text:
|Publication number
|Publication type
|Application number
|Publication date
|Jan 5, 1999
|Filing date
|Nov 1, 1995
|Priority date
|Nov 1, 1995
|Fee status
|Also published as
|Publication number
|Michel G. Awwad, Sonny Abraham, Mary E. White-Scharf, James A. Hope
|Original Assignee
|Biotransplant, Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (7), Non-Patent Citations (6), Referenced by (4), Classifications (24), Legal Events (7)
|External Links: USPTO, USPTO Assignment, Espacenet
Glycoproteins, particularly monoclonal antibodies of rat or mouse origin, are increasingly being used for the treatment and diagnosis of cancer, infections, and immunological diseases. When administered to the patient, an immune response is sometimes generated by patients against these antibodies. The response is defined by the difference between the reactivity of the pre- and that of the post-treatment serum or plasma with the infused antibody.
Regulatory agencies require the monitoring and quantification of human immune response to the administered glycoprotein in treated patients. However, pre-dose reactivity of human serum or plasma, in some cases, is high and may interfere with the monitoring and quantification of the specific immune response to the glycoprotein.
In most situations, and although most of the patients are naive with respect to exposure to these proteins, a pre-treatment serum reactivity with the therapeutic protein is typically detected. This reactivity can be attributed to the presence in human serum or plasma of "natural antibodies" reactive with the antibody and in particular with the carbohydrate portion of the administered glycoprotein. Since the pretreatment serum reactivity with the glycoprotein might either conceal or augment the real immune response, reduction of this reactivity is desired.
Awwad M, Strome PG, Gilman SC, and Axelrod HR, Modification of monoclonal antibody carbohydrates by oxidation, conjugation, or deoxymannojirimycin does not interfere with antibody effector function, Cancer Immunology Immunotherapy, 38:23-30 (1994) examined site-specific modification of monoclonal antibody (mAB) by periodate oxidation and subsequent conjugation to either a peptide linker/chelator or a cytotoxic drug. These mAB-associated carbohydrates were also modified metabolically by incubating hybridoma cells in the presence of a glucosidase inhibitor (deoxymannojirimycin) to produce high-mannose antibody. It was concluded that modification of mAB carbohydrates did not compromise their in vivo or in vitro biological functions and that, therefore, combination therapy using immunomodulators to enhance the effector functions of site-specific immunoconjugates could be seriously contemplated.
In accordance with one aspect of the invention it has been observed that the use of oxidized glycoprotein-associated carbohydrates results in a much lower pretreatment reaction with "natural antibodies" as compared to the unmodified forms, i.e., reduces the background level of reactivity of human serum to these glycoproteins so that it is less likely to interfere with an accurate determination of pre- and post-treatment patient reactivities. Modification can be achieved chemically, enzymatically or by the use of glycosylation inhibitors in tissue cultures producing the glycoprotein, e.g., monoclonal antibody. Alternatively, aglycosylated "glycoproteins" can be produced using molecular biology techniques.
Accordingly, the invention provides a method for determining the levels of immune responsiveness to a glycoprotein in an individual being treated therewith which comprises contacting a body fluid sample obtained from said individual prior to glycoprotein treatment with the glycoprotein that has been modified to have an oxidized carbohydrate portion;
contacting a body fluid sample obtained from said individual after glycoprotein treatment with the glycoprotein that has been modified to have an oxidized carbohydrate portion; and
observing the degree of difference in the specific binding response of the oxidized glycoprotein with the pre- and post-treatment samples. Preferably the oxidized carbohydrate portion of the modified glycoprotein is oxidized by the presence of aldehyde groups on the carbohydrate portion. It is particularly preferred that the oxidized carbohydrate portion of the modified glycoprotein has at least about three aldehyde groups.
Preferably the glycoprotein for which levels of responsiveness are being determined is an antibody, particularly a xenogeneic antibody or a monoclonal antibody. Preferably the level of responsiveness is measured by an in vitro assay of serum from said individual.
In another aspect the invention provides a composition for determining the levels of specific immune responsiveness to a glycoprotein in pre- and post- treatment samples in an individual being treated therewith which comprises:
the glycoprotein that has been modified to have an oxidized carbohydrate portion; and
means for observing the degree of difference in the specific immune response of the oxidized glycoprotein with the pre- and post- treatment samples. Preferably the composition is an in vitro assay composition and the oxidized carbohydrate portion of the modified glycoprotein is oxidized by the presence of aldehyde groups on the carbohydrate portion. It is particularly preferred that the oxidized carbohydrate portion of the modified glycoprotein has at least about three aldehyde groups. Preferably the glycoprotein for which levels of responsiveness are being determined is an antibody, particularly a xenogeneic antibody or a monoclonal antibody.
Conditions for the chemical as well as the enzymatic modification process vary with the level and type of glycosylation of a glycoprotein.
FIG. 1 shows reactivity of different sera from healthy blood donors with rat monoclonal antibody BTI-322. Positive control is hyper-immune serum obtained from chimpanzee immunized with BTI-322 (chimp). Concentration of serum used was 0.4%.
FIGS. 2A-2D show the A280 HPLC profiles of two batches of oxidized BTI-322 (FIGS. 2A and 2B), and that of the unoxidized BTI-322 Reference Standard (FIG. 2C) and molecular weight markers (FIG. 2D). The HPLC profile of Reference Standard BTI-322 shows that ≧95% of the preparation consisted of monomer. In addition to the monomer peak (84% or 70%), oxidized BTI-322 elution profiles showed a second peak which constituted about 13% or 27% of the sample, respectively. Probably, this peak consists of aggregated BTI-322. Based on the retention time, this fraction consists mainly of BTI-322 dimers.
