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|Publication number
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|Application number
|Publication date
|Jun 6, 1989
|Filing date
|Apr 20, 1987
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|Jul 1, 1981
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|John L. Toner
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|Eastman Kodak Company
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|Patent Citations (4), Non-Patent Citations (2), Referenced by (72), Classifications (67), Legal Events (5)
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This is a continuation-in-part application of U.S. Ser. No. 007,024, filed Jan. 27, 1987 which is a Rule 60 Divisional of U.S. Ser. No. 825,693 filed Feb. 3, 1986, now U.S. Pat. No. 4,637,988, which is a Rule 62 Continuation of U.S. Ser. No. 279,398 filed July 1, 1981, now abandoned.
The present invention relates to novel fluorescent labels and more particularly to fluorescent labels useful for the preparation of specific binding reagents comprising fluorescent labeled physiologically active materials.
In specific binding assays, sensitivity is of prime importance due to the generally low analyte levels that are measured. Radioimmunoassay sensitivity limits the assay to measurements of concentration of 10-12 M, and more often only in the 10-8 to 10-10 M range. In addition, radiolabels suffer from the drawbacks of short half-life and handling hazards.
In fluorescence spectroscopy assays, a sample containing a fluorescent species to be analyzed is irradiated with light of known spectral distribution within the excitation spectrum of the target fluorescent species. The intensity of the resulting characteristic emission spectrum of the fluorescent target molecules is determined and is related to the number of target molecules.
The sensitivity of fluorescence assays, although theoretically very high, is limited by the presence of background fluorescence. Background signal levels are picked up from competing fluorescent substances, not only in the sample, but also in materials containing the sample. This is an especially serious problem in quantitative measurements of species associated with the samples containing low concentrations of desired target fluorescent molecules such as found in biological fluids. In many situations, it is impossible to reduce the background sufficiently (by appropriate filtration and other techniques known in the art) to obtain the desired sensitivity.
Time resolution offers an independent means of isolating the specific fluorescent signal of interest from nonspecific background fluorescence. Time resolution is possible if the label has much longer-lived fluorescence than the background, and if the system is illuminated by an intermittent light source such that the long-lived label is measurable during the dark period subsequent to the decay of the short-lived background. Such techniques are described in greater detail in German Offenlegungsschrift No. 2,628,158 published Dec. 30, 1976.
The long-lived fluorescence (0.1-5 msec) of the aromatic diketone chelates of certain rare-earth metals, for example, europiumbenzoylacetonate and europiumbenzoyl-trifluoracetonate, has been known for some time. The chelating agent absorbs light and transfers it to the metal ion, which fluoresces. German OLS No. 2,628,158 describes the use of time resolution in fluorometric immunoassays (FIA) through the use of fluorescent labels whose emissions are long-lived as compared with those of species which produce background interferences in such assays. This publication also provides a useful discussion of the techniques of FIA and its advantage over other immunoassay techniques such as radioimmunoassay (RIA).
The fluorescent immunoreagents described in German OLS No. 2,268,158 comprise at least one member of the immune system, i.e. an antibody or an antigen, "conjugated" with a rare-earth chelate. Such "conjugation" can be achieved in one of two ways:
(1) by labeling, i.e. attaching the rare-earth chelate to the antigen as described in Fluorescent Antibody Techniques and Their Application by A. Kawamura, Ed., University Park Press, Baltimore, Maryland, 1969, and then adding antibody to the conjugated antigen whereby the antibody and antigen join in the usual fashion, or
(2) by covalent bonding of the antibody of the chelate via a chemical group which binds to both antibodies and the chelates.
The problem with immunoreagents of the type described in German OLS No. 2,268,158 is that the fluorescent labeling species, namely the rare-earth chelates, are quenched, i.e. their fluorescence is extinguished, when contacted with water. This problem, hereinafter referred to as an "aqueous stability" problem, is particularly serious because a principal use for fluorescent labeled immunoreagents is in the assay of aqueous biological liquids such as blood, serum, etc. If aqueous stability could be conferred on these materials, they would be useful as fluorescent labels, for these biological liquids, thus allowing increased fluorescence immunoassay sensitivity by the use of time resolution of signal from background.
Further rare-earth chelates previously used for fluorometric measurements have had undesirable properties such as a low quantum yield for emission, undesirable sensitizer extinction coefficients which result in insufficient fluorescence using small quantities of detectable species, low λmax which renders the determination subject to interference from other components in the sample which are usually in the low λmax range, poor water solubility (most biological fluids are aqueous) and poor stability of the chelate at low concentrations.
Highly fluorescent compounds have been discovered which are chelates of Eu+3 and a chelating agent comprising a nucleus which is a triplet sensitizer having a triplet energy greater than that of Eu+3 and at least two heteroatom-containing groups which form chelates (coordinate complexes) with Eu+3 and a third heteroatom-containing group or heteroatom which is in the sensitizer or appended to the sensitizer nucleus. The chelates are water-soluble, stable at low concentrations at pH of 7 to 10, highly sensitive, and have favorable molar extinction coefficients (10,000-40,000) and favorable λmax. Accordingly, the present invention provides a class of highly efficient, aqueous-stabilized fluorescent labels for physiologically active materials such as antigens, hormones, antibodies and enzymes.
The chelating agent has the structure ##STR1## wherein R represents hydrogen, alkyl, alkoxy, alkylthio, alkylamino, aryl, aryloxy or a heterocyclic group;
R1 represents hydrogen, alkyl, alkoxy, alkylthio, alkylamino, substituted or unsubstituted aryl or a heterocyclic group;
R2 represents carboxy, hydroxy carbonyliminodiacetic acid, methyleneimino diacetic acid, hydrazinylylidene diacetic acid or the salts of such acids;
n is 0 to 4; and
m is 0 to 1
provided that m can be 1 only when n is 0.
Alkyl refers to straight or branched alkyl of about 1 to 12 carbon atoms, preferably 1 to 6 such as methyl, ethyl, isopropyl, 2-ethylhexyl, decyl, etc.
The alkyl portion of alkoxy, alkythio, alkyamino has this same meaning.
Aryl and aryloxy refers to substituted or unsubstituted aryl or aryloxy of about 6 to 20 carbon atoms such as phenyl, naphthyl and phenanthryl including substituted derivatives thereof such as nitrophenyl, hydroxyphenyl, tolyl, xylyl, methoxyphenyl, methylthiophenyl, carboxyphenyl, 5,5-diphenyl-2,4-imidazolidinedione-3-ylmethylphenyl (a diphenylhydantoin substituent on a tolyl group) and the like.
Heterocyclic refers to substituted or unsubstituted heterocyclic groups having 5 to 6 nuclear carbon and hetero atoms such as pyridyl, methylpyridyl, nitropyridyl, methoxypyridyl, oxazolyl, imidazolyl, pyrazolyl, quinolyl, etc.
The alkyl, alkoxy, alkythio, alkyamino, aryl, aryloxy, and heterocyclic groups can be part of, or have appended thereto, proteins for use in biological assays, particularly haptens, enzymes, antibodies, and antigens for example the previously mentioned hydantoin substituent.
The present invention also provides a new class of specific binding reagents, such as antigens, enzymes, hormones and the like bearing these highly useful fluorescent labels.
The reagents are formed by adsorbing or covalently binding the fluorescent labeled antigens, haptens, antibodies, plant lectins, carbohydrates, hormones, enzymes and other such species-specific materials.
Eu+3 is complexed with the chelating agent comprising a nucleus which is a triplet sensitizer having a triplet energy greater than that of the lanthanide metal and at least two heteroatom-containing groups and a third heteroatom-containing group or heteroatom which is in or appended to the triplet sensitizer nucleus, each of said two heteroatom-containing groups appended to different carbon atoms of the triplet sensitizer nucleus, said heteroatom-containing groups forming coordinate complexes with Eu+3 and said groups being located in said chelating agent such that they and said third heteroatom or heteroatom-containing group are capable of forming a chelate structure with Eu+3.
The nucleus of the chelating agent is any triplet sensitizer having the requisite triplet energy. Examples of triplet sensitizers useful herein include heterocyclic and aromatic nitrogen-containing compounds such as quaterpyridines, quinquepyridines, terpyridines and phenanthrolines.
The terpyridines, quaterpyridines and quinquepyridines have the structure ##STR2## wherein R represents hydrogen, alkyl, alkoxy, alkylthio, alkylamino, aryl, aryloxy or a heterocyclic group;
R1 represents hydrogen, alkyl, alkoxy, alkylthio, alkylamino, substituted or unsubstituted aryl or a heterocyclic group;
R2 represents carboxy, hydroxy carbonyliminodiacetic acid, methyleneimino diacetic acid, hydrazinylylidene diacetic acid or the salts of such acids; and
n is 0 to 3.
The phenanthrolines have the structure ##STR3## wherein R1 represents hydrogen, alkyl, alkoxy, alkylthio, alkylamino, substituted or unsubstituted aryl or a heterocyclic group; and
R2 represents carboxy, hydroxy carbonyliminodiacetic acid, methyleneimino diacetic acid, hydrazinylylidene diacetic acid or the salts of such acids.
With europium, the triplet energy must be at least about 47 Kcal.
The complex contains any ratio of Eu+3 metal to chelating agent. In preferred embodiments, the mole ratio of Eu+3 to chelating agent is from about 1:1 to about 2:1. Especially preferred are complexes having a mole ratio of 1:1.
Eu+3 and the chelating agent are easily complexed by merely mixing an aqueous solution of the chelating agent with a Eu+3 salt in an aqueous solution of pH 7 to 10. The Eu+3 salt is any water soluble salt of the metal such as EuCl3 •6H2 O or other halogen salts. The chelate is generally prepared in aqueous solution at a pH of between 8 and 11 and preferably 8 and 9. The chelate optionally is mixed with buffers such as phosphate and borate to produce the optimum pH.
Especially preferred chelating agents are ##STR4## wherein R3 represents H, or alkyl of about 1 to 8 carbon atoms, and
R9 represents H, --OCH3, --NO2 ; and ##STR5## wherein R5 represents --CH3, NO2, or --OCH3 ; and ##STR6## wherein R9 represents H, --OCH3, --NO2.
