonium ion from L-cysteate and was purified to homogeneity by anion-exchange, hydrophobic-interaction and gel-filtration chromatography. The N-terminal amino acid sequence of this 39-kDa subunit confirmed the identification of the cuyA gene. The native enzyme was soluble and homomultimeric. The Km-value for L-cysteate was high (11.7 mM) and the enzyme also catalysed the D-cysteine desulphhydrase reaction. The gene cuyZ, encoding the putative sulphite exporter, was co-transcribed with cuyA. Sulphite was exported despite the presence of a ferricyanide-coupled sulphite dehydrogenase. CuyA was found in many bacteria that utilize cysteate.
- cysteate dissimilation
- desulphonation
- pyridoxal 5′-phosphate
- sequence comparisons
- sulphite exporter
L-Cysteate (2-amino-3-sulphopropionate) has been recognized as a natural product since 1946, when it was discovered as a product of the weathering of wool [1]. In the meantime, the compound has been found in Staphylococcus aureus and Bacillus subtilis [2,3] and shown to be an educt for the synthesis of the bacterial sulpholipid capnine [4], of algal sulpholipids [5] and possibly of another bacterial sulpholipid [6]; extracellularly, the compound is described as a component of spiders' webs [7]. This widespread occurrence of cysteate is mirrored by its biodiversity in microbial nutrition. The compound can serve as a source of sulphur for yeasts and bacteria; oxygenolytic desulphonation occurs in Escherichia coli [8,9]. Cysteate can serve as a sole source of nitrogen for bacterial growth (K. Denger and A. M. Cook, unpublished work); all the tested isolates degraded the compound apparently to biomass, CO2, ammonium and sulphate by the same pathway used by the organism for dissimilation of cysteate. Many aerobic and anaerobic bacteria have been found to dissimilate cysteate [10] (see below).
One pathway of cysteate dissimilation has been elucidated, in Paracoccus pantotrophus NKNCYSA [10]. Cysteate is transported into the cell and transaminated to 3-sulphopyruvate, which is subsequently reduced to 3-sulpholactate. SuyAB (3-sulpholactate sulpho-lyase) [EC 4.4.1.-, the full EC number has not yet been assigned] cleaves the C-sulphonate bond to yield sulphite and pyruvate. The results indicate that the last step in the pathway is excretion of sulphate, which is generated from sulphite by a cytochrome c-coupled sulphite dehydrogenase that we assume to have some phylogenetic relationship to the characterized bacterial sulphite dehydrogenase [SorAB (sulphite oxido-reductase)]. SorA is a molybdopterin-coupled oxidoreductase, SorB a cytochrome c [11]. The gene believed to encode the sulphate exporter, suyZ, is a paralogue of tauZ in the degradation of taurine and is co-transcribed with suyB [10].
Rein et al. [10] noted, however, that a different degradative pathway seemed to operate in most of their cysteate-dissimilating organisms. This pathway presumably involved a 35 kDa protein, which was strongly induced in cysteate-grown cells. The aim of the present study was to characterize this protein and its activity. However, each organism which Rein et al. used [10], had the disadvantage that its genome sequence is unavailable, and we were fortunate to obtain an alternative organism whose sequenced genome was available.
González et al. [12] detected that Silicibacter pomeroyi DSS-3T dissimilated cysteate, but it is clear from the genome sequence [13], that the bacterium does not encode orthologues of SuyAB. The organism, however, encodes a potential orthologue (SPOA0157) of SuyZ [10], which is located on the megaplasmid. As both the suyZ and tauZ genes are found immediately downstream of the gene whose product releases sulphite [10,14], we examined the gene upstream of SPOA0157. SPOA0158 was annotated as a 36.5 kDa protein in COG2515, which includes D-cysteine desulphhydrase [EC 188.8.131.52] and ACC (1-aminocyclopropane-1-carboxylate) deaminase [EC 126.96.36.199]. D-Cysteine desulphhydrase is a pyridoxal 5′-phosphate-coupled enzyme, which converts D-cysteine into pyruvate, and the sulphide and ammonium ions (e.g. [15,16]). We could thus hypothesize that the gene product from SPOA0158 catalyses the conversion of L-cysteate into pyruvate and the sulphite and ammonium ions (Figure 1).
We now report the presence of inducible CuyA (L-cysteate sulpho-lyase) in S. pomeroyi DSS-3. The enzyme was purified and characterized as a pyridoxal 5′-phosphate-coupled enzyme, and its corresponding gene cuyA identified. CuyA activity was found in some of the organisms used by Rein et al. [10] that did not contain SuyAB.
L-Cysteic acid (>99% purity), D-cysteine (99% purity) and L-cysteine hydrochloride (>99% purity), L-cysteine sulphinate (99% purity) and 1-aminocyclopropane-1-carboxylic acid (>99%) were purchased from Sigma–Aldrich. DL-3-Sulpholactate was synthesized by R. Gueta [10] (Department of Biology, The University, D-78457 Konstanz, Germany) as described in [17]. Oligonucleotides were synthesized by Hermann GbR Synthetische Biomoleküle (Denzlingen, Germany). Taq DNA polymerase and M-MuLV reverse transcriptase were obtained from Fermentas GmbH (St Leon-Rot, Germany) and they were used as specified by the supplier. RNase-free DNase was purchased from Qiagen. Chromosomal DNA was isolated from bacteria as described by Desomer in [18]. Total RNA was isolated using the E.Z.N.A. bacterial RNA kit (Peqlab Biotechnologie GmbH, Erlangen, Germany). A 50 bp DNA ladder (Fermentas) was used.
Organisms, growth, harvesting of cells and preparation of cell-free extracts
S. pomeroyi DSS-3T (DSM 15171) [12] and Desulfomicrobium norvegicumT (DSM 1741) were obtained from the German Culture Collection (DSMZ, Braunschweig, Germany). Bilophila wadsworthia RZATAU (DSM 11045) and Desulfovibrio sp. strain RZACYSA, which respire using L-cysteate as electron acceptor to yield sulphide and acetate, and Desulfovibrio sp. strain GRZCYSA (DSM 11493), which ferments cysteate to acetate, sulphate and sulphide, were from our own laboratory [19,20].
Cells of S. pomeroyi DSS-3 were grown aerobically at 30 °C in modified SBM-M (Silicibacter basal medium) [Dr J. R. Henriksen, personal communication (Department of Microbiology, University of Georgia, Athens, GA, U.S.A.)] which contained 200 mM NaCl, 50 mM MgCl2, 10 mM KCl, 10 mM CaCl2, 50 mM Pipes, 1 mM K2HPO4, 10 mM NH4Cl, 0.07 mM FeEDTA and trace amounts of vitamins and minerals cited in [21]: the sole added source of carbon and energy was 10 mM L-cysteate, unless otherwise stated. Precultures (3 ml) were grown in 30 ml screw-cap tubes in a roller. Growth experiments were done on the 100 ml scale in 500 ml Erlenmeyer flasks shaken in a water bath. Samples were taken at intervals to measure attenuance (at 580 nm), to assay protein and to determine the concentrations of sulphate, sulphite, carbon source and the ammonium ion. Similar cultures were used to generate small amounts of cells for enzyme assays or for molecular analyses. Cultures (1 litre) for protein purification were grown in 5 litre Erlenmeyer flasks on a shaker. Cells were harvested at D580 0.5 (about 150 mg protein/litre) by centrifugation (15000 g, 20 min, 4 °C), washed with 50 mM potassium phosphate buffer, pH 7.5 (containing 5 mM MgCl2) and stored frozen (−20 °C). The same buffer served as extraction buffer. Cell-free extracts free of nucleic acids were generated after disruption by at least five passages through a French pressure cell set at 140 MPa [22]. The membrane/particulate fraction was pelleted by ultracentrifugation (170000 g, 30 min, 4 °C) and the supernatant fluid was referred to as the soluble fraction.
B. wadsworthia RZATAU, Desulfovibrio sp. strain RZACYSA, Desulfovibrio sp. strain GRZCYSA, and D. norvegicum DSM 1741 were grown anoxically in carbonate-buffered, fresh-water medium (pH 7.2) with titanium(III)nitriloacetate as the reducing agent as described in [23]. The medium for B. wadsworthia RZATAU contained 50 mM formate as the electron donor, that for Desulfovibrio sp. strain RZACYSA and D. norvegicum DSM 1741 contained 20 mM lactate as the electron donor.
Analytical methods
L-Cysteate, L-cysteine sulphinate, taurine, and alanine were determined by HPLC after derivatization with fluoro-2,4-dinitrobenzene [19]. The ammonium ion was routinely determined by the improved Berthelot reaction (salicylate in place of phenol) [24] or by the specific reaction with glutamate dehydrogenase [25]. Sulphite was routinely determined as the fuchsin adduct [26] and occasionally measured by ion chromatography [27]. Sulphate was measured as the attenuance of a suspension of insoluble BaSO4 [28]. Pyruvate was routinely determined by the specific reaction of lactate dehydrogenase [29] and confirmed by MALDI-TOF-MS (matrix-assisted laser-desorption ionization–time-of-flight mass spectrometry) used in the positive-ion mode, after reaction with 1,2-phenylendiamine, by Dr K. Hollemeyer (University of the Saarland, Saarbrücken, Germany) [10].
Protein in whole cells was quantified by a Lowry-type reaction [30]. Protein in extracts was assayed by protein-dye binding [31]. Denatured proteins were separated in SDS/12% PAGE gels and stained with Coomassie-Brillant-Blue R250 [32]. The N-terminal sequence of a blotted protein was determined after Edman-degradation and HPLC separation at TopLab (Martinsried, Germany).
Enzyme assays
L-Cysteate sulpho-lyase was assayed routinely as the formation of sulphite after 2 min incubation at 30 °C. The reaction mixture (1 ml) contained 50 μmol Tris/HCl buffer (pH 9.0), 25 μmol L-cysteate and 0.1–0.8 mg protein, with which the reaction was started. Addition of up to 1 μmol pyridoxal 5′-phosphate to crude extracts or purified protein did not increase the activity and was thus omitted. On occasion, the reaction mixture was used to determine the formation of pyruvate or of the ammonium ion. Alanine dehydrogenase was assayed as reduction of NAD+ in Caps [3-(cyclohexylamino)propane-1-sulphonic acid] buffer, pH 10.0 [33]. 3-Sulpholactate sulpho-lyase was assayed discontinuously at 30 °C and pH 7.5 as the formation of sulphite (as described in [10]). Sulphite dehydrogenase [EC 220.127.116.11] was assayed photometrically with ferricyanide as electron acceptor at pH 7.5 [34]. Beef-heart cytochrome c could not replace ferricyanide.