FIGS. 3A and 3B show that aggregates detected by HPLC were also detected by SDS-PAGE under non-reducing (FIG. 3B) as well as reducing (FIG. 3A) conditions. Oxidized BTI-322, with the exception of an additional aggregate band, has a similar migration pattern and the same number of bands as the Reference Standard.
The development of specific binding assay techniques has provided extremely useful analytical methods for determining various substances of diagnostic, medical, environmental and industrial importance which appear in various liquid or solid (e.g., tissue) samples at very low concentrations. Specific binding assays are based on the specific interaction between a bindable analyte under determination and a binding partner therefor, i.e. analyte-specific moiety. The binding of the analyte-specific moiety and interaction of any additional reagents, if necessary, effect a mechanical separation of bound and unbound labeled analyte or affect the label in such a way as to modulate the detectable signal. The former situation is normally referred to as heterogeneous and the latter as homogeneous, in that the latter does not require a separation step.
Where either the analyte or its binding partner is a target antigen or hapten and the other is an antibody or specific binding fragment of such an antibody, the assay is known as an immunoassay.
In conventional label conjugate specific binding assay techniques, a sample of the liquid medium to be assayed is combined with various reagent compositions. Such compositions include a label conjugate comprising a binding component incorporated with a label. The binding component in the conjugate participates with other constituents, if any, of the reagent composition and the ligand in the medium under assay to form a binding reaction system producing two species or forms of the conjugate, e.g., a bound-species (conjugate complex) and a free-species. In the bound-species, the binding component of the conjugate is bound by a corresponding binding partner whereas in the free species, the binding component is not so bound. The amount or proportion of the conjugate that results in the bound species compared to the free species is a function of the presence (or amount) of the analyte to be detected in the test sample.
An alternative format for specific binding assays is the "sandwich" assay protocol in which the target analyte is bound between a first specific binding partner, which is fixed directly or through a linkage group to a solid matrix, and a second specific binding partner, which is associated with a signal generating or labeling system.
Additional immunoassay formats and protocols are also known in the art and are applicable to use in the present invention. Various binding assays using immobilized or immobilizable materials for the direct immobilization of one of the binding participants in a binding assay reaction, e.g., immobilized antigen or antibody, in order to accomplish the desired separation of the bound and free forms of a labeled reagent, have been proposed. In particular, a number of such binding assays have been described wherein an antibody to an antigen to be detected is bound to an immobilizing material such as the inner wall of a test tube or a plastic bead.
For example, in U.S. Pat. No. 4,243,749, a competitive binding assay is disclosed wherein a reaction is carried out in a test tube having a specific antibody to a hapten under determination insolubilized or immobilized on the inner wall of the test tube. The reaction includes a labeled hapten conjugate wherein the quantity of the labeled hapten conjugate which becomes bound to the test tube wall is inversely proportional to the amount of the hapten under determination.
Another of such binding assays is described by U.S. Pat. No. 4,230,683 which discloses a method employing a 6 mm polystyrene bead having antigen or antibody bound thereto wherein the antigen or antibody is reacted with a hapten-conjugated antibody to the antigen or antibody. The bound hapten-conjugated antibody is further reacted with labeled anti-hapten antibody in order to determine the amount of antigen or antibody in a test sample.
Still another of such binding assays is described by U.S. Pat. No. 4,228,237 which discloses a method for the detection and determination of ligands in a liquid medium using enzyme labeled avidin and a biotin labeled reagent in a specific binding process. In this method, the ligand to be detected is contacted with an insoluble phase containing a specific binding substance for the ligand.
In addition to the direct immobilization techniques heretofore described, indirect immobilization by marking or labeling a binding assay reaction participant to be immobilized with a first binding substance, and then adding an immobilized second binding substance, has been proposed.
For example, U.S. Pat. No. 4,298,685 discloses an enzyme immunoassay wherein a sample containing a biological substance under determination is mixed with antibodies to the biological substance tagged with biotin and with an enzyme-labeled form of the substance under assay. An immobilized form of avidin is then added wherein the avidin binds to the biotin-tagged antibody to immobilize the antibody-bound fraction of the enzyme-labeled reagent. Similarly, United Kingdom Patent Application No. GB 2,084,317A discloses an antigen-linked competitive enzyme immunoassay using avidin bound to a solid material and a biotin-labeled antigen. Radiolabeled and enzyme-tagged immunoassays require some type of separation step. Recently, a different approach was disclosed which does not require a separation step and therefore has been referred to as a homogeneous system, in contrast to a heterogeneous system in which separation is essential. U.S. Pat. No. 3,817,837 discloses a competitive binding assay method involving the steps of combining the liquid to be assayed with a soluble complex consisting of an enzyme as a labeling substance covalently bound to the ligand to be detected and with a soluble receptor, usually an antibody, for the ligand; and analyzing for the effect of the liquid to be assayed on the enzymatic activity of the enzyme in the complex.
Still further, the labeled reagent can include other conventional detectable chemical group. Such detectable chemical group can be any material having a detectable physical or chemical property. Such materials have been well-developed in the field of immunoassays and in general any label useful in such methods can be applied to the present invention. Particularly useful are enzymatically active groups, such as enzymes (see Clin. Chem. (1976)22:1243), enzyme substrates (see U.S. Pat. No. 4,492,751), prosthetic groups or coenzymes (see U.S. Pat. Nos. 4,230,797 and 4,238,565), and enzyme inhibitors (see U.S. Pat. No. 4,134,792); spin labels; fluorescers (see Clin. Chem. (1979)25:353); chromophores; luminescers such as chemiluminescers and bioluminescers (see U.S. Pat. No. 4,380,580); specifically bindable ligands (e.g., biotin and haptens); electroactive species; and radioisotopes such as 3 H, 35 S, 32 p, 125I, and 14 C. Such labels and labeling pairs are detected on the basis of their own physical properties (e.g., fluorescers, chromophores and radioisotopes) or their reactive or binding properties (e.g., enzymes, substrates, coenzymes and inhibitors).