The chelate is useful to label a variety of physiologically active materials by binding said materials to the chelate by adsorption or by covalent bonding. Among the physiologically active materials which are labeled in this fashion are enzymes and their substrates, antigens, antibodies, i.e. any substance which is capable, under appropriate conditions, of reacting specifically in some detectable manner with an antibody, carbohydrates, metabolites, drugs, other pharmacalogical agents and their receptors and other binding substances. Specific binding assay reagents are described in U.S. Pat. Nos. 3,557,555; 3,853,987; 4,108,972 and 4,205,058.
Techniques for performing such binding of physiologically active materials to the complexes are those well-known in the art and include simply mixing the materials together.
In specific binding assay methods, a compound having structural similarity to the analyte being determined is herein referred to as the ligand and the labeled compound as the ligand analog. Compounds which specifically recognize the ligands and ligand analogs and bond to them are referred to as receptors.
In performing one such type of assay, the ligand is placed in competition with the ligand analog for binding to the receptor. Unknown concentrations of the ligand are inferred from the measured signal of the labeled, ligand analog. The reaction proceeds as follows: ##STR7##
For illustrative purposes, the discussion which follows describes one particular type of specific binding assay technique, a competitive binding fluorescence immunoassay technique.
This system consists of antigen labeled with a fluorescent label of the present invention, unlabeled native antigen (in test sample) and specific antibody whereby there is competition between the unlabeled antigen and the labeled antigen for binding to the antibody.
The greater the concentration of unlabeled antigen from the test sample in the system, the less the labeled antigen will be bound by the antibody. If the concentrations of labeled antigen and antibody are fixed and the only variable is the level of unlabeled antigen, it is possible to determine the unknown level of unlabeled antigen by physically separating the antigen-antibody complex from the remaining free antigen (both labeled and unlabeled) and comparing the fluorescence of the labeled antigen, either free or bound, with a standard curve plotting of the values given by a range of known amounts of the antigen treated in the same manner.
Once prepared as described hereinabove, the fluorescent-labeled, physiologically active species is useful in fluorescent specific binding assays, particularly those which utilize temporal resolution of the specific detecting signal to distinguish from background as described in aforementioned German OLS No. 2,628,158. In this time-resolved mode (i.e. temporal resolution), the sample is excited in an intermittent fashion and information is accepted only during the dark cycle when the long-lived fluorescent label is still emitting strongly but when other sources of fluorescence have decayed. Discontinuous excitation is achieved in a variety of ways, including pulsed laser, mechanical chopping of a continuous excitation beam and moving the sample in and out of the excitation beam. Moreover, discontinuous excitation has the advantage of allowing the use of high radiant power without the absorption of a large amount of energy by the sample, thus diminishing the probability of sample photodegradation.
Examples of such fluorescent specific binding reagents described herein find utility are described in U.S. Pat. Nos. 3,988,943; 4,020,151; 3,939,350; 4,220,450 and 3,901,654.
In a preferred embodiment, the specific binding assay is carried out in a dry analytical element such as described in copending U.S. Pat. No. 4,258,001 granted Mar. 24, 1981 by Pierce and Frank. In this embodiment, the element contains a support and a spreading reagent layer comprised of polymeric beads, and optionally a registration layer. In some cases, the spreading layer is separate from the reagent layer. The spreading, reagent and registration layers optionally comprise the polymeric bead structure. The polymeric beads of the reagent layer have receptors such as antibodies adsorbed to their surfaces.
The chelate label of the present invention is placed above, below, or in the reagent layer in a manner that prevents the specific reaction from occurring prior to sample wetting, or it is spotted onto the element concurrently with or subsequent to the sample. It is only necessary that the labeled ligand analog permeate the element upon wetting subsequently to compete with the unknown amount of ligand in the sample in the formation of the ligandreceptor complex. The assay is performed by fluorimetrically determining the amount of free labeled ligand analog present or the amount of bound labeled ligand analog-receptor complex.
The following non-limiting examples will serve better to illustrate the successful practice of the instant invention.
The syntheses of terpyridines, quaterpyridines and quinquepyridines are similar. The 1 H-NMR spectra, the mass spectra, and the infrared spectra were consistent for the desired products in each of the following examples. ##STR8##
A mixture of compound 1 wherein R1 is methoxy (7.96 g, 34.9 mmol) and the bispyridinium salt 2 (10.0 g, 17.5 mmol) was refluxed for 18 hours in 600 mL of MeOH with 15 g of NH4 OAc according to the procedure of F. Kronke, Synthesis, 1, 1-24 (1976). The solution was filtered and the solid was triturated with hot dilute HCl. The resulting terpyridine diacid was collected by filtration, washed with MeOH then Et2 O (diethyl ether) and dried to give 4.74 g of product as white powder (51%).
Anal. Calcd. for C31 H23 N3 O6 •H2 O: C, 67.5; H, 4.6; N, 7.6. Found: C, 67.8; H, 4.4; N, 7.5.
Terpyridine compounds 13, 14 and 15 were produced from compound 3 in which R1 was --CH3, according to the following reaction scheme. THF is tetrahydrofuran. ##STR9##
Other representative examples of useful terpyridines are presented in Table 1.
TABLE 1__________________________________________________________________________ ##STR10## (1)wherein R4 represents H, SCH3, OCH3, CN, NO2,CO2 CH3, CH3, etc.; ##STR11## (2)wherein R5 represents CH3, OCH3, NO2, etc.; ##STR12## (3)wherein R5 represents CH3, OCH3, NO2, etc; ##STR13## (4)wherein R3 represents H, alkyl of 1 to about 8 carbon atoms;andR9 represents H, OCH3, NO2, etc.; ##STR14## (5) ##STR15## (6) ##STR16## (7)wherein R7 represents H, OCH3, NO2, etc.; ##STR17## (8)wherein R8 represents H, OCH3, NO2, etc.__________________________________________________________________________
Quaterpyridine compounds were produced according to the following reaction scheme. Bu is butyl. ##STR18##
6,6'-Diacetyl-2,2'-bipyridine was synthesized by the method of J. E. Parks, B. E. Wagner and R. H. Holm, J. Organomet Chem. 56, 53-66 (1973). The bispyridinium iodide was generated by refluxing 2.00 g of the diacetylbipyridine (8.33 mmol) in 50 mL of pyridine with 4.23 g of I2 (16.7 mmol) for 1 hour. The solution was cooled and filtered, and the solid was washed first with pyridine then with Et2 O (diethyl ether) to give, after drying, 4.46 g of the desired product as a sand-colored solid (82%).
Anal. Calcd. for C24 H20 I2 N4 O2 : C, 44.3; H, 3.1; N, 8.6. Found: C, 44.4; H, 3.2; N, 8.8.
The bispyridinium salt (3.00 g, 4.62 mmol) and 1, with R10 being methyl, (1.75 g, 9.23 mmol) were combined and refluxed together with 20 g of NH4 OAc in 500 mL of MeOH for 16 hours. The solution was cooled and filtered. The solid was worked up by the method used for 3 (Example 1), then recrystallized from N,N-dimethylformamide (DMF) to give 0.80 g of the titled compound as a cream-colored powder (30%).
Anal. Calcd. for C36 H26 N4 O4 •H2 O: C, 72.5; H, 4.7; N, 9.4. Found: C, 72.4; H, 4.4; N, 9.3.
Quaterpyridine compounds made by the procedure in Example 2 includes: R11 =--CH3 and --OCH3.
Quinquepyridine compounds were prepared according to the following reaction scheme: ##STR19##
Compound 16, available by the previously cited Parks et al method, (18.1 g, 74.2 mmol) was dissolved in 400 mL of Et2 O which had been freshly distilled from LiAlH4. With the temperature maintained at -50� C., 35.3 mL of 2.1 M n-BuLi (74.2 mmol) was added dropwise with stirring under N2. After stirring an additional 0.5 hour at -50� C., dry CO2 (g) was bubbled through the solution and the temperature was allowed to rise to 22� C. After 2 hours, the mixture was extracted with dilute aqueous NaHCO3. The organic phase was washed with water and the combined aqueous phase was acidified to pH 2 with HCl then extracted with CH2 Cl2. The CH2 Cl2 phase was washed twice with water then dried over Na2 SO4. Filtration followed by addition of ligroin, and solvent removal gave a yellow solid. Recrystallization from cyclohexane gave 17 as an off-white solid (7.2 g, 46%).
Anal. Calcd. for C10 H11 NO4 : C, 57.4; H, 5.3; N, 6.7. Found: C, 57.2; H, 5.3; N, 6.7.
Compound 17 (6.1 g, 29.2 mmol) was refluxed in 100 mL of 3N HCl for 2 hours under N2. The resulting solution was treated with NaHCO3 to pH 2 then extracted 3 times with CH2 Cl2. The combined organic phase was dried over Na2 SO4, filtered, and evaporated to give 4.4 g of 18 as an off-white powder (91%).
Anal. Calcd. for C8 H7 NO3 : C, 58.2; H, 4.3; N, 8.5. Found: C, 58.0; H, 4.3; N, 8.4.
Compound 18 (0.50 g, 3.0 mmol) was refluxed with 2-furaldehyde (0.29 g, 3.0 mmol) in 25 mL of MeOH with 0.2 g of KOH under N2 for 1.5 hours. The solution was poured into 100 mL of aqueous NaHCO3 and extracted with CH2 Cl2 which was discarded. The aqueous phase was acidified and extracted twice with CH2 Cl2. The organic phase was worked up by the procedure used for compound 18 to give chalcone 19 as a light green foam which was pure enough to continue with.
Chalcone 19 (0.60 g, 2.5 mmol) and 2 (0.71 g, 1.2 mmol) were combined and refluxed together with 5 g of NH4 OAc in 100 mL of MeOH for 16 hours. The solution was cooled and filtered and the resulting solid was worked up by the procedure used for the terpyridine diacid 3 in which R10 is --OCH3, to give 20 as a tan powder (0.40 g 53%).
Anal. Calcd. for C35 H21 N5 O6,•1/2 H2 O: C, 68.2; H, 3.6; N, 11.4. Found: C, 68.3; H, 3.5; N, 11.5.
A 10-6 M solution of the quinquepyridine 20 and EuCl3 •6H2 O was highly luminescent under long wavelength UV light.
Stock 10-5 m solutions of terpyridine. Eu+3 complexes were prepared by weighing the appropriate amount of the terpyridine into a 1 l volumetric flask; adding 1.00 mL of a stock 10-2 M EuCl3 solution and then diluting with pH 10 borate buffer. For luminescence vs. concentration studies, the dilutions were carried out with 10-5 M Eu+3 solutions in pH 10 borate buffer. The constant Eu+3 concentration forces the terpyridine to remain complexed at low concentrations (Ka=about 5�106 M-1).
Solutions of the quaterpyridine diacid Eu+3 complexes were prepared so that the Eu+3 concentration was maintained at 10� the concentration of the quaterpyridine diacid.
For solutions of the terpyridine tetraacid•Eu+3 chelates, the Eu+3 concentration was maintained at twice the ligand concentration.