Purification of CuyA
A three-step purification protocol was used. The soluble fraction was diluted with water (1:2.5) and loaded on to an anion-exchange column (Mono Q 10/10; GE Healthcare) equilibrated with 20 mM potassium phosphate buffer (pH 7.5); the flow rate was 2 ml/min, and proteins eluted with a gradient to 0.5 M Na2SO4 in buffer as described in [35]. CuyA eluted at about 100 mM Na2SO4. Active fractions were pooled, concentrated using Vivaspin concentrators (10 kDa cut-off; Sartorius), rebuffered in 20 mM potassium phosphate buffer (pH 7.5), containing 1.7 M ammonium sulphate, and subjected to hydrophobic interaction chromatography on phenyl Superose HR (5/5 column; GE Healthcare). A linear decreasing gradient of ammonium sulphate in 20 mM potassium phosphate buffer (pH 7.5) was applied, and CuyA eluted at 0 mM ammonium sulphate. Concentrated active fractions were loaded on to a gel filtration column (Superose 12 HR 10/30; GE Healthcare) and eluted at a flow rate of 0.4 ml/min with 50 mM potassium phosphate buffer (pH 7.5) containing 150 mM Na2SO4.
RT (Reverse-transcriptase)-PCR
The reverse PCR primers listed below were used for the RT-PCR reactions. PCR was performed as described previously in [36]. The PCR primers were as follows: for cuyA, primer combination cuyAF (5′-atctggatcaagcgcgacgactg-3′) and cuyAR (5′-gcatctcgaaagcgcagttgaca-3′), and for cuyZ, primer combination cuyZF (5′-gacactgttgcggttcggggtc-3′) and cuyZR (5′-ccgcagatagccaccgacccac-3′). Positive controls for RNA integrity after isolation were done using the 16S rRNA-specific primers 27F and 533R [37]. For the combination cuyAZ, primer combination cuyAF and cuyZR was used. RT-PCR products were visualized on 1.5–2% agarose gels.
Software for sequence analyses and accession numbers
Sequence analyses of the genome (accession number NC_003911) and the megaplasmid (accession number NC_006569) of S. pomeroyi DSS-3 were performed using the BLAST algorithm on the National Center for Biotechnology Information website (http://www.ncbi.nlm.nih.gov/). Sequence data were manipulated with different subroutines from the LASERGENE program package (DNASTAR). Transmembrane regions were predicted using the program TMHMM at the Center for Biological Sequence Analysis (CBS; http://www.cbs.dtu.dk/services/).
Growth of S. pomeroyi DSS-3 with sulfonates
Strain DSS-3 grew exponentially in L-cysteate-SBM-M with a μ (specific growth rate) of 0.07 h−1 (Figure 2A). The utilization of cysteate was concomitant with growth and was quantitative (Figure 2B), and approx. 80% of the sulphonate moiety was recovered as sulphite, the remainder as sulphate (results not shown). Some 80% of the cysteate-nitrogen, as the ammonium ion, was excreted into the growth medium concomitantly with growth (Figure 2B); the balance of the cysteate-nitrogen was calculated to be in cell material. The molar growth yield was 5.0 g of protein per mol C (a value which indicates quantitative utilization of the carbon source [38]), so, together with the growth rate, a specific degradation rate for cysteate of 1.3 mkat per kg of protein could be calculated.
Strain DSS-3 grew quantitatively with taurine or isothionate and excreted solely sulphate (and the ammonium ion, if appropriate) (results not shown). Cysteine sulphinate was a good growth substrate (with excretion of 20% sulphite and 80% sulphate); D-cysteine, L-cysteine and 1-aminocyclopropane-1-carboxylate were not utilized. The organism grew slowly with a poor yield (in 21 days compared with 3–5 days for other substrates) in DL-sulpholactate-SBM-M, and excreted sulphate (10% of the theoretical yield), but no sulphite. The growth medium interfered with the ion chromatographic analysis of sulpholactate, so we were unable to determine its fate.
CuyA in crude extracts of S. pomeroyi DSS-3
Soluble extracts of cysteate-grown cells of strain DSS-3 catalysed the hypothesized CuyA reaction, whereas extracts of acetate- or taurine-grown cells did not. The specific activity of CuyA in the crude extract (7.7 mkat per kg of protein; Table 1) was higher than the specific degradation rate (1.3 mkat (kg protein)−1) in growing cells and thus was sufficient to support the growth rate of the organism. The reaction products pyruvate (three independent methods), ammonium (two independent methods) and sulphite (two independent methods) were thoroughly identified. The pH optimum for the enzyme in crude cell extract was determined to be 8.8–9.0 when using a 50 mM Tris buffer in the pH range 7.5–9.0, whereas in sodium carbonate buffer pH 9.0, in Caps buffer at pH 9.5 or in 50 mM potassium phosphate buffer at pH 7.5, a lower activity was obtained.
The proteins in soluble extracts from cysteate-grown and acetate-grown cells were separated by SDS/PAGE (Figure 3). The major difference was the presence in cysteate-grown cells of two strongly-induced bands at 39 kDa and 42 kDa. The 39 kDa protein was shown to represent the cysteate sulpho-lyase (see below). The 42 kDa protein was isolated and determined to be alanine dehydrogenase, whose high activity was observed in crude cell extracts (Table 2). SuyAB activity was not detected (Table 2), which excluded the possibility that the 42 kDa protein represented SuyB (a 42 kDa protein [10]).
Purification and some properties of CuyA from S. pomeroyi DSS-3
Proteins in the soluble fraction of cysteate-grown cells were separated on an anion-exchange column. Two peaks of activity, measured as production of sulphite from L-cysteate, were observed. The minor peak eluted non-reproducibly at low salt concentrations often in the apparent absence of protein, whereas the major peak eluted reproducibly at about 100 mM Na2SO4 in the presence of protein. We worked with the major peak only, which catalysed the CuyA reaction and contained the 39 kDa protein (Figure 3). Further separation on hydrophobic-interaction and gel-filtration columns yielded an effectively homogeneous protein with an apparent molecular mass of approx. 39 kDa (Figure 3). Unit stoichiometry of all three products from the reaction, pyruvate, ammonium and sulphite ions was observed (Figure 4). The Km value was determined to be 11.7±2.1 mM. Determination of the stoichiometry of substrate disappearance and product formation was impractical, because 25 mM cysteate had to be added in the assay and during the linear phase of the assay [maximally 8 min (Figure 4)], and a turnover of only 0.3 mM substrate was obtained based on the quantification of the end products.
The specific activity of CuyA from S. pomeroyi did not apparently rise during purification, although the enzyme was effectively homogeneous after the third step of the purification. This might be due to inhibition by sulphate ions, which were present in all separated fractions, although desalting improved the specific activity only slightly (results not shown). The cofactor pyridoxal 5′-phosphate was tightly bound to the enzyme and its addition did not enhance the specific activity. CuyA from S. pomeroyi DSS-3 could be stored for at least 2 months at 4 °C without significant loss of activity. Frozen samples lost about 90% of their activity on thawing.
A UV–visible spectrum (pH 9.0) of the purified enzyme, which was yellow, gave maxima at 278 and 412 nm and a minimum at 260 nm, which supported the presence of bound pyridoxal 5′-phosphate, whose spectrum in the same buffer had maxima at 278 and 411 nm and a minimum at 260 nm.
D-Cysteine was converted into sulphide and pyruvate (and presumably the ammonium ion) by CuyA from S. pomeroyi, but at 15% of the rate with L-cysteate; L-cysteine was not a substrate. Neither L-cysteine sulphinate nor ACC was a substrate for the enzyme.
The N-terminal amino-acid sequence of CuyA was unambiguously MHLARYP, which was identical with the deduced N-terminal sequence of SPOA0158. This was the gene we predicted (see the Introduction). The deduced molecular mass of this protein, however, was 36.5 kDa, whereas we routinely observed 39 kDa for the denatured protein (Figure 3). Altered preparation procedures for SDS/PAGE did not significantly alter this value. The molecular mass of the native enzyme was estimated by gel filtration chromatography to be 100±6 kDa, indicating a homomultimeric enzyme.
Sulphite dehydrogenase in crude extracts of S. pomeroyi DSS-3
The almost quantitative excretion of sulphite during growth with cysteate (Figure 2) was initially attributed to the absence of a sulphite dehydrogenase, because no convincing homologue of SorA [11] was found on the genome of S. pomeroyi DSS-3. However, the excretion of sulphate during growth with taurine caused us to assay extracts for sulphite dehydrogenases. No cytochrome c-coupled activity was detected, but inducible ferricyanide-coupled oxidation of sulphite was detected in crude cell extracts of both cysteate-grown (3.2 mkat per kg of protein) and taurine-grown (3.8 mkat per kg of protein) cells of S. pomeroyi DSS-3 (Table 2). The level in crude cell extracts of acetate-grown cells was below the detection limit (<0.03 mkat (kg protein)−1). The dehydrogenase was found in the soluble fraction of the extract after ultracentrifugation (7.0 mkat (kg protein)−1) and an apparent Km value of 10.7±3.7 mM was observed.
Genes involved in cysteate degradation and their transcription in S. pomeroyi DSS-3
CuyA, encoded by cuyA, was identified as SPOA0158, which is located on the megaplasmid present in S. pomeroyi DSS-3. The locus is annotated as a 36.5 kDa protein in COG2515, which includes D-cysteine desulphhydrase and ACC deaminase (Supplementary data, Figure S1 http://www.BiochemJ.org/bj/394/bj3940657add.htm). Directly downstream of cuyA, a potential orthologue (SPOA0157) of SuyZ [10] is found. A potential LysR-type regulator (SPOA0159), which we termed cuyR, is located upstream of cuyA, on the complementary strand (Figure 1B). A chromosomal paralogue of cuyA (SPO2657) was obviously not involved in cysteate metabolism.