BTI-322, a rat anti-human CD2 antibody, currently is being used in clinical studies.
An assay was developed to measure human anti-rat antibody response (HARA) in patients treated with BTI-322, a rat IgG2b anti-CD2 antibody. However, serum harvested from patients before infusion with the antibody and from blood donors that have not been exposed previously to rat IgG showed reactivity with BTI-322. This pre-dose reactivity may interfere in the monitoring and quantification of HARA. This study was conducted to determine whether modification of antibody-carbohydrates by oxidation can facilitate monitoring and quantification of HARA by reducing the pre-dose antibody binding activity.
The term "hyper-immune animals" relates to chimpanzees that were injected and then boosted, 9 months later, with BTI-322.
The term "pre-immune serum" refers to serum harvested from animals (Chimpanzees) before infusion of BTI-322.
The term "pre-dose serum" refers to serum from patients before infusion of BTI-322 or from healthy blood donors.
Antibodies: The following antibodies were used in conjunction with the ELISA assay: BTI-322 rat monoclonal anti-human CD2 (BioTransplant, Inc.); unmodified and HRP-conjugated rabbit immunoglobulin to rat immunoglobulin (Dako); HRP-conjugated donkey anti-human IgG antibody (Accurate); mouse anti-rat IgG2b antibody (Imex, MARG2b-3); and a cocktail of HRP-conjugated mouse anti-rat κ chain antibodies (Imex, MARK-1/3-PO).
Biochemicals: Oxidation was achieved by sodium m-periodate (Sigma). The efficiency of oxidation was determined by para-nitrophenyl hydrazine (PNPH, ICN) reaction. O-phenylenediamine Dihydrochloride (OPD, Sigma) at a concentration of 0.091 mg/mL in phosphate citrate buffer with urea hydrogen peroxide (Sigma) was used as an ELISA color development reagent. TWEEN� 20 brand of polyoxyethylene sorbate surfactant (manufactured by ICI) (Sigma) in PBS (Sigma) was used as an ELISA washing buffer and 1% BSA (Sigma) in PBS-TWEEN� brand of polyoxyethylene sorbate surfactant (manufactured by ICI) was used to block ELISA plates. Tris buffer (Sigma) was used for storage of oxidized antibody.
HARA Assay: Briefly, 96-well plates (certified Maxisorb Nunc-Immuno plates) were coated with 50 μL of 5 μg/mL of unmodified or oxidized BET-322. A decreasing concentration of pre- and hyper-immune chimp sera as well as human serum was loaded onto the coated plates. Bound chimp and human gamma-globulins were detected by donkey anti-human IgG. Color development was then achieved by OPD.
Oxidation: BTI-322 (Lot No. S082/S3), at a concentration of ≧5 mg/mL, was incubated at 770-fold molar ratio with sodium m-periodate. The oxidized antibody was then desalted on a prepacked PD-10 gel filtration column (Pharmacia Biotech). The efficiency of oxidation was determined by measuring the number of aldehyde groups generated. This was achieved by reacting para-nitrophenylhydrazine (PNPH) with the aldehyde groups on the oxidized antibody and then determining the molar ratio of pNPH to BTI-322 by spectrophotometry at 390 nm and 280 nm, respectively, as described in Shrines RL, Fuson RC, and Curtin DY, The systemic identification of organic compounds, a laboratory manual, 5th ed. Wiley, New York, P 126(1967). Molarity was calculated using 208000 and 8100 as the values for the molar extinction coefficients of BTI-322 and pNPH, respectively. A molecule of unmodified (native) and oxidized BTI-322 typically contained less than 1 and ≧3 aldehydes groups, respectively. Using an ELISA, it was shown that the reactivity of pre-treatment serum with oxidized BTI-322 was substantially reduced as compared with that of native BTI-322.
Characteristics and Stability of Oxidized BTI-322:
The biochemical stability of oxidized antibody was compared to that of the unmodified BTI-322 by comparing the HPLC (size exclusion) elution profiles and SDS-PAGE migration patterns of these antibody forms were compared. Briefly, 50 μL of oxidized and unoxidized BTI-322 were loaded onto Water HPLC (Millipore) and eluted with PBS. The A280 HPLC elution profiles of the different forms of BTI-322 were compared.
The migration patterns on SDS-PAGE were produced by loading about 5 μg of oxidized or unoxidized BTI-322 under reducing and non-reducing conditions onto 4-20% gels (BioRad). Gels were stained with Coommasie Blue and the migration patterns were then compared.
To assess the functional stability of the oxidized antibody, the reactivity of soluble CD2 as well as the anti-BTI-322 reactivity of Chimpanzee hyper-immune serum, rabbit polyclonal and mouse monoclonal antibodies with the oxidized form was compared to that of the unoxidized form.
The immunoreactivity of BTI-322, i.e., its reactivity with CD2, was assessed using plates coated with recombinant soluble human CD2 (50 μL/well of 5 μg/mL). These plates were loaded with a graded dose of oxidized and unoxidized BTI-322. Captured antibodies were then detected by rabbit immunoglobulin to rat immunoglobulin conjugated to HRP (DAKO). Color development was achieved using the substrate o-phenylenediamine dihydrochloride (OPD). Reactivity of the two forms of BTI-322 to rabbit polyclonal antibody was determined in an assay similar to that of immunoreactivity except that the capture antibody was unconjugated rabbit immunoglobulins to rat immunoglobulins.