The luminescence properties of the chelates are presented in Tables 2, 3 and 4.
TABLE 2__________________________________________________________________________Luminescence Properties of the Ter- andQuaterpyridine Eu+3 ChelatesaCompound ε (M-1 cm-1)* Φ*** Irel**__________________________________________________________________________Table I, compound 1; 13,000 (340 nm) 0.08 9.11R4 = --CH3Table I, compound 1; 16,000 (340 nm) 0.07 12.5R4 = --OCH3Example 2, compound 12a; 8,500 (350 nm) 0.16 21.5R11 = --CH3Example 2, compound 12b; 16,000 (350 nm) 0.14 35.7R11 = --OCH3Example 1, compound 15**** 14,000 (339 nm) -- 18.5__________________________________________________________________________ a Diacid = 10-6 M, [Eu+3 ] = 10-5 M. *Refers to the extinction coefficients. **Refers to relative luminescence intensities. **Refers to the relative emission quantum efficiencies. ****Contains some diacid impurity.
TABLE 3______________________________________Luminescence vs. Concentration for theEu+3 Chelate with Compound 1 ofTable I (R4 = --CH3)Concentration Luminesenceof Chelate (M) pH at 614 nm______________________________________10-6 10 11.465 � 10-7 10 7.4110-7 10 0.865 � 10-8 10 0.353 � 10-8 10 0.2310-8 10 0.075 � 10-9 10 0.03______________________________________
TABLE 4______________________________________Luminescence vs. Concentration for theQuaterpyridine � Eu+3 Chelate(Example 2, Compound 12b; R11 = --OCH3)Concentration Luminesenceof Chelate pH at 616 nm______________________________________10-5 10 36510-6 10 35.664 � 10-7 10 14.262 � 10-7 10 6.4610-7 10 2.614 � 10-8 10 0.982 � 10-8 10 0.4210-8 10 0.244 � 10-9 10 0.09______________________________________
A series of phenanthroline chelating agents were prepared using the following series of reaction schemes. In the chemical structures tBu refers to tertiary butyl. ##STR20##
A mixture of 6.3 g (0.03 mol) of 2,9-dimethyl-1,10-phenanthroline, 6 mL of 30% H2 O2 and 30 mL of trifluoroacetic acid (TFA) was refluxed for 3 hours. The reaction was followed by 1 H NMR analysis of the reaction mixture. The volatiles were removed using a rotary evaporator initially and then a vacuum pump to yield a mono N-oxide isolated as a golden syrup that crystallized from Et2 O as the TFA salt. 1 H NMR (CF3 CO2 H) 2.9 (s, 3H); 3.1(s, 3H); 7.95 (ABq, 1H); 8.1 (ABq, 1H): 8.2 (s, 2H); 8.35 (ABq,1H); 8.9 (ABq,1H).
The N-oxide was dissolved in 90 mL of Ac2 O and then added over 20 min to 250 mL of refluxing Ac2 O. Ac2 O is acetic anhydride. After refluxing for an additional 30 minutes, the dark solution was cooled and the Ac2 O removed in vacuo. The crude residue was purified by chromatography on silica gel using CH2 Cl2 to elute 2-acetoxymethyl-9-methyl-1,10-phenanthroline (21). 1 H NMR (CDCl3) 2.06 (s, 3H); 3.82 (s, 3H); 5.53 (s, 2H); 7.4 (ABq, 1H J=9 Hz); 7.57 (ABq, 1H, J=9 Hz); 7.62 (s, 2H); 8.0 (Abq, 1H, J=9 Hz); 8.13 (ABq, 1H, J=9 Hz).
The acetate (21) was dissolved in 60 mL of 31% HBr in HoAc and refluxed for 4 hours. After cooling, all the solvent was removed in vacuo. The resulting black residue was suspended in a mixture of CH2 Cl2 and dilute aqueous NaHCO3. Four 200 mL CH2 Cl2 extracts removed the desired bromide from the insoluble material floating at the solvent interface. After drying over Na2 SO4 and removal of the solvent, the dark residual solid was relatively pure phenanthroline (22) containing a trace amount of 2,9-dimethyl-1,10-phenanthroline. The dark impurities were removed by chromatography on silica gel using CH2 Cl2 as the eluent which gave 3.5 g (42%) of 22. 1 H NMR (CDCl3) 2.9 (s, 3H); 4.94 (s, 2H); 7.43 (ABq, 1H, J=9 Hz), 7.6 (s, 2H); 7.8 (ABq, 1H, J=9 Hz); 7.97 (ABq, J=9 Hz); 8.1 (ABq, 1H, J=9 Hz).
A mixture of 5.0 g (38 mmol) of iminodiacetic acid, 4.5 mL (50 mmol) of HClO4 and 100 mL of tert-butyl acetate was stirred for about 20 minutes at 20� C. until the reaction became homogeneous. After standing for 3 days, the solution was poured into 10% aqueous NaHCO3 and extracted three times with 50 mL portions of CH2 Cl2. After drying over Na2 SO4 and removal of solvents using an aspirator, the resulting oil was a mixture of di-tert-butyl iminodiacetate and tert-butyl acetate. The crude product was dissolved in 50 mL of CH2 Cl2 and extracted at 0� C. with 100 mL of 6 M HCl; the aqueous phase was extracted twice with CH2 Cl2 and then poured into ice water saturated with NaHCO3. The resulting alkaline solution was extracted three times with CH2 Cl2. After drying over Na2 SO4, the volatiles were removed to yield 26, isolated as a yellow oil (2.5 g, 27%) which upon standing at -5� C. solidified, 1 H-NMR (CDCl3 1.4 (s, 18H); 2.0 (s, 1H); 3.23 (s,4H). Mass spec m/e 245 (M+), 133.
A mixture of 900 mg (3.6 mmol) of di-tert-butyl iminodiacetate, 800 mg (2.8 mmol) of bromide (22) and 0.5 g of diisopropylethylamine were stirred in 8 mL of dry DMF at 25� C. for 45 hours. The solution was poured into dilute NaHCO3 in deionized H2 O and extracted 3 times with CH2 Cl2. After drying over Na2 SO4 and solvent removal, by 1 H NMR the residue was a 1:1 mixture of 23 and di-tert-butyl iminodiacetate. The material was chromatographed on florisil; a mixture of 40 percent CH2 Cl2 in cyclohexane was used to elute the di-tert-butyl iminodiacetate followed by a mixture of the diacetate and 23; CH2 Cl2 eluted 200 mg of pure 23. 1 H NMR (CDCl3) 1.3 (s, 9H); 2.7 (s, 3H);3.35 (s, 4H); 4.3 (s, 2H); 7.4 (ABq, 1H); 7.5 (s, 2H); 7.6 (ABq, 2H); 8.05 (ABq, 1H); 8.05 (m, 2H). Mass spec (field desorption) m/e 451 (M+).
A mixture of 200 mg of 23 and 5 mL of trifluoroacetic acid was allowed to stand at 20� C. for 1 hour. In order to monitor the reaction by NMR the solvent was removed under vacuum. A 1 H NMR spectrum of the residue residue redissolved in TFA revealed the t-butyl esters had been removed.
A mixture of 6 g (0.03 mmol) of tert-butyl bromoacetate in 15 mL of CH3 CN was added over 30 minutes to a 20� C. stirred solution of 30 g of 2-aminoethanol in 30 mL of CH3 CN. After stirring for 3 hours, the solvent was removed using a rotary evaporator.
The residue was poured into H2 O and extracted six time with CH2 Cl2. After drying over Na2 SO4 and removal of the CH2 Cl2, 3.1 g (60%) of 10 was obtained as a water-white liquid. 1 H NMR 1.35 (s, 9H); 2.1 (broad s, 1H); 2.67 (t, 2H); 3.18 (s, 2H); 3.5 (t, 2H).
A solution of 3.1 g (18 mmol) of the glycinate from Part A, 4.0 g (11 mmol) of bromide 22 and 2.5 g (19 mmol) of diisopropylethylamine in 15 mL of CH2 Cl2 was stirred for 24 hours at 30� C. The solution was poured into dilute NaHCO3 in deionized H2 O and extracted 3 times with CH2 Cl2. After drying over Na2 SO4 and removal of the solvent, 3 g of residue was obtained which was essentially clean 24. Chromatography on florisil required 1-10% MeOH/CH2 Cl2 to elute pure 24. 1 H NMR (CDCl3) 1.44 (s, 9H); 2.83 (s, 3H); 2.94 (t, 2H); 3.4 (s, 2H); 3.6 (t, 2H); 7.4 (ABq, 1H); 7.6 (s, 2H) 7.7 (ABq, 1H); 8.0 (ABq, 1H); 8.1 (ABq, 1H).
To a mixture of 100 mg of 24 and 0.3 g of triethylamine in 2 mL of CH2 Cl2 was added 0.2 g of propionyl chloride. The reaction mixture was stirred for 15 hours, poured into dilute NaHCO3 in deionized H2 O and extracted 3 times with CH2 Cl2. After drying over Na2 SO4 and removal of the solvent, the residue was chromatographed on florisil using CH2 Cl2 to elute 110 mg of 25. 1 H NMR (CDCl3 ) 1.03 (t, 3H); 1.43 (s, 9H); 2.3 (q, 2H); 2.87 (s, 3H); 3.0 (t, 2H); 3.35 (s, 2H); 4.16 (t, 2H); 4.33 (t, 2H); 7.43 (ABq, 1H, J=9 Hz); 7.65 (s, 2H); 7.97 (ABq, 1H); 8.03 (ABq, 1H); 8.13 (ABq, 1h). Mass spec (FD) m/e 437.
As described by C. J. Chandler, L. W. Deady, and J. A. Reiss, J. Heterocyclic Chem., 18, 599 (1981), a mixture of 9.0 g (43.2 mmol) of 2,9-dimethylphenanthroline and 22.5 g (0.2 mol) of selenium dioxide were refluxed in 600 mL of dioxane containing 4% H2 O for 2.5 hours. The hot dark solution was filtered through Celite�, cooled, and the resulting solid was collected by filtration. After air drying, 8.0 g (80%) of 26 isolated as a yellow solid was obtained. 1 H NMR (DMSO-d6) 8.2 (s, 2H); 8.32 (ABq, 2H); 8.7 (ABq, 2H) 10.35 (s, 2H).
To a mixture of 2.0 g of dialdehyde 26 in 40 mL of DMF and 5 Ml of ethanol was added 800 mg of NaBH4. The solution was stirred 6 hours at 30� C. then 5 mL of acetone was added. The solvent was removed under vacuum and the residue recrystallized from 50 mL of H2 O to yield 1.7 g of diol (27). 1 H NMR (DMSO-d6) 4.7 (s, 4H); 5.45 (broad s, 2H); 7.6 (ABq, 2H); 7.7 (s, 2H); 8.25 (ABq, 2H).
A mixture of 250 mg of diol 27 in 10 mL of 31% HBr in HOAc was refluxed for 3 hours. The solvent of the resulting orange solution was removed in vacuo and the residue was taken up in CH2 Cl2 and dilute aqueous NaHCO3. Due to low solubility, 3 CH2 Cl2 extractions were necessary to leach all the dibromide 28 from the insoluble material. After drying over Na2 SO4 and removal of the solvent, 220 mg (˜60%) of 28 was obtained as a dark solid which was pure by NMR. Chromatography on silica gel using CH2 Cl2 eluted 28 as a white solid. 1 H NMR (CDCl3) 4.87 (s, 4H); 7.63 (s, 2H); 7.83 (ABq, 2H, J=8 Hz); 8.13 (ABq, 2H, J=8 Hz).
A mixture of 220 mg (0.57 mmol) of dibromide 28, 400 mg (1.6 mmol) of di-tert-butyl iminodiacetate, 0.5 g of diisopropylethylamine in 5 mL of 1:1 CH2 Cl2 /CH3 CN was stirred for 15 hours at 25� C. The solution was poured into deionized H2 O/Na2 HCO3 and extracted three times with CH2 Cl2. After drying over Na2 SO4 and solvent removal, the residue was chromatographed on florisil. Residual iminodiacetate and tetraester were eluted with 20-50% CH2 Cl2 in cyclohexane; 5% MeOH/CH2 Cl2 eluted 120 mg of clean 32. 1 H NMR (CDCl3) 1.30 (s, 18H); 3.42 (s, 8H); 4.23 (s, 2H); 4.33 (s, 2H); 7.60 (s, 2H); 8.05 (ABq, 4H).
Following the procedure of Chandler et al, referred to in Example 10, a solution of 0.6 g (2.5 mmol) of bisaldehyde 26 in 12 mL of 80% nitric acid was refluxed for 3 hours. Upon cooling, the reaction was poured onto ice and the diacid 29 was collected by filtration. Upon air drying, 0.5 g (75%) of 18 was isolated 1 H-NMR (DMSO-d6) 8.20 (s, 2H); 8.45 (ABq, 2H); 8.75 (ABq, 2H).
A mixture of 0.5 g of dicarboxylic acid 29 was refluxed in 5 mL of thionyl chloride for 5 hours. The suspension slowly went into solution as the reaction proceeded. The volatiles were removed using a rotary evaporator; the residue was twice dissolved in benzene and stripped to dryness.
1 H NMR (CDCl3) 8.05 (s, 2H); 8.45 (s, 4H).
To a 20� C. mixture of 400 mg (4 mmol) of Et3 N and 0.5 g (3 mmol) of tert-butyl N-(2-hydroxyethyl)glycinate in 3 mL of CH2 Cl2 was added 0.3 g (1 mmol) of the bisacid chloride 30. The solution was stirred overnight, poured into Na2 HCO3 /H2 O, and extracted three times with CH2 Cl2. After drying over Na2 SO4 and removal of solvent, the residue was chromatographed on florisil. The product came off mainly with CH2 Cl2 -5% MeOH/CH2 Cl2 but was contaminated with residual glycinate. This material was slurried in 4 mL of Et2 O, 1 mL of petroleum ether was added and the solution decanted from the clean product 20. 1 H NMR (CDCl3) 1.13 (s, 9H); 1.4 (s, 9H); 3.79 (m, 8H); 4.2 (s, 2H); 4.3 (s, 2H); 7.83 (s, 2H); 7.87 (ABq, 1H); 8.35 (m, 3H). Mass spec (FD) m/e 582.