The gene product of cuyZ was predicted to be a sulphate exporter (see Introduction section), whereas excretion of sulphite was observed (Figure 2). RT-PCR-data showed that cuyZ was transcribed in cysteate-grown cells (Figure 5), but not in either taurine-grown (Figure 5) or in acetate-grown cells. A 1358 bp amplicon spanning cuyAZ was also observed (results not shown), so it was evident that the two genes were co-transcribed during growth with cysteate.
CuyA activity in other bacteria
Rein et al. [10] postulated that the inducible, 35 kDa protein in extracts of many cysteate-utilizing bacteria represented a new desulphonative enzyme. The results from the present study indicate that the enzyme could be CuyA. Cell extracts from B. wadsworthia RZATAU and from Desulfovibrio spp. strains GRZCYSA and RZACYSA were examined. Each organism contained inducible CuyA activity. CuyA from B. wadsworthia RZATAU was purified 10-fold to near homogeneity on the anion-exchange column (results not shown): this was the 35 kDa protein (Table 1).
cuyA-like genes in genome projects
We found cuyA-like genes in the sequences from several genome projects (see Supplementary data, Figure S1 http://www.BiochemJ.org/bj/394/bj3940657add.htm). These genes are often designated as COG2515, whose most prominent characterized protein is ACC deaminase. The deduced protein sequences had <62% identity with CuyA. The E. coli cysteine desulphhydrase (YedO, also known as DcyD) had 45.5% sequence identity with CuyA, the characterized ACC deaminases <40% identity. One complete CuyA homologue from the Sargasso Sea metagenome project [39] had 64% identity with CuyA (see Supplementary data, Figure S1), while several fragments (single reads) had similar sequence identities (results not shown).
The relatively high level of identity of some orthologues led us to the hypothesis that these proteins could also encode CuyAs. However, neither Desulfotalea psychrophila LSv54 (two paralogues sharing 51.2% and 46.4% sequence identity with CuyA, respectively) nor Caulobacter crescentus CB15 (59.7% sequence identity to CuyA) grew with cysteate [Dr R. Rabus, personal communication (Max Planck Institute for Marine Microbiology, Bremen, Germany); Dr A. Sporman, personal communication (Stanford University, Palo Alto, CA, U.S.A.), so we presume that the products of these cuyA-like genes are other enzymes in COG2515, such as D-cysteine desulphhydrase or ACC deaminase.
The data, which indicated that S. pomeroyi grew with L-cysteate as a sole source of carbon and energy [12], have been confirmed in medium in which the mass balances for carbon, nitrogen and sulphur can be calculated (Figure 2). A key hypothesis, for the cysteate degradative pathway involved in growth, was a novel desulphonative enzyme, CuyA (see the Introduction and Figure 1). This hypothesis has been confirmed experimentally (Figure 4) with a pure enzyme (Figure 3), as was predicted based on the identity of the corresponding gene, SPOA0158. The presumed reaction mechanism of β-elimination of sulphite followed by deamination [40], though a novel desulphonation reaction, falls within the standard reaction range of enzymic transformations involving pyridoxal 5′phosphate [41]. The enzyme thus carries out a lytic reaction that is not a hydrolysis, and can be attributed to the lyases [EC 4], specifically the 'carbon-sulphur lyases', EC 4.4, which has only one subgroup, EC 4.4.1. We propose that the enzyme (EC number has not yet been assigned) could have the systematic name 'L-cysteate sulpho-lyase (deaminating)' and the common name 'L-cysteate sulpho-lyase'. The fact that the enzyme can catalyse the D-cysteine desulphhydrase reaction does not alter the nomenclature, because the organism does not dissimilate D-cysteine, and the biological function is L-cysteate sulpho-lyase.
Native CuyA may be trimeric, based on the facts that the N-terminal sequence analysis gave a reliable start codon, on a reliable subunit molecular mass of 36.5 kDa (derived) to 39 kDa (observed), and on a value of 100 kDa for the native enzyme derived from gel filtration, which, unsupported, is not a robust method [42]. The first bacterial ACC deaminase also appeared to be trimeric, based on gel filtration data [43], whereas the enzyme of the yeast Williopsis (Hansenula) saturnus is a dimer, based on both chromatographic and crystal structure data [44]. The L-cysteine desulphhydrase from E. coli is described as being a dimer (gel filtration and ultracentrifugation) [45]. A conservative conclusion is that CuyA is homomultimeric.
CuyA is induced to about 10% of soluble cell protein (e.g. Figure 3). This high level of expression is typical of all the organisms in our studies (results not shown) and elsewhere [46]. The enzyme has a poor Km value, 11.7 mM, which might contribute to the requirement for a large amount of enzyme. We thus speculate that a powerful pump is required to supply cysteate at adequate concentrations in the cell. It has been noted that ACC deaminases have high Km values [47]. The high level of induction may involve the hypothetical LysR-type transcriptional regulator, cuyR, encoded upstream of cuyA in the opposite orientation (SPOA0159; Figure 1B). The spacing between cuyA and cuyR would allow this type of regulation of cuyAZ, and three possible LysR-boxes [48,49] are located in the promoter region between cuyA and cuyR, so there is circumstantial evidence for transcriptional regulation by CuyR.
CuyA was first hypothesized from data in the genome of the α-proteobacterium S. pomeroyi DSS-3, but it is also found in several δ-proteobacteria (Table 1). We thus presume the enzyme to be widespread, although it is not represented in currently accessible genome sequences (Supplementary data, Figure S1 http://www.BiochemJ.org/bj/394/bj3940657add.htm). We also presume that CuyA takes part in a degradative pathway for 3-sulpholactate, because all tested organisms which encode CuyA degrade 3-sulpholactate (Table 1). We hypothesize that sulpholactate is converted into cysteate prior to desulphonation [40], a pathway implicit in the synthesis de novo of the bacterial sulpholipid, capnine [4,50].
The cuyAZ-genes are co-transcribed (Figure 5), analogous to the suy(A)BZ genes in the degradation of L-cysteate in P. pantotrophus NKNCYSA [10], so they presumably represent an operon; the up- and downstream genes are on the complementary strand. CuyZ is deduced to have a leader peptide and nine membrane-spanning helices. We again [10] presume, that the only metabolic function, which is equally important to the cell as desulphonation (CuyA), is the excretion of the excess inorganic sulphur oxyanion (CuyZ) to maintain homoeostasis in the cell.
The relatively simple desulphonation reaction, to cleave the stable C-sulphonate bond and release pyruvate into the amphibolic pathways to allow energy conservation and growth (Figure 1), thus represents only a small portion of the solution to the metabolic problem of sulphonate dissimilation. The nature of cysteate uptake is still unknown. The sulphite (and ammonium) ions must be safely disposed of [10,14]. Sulphite is usually converted into and excreted by aerobes as sulphate, sometimes with a significant portion of sulphite [10,34,51–53]. The excretion of sulphite, with some sulphate, is novel, though we recently detected it in Paracoccus versutus N-MT utilizing N-methyltaurine [53a]: a dendrogram of sequences of putative exporters does not show an especially close similarity of CuyZ and TauZ (from strain N-MT) (Supplementary material, Figure S2 http://www.BiochemJ.org/bj/394/bj3940657add.htm). We also detected an induced sulphite dehydrogenase in strain N-MT [53a]. We speculate that the presumed sulphite-exporter, CuyZ (TauZ in strain N-MT), has a higher affinity for sulphite than the sulphite dehydrogenase (Km=0.7 mM). Bacterial sulphite dehydrogenases, especially the ferricyanide-coupled enzymes [34], are poorly characterized, as is their regulation, while the sulphate and sulphite exporters have only just been recognized [10,14].
Given the wide range of amino acids degraded by S. pomeroyi DSS-3 [12,13], we presume that excretion of ammonium occurs under many nutritional conditions and independent of any specific amino acid. This general process, though recognized for many decades, does not seem to have been explored in this context, though the structure of an AmtB (ammonia/methylammonia transport) protein was recently established [54], and in S. pomeroyi DSS-3, amt-1 (SPO 2093; closest orthologue to E. coli amtB) is expressed under all conditions tested (A. K. Gorzynska, K. Denger, A. M. Cook and T. H. M. Smits, unpublished work).
We are grateful to Ms Andzelika K. Gorzynska for the RT-PCR experiments, to Professor M. A. Moran (Department of Marine Sciences, University of Georgia, Athens, GA, U.S.A.) for advice on growing S. pomeroyi, to Dr R. Rabus (MPI Bremen) and to Dr A. Spormann (Stanford University, Palo Alto) for testing the growth of D. psychrophila and C. crescentus with cysteate, and to Dr H. B. F. Dixon (Department of Biochemistry, Cambridge University, Cambridge U.K.) for valuable discussions. This work was supported by the University of Konstanz. The International Association for the Exchange of Students for Technical Experience (IAESTE) funded Ms Gorzynska's visit.
Abbreviations: ACC, 1-aminocyclopropane-1-carboxylate; Caps, 3-(cyclohexylamino)propane-1-sulphonic acid; CuyA, L-cysteate sulpho-lyase; L-cysteate, 2-amino-3-sulphopropionate; RT, reverse transcriptase; SBM-M, modified Silicibacter basal medium; SorAB, sulphite oxido-reductase; SuyAB, 3-sulpholactate sulpho-lyase
- The Biochemical Society, London