Reactivity with mouse monoclonal antibodies was determined by using mouse anti-rat IgG2b (MARG2b-3, IMEX) to capture the two forms of BTI-322 and HRP-conjugated mouse monoclonal antibodies to rat κ chain to detect captured antibodies. Color development was achieved, as usual, using OPD.
Human and Chimpanzee Serum: Chimpanzee serum was collected from 2 animals 14 days after being boosted with BTI-322, approximately 9 months after the priming dose. Day 14 sera were pooled and used as positive control in every experiment on every plate. Chimpanzee serum was diluted at first 50-fold, starting concentration, followed by 7 five-fold dilutions. Dilution was achieved in PBS-TWEEN brand surfactant (0.5%).
Human serum was collected from healthy blood donors that were naive to rat immunoglobulin. In other words, these donors had no documented previous exposure to rat immunologlobulin. The reactivity of human serum with BTI-322 was assessed using HARA assay (see above).
Data analysis: ELISA data were analyzed by comparing averages of triplicates of optical density readings. HPLC data were analyzed by comparing the A280 elution profile of unmodified BTI-322 to that of the oxidized form. This involved comparing the proportions of monomer in the different preparations. SDS-PAGE data were analyzed by comparing the number of bands and the migration pattern of the different BTI-322 forms under reduced and non-reduced conditions.
Reactivity of Human and Chimpanzee Serum with BTI-322:
Reactivity of Pre- and Hyper-Immune Chimpanzee Serum with BTI-322. Antibodies generated against BTI-322 following infusion into patients must be monitored and quantified. Since patients' sera were not available, pooled hyperimmune sera from 2 chimps that were injected with BTI-322 and then boosted about 9 months later were used as positive control. Table 1 shows the reactivity in a HARA assay of pre- and hyper-immune chimpanzee sera with BTI-322. Optical density readings obtained using hyper-serum were significantly higher than those obtained using pre-immune chimp serum. This indicates that hyper-immune serum contains anti-BTI-322 antibodies. Note that serum harvested from chimpanzees before injecting BTI-322 (pre-immune) had some reactivity with BTI-322. The addition of BTI-322 to hyperimmune chimpanzee serum reduced detectable HARA to the pre-dose level. This demonstrates the specificity of the interaction of chimpanzee serum with BTI-322.
TABLE 1______________________________________Reactivity of Pre- and Hyper-Immune Chimpanzee Serum with BTI-322 Pre- ImmuneSerum Hyper- ChimpConcentration Immune Dose of BTI-322 (μg/mL) Pre-(%) Chimp 80.5 8.7 0.87 dose______________________________________0.4 0.715 0.379 0.544 0.639 0.2030.08 0.642 0.305 0.35 0.547 0.2970.016 0.495 0.135 0.203 0.251 0.2850.0032 0.241 0.03 0.08 0.099 0.1750.00064 0.097 0.019 0.021 0.036 0.060.000128 0.037 0 0.025 0.013 0.024______________________________________
Reactivity of healthy donor sera with BTI-322:
Similar to chimpanzee pre-immune serum, sera harvested from healthy blood donors with no documented previous exponsure to rat immunoglobulin showed varied level of reactivity with BTI-322 (FIG. 1).
Oxidation of BTI-322: At least three different batches of oxidized antibody were produced by incubation of BTI-322, in complete darkness, with sodium m-periodate for 1 hour. Oxidized antibody was then desalted on prepacked G-23m SEPHADEX� brand of polyglucose polymer crosslinked with epichlorohydrin in columns (manufactured by Pharmacia, Inc.). After two rounds of oxidation, the molar ration of PNPH to oxidized BTI-322 was 3, 4 and 10 as compared to 0.6 of un-oxidized control BTI-322. These results indicate that whereas no free aldehyde groups could be found in un-oxidized BTI-322, 3-10 groups could be found in oxidized antibody. These results indicate that oxidation was successful.
Biochemical Stability of Oxidized BTI-322
FIGS. 2A-2D shows the size exclusion profiles (A280) of batches of oxidized BTI-322 (FIGS. 2A and 2B) and that the Reference Standard. HPLC profile of Reference Standard (FIG. 2C) shows that ≧95% of the same consisted of monomer. In addition to the monomer peak, oxidized BTI-322 ion profiles showed a second peak which constituted about or 27%, respectively of the two preparations. Probably, this peak consists mainly of BTI-322 aggregate. Based on the retention time, this aggregate consists mainly of BTI-322 dimers.