A mixture of 3.0 g (11.2 mmol) of diacid 29 and 100 mL of thionyl chloride was refluxed for 16 hours, cooled, and stripped under vacuum. The crude bis(acid chloride) 30, without further purification, was suspended in 200 mL of dry THF to which was added 6.03 g (24.6 mmol) of di-t-butyl iminodiacetate, followed by 3.4 mL (24.6 mmol) of triethylamine. After stirring at 200� C. for 2 hours, the solution was filtered. The filtrate, after concentration, was chromatographed on florisil using a gradient from 25% EtOAc/CH2 Cl2 -EtOAc to elute 3 g (37%) of 33 as a clean oil which formed a stiff foam. 1 H NMR (CDCl3) 1.13 (s, 18H); 1.53 (s, 18H); 4.37 (s, 4H); 5.07 (s, 4H); 7.8 (s, 2H); 8.17 (ABq, 2H, J=8 Hz); 8.27 (ABq, 2H, J=8 Hz). Mass spec (FD) m/e 723 (M+1).
Anal. Calcd. for C38 H50 N4 O10 : C, 63.1; H, 7.0; N, 7.8. Found: C, 62.8; H, 7.0; N, 7.9.
A mixture of 800 mg (1.1 mmol) of 21 in 25 mL of TFA stood for 1 hour at 20� C. Upon removal of the solvent under vacuum and washing of the residue with Et2 O, 0.54 g of product (98%) was obtained, isolated as a white powder. 1 H NMR (CDCl3 /DMSO-d6) 4.32 (s, 4H); 4.84 (s, 4H); 8.05 (ABq, 2H, J=8 Hz); 8.1 (s, 2H); 8.57 (ABq, 2H, J=8 Hz). Anal Calcd. for C22 H18 N4 O10 1/2 H2 O: C, 52.1; H, 3.8; N, 11.0. Found: C, 52.1; H, 3.9; N, 11.0.
A mixture of 200 mg (0.9 mmol) of 9,10-phenanthroline-2-carboxylic acid in 10 mL of thionyl chloride was refluxed for 6 hours. Although the reaction never became homogeneous, the volatiles were removed using a rotary evaporator. The residue was suspended in benzene and stripped to dryness. The crude product, dissolved in 10 mL of CH2 Cl2, was combined with 240 mg (1 mmol) of di-tert-butyl iminodiacetate followed by 300 mg of triethylamine. After stirring at 30� C. for 24 hours, the reaction was poured into NaHCO3 /deionized H2 O and extracted three times with CH2 Cl2, dried over Na2 SO4 and evaporated. The residue was chromatographed on florisil using 1-5% MeOH/CH2 Cl2 to elute 100 mg (24%) of the desired amide 38. 1 H NMR (CDCl3) 1.1 (s, 9H); 1.37 (s, 9H); 4.2 (s, 2H); 4.63 (s, 2H); 7.5 (d, 1H); 7.65 (s, 2H); 8.0 (d, 1H); 8.03 (d, 1H); 8.13 (d, 1H); 9.0 (m, 1H). Mass spec (FD) m/e 451.
Hydrolysis proceeded quantitatively by the procedure used for compound 39, Table 5, to give diacid compound 44, Table 5. Other similarly produced compounds are also presented in Table 5.
TABLE 5______________________________________Chelating Agents______________________________________ ##STR21## 39 ##STR22## 40 ##STR23## 41 ##STR24## 42 ##STR25## 43 ##STR26## 44______________________________________
Stock 10-4 M solutions of the phenanthroline chelating agents were prepared by weighing the appropriate amounts into 1L volumetric flasks, adding 10 mL of 10-2 m EuCl3 solution and diluting with pH 10 borate buffer. In the case of compound 43, Table 5, the solution reached constant luminescence immediately. With compound 39, Table 5, it was necessary to allow the solution to stand for eight days to reach constant luminescence. Relative quantum efficiencies (Φ), absorption maxima and luminescence of the chelates prepared in this example are presented in Tables 6 and 7.
TABLE 6______________________________________Quantum Efficiencies and Absorption Maxima for SomeEu+3 Chelates in pH 10 Borate Buffer ΦEu+3 Chelate λmax______________________________________ ##STR27## 0.02 29139, TABLE 5 ##STR28## 0.04 27742, TABLE 5 ##STR29## 0.06 27743, TABLE 5______________________________________
TABLE 7______________________________________Luminescence vs. Concentration for the Eu+3Chelates of Phenanthroline Tetraacids 39 and 43Tab1e IIChelating Agent -Log[L]a Log Ib______________________________________39 4.1 -0.4839 5.2 -1.5043 4.2 1.3643 5.2 0.4243 6.2 -0.6043 7.2 -1.52______________________________________ a [Eu+3 ] = [L]- b λexcitation 330 nm, λemission 614 nm in pH 8.5 borat buffer
The invention has been described in detail with particular reference to preferred embodiments thereof, but it will be understood that variations and modifications can be effected within the spirit and scope of the invention.
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|International Classification
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text:
In the garden
By Patty Campbell, Special to the Citizen 06/18/12
Tomatoes, say the folks at the Burpee Seed Company, are America's favorite vegetable. Fresh, vine-ripened tomatoes are delicious. The lycopene they contain is good for your heart. And, provided they are properly cared for, a standard tomato plant can yield 10 to 15 pounds or more of fruit.
In Central Virginia tomatoes are usually transplanted into the garden in early May, but even now you have time to plant tomatoes since the growing season stretches into early October.
Once tomato plants are established in the garden, good care involves keeping the plant's foliage off the ground, fertilizing, and providing adequate water consistently.
Even if you've staked or caged your tomatoes, once plants are about three feet tall you may need to remove all the growth from the bottom sin to ten inches. This improves air circulation and helps fight diseases such as early blight.
And you may want to sucker your plants. Suckering involves pinching out the shoots that develop in the crotch joints of branches. They don't produce fruit and can take energy from the rest of the plant. According to the Extension Service, gardeners using stakes usually sucker, those using cages don't.
And now that the ground has warmed up, apply a layer of mulch around your plants.
"Mulching is a very good cultural practice," says Henrico Extension Agent Lisa Sanderson. "Mulching helps keep soil that can contain pathogens detrimental to the plant from splashing up on it, reduces weeds, and helps ensure adequate moisture." Shredded newspaper covered with grass clippings makes good mulch that will decay over the growing season and can be tilled in to the soil in the fall, adding organic matter.
Tomatoes need moisture. While plants are developing, water them deeply and regularly, up to twice weekly depending on rain. Watering early in the day and using soaker-hose or drip irrigation saves water and helps ensure water won't contribute to fungus problems.
When plants start setting fruit, inconsistent or inadequate watering can cause tomatoes to crack or get blossom end rot, a dark spot at the blossom end of the fruit that can cover half the tomato. Blossom end rot can also result from a calcium deficiency.
Tomatoes often require fertilizer. "We recommend fertilizing in response to a soil test report," says Sanderson. She recommends incorporating organic matter and a fertilizer when you plant your tomatoes.
"Read the label and follow the instructions," says Sanderson. "Over-fertilizing doesn't benefit the plant, and it contaminates groundwater."
A number of different diseases can affect your plants, but many of the hybrids offer resistance to some of these diseases. Good pictures are available on the web to help you identify problems. Call the Master Gardener Helpline at 501-5160 to discuss possible remedies which can include treatment or eradication.
Spider mites, stinkbugs and beetles can also cause problems. Before using chemical pesticides, check with the Extension Office for current recommendations.
Some bugs, like the tomato hornworm, can be removed by hand. "But don't remove a hornworm that has white cocoons on its back," says Sanderson. "Those are the pupa of brachonid wasps that have already eaten the hornworm on the inside, and the wasps are beneficial insects in your garden!"
FirstComp Insurance Company, Markel Corp. named top insurance carriers
Citizen Staff Reports 07/26/2017 BusinessGeneral News
Two Glen Allen-based insurance carriers have been named among the top such companies in the nation.
Markel Corp. and FirstComp Insurance Company (now owned by Markel) are among 38 carriers recognized on the Insurance Business America list of the top insurance carriers in the country. > Read more.
Walmart honors Citizen's 'Top Teachers' during grand opening
Citizen Staff Reports 07/25/2017 BusinessEducationGeneral News
The much-anticipated new Walmart Supercenter in Eastern Henrico opened to busy crowds last week, and company officials welcomed a special group of shoppers, too. > Read more.
'It's such a blessing'
By Ashlee Korlach, Special to the Citizen 07/24/2017 FeaturesGeneral News
On Independence Day, a Brazilian-born Henrico resident took the Naturalization Oath of Allegiance on the steps of the Virginia Historical Society and felt peace finally settle around her.
Jeanette Hamlett's journey to U.S. citizenship is a story of faith and love, which she told with her husband, John Hamlett, by her side. > Read more.
Preschoolers give pillows to families in need
By Ashley Luck, Special to the Citizen 07/24/2017 FeaturesGeneral News
A group of preschoolers from Chesterbrook Academy Preschool donated 56 pillows to local families in need at the Housing Families First shelter. The shelter is always in need of pillows and linens as, since families take the donated ones with them to their new homes.
Housing Families First provides shelter and support for homeless families and assist them in finding a permanent housing solution. The shelter serves people year-round, said Terri Iguina, operations and volunteer manager at Housing Families First. > Read more.
Dairy Queen's Blizzard sales July 27 to benefit Children's Hospital of Richmond
Citizen Staff Reports 07/24/2017 BusinessGeneral News
Dairy Queen's 13th Annual Miracle Treat Day – Thursday, July 27 – will raise fund to benefit sick and injured children being treated at Children's Miracle Network hospitals throughout the United States. Locally, $1 or more from every Blizzard Treat sold at participating locations will be donated to the Children's Hospital of Richmond at VCU. Last year, the event raised more than $14,400 for the hospital. > Read more.
Dairy Queen's Blizzard sales July 27 to benefit Children's Hospital of Richmond
Crime Stoppers' Crime of the Week: July 24, 2017
Henrico Police locate missing man
Business in brief
Mother, son work to raise funds for Make-A-Wish
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Jul. 20, 2017Current edition | Archive
Restaurant Watch
Weekend Top 10
'Five Flights' opens July 20 at CAT Theatre
Henrico actress lands role in Christian TV series
All's fair in food and fun
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Obituary – Hoyle S. "Chick" Walters
Obituary – Janet Sue Vest
Obituary – Barbara Jean (Chilton) Crider
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Glen Allen woman raising companion dog
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CalendarWoody Woodworth & The Rebel City will perform at 8 p.m. at The Tin Pan, 8982 Quioccasin Rd. Woodworth is a singer-songwriter that hails from the Rebel City of Richmond, Va. His Americana style is a melting pot of alt-country, rock and roll, bluegrass, folk and blues. Also performing is Drew Gibson, who released his debut album "Letterbox" in 2007. The self-taught finger-style guitarist has garnered attention for his intensely personal songs and his stagecraft as a live performing storyteller. His most recent album is "1532." Doors open at 6 p.m. Tickets are $10. For details, call 447-8189 or visit http://www.tinpanrva.com. Full text