text:
COLUMBUS, Ohio (AP) -- Officials say 25 motorcycles are among more than 175 surplus vehicles up for auction by the state.
The auction on Saturday at the General Services Division of the Ohio Department of Administrative Services also includes cars, vans and trucks. All vehicles are sold as-is with no warranties and must be paid for in full on the day of the auction.
Surplus state property may be damaged or is at the end of its government use. It also may include items seized by law enforcement officials
Surplus property auctions allow the public to purchase items at reduced prices and enable the state to recoup part of its initial investment.
State officials say miscellaneous property and vehicle auctions generated more than $3.9 million during fiscal year 2013.


text:
Accurate temperature
Floats in water
The Philips AVENT Digital Bath and Bedroom Thermometer is a convenient way to monitor the temperature both in baby's room and in the bath. It has also been designed and tested as a safe, fun bath toy.
Fast and reliable measurement
Accurate temperature readings for bath or bedroom
Safe to play with
Complies with all relevant toy and safety standards
The products that comply with the toy standard have been thoroughly tested to ensure they meet these norms and are totally safe.
-Operating temperature range 10-45 DegC
-Accuracy +/-1 DegC
-Complies with EN 71, ASTM F963-96a Technical
-Battery type LR44
-Removable/replaceable Yes
-Number of batteries 2
Customer Reviews
Orders within Australia – Regular Post
Orders will be delivered by Australia Post.
$8.95 for orders weighing less than 3kg
$13.00 for orders weighing more than 3kg
Delivery is FREE if your order is over $149 and weighing less than 3kg.
Extremely heavy parcels may incur additional postage costs. We will contact you prior to dispatch if this is applicable to your order.
Orders within Australia – Express Post
Express Post is available for orders under 500g at a cost of $11.95 and between 501g – 3kg for $19.95. If your order qualifies for Express Post, this will be offered at checkout.
Orders outside Australia
International orders will be dispatched via Airmail. Please allow up to 14 days for international Airmail delivery.
Shipping costs depend on the order weight and destination country. Once you have added the products you require to your shopping basket, select your country from the dropdown list and the shipping cost will be displayed.
Returns Policy
We hope that you will be delighted with your order. However, if for any reason you are not completely satisfied then please contact us within 7 days of receipt of your parcel, and our customer service team will be pleased to assist you with organising a return authorisation. All returned merchandise must arrive back to us in their original, unopened and unused condition.
Payment Options
We accept credit cards and PayPal:
* Visa
* MasterCard
* American Express
* PayPal
For your peace of mind, credit card information does not pass through or stored on our servers. Once you pay for your order, your credit card details are encrypted and processed through the our PCI-Compliant Payment Gateway.