FIG. 2A shows the A280 Waters HPLC profile of a first batch of oxidized BTI-322, where the reported test sample analysis was as follows:
______________________________________Analysis of Test Sample Retention Area# Time (min) Height (uV) (uV*sec) % Height % Area______________________________________1 6.350 3379.133 123771.44 10.32 13.132 7.350 28286.467 796564.96 86.41 84.513 9.200 434.333 12136.60 1.33 1.294 11.067 635.714 10054.91 1.94 1.07______________________________________
FIG. 2B shows the A280 Waters HPLC profile of a second batch of oxidized BTI-322, where the reported test sample analysis was as follows:
______________________________________Analysis of Test Sample Retention Area# Time (min) Height (uV) (uV*sec) % Height % Area______________________________________1 6.383 39067.141 1801700.26 17.79 27.042 7.333 174910.418 4684104.09 79.64 70.293 9.183 1632.482 89786.63 0.74 1.354 11.133 2938.524 71887.16 1.34 1.085 11.500 1077.771 16252.91 0.49 0.24______________________________________
FIG. 2C shows the A280 Waters HPLC profile of an unoxidized BTI-322, where the reported test sample analysis was as follows:
______________________________________Analysis of Test Sample Retention Area# Time (min) Height (uV) (uV*sec) % Height % Area______________________________________1 0.783 571.989 22053.38 0.26 0.412 5.600 1274.850 32016.81 0.57 0.593 6.400 2477.362 83736.27 1.11 1.554 7.350 215841.157 5179856.91 96.40 95.735 8.983 1385.577 58815.86 0.62 1.096 9.833 633.184 6946.22 0.28 0.137 10.017 513.864 3424.26 0.23 0.068 10.150 493.449 12107.64 0.22 0.229 10.833 400.073 8396.61 0.18 0.1610 11.317 303.320 3415.37 0.14 0.06______________________________________
FIG. 4D shows the A280 Waters HPLC profile of molecular weight standards, where the reported test sample analysis was as follows:
______________________________________Analysis of Test Sample Retention# Time (min) Height (uV) Area (UV*sec) % Height % Area______________________________________1 5.650 483553.045 21972819.63 11.00 18.562 6.0503 7.517 615030.303 29158589.10 13.99 24.634 8.433 658785.225 17680701.34 14.99 14.945 9.0206 9.983 1021040.037 20970629.68 23.23 17.727 11.167 1598440.282 28259944.11 36.37 23.878 11.9009 13.293 17794.155 325114.14 0.40 0.27______________________________________
FIGS. 3A and 3B show the SDS-PAGE migration pattern of oxidized Lanes 8, 7, 6 and 5!, unoxidized Lanes 3 and 4 (Reference Standard)!, sample buffer (Lane 2), and molecular weight standards (Lane 1) under reducing (FIG. 3A) and no-reducing (FIG. 3B) conditions. Under non-reduced as well as reduced conditions, oxidized BTI-322 has an identical migration pattern and the same number of bands as the Reference Standard.
The above results indicate that oxidation did not alter significantly the biochemical characteristics of BTI-322.
Functional Stability of Oxidized BTI-322:
To test whether oxidation has affected the capacity of BTI-322 to bind CD2, a graded concentration of Reference Standard and the different batches of oxidized BTI-322 were loaded onto plates coated with recombinant soluble human CD2. Captured antibody was then detected using HRP-conjugated rabbit anti-rat immunoglobulin. Since equivalent optical density readings (Tables 2 and 3) were observed for oxidized and un-oxidized antibody, oxidation, therefore, did not affect binding of BTI-322 to CD2.
TABLE 2______________________________________Reactivity of Oxidized BTI-322 (Batch 1) with CD2BTI-322 Concentration Optical Density of BTI-322(ng/ml) Oxidized Reference______________________________________10,000 0.483 0.3975,000 0.5236 0.4412,500 0.499 0.4141,250 0.471 0.387625 0.452 0.356312.5 0.376 0.312156.25 0.324 0.28478.125 0.197 0.25839.063 0.194 0.20519.531 0.213 0.1999.766 0.204 0.1874.883 0.194 0.197Blank 0.186 0.186______________________________________
TABLE 3______________________________________BTI-322 Concentration Optical Density of BTI-322(ng/ml) Oxidized Reference______________________________________10,000 0.476 0.4215,000 0.442 0.3772,500 0.424 0.3621,250 0.389 0.318625 0.322 0.3312.5 0.269 0.13156.25 0.221 0.13878.125 0.149 0.16239.063 0.053 0.07819.531 0.048 0.044Blank 0.189 0.189______________________________________
To test whether oxidation modified the epitopes recognized by polyclonal rabbit immunoglobulin to rat immunoglobulin, plates coated with the polyclonal antibody were loaded with graded concentrations of both forms of BTI-322. Equivalent optical density readings were obtained using both forms of BTI-322 (Table 4). This indicates that oxidation did not change the epitopes that are recognized by polyclonal rabbit anti-rat antibodies.
TABLE 4______________________________________Reactivity of Polyclonal RabbitAnti-rat Antibody with Oxidized BTI-322 (Batch 1)BTI-322 Concentration(ng/ml) Oxidized Reference______________________________________10,000 0.656 0.6645,000 0.681 0.6742,500 0.609 0.6421,250 0.647 0.633625 0.661 0.658312.5 0.607 0.637156.25 0.594 0.64978.125 0.49 0.56239.06 0.389 0.36119.53 0.305 0.2340 0.057 0.057______________________________________
The effect of oxidation of BTI-322 on specific epitopes recognized by mouse monoclonal antibodies to rat IgG2b and rat κ chain was determined. MARG2b-3 (anti-rat α-2b) coated plates were loaded with oxidized and un-oxidized BTI-322 and the captured antibodies were detected by MARK-1/3-PO anti-rat κ). Results in Table 5 show that the optical density readings of the two forms of antibodies were equivalent. This indicates that oxidation did not modify epitopes recognized by mouse monoclonal antibodies.
TABLE 5______________________________________Reactivity of Mouse MonoclonalAntibodies with Oxidized BTI-322BTI-322 Concentration(ng/ml) Reference Oxidized______________________________________10,000 0.496 0.5125,000 0.469 0.4952,500 0.475 0.4561,250 0.460 0.442625 0.416 0.463312.5 0.388 0.403156.25 0.374 0.31578.125 0.292 0.25639.06 0.187 0.16219.53 0.115 0.0960 0.070 0.07______________________________________
The effect of oxidation on the specific anti-BTI-322 reactivity in hyperimmune Chimpanzee serum was tested by performing a HARA assay using plates coated with either un-oxidized or oxidized BTI-322. Results in Table 6 show that optical density readings obtained using a serial dilution of hyperimmune Chimpanzee serum in both plates were equivalent. This indicates that oxidation did not have an effect on the specific anti-BTI-322 reactivity.