subset: commoncrawl


text:
How to Use deceptive shooting by crunching the depth of field
Mark and Andrew demonstrate how to crunch the depth of field to achieve two camera tricks. By crunching the field of view it makes the action closer than it really is. Often used when filming fight scenes or being "hit by a car" scene.


text:
Surya Maeva Rug collection
Surya Maeva Rug collection
We love all 4 rug in the Surya Maeva Rug collection. Surya did an excellent job of putting together a grouping of beautiful and stylish rug that are inviting and comfortable using today's fashionable color palettes. The 60% Wool, 40% Viscose material and 1/4 inch thickness give charm and comfort to any hard surface floor.
Considering free shipping and a 30-day return policy, the Surya Maeva Rug collection is irresistable.
Incredible Rugs and Decor is currently rated...


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Cool. Classic. Coastal. Fun and funky .925 sterling silver jewelry with a little beach attitude!
We showcase handpicked artisan-crafted designs from around the world with a focus on designs from the United States, Mexico, Thailand, Dominican Republic and Israel.
Classic silver designs are featured along with unique one-of-a-kind items made from natural stones and gemstones. Equestrian, animals, nature & coastal-inspired collections.
Born from a love of the ocean and turquoise water
Are YOU a Silvergirl?
If you're a fan of fine sterling silver jewelry then you are going to LOVE our collection. Take a look and you will find unique, wearable art from around the world featuring .925 sterling with natural stones, gemstones and freshwater pearls. We feature the classic sterling pieces that you expect alongside cool, new designs in beach, sea life, nautical collections and much more!
Silvergirl Sterling proudly features jewelry from Israel, Bali, Indonesia, Peru, India, Mexico, Thailand and the Dominican Republic and talented artisans from right here in the United States. Come see us in person in the Baltimore/Washington metro area at a local event or shop online.
Be sure to LIKE us on Facebook and join our email list for all the latest news and specials...
Shops and Boutiques
The Silvergirl Sterling is featured in fine boutiques in the United States and throughout the Caribbean. After successful gift shows in January and July in Atlanta and Orlando, we had a fun detour to visit shops (existing and new) on the Jersey Shore. It has been great meeting so many of you in person to introduce our collection and you will be seeing more loaded on the site every week.
Are you a shop owner that would like to carry our collection? Visit our Wholesale page for information on registering as a wholesale account.
Silvergirl on Social Media
Be sure to visit Silvergirl Sterling on social media! You don't want to miss new product announcements, events, specials and pop-up sales!