text:
I want to erase your footprints
from my walls. Each pillow
is thick with your reasons. Omens
fill the sidewalk below my window: a woman
in a party hat, clinging
to a tin-foil balloon. Shadows
creep slowly across the tar, someone yells, "Stop!"
and I close my eyes. I can't watch
as this town slowly empties, leaving me
strung between bon-voyages, like so many clothes
on a line, the white handkerchief
stuck in my throat. You know the way Jesus
rips open his shirt
to show us his heart, all flaming and thorny,
the way he points to it. I'm afraid
the way I'll miss you will be this obvious.
I have a friend who everyone warns me
is dangerous, he hides
bloody images of Jesus
around my house, for me to find
when I come home; Jesus
behind the cupboard door, Jesus tucked
into the mirror. He wants to save me
but we disagree from what. My version of hell
is someone ripping open his shirt
and saying, Look what I did for you. . .
Poet: Nick Flynn
Poem: Emptying Town
Volume: Some Ether
Year: Published/Written in 2000
Poem of the Day:
Oct 17 2013
Are you looking for more information on this poem? Perhaps you are trying to analyze it? The poem, Emptying Town, has received 42 comments. Click here to read them, and perhaps post a comment of your own.
More Poems
(9 poems)
Nick Flynn books


text:
Bachelor of Business Administration in Management
The major in Management enables students to acquire skills and master concepts which lead to effective management and utilization of human and physical resources. Students will be able to contribute to strategic planning, budgeting, job analysis, personnel evaluation, production scheduling, and quality control. Students will also become proficient at organizing and motivating personnel.
General Requirements
English I and II
Music, Art, or Fine Arts
Laboratory Science (4 hrs. with lab)
Science (3 hrs. lab or non-lab)
Computer Science
Philosophy or Religion
Health & Wellness
Cross Cultural
Psychology, Sociology,Political Science
(6 credit hours from 2 of these 3 disciplines)
Major Requirements
Principles of Management
Principles of Marketing
Business Law I or II
Quantitative Analysis for Business Decisions
Principles of Financial Accounting
Principles of Managerial Accounting
Principles of Business Finance
Principles of Macroeconomics
Principles of Microeconomics
Management Information Systems
Accounting Information Systems
Business Strategy & Policy
Operations Management
Organizational Behavior
Human Resources Management
Four Management Electives (12 hours)
General Electives (as needed)


text: One of the best vibrators, and this is why; a powerful motor in the head that rotates and vibrates for G-Spot stimulation, a specially designed clitoris tickler that flutters and stimulates when vibration is turned on, soft fringes for vaginal stimulation, a tickler that encompasses the entire vulva area, and the shaft and clitoris tickler turn on separately from each other. There are two controls that independently control the tickler and the shaft, resulting in an infinite variety of sensations! Rock Your World is a very special edition to the Rabbit Sex Toy series!


text:
Items 1 to 2 of 2 on page 1 of 1
in stock $8.57
P.O.D Records: 3 Years Part 1 (140 gram vinyl 12")
Cat: POD 012 . Rel: 13 Mar 17
Deep House
Janeret - "First Light" (8:32)
D'julz - "Organic" (6:28)
Djebali - "Aliahs" (6:38)
Politics Of Dancing - "One & Only" (7:14)
Review: Amongst the hustle and bustle of the Parisian house music scene Politics Of Dancing have been doing great things for the past three years. Now S.M.A.L.L and Paco are celebrating what they have achieved so far with a 12" that welcomes some highly respected characters to the label alongside familiar favourites. Janeret is on killer form with "First Light", using shimmering dub chords to course their way through a rock solid house beat in a simple but utterly effective style, before D'julz demonstrates his instinct for a killer hook with the organ-led fun and games of "Organic". Djebali keeps things tight and funky on "Aliahs", before the label bosses themselves round the record off with the irrepressible pump of "One & Only".… Read more
Played by: Mihalis Safras, Paul Starey, Graham Sahara, Anna Wall, 2VILAS, 25 Places, Michael Walsh, Tuccillo, Politics Of Dancing
in stock $9.64
Down To Business Vol 1 (12")
Cat: VES 006. Rel: 13 Mar 17
Minimal/Tech House
Purveyors Of Fine Funk - "Depth Of Bass" (6:01)
James Barnsley - "She Not Fast" (6:03)
Politics Of Dancing - "Queens Road" (6:49)
Rhymos - "Jazz Hand" (7:58)
Played by: DJ Mau Mau, Graham Sahara, Politics Of Dancing
Items 1 to 2 of 2 on page 1 of 1


text:
Since Hazrat Mahdi (as) will have a wide stomach, his thighs will naturally be broad, as well.
His [Hazrat Mahdi's (as)] brow is open... his stomach large, and the space between his thighs is open. ... (Mari' bin Yusuf Karami Hanbali, Fawaid Fawaid al-Fikr fil Mahdi al-Montazar, p. 13)
THERE IS A SPACE BETWEEN HIS THIGHS... (Mar'i ibn Yusuf al-Maqdisi "Fera Idu Fevaidi'l Fiqr Fi'l Imam al-Mahdi al-Muntadhar)


text:
Special Offer for Philly.com Readers / Get The Philadelphia Inquirer and Daily News
Special Offer for Philly.com Readers
Your Daily Exhale (Sheffield's agent, Kyle Drabek, Explaining Jenkins and more)
0 CommentsOne of the big requirements of this job is the ability to process a massive amount of information in a minimal amount of time, all while blogging... Read more
Park named 5th starter; Happ will compete for bullpen spot
0 CommentsSorry, but something got lost in translation between here and Clearwater. Chan Ho Park has been named the Phillies' fifth starter. But lefthander... Read more
Phillies release Geoff Jenkins; Have interest in Sheffield
0 CommentsThe Phillies have released outfielder Geoff Jenkins and are hoping to sign Gary Sheffield, who was released by the Detroit Tigers today. General... Read more
My thoughts on the Park/Happ Conundrum
0 CommentsI want you all to take a few seconds on click on this link and read the following story, because it does a good job of accentuating the uniqueness... Read more
Park not eager for relief role
0 CommentsThe modern professional athlete is well schooled in limiting his remarks to platitudes. So Chan Ho Park's answer to a question on whether... Read more
2009 Phillies Odds, Part I; Plus Cole Hamels' line
0 CommentsFigure this is a good way to kill some time at work. The following are some Phillies-related Over/Unders courtesy of BoDogLife.com. Log your... Read more
Hamels struggles
0 CommentsPhillies ace Cole Hamels gave up 10 hits and three earned runs in four innings of work against the Yankees' Triple A team today. Hamels struck... Read more
Carrasco, Mayberry optioned to minors
0 CommentsThe Phillies have optioned pitcher Carlos Carrasco and outfielder John Mayberry Jr. to the minor leagues. Carrasco's demotion crystallizes... Read more
Amaro's Offseason
0 CommentsI have a confession to make: I missed the second half of the Villanova game. I wanted to wake up early today and get some work done on my season... Read more
Phillies acquire lefthanded reliever
0 CommentsThe Phillies have acquired lefthander reliever Jack Taschner from the San Francisco Giants in exchange for catcher Ronny Paulino, multiple... Read more