In addition, results in Table 6 also demonstrate that oxidation of BTI-322 can cause 5-20 fold reduction in the reactivity of BTI-322 with serum from an individual (H23) that is naive to rat gamma-globulin.
TABLE 6______________________________________Reactivity of Chimpanzee serum with oxidized BTI-322Serum ReferenceConcentration Standard Oxidized Oxidized(1%) H23 Chimp H23 Chimp H23 Chimp______________________________________2 0.307 ND 0.179 ND 0.196 ND0.4 0.306 ND 0.123 ND 0.135 ND0.08 0.205 0.543 0.058 0.520 0.063 0.5370.016 0.097 0.376 0.019 0.326 0.020 0.3470.0032 0.037 ND 0.002 ND 0.005 ND0 0.054 0.054 0.054 0.054 0.054 0.054______________________________________
In summary, neither the biochemical nor the functional stability of BTI-322 were altered by oxidation. HPLC profile and SDS-PAGE migration patterns of un-oxidized and oxidized BTI-322 were equivalent. Binding of BTI-322 to its antigen (recombinant soluble human CD2) was not modified by oxidation. Binding of mono- and polyclonal antibodies to rat determinants of BTI-322 was not modified by oxidation. Binding of specific chimpanzee anti-BTI-322 antibody to BTI-322 was not modified by oxidation. Oxidation can cause a reduction of the pre-dose reactivity of human serum with BTI-322. The reactivity of the oxidized antibody with serum from healthy blood volunteers was then determined. The results demonstrate that modification of antibody-associated carbohydrates by oxidation can cause 5-20-fold reduction in the level of the pre-dose reactivity of human serum with BTI-322.
|Cited Patent
|Filing date
|Publication date
|Mar 9, 1982
|Jun 9, 1987
|Cytogen Corporation
|Antibody conjugates for the delivery of compounds to target sites
|Aug 31, 1984
|May 3, 1988
|Cytogen Corporation
|Antibody-metal ion complexes
|Aug 1, 1986
|Jan 17, 1989
|Ortho Pharmaceutical Corporation
|Compositions and methods using a monoclonal antibody to a human T cell antigen
|Sep 13, 1984
|Sep 19, 1989
|Cytogen Corporation
|Antibody-therapeutic agent conjugates
|May 20, 1987
|Oct 23, 1990
|Quash Gerard A
|Oxidized viruses or viral antigens and utilization for diagnostic prophylactic and/or therapeutic applications
|Jun 7, 1988
|Oct 30, 1990
|Cytogen Corporation
|Radiohalogenated compounds for site specific labeling
|Feb 25, 1987
|Oct 15, 1991
|The Center For Molecular Medicine And Immunology
|Diagnostic and therapeutic antibody conjugates
|Awwad et al. "Modification of monoclonal antibody carbohydrates by oxidation conjugation or deprymannoyrimycin does not interfere with antibody effector functions" Cancer Immunol. Immunotherap (1994) 38:23-30.
|Awwad et al. Modification of monoclonal antibody carbohydrates by oxidation conjugation or deprymannoyrimycin does not interfere with antibody effector functions Cancer Immunol. Immunotherap (1994) 38:23 30.
|Buisy et al., "Minor human antibody response to a mouse and chimeric monoclononal antibody after a single I.V. Infusion in ovarian carcinoma patients . . . " Cancer Immunol. Immunotherap (1995) 40:24-30.
|Buisy et al., Minor human antibody response to a mouse and chimeric monoclononal antibody after a single I.V. Infusion in ovarian carcinoma patients . . . Cancer Immunol. Immunotherap (1995) 40:24 30.
|Lin et al., "A flow cytometric method for the detection of the development of antibody to Orthoclone OKT3" J. of Immunol. Methods (1989) 121:197-201.
|Lin et al., A flow cytometric method for the detection of the development of antibody to Orthoclone OKT3 J. of Immunol. Methods (1989) 121:197 201.
|Citing Patent
|Filing date
|Publication date
|Oct 25, 2002
|Jul 3, 2003
|Large Scale Biology Corporation
|Endothelial cell derived hematopoietic growth factor
|Feb 25, 2004
|Jan 13, 2005
|Neorx Corporation
|Humanized antibodies that bind to the antigen bound by antibody NR-LU-13 and their use in pretargeting methods
|Jul 31, 2003
|May 12, 2005
|Neorx Corporation
|Methods of use of antibodies with reduced immunogenicity or toxicity
|Mar 19, 2008
|Apr 9, 2009
|Poniard Pharmaceuticals, Inc.
|Methods of use of antibodies with reduced immunogenicity or toxicity
|U.S. Classification
|435/7.1, 435/7.9, 435/7.93, 436/518, 530/387.5, 435/7.95, 435/7.94, 435/962, 436/548, 436/517, 435/7.92
|International Classification
|Cooperative Classification
|European Classification
|Nov 1, 1995
Owner name: BIOTRANSPLANT, INC., MASSACHUSETTS
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:AWWAD, MICHEL G.;ABRAHAM, SONNY;WHITE-SCHARF, MARY E.;AND OTHERS;REEL/FRAME:007754/0947
Effective date: 19951031
|Apr 6, 1999
|Certificate of correction
|Jul 3, 2002
|Fee payment
Year of fee payment: 4
|Jul 23, 2002
|Maintenance fee reminder mailed
|Jul 26, 2006
|Maintenance fee reminder mailed
|Jan 5, 2007
|Lapse for failure to pay maintenance fees
|Mar 6, 2007
|Expired due to failure to pay maintenance fee
Effective date: 20070105


text:
Atlanta Falcons Playoffs StripsFri, January 4, 2013 - TBAGeorgia Dome in Atlanta, GA
We are sorry, the date for this event has already passed. This event took place on 1/4/2013. Click here to view upcoming events for Atlanta Falcons.