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1225 Town Center Dr, Pflugerville, TX 78660
- Today
Regular Hours
- Mon - Fri
10:00 am - 6:00 pm
- Sat
10:00 am - 5:00 pm
- Sun
More info
Extra Phones
Primary Phone: (512) 990-2686
Phone: (512) 251-2450
Primary Phone: (512) 251-2450
Primary Phone: (512) 252-8283
Phone: (512) 252-8283
Phone: (512) 990-2686
TollFree: (855) 887-6796
Other Information
Parking: Lot, Free
Wheelchair Accessible: Yes
Add Your Review
I am not happy with the way that they run the office... My brother and I were going to rent from here with the assumption that our mom was going to cosign for us... and we told them this from the get go. We filled out the application and paid the admin fee of $150 since the app fees were waived. They then had us sign the lease agreement for a move in date of the 28 of January. We told them that our mom was out of town until Sunday night and that we would talk to her about it when she got home. they told us that was fine and should anything happen to give them a call. Well mom didn't feel comfortable with the length and amount of the rent. so we called them first thing this morning the 27th and we were told that they could give us and 8 month lease at the same price. Mom still said she would rather not cosign. We then called them to tell them this and that without mom cosigning we would NOT qualify with out her. They then tell us that they have still not received any of our apps back, and if they came back and we did qualify there is no way for us to get out of this lease (they should not have made us sign the lease without us being approved). We then started rereading all of our papers, and found out that if we don't move in on the 28th it counts as a "unlawful early move out" and we would have to pay a fine. How can we unlawfully move out if we haven't even been approved to move in yet? We are supposed to move in tomorrow but after everything that has happened this is not a place that I am wanting to live at all... They are all very money hungry and I i wouldn't recommend any one move here.
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Parquet Wood Plank Natural Wallpaper
- Recommended for bedroom, hallway & living areas
- Paste the paper
- Covers minor cracks & imperfections
- Peelable
- Wipe clean with a damp cloth to remove marks caused by everyday life
- Available in-store only
Product info
Features and benefits
This statement wallpaper gives the effect off parquet wooden planks. Ideal as a conversation starter, or for use on all four walls to create a rustic effect in your chosen space.
Tech specs
|Colour description :
|Style :
|Brick, tile & natural
|Range :
|Parquet wood plank
|Surface finish :
|Texture :
|Sizes :
|(H)10m x (W)520mm
|Coverage :
|Up to 5.2 m²
|Application :
|Paste the paper
|Pattern repeat :
|Straight match every 53 cm
|Brand :
|Fine Decor


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Tuesday, August 25, 2009
The Search Agency Ties With CyberDefender
Santa Monica-based The Search Agency, a provider of search engine optimization, search engine marketing, and other related online marketing services, has signed on Los Angeles antivirus software company CyberDefender as a customer. Financial details of the deal were not disclosed. According to CyberDefender, The Search Agency will manage the company's search marketing campaigns, including search engine marketing and search engine optimization. The Search Agency is backed by private equity firm JMI Equity.


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the Foundation University Islamabad NTS Admission Test 2015 Roll no. slip. Foundation University Islamabad NTS Admission Test Date is Saturday, 23rd January 2016.
Now all candidates can check or download their Foundation University Islamabad NTS Admission Test 2016 roll no. slip Download Online from the official website of NTS or website www.pakresult.com So please stay with us & keep visiting our our web site & Join us on our Facebook page for further Information.
NTS Test Date: Saturday, 23rd January 2016.
About FUI:
The FUI's mission is to inspire creative inquiry and research to foster personal and professional development of its students. The FUI is committed to provide equitable access to holistic education in diverse disciplines to produce valuable human resource for the local and the global communities.


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Subscribe to our notifications so you don't miss out on our new sales!
Barbie 120x150cm A Fashion Fairytale Polar Fleece Throw
Keep the kids warm & cozy with this gorgeous throw for $14.99!
120cm x 150cm size | Perfect for jazzing up their bedroom decor!
Getting your kids to hit the hay is a cinch with this gorgeous polar fleece throw! Featuring a fun print that's bound to add a splash of colour to any room, it's perfect for cuddling up with whilst watching a spot of telly or reading bedtime stories.
- Kids' bedding
- Style: Barbie A Fashion Fairytale Polar Fleece Throw
- Colour: Pink
- Dimensions (approx. cm): 120 x 150
- Materials: Polyester
- Machine washable
- Country of origin: China


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Mrs Dunderhi plans to stay home next time...
View Single Post
Officially Welcomed to the 'Fest
Location: in a van down by the river
Join Date: Sep 2007
Posts: 3,676
Mein Auto: 997TT/335xi/R500
View My Garage
I cross shopped the R8, F430, and the Gallardo before deciding on the 997 911 Turbo. If you can, I would highly recommend the Turbo. Best car I have ever owned, hands down. Unfortunately with 3 small kids and the role as family man, it was time to move to a more family friendly ride, but I miss my 911 every day. I told my wife that I am buying a 997.2 Turbo S on my fortieth birthday. She agreed, but it's going to be a long four year wait.
Current: 2015 Nissan GTR Premium, 1973 911 with 930 Motor (340 hp 2200 lbs), 2016 Lexus LX570
Gone but not forgotten: 2012 M-Sport 550i X-drive Dinan Stage 2, 2007 911 Turbo, 2005 911 Carrera S, 07 BMW M Coupe, 07 335 XI, 05 Mercedes R500
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Add getz to Your Ignore List


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Main Menu / Inline Skates
Main Menu / Roller Skates
Main Menu / Scooters
Main Menu / Ice Skates
Main Menu / Protective
$499.95 $399.95
$499.95 $399.95
The Hybrid boot combines the classic look of Bont's 80's quad boots with all the modern advancements in boot technology such as the use of heat mouldable resins and new layup techniques.
The Hybrid Fibreglass used a heat mouldable fibreglass base with a long lasting 1.5mm Australian leather outer skin and powerstrap to hold is boot securely to your foot.
We carry a huge stock of Bont boots, but it isn't possible to carry all colours in all sizes, all the time. We will confirm availability within 24 hours of placing your order, offer options if your colour choice isn't available and confirm a delivery date if we have to order from Bont.


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If an item you ordered from TextbookRush does not meet your expectations due to an error on our part, simply fill out a return request and then return it by mail within 30 days of ordering it for a full refund of item cost.
Learn more about our returns policy
Description: If you were ill and doctors couldn't help you, would you have 20 years to wait before medical science discovered the true cause of your suffering and the way to get better? What would it be worth if someone else were able to help you recover and heal--now?Anthony William, the one and only Medical Medium®, has helped tens of thousands of people heal from ailments that have been misdiagnosed or wrongly treated or have baffled medical communities. He's accomplished all this by listening to a divine voice--called Spirit of the Most High--that literally speaks into his ear, telling him what is at the root of people's pain or illness and what they need to do to restore their health.Anthony has been called the Edgar Cayce of our time. He's also been compared to Padre Pio. What he does is several decades ahead of scientific discovery. His methods achieve spectacular results, even for those who have spent years and many thousands of dollars on all realms of medicine before turning to him. Now, in this revolutionary book, he gives readers access to all that he has learned in over 25 years of bringing people's lives back: a massive amount of healing information, much of which has never appeared anywhere before.Medical Medium reveals the root causes of diseases that medical communities either misunderstand or fail to understand at all, conditions that wreak havoc with the lives of countlesspeople around the world. And it explores all-natural solutions--not merely relief of symptoms, butcritical information that can address core causes and completely restore a person's health--fordozens of the illnesses that plague us, including Lyme disease, diabetes, adrenal fatigue,Candida, chronic fatigue syndrome, lupus, multiple sclerosis, anxiety, depression, ADHD andOCD, rheumatoid arthritis, fibromyalgia, shingles, and leaky gut syndrome. Beyond the body,Anthony brings his wisdom to bear on soul, heart, and spirit as well, offering readers uniqueinsights on how to find inner peace and live longer, healthier, richer lives.
Limited time offer:
Get the first one free! (?)
All the information you need in one place! Each Study Brief is a summary of one specific subject; facts, figures, and explanations to help you learn faster.
List price: $37.99Copyright year: 2015Publisher: Hay House, IncorporatedPublication date: 11/10/2015Binding: HardcoverPages: 392Size: 6.33" wide x 9.27" long x 1.31" tall Weight: 1.716Language: English
*A minimum purchase of $35 is required. Shipping is provided via FedEx SmartPost®
and FedEx Express Saver®. Average delivery time is 1 – 5 business days, but
is not guaranteed in that timeframe. Also allow 1 - 2 days for processing. Free shipping is eligible only in the continental United States and excludes
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orders are not eligible for free or discounted shipping.
Learn more about the TextbookRush


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How to find the best home improvement ideas for next summer?
May 18, 2016
Summer is a very hot season that people need to make sure that indoor ventilation and temperature regulation is perfect at all times. Below are some the ways on how you can get the best home improvement ideas for summer.
#1. Watch YouTube videos about summer home improvement.
This is the best way to ensure that you get exact procedures that would make you prepare your house for summer. It makes you to understand the cost, who to hire for your home improvement task and how best it can be done at all times.
#2. Read magazines
There are so many magazines and blogs online that talk about home improvement for the various seasons of the year. You can ensure that you grad one for yourself to find out the best home improvement that could be excellent for your home. The magazines does not have to be soft, you can subscribe to online magazines, they are not different and infect it would be easy for you to read them from your smartphone, desktops or IPad. Blogs can be followed to ensure that every idea that they post, you are there to learn what it says or talks about summer home improvements.
#3. Ask the experts directly.
You can decide to check the renovators and home designers. They have the perfect knowledge concerning all the home improvements methods at all times. Home interior designers are located in almost every town and they can assist a person to get the best ideas ever so that it becomes easy to pan for your home improvement. You can also consult with experts online, like Gareth from PortableACNerd, if you're planning to get an AC this summer. This is the best thing to ensure that you get typical plan for your home, which is going to directly improve your home to become more than luxurious.
#4. Google the keywords
There are so many websites dealing with home design which is a clear indication that there is information you can just copy from the websites.