text:
The Party Animal
Juicy Couture Juicy Couture Rollerball: There are two major downsides to dancing the night away: Your feet take a beating, and your perfume always wears off by midnight. While the former is inevitable, prevent the latter with this handy rollerball.
theBalm Hot Mama: A two-in-one blush and shadow with a subtle shimmer that catches the light in an oh-so-radiant way.
Jouer Moisturizing Lip Gloss in Birchbox Pink: While we might be biased about this sheer fuchsia shade, one thing is true: Nothing says "party" quite like bright pink lips.
Triple C iPhone Case: Don't let your bag hog all the fun. This bright floral case is equal parts playful and stylish.
Supersmile Powdered Mouthwash: This spill-proof mouthwash is totally genius. Just tear and add water to the pouch to create a single-dose rinse.
Boscia Green Tea Blotting Linens: Keep shine from ruining cute Kodak moments with these green tea infused blotting papers.
The Chic Classic
Oscar de la Renta Rollerball: This romantic floral fragrance would dress up even a T-shirt and yoga pants.
Beauty is Life Hot Red Lipstick: Get noticed with this stop sign-red hue.
stila All Over Shimmer Liquid Luminizer: While caffeine is a great pick-me-up, nothing makes you look fresh-faced quite like strategically placed highlighter.
Tatcha Aburatorigami Blotting Papers: The 24-carat gold flakes in these blotting papers really up the fancy ante.
The Luxe Lady
Kate Spade Twirl Rollerball: This floral scent strikes the ultimate balance between sophisticated and playful. We keep it in our day-to-day handbag too.
ModelCo Shine Gloss: Not only will this long-lasting gloss stay on through multiple toasts, but you don't even have to head to the ladies' room to reapply—it comes with its own mirror!
YoungBlood Mineral Perfecting Powder: When it comes to translucent powder, less is more. A light dusting will keep your skin matte without disturbing your makeup.
Chantecaille Les Pétales Lumière de Rose: Part highlighter and part shadow, this velvety two-in-one gives skin a gilded look.
Minteas: These petite mints freshen breath and also provide an antioxidant boost, thanks to green tea extract.
Baggu Wedge Pouch: This keychain doubles as another storage compartment.


text:
15 Books to Empower Tween Girls
Learn about good books that will empower young girls, tweens and teens. Shop books for girls on Overstock.com.
Eric Dodge is a professor of economics and chair of the Department of Economics and Business Administration at Hanover College in Hanover, Indiana.
Get ready for your AP exam with this straightforward and easy-to-follow study guide, updated for all the latest exam changes!
5 Steps to a 5: AP Macroeconomics features an effective, 5-step plan to guide your preparation program and help you build the skills, knowledge, and test-taking confidence you need to succeed. This fully revised edition covers the latest course syllabus and provides model tests that reflect the latest version of the exam.
Inside you will find:
|STUDY AIDS / Advanced Placement
|Publisher Date
|2013-07-09 00:00:00.0
|Copyright Year
|11.0 in
|8.75 in
|0.75 in
|Unit weight
|0.95 lb
|Series Name
|5 Steps to a 5
|Dodge, Eric R., Ph.D.
|Country of Origin
|United States
Learn about good books that will empower young girls, tweens and teens. Shop books for girls on Overstock.com.
Best Books to Give for Mother's Day from Overstock.com. If your mom is a bookworm, use these tips to find her a literary Mother's Day gift.
How to Create Your Own Reading List from Overstock.com. Learn how to choose books that you'll enjoy and create a useful, engaging, and fun reading list.
FAQs about Banned Books from Overstock.com. Celebrate the freedom to read! Here are a few of the most frequently asked questions about banned books.
Twilight: Beyond the Novels from Overstock.com. Our guides provide customers with information about Twilight, the novel.
Pulitzer Prize Literature Guide from Overstock.com. Looking for something to read? Here is a list of notable Pulitzer Prize-winning books.
How to Start a Book Club from Overstock.com. Our guides provide customers with information about how to start a book club.
10 Fun Facts about Disney from Overstock.com. Disney has a long, exciting history, and here are some fun and lesser known facts.
Tips on Selecting the Perfect Entertainment Gift from Overstock.com. Our guides provide customers with tips on selecting the perfect entertainment gift.
This date is subject to change. In order to assure you receive Overstock.com's low price on this item, your credit card will be charged upon order placement. The item will ship immediately upon release.
Items must be returned in new or unused condition and contain all original materials included with the shipment. More Details
FINAL SALE EXCLUSION: Items marked as FINAL SALE are not returnable unless the problem you experience is the result of our error.
For your protection, all orders are screened for security purposes. If your order is selected for review, our Loss Prevention Team may contact you by phone or email. There may be a two business day delay to process your order.
** Most Oversize orders are delivered within 1-4 weeks. Some orders may take 6 weeks to be delivered.
5 Steps to a 5 AP Macroeconomics, 2014-2015
Today $19.93


text:
Michigan Valley Park
(785) 453-2201
5260 Pomona Dam Rd, Vassar, KS 66543
5260 Pomona Dam Rd, Vassar, KS 66543
Michigan Valley Park Campground has great appeal to family campers, sportsman, and water sports enthusiasts. The reservoir covers 4000 acres at normal multi-purpose pool and offers some excellent fishing and water sports opportunities and has 95 campsites with electric and water hookups available. Other amenities include a shower building, large playground, basketball court, swimming beach, trailer dump station, reservable picnic shelters, and nearby boat launching and marina facilities, handicapped accessible fishing dock and numerous bank-fishing areas.
Dining Types
- American:
- Drinking Water:
- Dump Stations:
- Showers:
State Parks
- Marina:
- Primitive Camping:
- Lodging Nearby:
- Trails:
- Equestrian Trails:
- Playgrounds:
- Fishing:
- Electric And Water:
- Bathhouse:
- Beach:
- Boating:
- Boat Ramp Lanes:
- Picnic Areas:


text:
Now you have me curious.<quoted text>
As is often the case here, after I TOTALLY shred one of the left's bullsh** lies, I don't get any debate back.
I don't get any attempt to bolster their argument with any statistics or facts or rational argument.
Instead I just get a pile of personal attacks.
That's what these left wing nut losers do - they post lies, I refute the lies, and they all start making personal attacks.
This is how I know I've won.
I've so thoroughly destroyed their lies that they have nothing left.
When you nailed yourself to the cross, it's easy to see how you secured your feet and one arm. Who drove in the nails into the other arm?
Tell us how you did it? Did you pay someone off the free market?
Did your boss do it?
Did some GOP politician lie to you that if you do this, he'd promise to take care of your children but crossed his fingers behind his back?
I'd put a dollar down that it was your mom.


text:
A "Reading Rainbow" Feature Title Zum, zum, buzz.... zum, zum, buzz... What's that strange buzz coming from the double bass? Berlioz has no time to investigate, because he and his bear orchestra are due at the gala ball in the village square at eight. But Berlioz is so worried about his buzzing bass that he steers the mule and his bandwagon full of magicians into a hole in the road and gets stuck. Time is running out, and if a rooster, a cat, a billy goat, a plow horse, and an ox can't rescue the bandwagon, who can? As the suspense mounts, intricate borders reveal the village animals making their way to the square one by one. When the clock chimes eight, the animals, ready to dance, have filled the square-but there's no sign of Berlioz. Jan Brett's glorious illustrations invite the eye to linger over exquisite details and humorous nuances that enhance the story. This delightful cumulative tale is one that will be looked at again and again.
Berlioz the bear and his fellow musicians are due to play for the town ball when the mule pulling their bandwagon refuses to move. A strange buzzing in Berlioz's double bass turns into a surprise that saves the day.


text:
Friday & River makes hand-crafted leather goods in San Diego, California that feature well-considered design and products that develop a patina over time.
Made from Tan Horween Leather with a rich brown color, this handsome key chain will keep your keys where you can find them- attached to you. Features a marine trigger snap and a key ring loop, both Nickel-plated for a modern look that will wear well with age.
Measures 5 3/4" in length.
We offer the following shipping options within the United States:
- FREE SHIPPING on orders over $100 via USPS PRIORITY MAIL
- $5.95 via STANDARD GROUND SHIPPING
- $9.95 via USPS PRIORITY MAIL
We ship to over 50 countries internationally. For a complete list of countries and rates, please visit our Shipping Page.
If for any reason you are not happy with your purchase, you can return it to us within 21 days of delivery for an exchange or refund. All items must be returned in new, unworn, unwashed condition, in their original packaging, and with tags attached.
Grooming products and products marked "Final Sale" are not returnable.
For more information please see our complete Returns Policy.