Still unsure? Check out our user reviews for:
Have a question? Need help? Call us at 800.477.5285

subset: commoncrawl


text:
guest is using Toluna to create cool content and you can too by signing up now.
! You are using a non-supported browser Your browser version is not optimised for Toluna, we recommend that you install the latest version Upgrade
guest64 months ago
I think ME is the most profitable engineering branch. In every industry there is requirement of ME engineers and all top post are occupied by them.Carrier in this field will become growth in future. You will control project work of any kind of industry. In india there are huge requirement of me engineers.If you handle project works then you conside that profit will show itself. All the vendors and contractors will follow you.
0 comments
guest64 months ago
Master of science in computer science is the most profitable branch in recent times. The exposure the students get when they are out from the course is huge. If done properly then the placement offer get huge package from starting. Also with new technology, Hardware & software innovation, people with genius mindset creates a life from the field which is not their in other branch of engineering
0 comments
guest64 months ago
i think electronics and communication is most profitable thing because it has a broad spectrum and and the boom is not onn and off . u can find jobs offen and easy to find.the subject is vast and wide ,entry into that can fetch u nice placements in that. one can easy 2 earn money as the minimum wages are hiking up in this field. i think its the best
0 comments
guest65 months ago
Computer Science Engineering is the most profitable branch as the knowledge in this field is used in all type of profession and all types of business. The demand of experts in Computer Science Engineering is every where around the word weather it is private sector or government sector. The demand of such trained people are increasing day by day.
0 comments
guest65 months ago
Mechanical Enginn is the most profitable of all the branches in Enginn.
0 comments
You're almost there
In order to create content on the community
Complete your registration
Verify your Email / resendNo thanks, I'm just looking
Your Facebook token has expired, you need to reconnect your Toluna account with Facebook or disconnect the two accounts for now.
It's all about you!
Sign up on Toluna or
Log in to Toluna orFacebook Connect What is my password?
Please enter correct Toluna credentials.


text:
|Product description
パンラクミン 350 tablets are antidiarrheal agents containing acid-resistant coagulans ( active ingredients ラクボン ) 4.5億 pieces. With a proliferation of harmful bacteria, such as rot, intestinal environment, improves constipation and loose stools, bloating.
ラクボン fungi spores in the acid and heat to very strongly stand up well to save, not affected by stomach acid reaches in the intestines.
Beating ラクボン fungus spores reaches the intestines begin to breed, but support component タカヂアスターゼ N1 of the digestive enzymes and biotin promotes ラクボン bacterial growth and increases the effective.
|Efficacy and effects
Intestinal (bowel condition), constipation and loose stools
Bloating, indigestion, digestion enhancement
Backrest, chest dysphasia, loss of appetite, eating too much
|Dosage and administration
The following amount of meal, taking leave or かみくだいて, please take.
Adults (over 15 years). 1 1 3 Tablets, 3 times
11-15 Years: 1 2 tablets 1, 3 times
Over 5 years old and less than 11 years old. 1 Times 1 tablet 1, 3 times
Under 5 years old. Please do not take
|Coagulans ( ラクボン active ingredients ): 45 mg / タカヂアスターゼ N1: 135 mg / biotin (vitamin H): 18 μ g additives: TALC / stearic acid Mg / chloride Na / hydroxypropyl cellulose / fragrances / vanillin and ethylvanillin / white sugar and cellulose and lactose
|Content amount
|350 Tablets
|一三 Sankyo healthcare
|Ads responsibility
Fuzhou co., KIA
|Made in Japan, pharmaceutical products
This product cannot be shipped to United States from the store.
Please contact the store for further information.


text:
Change Location
ACE Fit Share
Use a combination of these core back exercises to strengthen your lower, upper or middle back areas. Whether you're interested in barbell deadlifts or reverse extensions, we have back strengthening exercises for every level. These exercises are also featured in convenient workout routines that include proper warm-up and cool-down techniques, which greatly reduces the risk of injury.
Fit Life Articles
If you find yourself not making any gains or simply want a different exercise program, here are six ways using heavy weights can help you make the changes you want to see in your body.
If you notice that the same parts of your body are always a little too sore after a workout or a day full of physical activity, you might have muscle imbalances. The good news is that the right exercise program can help improve your muscle strength and enhance joint range...
Eating enough protein is essential for both maintaining and losing weight, as well as for helping muscles repair after workouts. But exactly how much protein do you need? Learn why you may be underestimating your protein needs and how you can get more protein in your diet....


text:
Three dozen bouquets graced the steps of the Old Dutch Church in Uptown Kingston Thursday, put there by residents as a show of sympathy and support for the Dutch nation which lost dozens of its citizens when Malaysian Airline Flight 17 was shot down. "If you can get people to pause, hopefully something good can come out of it," Sean Pellegrine of Kingston said after he placed one of the bundles of flowers on the steps.
Location, ST | website.com


text:
Jake ZukowskiShared publicly -
Our São Paulo cab driver told us that favela is a politically incorrect term and that we should be using morador da comunidade or just comunidade. Makes me remember how important language is. I wouldn't want a foreigner coming to my city and asking about the "ghettos" when talking about the poor communities.