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Magnificat to Perform Charpentier's Nativity Pastorale
Magnificat is pleased to announce performances of the Marc-Antoine Charpentier's Nativity Pastorale on the weekend of Dec. 7-9. The program will feature the Pastorale sur la naissance de Nostre Seigneur, one of Charpentier's most brilliant and moving works. In this exhilarating blend of dramatic dialogues and ensembles, instrumental dances, and exquisite choral writing, Charpentier's displays his imagination and technical mastery and his extreme sensitivity to poetic imagery. Magnificat's program will include Charpentier's settings of several of traditional French carols, or noëls, that are, by turns, charming, poignant, and amusing. Tickets are available online at magnificatbaroque.tix.com.
On a sabbatical after celebrating Magnificat's 20th anniversary season, Artistic Director Warren Stewart will return to lead an ensemble featuring singers Catherine Webster, Jennifer Paulino, Clara Rottsolk, Clifton Massey, Paul Elliott, and Peter Becker, together with Vicki Boeckman and Louise Carslake, recorder; Rob Diggins and Jolianne von Einem, violin; John Dornenburg, viola da gamba and Jillon Stoppels Dupree, harpsichord.
Read more…


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Your child may be growing faster than you can keep
track of, but his personality? That's pretty much set.
A new study published in the journal Social Psychological and
Personality Science has found that we tend to remain recognizably
the same person from childhood to adulthood. "Our personality does
seem to stay with us," says lead author Christopher Nave, a
doctoral candidate at the University of California Riverside.
Nave and his colleagues used data collected on
schoolchildren in the 1960s, where teachers had provided feedback
about the personalities of their students. More than 40 years
later, the researchers tracked down a large number of the original
participants to bring them in for a personality interview. "We saw
that the way teachers perceived their students predicted how they
were behaving decades later," Nave says.
And these findings may mean you can get a sneak peak into
your child's future. Children who were rated "verbally fluent,"
defined as unrestrained talkativeness, grew up to be dominant and
socially adept by the time they were middle-aged. "Adaptable"
students tended to become cheerful and curious, while "impulsive"
kids were later talkative and loud. And humble kids-those whom
teachers said had a tendency to "self-minimize"-were linked with
feelings of insecurity and humility as adults.
The findings, Nave says, mean that while people's
personalities can change over time, it may take more work than
previously thought. He suggests promoting a blend of different
behaviors in your children, as well as asking teachers for
feedback. While parents tend to promote being humble, for instance,
you may want to step in if the teacher says it's happening to an
extreme degree. On the other hand, talkative children-even to the
point of almost disturbing the class-tended to find appropriate
outlets and succeed in the long run.
See more of Laura's stories here.
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A collapsible promotional water bottle that comes with an aluminum foil coating and fun design.
Your logo will be a will noticed with this unique mood paw print custom pencil.
Your company logo will bloom as big as this fun novelty promotional pen.
Perfect when you need to promote on a budget, these custom closeout items will give your company the coverage you desire at great low costs.


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Denise Green was the line producer at HiT Entertainment for Hero of the Rails, Misty Island Rescue as well as the thirteenth and fournteenth seasons. She also worked alongside Ian McCue in the fifteenth season.
- She, Andrew Brenner, Oliver Davis, Marion Edwards and Salty's Lighthouse actor Ian James Corlett all worked on "The Cramp Twins".


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I am new to whitetail, and will be heading to southern illinois for the first week of december. I have heard that hunting funnels will probably be very affective. The only problem I have is trying to figure out where and what they are. I hunt mule deer and elk all the time, so I havn't really had to worry about it. Any info would be great.
- Home
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Valencia Cove, a new construction, 55 Plus luxury gated community in Boynton Beach, Florida. This new gated community is dedicated to the demands of the over 55 population with a myriad of clubs, activities and amenities. 18 models are available to choose from with 10 showcase decorated models. Kathleen Fineman and Seymour Fineman studied the options available to customize your Valencia Cove home. Valencia Cove has everything but golf & country club fees: lighted tennis and bocce courts, pickle ball court, 3 heated pools, luxurious spacious Clubhouse with 4 card rooms, indoor/outdoor cafe, internet/computer center, arts & crafts room complete with a kiln, jogging/running track, fitness center, massage room, Lifestyle Director, 822 homes with 2 to 3 car garages, gated entry, low HOA fees, and numerous social & entertainment activities. Valencia Cove is very close to the Delray Marketplace for shopping, dining and entertainment, convenient to the airports and minutes from Boca Raton and Delray Beach, Florida.
The Finemans know the community, met extensively with GL Homes and completed the entire selection, design and options program in depth with the GL Design Team. They've worked with purchasers including a family member whose Valencia Cove home is near completion. Kathy & Seymour are Valencia Cove experts who help in lot and design selection to provide their Buyers with optimal view, convenience and cost savings. They provide as much assistance & research as you'd like concerning color selections, options, layout and customization of your home while in the planning stages, as well as information about what's available outside of GL's standard options. Since GL Homes pays the commission, there is NO COST TO YOU as long as the Finemans are registered as your Realtor doing your initial visit to Valencia Cove. Kathy and Seymour Fineman can describe the options to you as you stroll through the model homes, such as what walls are removable, what's standard in the model and what's not.
If you're new to the area, our database of service providers from handymen, carpenters & tile installers to cleaning ladies and babysitters (for grandchildren or pets) will help you settle in quickly and effortlessly. Our family members have purchased in Valencia Cove, so we really know the community and the residents who call Valencia Cove home. The Finemans can arrange for you to see homes under construction and just completed so you know what to expect along the GL building journey.
The Finemans provide additional services new home buyers. We assist in lot & options selections, acquiring service people for contracted work outside of GL options, visit and photograph the house during the construction journey providing a record of the construction and a blueprint for future workmen, attend with or without you the various inspections such as pre-drywall and walk-throughs, and are with you each step of the way to answer or find the answer to each of your questions and concerns.
Call Kathy Fineman or Seymour Fineman, your Valencia Cove experts to tour this over 55 community, walk through model homes and discuss your options. Kathleen Fineman is an SRES, a Senior Real Estate Specialist as recognized by the National Association of Realtors and many Senior organizations. Her expertise is making a home comfortable for Seniors now and as they age in place. Remember, it's not only about quality of life now, but for the rest of your life ... and most of us prefer to age in place comfortably and safely.
Call Kathy Fineman # 917-709-4957 to arrange a time to view Valencia Cove now. The lots are going fast at Valencia Bay and Valencia Cove where the facilities are completed.


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One of the key aspects of bringing out a given wine's fullest potential is to serve it alongside a meal that excellently pairs with the wine's flavors and complexity. This element of wine consumption is just as important in Champagne enjoyment as it is with any other wine.
Pairing Champagne with cuisine involves careful choices and a basic knowledge of what flavors match and what flavors don't complement each other.
Champagne and Cuisine Matching Basics
When going forward with planning a meal you intend to serve with Champagne, first consider the characteristics of the wine itself.
Champagne is entirely capable of being a full-bodied wine, though it is not a "big" wine in the manner of certain red wines–Malbec, Zinfandel or Cabernert Sauvignon, for example. The dishes that red wines serve as an excellent complementary beverage to, such as red meat, hearty red-sauce pasta dishes and spicy foods, won't work with Champagne. The white wine's effervescence and floral, light fruity tastes will be overpowered by such foods.
Pick lighter dishes to complement a Champagne, such as various seafood items, chicken and poultry dishes, and pastas with lighter sauces or butter rather than heavy red, marinara-style sauces.


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The world is your oyster
Global experiences don't just add stamps to your passport. They also boost your resume.
Culture. Language. Religion. Climate. Laws. Currency. All of these affect how business is conducted in disparate parts of the world. That's why employers are looking for grads with the cultural competency and cross-cultural understanding needed to navigate foreign markets.
And Tippie gives you countless ways to acquire these skills. Apply for a global internship in one of 11 destinations. Spend two weeks studying international business in London or Beijing. Study leadership in Northern Italy for an entire semester. Or build lifelong global connections here on campus. We've got it all and it's yours for the taking.
We offer study abroad programs around the globe, from two weeks to two semesters long.
Get hands-on experience in an international workplace with an internship that is customized to your professional interest.
Find out what kind of cultures interest you without leaving PBB.
From Iowa to international
When he got to France, Torrance Windham (BBA15) took the metro into Paris. There were a lot of connections—very busy and all underground. He hadn't even seen the city yet, but there were a lot of people.
"I had no idea that Paris was going to look the way it did. Everything was amazing!"
Then he walked up the stairs of the Odeon metro stop, and he was taken aback.Read Story


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12 November 2008,
New York
Price realised USD 2,210,500
Andy Warhol (1928-1987)
Statue of Liberty
signed and dated 'Andy Warhol 86' (on the overlap)
synthetic polymer and silkscreen inks on canvas
72 x 72 in. (183 x 183 cm.)
Painted in 1986.
New York +1 212 636 2000
London +44 (0)20 7839 9060
Hong Kong +852 2760 1766
Shanghai +86 21 6355 1766
Robert Manley
Tel: +1 212 636 2381
In 1986, the year that marked the centennial celebration of the famous Statue of Liberty's arrival in the New York harbor, Andy Warhol turned the celebrated Lady Liberty into a majestic Pop icon. Already one of the central images of American culture, Warhol claimed this symbol of freedom and the American Dream as his own goddess of Pop.
Statue of Liberty is painted in Warhol's classic Pop style, using a silkscreen based on a photograph of the monument, cropping her face in a square close-up format, just has he had isolated famous silver screen idols such as Marilyn Monroe and Liz Taylor. The present painting's bright candy colors also evoke the glamor with which he teated these stars. He then superimposed this portrait with a colorful camouflage pattern that had become one of his signatures in the late 1980s. His interest in camouflage was in part linked to his interest in street fashion among the young generation of graffiti-inspired artists such as Keith Haring and Kenny Scharf, as well as camouflage's quality of readymade abstraction. Warhol superimposed the same type of colorful camouflage on other familiar images such as Leonardo da Vinci's Last Supper, and also employed it in an important series of self-portraits that he created the same year he executed the present work.
This large-scale work is part of the original series of only ten paintings of the Statue of Liberty that Warhol created for an exhibition at the Galerie Lavignes-Bastille in Paris. The Statue of Liberty was a fitting subject for Warhol's exhibit in France, as the statue, designed by French sculptor Frederic-Auguste Bartholdi, was created as a gift from France to the United States, in honor of their friendship that dated from the time of the American Revolution. Warhol made humorous reference to this legacy by including in the lower left corner the label for a cookie manufacturer, which reads "Fabis: les bon biscuits" underneath its logo of French and American flags. Long fascinated with the power of brand labels, Warhol amusingly contrasts this commercial logo with the statue that also serves as a kind of logo for the U.S.
Warhol had previously paid homage to the Statue of Liberty in one of his formative Pop silk-screens of 1963, where he repeated the statue twenty-four times. He therefore came full circle in the present work, creating in his mature style a large-scale homage to an image that had powerful and lasting resonance in American popular culture.
Galerie Lavignes-Bastille, Paris
Acquired from the above by the present owner
Paris, Galerie Lavignes-Bastille, Andy Warhol: 10 Statues of Liberty, April-May 1986.
As initiatives to rescue 'lost' modernist works are put in place, Chris Mugan discovers how sculpture is 'having a ..
A selection of accessibly priced works by the art world's greatest names, offered in two online auctions in July
From Campbell's Soup Cans to the Velvet Underground, and everything in between — a guide to the Pop master
From Hockney to Baldessari, Ruscha to Thiebaud — how the countercultural figures who flocked to California in the Sixties..
Christie's specialists, a gallerist, a conservator and an expert consultant share their inside knowledge with Holly Black
An expert guide to one of the most important events in the art-world calendar, and an index of in-depth features on Venice
Leaving behind Dubai's high-rise heart, Andrew Graham-Dixon goes in search of its low-slung soul