text:
Home » Finance » Human Resource Department » Human Resource Forms
Human Resource Forms
Click on the desired title to access the form. The PDF format forms can be viewed and printed using the free Adobe Acrobat Reader. If you do have the reader, you may download from the link below.
Delta State University is committed to a policy of equal employment and educational opportunity. Delta State University does not discriminate on the basis of race, color, religion, national orgin, sex, age, disability, or veteran status. This policy extends to all programs and activities supported by the University.
Send questions and comments regarding this site to Rachel Becker.


text:
lead paint
In the first of two reports on toy safety, NewsHour economics correspondent Paul Solman looks at how the recent controversy over lead levels in toys is affecting consumer shopping this holiday season. Continue reading
Reporter Loretta Tofani spent fourteen months in China researching working conditions in Chinese factories. Tofani details her investigation and the risks some Chinese workers face in the manufacturing sector. Continue reading


text: Business & EconomyRussian tycoon in fresh bid for freedom Mikhail Khodorovsky has appealed against the verdict that extended his jail time. 24 May 2011 01:16 GMT | Business & Economy, Russia The jailed Russian billionaire Mikhail Khodorovsky has launched a new bid for freedom. Once Russia's richest man, Khodorkovsky was jailed in 2005 for tax evasion. He'd been up for release this year, but a second trial extended his jail time. He's now appealing that verdict. Al Jazeera's Neave Barker reports from Moscow.Source: Al Jazeera


text:
Discounted EHRs Offered To Doctors In N.C. Allscripts has joined forces with Blue Cross and Blue Shield of North Carolina to expand the reach of its MyWay electronic health record tool.
HHS Offers $13.2 Million For Behavioral Health IT Organizations serving people with mental and substance use disorders get grants to improve IT capabilities.
Will Your EHR Land You In Court? Legally, EHRs are double-edged swords: They protect clinicians from malpractice litigation but also put them at greater risk.
Health IT Ready To Tackle ACO's Challenges The feds have yet to explain exactly what's needed to participate in an Accountable Care Organization, but many providers are ready jump on the bandwagon anyway.
Sage Practice Management Software Sold At Big Loss Misguided development doomed the practice management/electronic health record system company, says an analyst.
Health Searches Scare You? Medify's Personalized Answer Search on a health term and you often find data on the extremes, either too generic or too complex. Medify database aims to fix that.
IT Best Practices Can Sidestep EHR Snafus This guide to electronic health record basics isn't rocket science, but it's required reading for healthcare IT pros.
How To Fix The Personal Health Record Mess So few Americans are interested in creating a digitalized PHR that Google had to close its service. But there are ways to get the public interested.
Mobile Diabetes App Makes The Grade WellDoc's DiabetesManager lowered long-term blood glucose levels in a recent randomized trial, but didn't affect clinical symptoms.
Cloud Apps In The Real World Not every clinical app is right for the cloud, but these two case studies offer some insights into what does work.
How EHRs Feed The Clinical Research Pipeline Natural language processing programs can now data mine e-records to help locate the best candidates for clinical trials. Several major healthcare organizations have taken notice.
New Software Helps Docs Identify, Contact At-Risk Patients Phytel's suite of care management tools uses e-medical record data to identify gaps in patient care.
AmeriHealth Taps NaviNet For Mobile Care Management Pilot study will send patient information to physicians' smartphones in the exam room, where critical clinical decisions are made.
Want Career Advancement? Try Health IT The demand for health IT skills and experience is fueling career advancement and job opportunities at all levels, according to recruiters and hiring managers.
Is The Cloud Safe For Health Apps? Some healthcare providers are still hesitant to put patient data and clinical apps in the cloud. Here's how they cope with their angst.
Can Health IT Fix The Evidence Gap? It's impossible for clinicians to stay up to date on every medical innovation. But certain technologies and practices can help them stay current.
More Patients Collect Prescriptions In Integrated Healthcare Systems Healthcare systems that link pharmacy data with electronic health records see better compliance with patients picking up their medications, reports Kaiser Permanente study.
Xbox Kinect Helps Monitor Seniors' Health Doppler radar technology is also used in research to help detect oncoming illnesses, fall risk in older adults.
Do EHRs Improve Clinical Outcomes? An Objective View The evidence to suggest that electronic health records really make a difference has not been overwhelming--until now.
Feds Inject $12 Million Into Rural Health IT Forty grant recipients will get about $300,000 each to purchase technology, install broadband networks, and train staff members.
How Penn Medicine Mines E-Records For Clinical Trial Prospects It shows a hospital wringing value out of an electronic record system that took years to implement. One in a series of profiles of InformationWeek 500 innovators.
Health Monitoring Data Supplements Telehealth Capabilities American Well and Numera partner to help doctors better track chronically ill patients and initiate early intervention when needed.
Telemonitoring Pilot Attacks Diabetes Health information exchange's project aims to keep diabetic patients out of the ER; if it works, the technology could pay for itself.
10 Healthcare IT Innovators: InformationWeek 500 InformationWeek 500 healthcare companies and hospitals are using technology in inventive ways to improve care. These innovations include a unique virtual reality system for burn victims, "Smart Rooms," and a customized program to track application and system performance problems. Take a quick peek at 10 of these IT initiatives in our slideshow.
EHR Vendors Pan Medicare Quality Reporting Approach Plan to require submission of patient-level, rather than aggregate, data would cost too much and be too complex, say Electronic Health Record Association members.
HIE Vendor Market Poised For Shakeup Among dozens of vendors selling technology products and services to health information exchanges, no clear player dominates, study says.
Most Connected Hospitals List Ranks EMR Achievers U.S. News and World Report turns to Healthcare Information and Management Systems Society's metrics to compile first-ever list.
VA, DOD Choose Open Source To Combine EHRs Two federal departments launch open-source community to create a single EHR that combines military and VA records.
NYC Program Shows EHRs Can Boost Preventive Care Program led by Farzad Mostashari, before he became national health IT coordinator, supports entering structured data into an EHR to monitor a population's health.
EHRs Produce Better Clinical Outcomes In Cleveland Large study finds greater adherence to healthcare standards and improved outcomes for diabetics when physicians use electronic health records.
Smart Ways To Deal With Dumb Clinical Alerts Clinical alerts may help reduce dangerous drug interactions, but sometimes they cause more problems than they solve. Experts describe best practices to make them work better.
Athenahealth's EHR Tools Boost Quality Of Care, Revenue The insurer has developed a cloud-based "accelerator" program that may help clinicians more accurately report patient data, meet National Committee for Quality Assurance standards, and improve their bottom line.


text:
TAMPA — C Zenon Konopka, all 6 feet, 215 pounds of him, said his plan on Thursday was to challenge the largest player he could to a fight.
Evgeny Artyukhin, 6 feet 5, 254 pounds, said sure.
The multipunch dustup, two minutes and nine seconds into a 60-minute scrimmage at the St. Pete Times Forum, set the tone for two more fights and board-bending checks.
Konopka, 27, trying to win a job, was the centerpiece, fighting twice and hitting often to catch the eye of coach Barry Melrose.
"I don't know what kind of analogy you want to use, but I have a bunch of chips, and before camp is over, I want all my chips in," Konopka said. "I'll do anything to make the team, and that's the bottom line."
"He came here to make the team. That's all this kid is," Melrose said. "He doesn't care whether he takes a veteran's job. I love that about him."
Konopka fought Artyukhin even and wrestled with D Matt Smaby after an open-ice hit on Vinny Prospal. F Adam Hall fought D Radek Smolenak, and Artyukhin's hit on 6-7 D Vladimir Mihalik made the boards lean.
It was good stuff to Melrose, who wants to "create a culture here of physical play and guys who play hard."
That is Konopka, who said only captain Vinny Lecavalier and RW Marty St. Louis are "off limits" as far as being hit while he tries to avoid a trip to AHL Norfolk.
"Anyone else, when I line up against them, they're my enemy. That's the way it has to be," he said. "I'm not worried about repercussions. I don't want to sit in a hotel room in Norfolk saying I should have done this or I should have done that."
Konopka, with 24 goals, 55 points and 194 penalty minutes in 62 games last season for AHL Syracuse, said he has no hard feelings after a fight.
"It's forgotten until we strap them on (today)," he said. "I'm not here to make friends. It would be nice, but right now, everything I've been working for for years is in this camp."
Scrimmage report: The White scored a 2-0 victory on goals by Paul Szczechura and Gary Roberts. Mike Smith and Karri Ramo made 17 saves each. … C Chris Gratton teamed well with Roberts and Mark Recchi. … Smaby and D Andrej Meszaros were solid. … No. 1 draft pick Steven Stamkos said the speed of the game was not a problem.
Melrose's report: The coach said he liked Stamkos with Ws Ryan Malone and Radim Vrbata: "They gave us a lot of reasons to think they could be starting the season together."
Melrose liked Mihalik, who he said has made "great strides," and St. Louis: "That guy is awesome. You can argue he's the most fun guy to watch in the NHL."
But he said, "Some guys look like they're here on their way to Norfolk, and I'm disappointed with that, because if you look at our lineup, there are openings, especially on defense."
Odds and ends: The roster for Saturday's game at Pittsburgh has not been revealed, but Melrose said it will be young. … Gratton said his surgically repaired left hip held up well.
Damian Cristodero can be reached at firstname.lastname@example.com.


text:
About 900 elementary school students in Herriman were evacuated late Tuesday morning when smoke was detected above an equipment room.
Jordan School District spokeswoman Sandra Riesgraf said the students were taken outside Butterfield Canyon Elementary, 6860 W. Mary Leizan Lane (13825 South), as a precaution while Unified Fire Authority crews responded to the 10:32 a.m. call.
About half an hour later, the emergency was over. Firefighters found no flames, determining a small amount of smoke drifting from the second story above the school's equipment room was coming from an overheated air-conditioning motor.


text:
Have Cat Questions? Ask a Cat Vet Online.
How long has this been going on?
Does your cat ever get outside?
Do the nails on that foot look normal?
Dr. M
What exactly has grown? The pad or something on it that you can see?
Or the area that the blood is leaking from?
With the fact that the pad is warmer than the rest I would think he has an infection. Is the infection just from him tearing/catching the pad on something or is there a serious problem under the skin? The fact that he doesn't go outside makes some injuries less likely.
I don't think it is an emergency but I would take him to your veterinarian to have the foot checked. They may want to at least start him on some antibiotics. It is important to make sure something more serious is not going on.
If I can answer anything else please let me know.


text:
Definition of premise
1a : a proposition antecedently supposed or proved as a basis of argument or inference; specifically : either of the first two propositions of a syllogism from which the conclusion is drawnb : something assumed or taken for granted : presupposition
2 premises also premisses plural : matters previously stated; specifically : the preliminary and explanatory part of a deed or of a bill in equity
3 premises also premisses plural [from its being identified in the premises of the deed] a : a tract of land with the buildings thereonb : a building or part of a building usually with its appurtenances (such as grounds)
Examples of premise in a Sentence
Called behavioral ecology, it starts from the premise that social and environmental forces select for various behaviors that optimize people's fitness in a given environment. Different environment, different behaviors—and different human "natures." —Sharon Begley, Newsweek, 29 June 2009
Although the Voting Rights Act served, in some measure, to formalize the notion of racial representation, its consequences undermined its premise—that a transparency of interests existed between the representative and the represented. —Henry Louis Gates, Jr., New Yorker, 24 Oct. 1994
Thirty years ago the modesty of the general expectation was still consistent with the original American premise of self-government. —Lewis H. Lapham, Harper's, November 1992
They were asked to leave the premises.
The company leases part of the premises to smaller businesses.
The premises were searched by the police.
He disagreed with her premise.
the basic premises of the argument
a theory based on the simple premise that what goes up must come down
Origin and Etymology of premise
in sense 1, from Middle English premisse, from Anglo-French, from Medieval Latin praemissa, from Latin, feminine of praemissus, past participle of praemittere to place ahead, from prae- pre- + mittere to send; in other senses, from Middle English premisses, from Medieval Latin praemissa, from Latin, neuter plural of praemissus
First Known Use: 14th centurySee Words from the same year
Other Logic Terms
Examples of premise in a Sentence
Niebuhr … adhered to a form of liberalism more premised on a realistic assessment of human nature than Rauschenbusch's naïve progressivism was. —Alan Wolfe, New York Times Book Review, 21 Oct. 2007
Fears of a nuclear holocaust were fueled by President Nixon's "madman" theory of diplomacy. The madman theory was premised on the assumption that if the Soviets thought that Nixon was crazy enough to drop the bomb, they would leave us alone. —Will Manley, Booklist, 1 & 15 June 2006
let us premise certain things, such as every person's need for love, before beginning our line of reasoning
Origin and Etymology of premise
see 1premise
First Known Use: 1526See Words from the same year
Other Logic Terms
PREMISE Defined for English Language Learners
Definition of premise for English Language Learners
premises : a building and the area of land that it is on
: a statement or idea that is accepted as being true and that is used as the basis of an argument
PREMISE Defined for Kids
Definition of premise for Students
1 : a statement or idea taken to be true and on which an argument or reasoning may be based
2 premises plural : a piece of land with the buildings on it
Learn More about premise
Thesaurus: All synonyms and antonyms for premise Spanish Central: Translation of premise Nglish: Translation of premise for Spanish speakers Britannica English: Translation of premise for Arabic speakers Britannica.com: Encyclopedia article about premise
Seen and Heard
What made you want to look up premise? Please tell us where you read or heard it (including the quote, if possible).


text: El Peluquero Feliz is a barber shop in Little Havana, Vice City, and it appears in Grand Theft Auto: Vice City Stories. The shop is inaccessible to the player, and plays no role in the storyline in either of its appearances. El Peluquero Feliz is Spanish for "The Happy Barber".


text:
Palo Alto Software is a proud partner with the academic community. As part of our commitment to helping people succeed in business, we are eager to support educational activities in business planning, marketing planning, and entrepreneurship in general.
Business Plan Pro will help ensure you complete a compelling business plan be it for a course project or business plan competition.
With our special discount pricing for students, we offer the tools you need to succeed in your class without breaking the bank.
To place an order, email us and a sales representative will call you to process the order. Make sure to enter your student email address to verify you are a student.
We know that not all students can afford software even with the special student discount prices. To that end we also sell software licences to Universities so you can access our software for free from your computer lab. All you need to do is email us and let us know the name of your academic institution, the course name and the course administrator and we'll do the rest.


text:
- Home
- News
- Sports
- Business
- Entertainment
- Lifestyles
- Opinion
- Driveway
- Impress
All-Stars golden in Whistler
The Langley All-Stars warmed up for this weekend's provincial championships with a dominating performance.
The All-Stars, a team of 11, 12 and 13-year-olds, were competing at the Sea to Sky Invitational over the weekend in Whistler.
And the Langley squad went a perfect 5-0, out-scoring their foes by a combined 44-6.
The All-Stars cruised through the round robin with a 5-1 win over Whistler, 11-3 over Dunbar and 7-1 against Squamish.
In the semifinals, they hammered the hosts 11-1 and then shut-out Hastings 10-0 in the championship final.
Langley faces Whalley in a best-of-three series this weekend in Whalley with the winner advancing to the national championships in Calgary.
The team consists of Ryan Bitschy, Brayden Cameron, Mattias Dornblut, Shone Fyke, Daniel Hoodikoff, Parker Kubilius, Kristian Lyon, Jacob McDermott, Matthew Myers, Kyle Pfeifer and Cheylon Smith.
Scott Hoodikoff is the head coach with Jeff Lyon and Lonnie Cameron the assistant coaches. Wyatt Lyon is the bat boy.