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Criar um Site
Budget $750-1500 USD
Hi there! I am looking for some one or company that can develop a website (plataform) just like this web site [url removed, login to view] . The idea for this site, is to offer a place for architects to sell their house plans (projects). Each architect should be able to have an account on the site, where they can upload their projects with complete information (just like [url removed, login to view]) and after approvement by us (site) the project is ready for sale. The payments can be through PayPal, and after the client received everything that he paid for, the site keeps a percenteged and the architect gets the other percenteged (just like is in freelancer.com. Any questions, please, feel free to contact me. Thanks, Rafael
63 freelancers are bidding on average $1623 for this job
__Still waiting for FEEDBACK. Please feel free to discuss issues__ Hello Rafael, Greetings from phpMaestro!! We have reviewed your project requirements along with your shared reference "houseplans" and have pe More
Let's discuss more about project to finalize the proper scope with estimated cost and time so ping me over the freelancer chat.....I will share some demo as well for recent works if you want to check. You can also c More
Hi Rafael, I have visited this website www.houseplans.com to check all the features & functionality. Let me first confirm you that our team can deliver this kind of website with much better layout/design but wonderi More
Hello, We have checked "www.houseplans.com" throughout and got fair understanding of your requirements that will do proficiently for you. We are professional full stack web developers with hands on experience in More
Hello...Nice to see your project posted here. I have reviewed your sample website and i have few things to discuss...Let me know when you are available we can discuss over chat...Ty...
Hello there, Greetings! We have gone through the complete job description and are pretty sure to assist you with this task as we are expertise team of developers & designers working on Web & App Development/Desig More
Hello Rafael, I understand you need a website where architects can sell their house plans, after approval of admin architect design will be listed on website and website will get commission upon each sale. I am wil More
Hello Sir,I just checked short description of project,I'm very interested to your project and sure give to you best quality of project,Can you please take short time to me for discuss about your project thanks
Dear client, Thank you for allowing me to post bid on your project. We have an expert team of professional blog, web application, responsive and mobile website developers in India.We have many satisfied clie More
Attention Please: Before HIRING US for the project, we can provide you FREE 03 Mock-up Designs for website landing pages. Hi Rafael Hope you are doing well We've gone through the links (www.houseplans.com) and More
Dear Employer, Greetings from fortranPRO. I have worked on similar projects to what you are looking for, and I am 100% confident that I can exceed your expectations. I have attached samples of work very simi More


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LOCAL groups across Cumbernauld are being given the chance to comment on the future of Cumbernauld Community Park.
Community organisation the Friends of Cumbernauld Community Park has commissioned a public consultation, to be carried out by an independent research company, on the future developments people wish to see in the park.
This consultation will explore a number of possible directions the park could take in future, and should help the Friends find the best uses for the park and the most popular amenities.
A spokesman said: "The Friends group will be holding a scoping meeting with the consultant shortly and hope to start the process after the summer with the intention of completing the survey and issuing a report by the year end.
"The consultation will address what the people of Cumbernauld want to have in the Community Park - wildlife habitats, paths, playing fields, community buildings, public art, events... all options are on the table at this time.
The next stage after this report will be to determine what is possible and what can be funded. Work will then be done on design development of a timeline for the implementation of the aspirations of Cumbernauld's residents in the Community Park."
The Friends plan to organise public meetings to discuss these developments - see the News and Chronicle for details. Further information on Cumbernauld Community Park can be found at www.cumbernauldcommunitypark.org.uk.
There are also plans to put the consultation questionnaire online to allow more people the opportunity to offer their views. The findings of the consultation will also be published online.


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Learning modules
- Lynn presents a problem she's observed and asks students to model their thinking.
Designing an Animal HabitatWatch as staff model the engineering design process when working with youth to design and build an animal habitat.
Real-World Modeling of the Engineering ProcessWatch as youth are guided through an actual engineering challenge with real engineers.
Making Learning Fun
Youth look to you as an example of what they should do and how they should look at the world. Do you model scientific thinking and share your thought processes and reflections? Not sure how? Learn ways to model scientific and engineering processes in your daily work.
- Start every STEM activity with a problem to solve or question to answer.
- Refer to a chart or poster with the science inquiry and design process throughout the STEM activity.
- Ask questions that support the science inquiry or design process. For example: what do you already know? or how does the test influence your re-design?
Professional Development Resources
Making Learning FunCategory: Coaching ResourcesHelp youth apply science and engineering practices as they investigate questions, solve problems and develop explanations15 minutes
Inventors: Using Design ProcessCategory: Training Resources
Frontline staff and volunteers will facilitate opportunities for youth to apply the practices of engineering as they investigate questions and sol1 hour, 30 minutes
InquiryCategory: Training Resources
Help staff facilitate opportunities for youth to apply the practices of science as they investigate questions and develop explanations.1 hour, 30 minutes
Prompting QuestionsCategory: Meeting Resources
Work with staff to support youth construct explanations for phenomena, model scientific and engineering processes and remind youth about the steps30 minutes
Success and ChallengesCategory: Meeting Resources
With this virtual meeting guide, frontline staff and volunteers will facilitate opportunities for youth to apply the practices of science as they15 minutes
Start with a Question!Category: Meeting Resources
Frontline staff and volunteers will have the opportunity to practice designing an inquiry-based activity.30 minutes
Imagination is the AnswerCategory: Coaching Resources
Frontline staff and volunteers will be virtually able to provide opportunities for youth to create preliminary designs (written, drawn, or verbal)15 minutes
Finding Out TogetherCategory: Coaching Resources
To encourage frontline staff and volunteers to be open with youth, letting them know that sometimes they do not have all the answers and seeking s30 minutes
Wonderwise: Nutty Investigations
Youth develop an understanding of the work of a scientist, learn to use simple equipment to gath30 minutes


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| Aurora
|Light Albion
|Welcome to the town of Aurora, but beware the dangerous desert wastelands beyond.
|Level Information
Sand Furies
Skeleton Hobbes
|Boss Fight
|Tomb: Sentinel
|River: Boat Racing
|Medal Scores
|World Map Board
|Previous Level
|Next Level
|The Credits
Aurora is a level in Fable Heroes. It is based on the regions of Aurora in Fable III, specifically the City of Aurora and Shifting Sands regions.
Level Breakdown Edit
- Power-Up Chest!: A Power-Up Chest sits just beyond the start line.
- Minions: These forms of the darkness first appear on a stone bridge.
- Power-Up Chest!: A chest lies in a doorway behind a collection of urns.
- Sand Furies: These tribal Aurorans appear as you pass underneath a large stone arch. There is a Power-Up Chest nearby.
- Mini-Inions (2 waves): A group of small minions appears near the skeleton of a large animal. The second wave consists of normal sized minions. A Power-Up Chest sits behind them.
- Sand Furies: Another group of sand furies appears next to a Power-Up Chest.
- Skeleton Hobbes: These appear near a Power-Up Chest in an oasis.
- Good or Evil?: Open one of the two chests to assign a random power-up to one of your puppets.
- Break Time: Destroy the urn to activate the coin cloud and proceed.
- Sand Furies: Ahead lies another pack of sand furies and another Power-Up Chest.
River Edit
Take the path on the left to reach the river: Puppets, start your engines! Tap the buttons shown over character's head as fast as you can!
Boat Racing Edit
Set sail and try to beat your fellow Heroes in this boat race!
The aim is to be the first puppet to reach the end of the course. Mash the buttons as they appear over your puppet and try to be the first to the end. The quicker you mash the buttons, the faster you go down the track. The winner receives 5000 coins, second place gets 3000, third gets 1500, and last place gets 1 coin.
Tomb Edit
Take the path on the right to reach the tomb. There are two Power-Up Chests to open and a group of mini-inions, and then full-sized minions, to kill before reaching the boss fight.
Sentinel Edit
The ancient Sentinel statue has come to life! What a piece of work...
The sentinel attacks with a punch of his right hand during the first third of the fight. Once a third of his health has gone, he will close his wings and summon a group of minions. When they are defeated, the sentinel will activate again, and will have gained a new attack. He sweeps his staff across in front of him, knocking down any puppet that gets in the way. Once another third of his health is gone, he closes up and summons minions again. During the final third, he gains a third attack where he slams his staff down on the ground. The two Power-Up Chests respawn after upwards of 75 seconds.
Scores Edit
Notes Edit
- In addition to barrels and crates, Aurora features urns that can be destroyed for coins.


text: Help, I need a list of what to check for the following problem. I have a Craftsman LT 1000 with a 17 horse Briggs and Stratton Engine and Auto trans. I have encountered the following problem the last two times I mowed. After mowing about 1 hour, the engine starts sputtering and then dies. I know the mower has been neglected this year. After sitting for about 30 minutes, the mower will restart and run another hour before the same symptoms return. I plan on replacing the plug, air filter and fuel filter as a start. But could the problem, as it starts and runs fine for a while, be more serious? The mower is about 5 or more years old. Any assistance would be greatly appreciated.