Add a comment...


text:
The Australian Federal Police will not investigate fresh claims a baggage handler may have stashed four kilograms of cannabis in Schapelle Corby's bag until an official complaint is made by her defence team.
Last night a woman, going by the name of "Sue", gave Nine News a sworn statement that a baggage handler had hidden a large bag of marijuana in luggage at the time Corby went through Brisbane Airport.
More News Videos
Schapelle Corby case could be reopened
Spotlight on the Northern Territory
Rudd rejected
Australia's first whole genome testing
Lamborghini driver kiddin' around
Karl Stefanovic uses transphobic slurs
Jetstar and AirAsia planes near miss
Calls to stop animal testing at university
Schapelle Corby case could be reopened
Claims by an anonymous woman that a baggage handler dumped a stash of marijuana in Schapelle Corby's bag.
The woman told Nine her former boyfriend, a baggage handler, had told her of a work colleague who had brought a large bag of marijuana to work and later stashed it in a bag.
The broadcast caused Corby's Australian lawyer, Kerry Smith-Douglas, to call for another review into the case.
But an AFP spokeswoman said this morning pursuit of the latest claims rested with Ms Smith-Douglas.
''It is a matter for Ms Corby's defence team as to how they wish to pursue these claims in defence of their client,'' the spokeswoman said.
''If people have information regarding criminality at any of the 11 Australian airports the AFP has responsibility for they are encouraged to come forward.''
Corby is serving a 20-year term for smuggling more than four kilograms of marijuana to Bali in October 2004.
Sue, who claimed to be a clinical counsellor with a remote area health service, told Nine she had been in a relationship with a baggage handler in October 2004 - the month Corby went to Bali.
She said her boyfriend had told her about the day a fellow employee came to work with a "big bag" of cannabis.
"When the supervisor came down, he got the shakes," she said.
"The first thing he did was look for somewhere to hide it and he grabbed one of the bags that was behind him and hid it in there."
Sue said she had come forward after all these years because her conscience had weighed heavily on her. She has given Nine the name of her former boyfriend.
Ms Smith-Douglas last night told Nine Sue's statement should trigger another judicial review of the case in Indonesia.
"I think [Sue's] statutory declaration is extremely important because it is from an individual who is not known to the Corby family whatsoever," she said.
- with Cameron Atfield


text:
Jae C. Hong, Associated Press
CULVER CITY, Calif. — YouTube is enlisting Hollywood's help to reach a generation of viewers more familiar with smartphones than TV remotes.
The online video giant is aiming to create 25 hours of programming per day with the help of some of the top names in traditional TV. The Google-owned site is spreading its wealth among producers, directors, and other filmmakers, using a $100 million pot of seed money it committed last fall. The fund represents YouTube's largest spending on original content so far.
YouTube believes it is laying groundwork for the future. While the number of traditional TV watchers has leveled off in recent years, more and more people are watching video on mobile phones, tablets and computers, especially the 18- to 34-year-old age demographic that advertisers covert.
The idea is to create 96 additional YouTube channels, which are essentially artists' home pages, where viewers can see existing video clips and click "subscribe" to be notified when new content goes up.
Well-funded videos by a select roster of stars are likely to be more watchable than the average YouTube fare of cute cats and webcam monologues. YouTube is betting that a solid stream of good content will attract more revenue from advertisers, bring viewers back frequently and bolster its parent company's fledgling Web-connected-TV platform, Google TV.
The cash has enticed some of TV's biggest stars, including "Fast Five" director Justin Lin, who directs episodes of "Community," ''CSI" creator Anthony Zuiker and Nancy Tellem, the former president of CBS entertainment.
Zuiker is teaming up on a horror series for YouTube after observing his own family's behavior. His three pre-teen sons spend more time on phones, iPads and computers than watching TV these days.
"We want to jointly take the risk with YouTube and roll the dice on the future," Zuiker says. "The old regime is going to falter because everybody thinks the TV is the only device that really counts, and that's just not the case."
For producers, it's a chance to create shows that are completely free of meddling from major studios. They can also stay relevant with a younger crowd whose viewing is moving increasingly online.
Several new channels such as the extreme sports-focused Network A and Spanish-language Tutele have launched already. YouTube hopes to have them all up and running by this summer.
"This was really about galvanizing the ecosystem at large," says Alex Carloss, global head of original programming for YouTube. "We see the portfolio (of funded channels) really representing the best of TV meeting the best of the Web."
YouTube isn't the only Web video service that has started to pay for original content. Netflix Inc. recently launched the original series "Lilyhammer," while Hulu premiered "Battleground." But YouTube videos tend to be under 10 minutes, instead of fitting into traditional half-hour or hour-long TV slots. And aside from a few guidelines, ultimate control is given over to the artist, including what is uploaded and when new episodes appear.
YouTubers also get away with far edgier stuff than the middle finger that rapper M.I.A. flashed during the Super Bowl halftime show.
Although YouTube's entire investment is less than half of what some studios spend on one blockbuster movie, about a third of the new channels were awarded to scrappy YouTube veterans who already know how to make it big online while keeping production costs low.
YouTube expects to recoup what it spends on the grants by sharing ad revenue the new videos generate.
At Maker Studios, which received money for three new channels, the funds have turbo-charged an already teeming operation that has about 160 full-time staff spread across several buildings crammed with props and computers in the west Los Angeles suburb of Culver City.
- Verizon buys Yahoo for $4.83B, marking end of...
- Historic solar flight marks first...
- Judge set to decide on $15B Volkswagen...
- Former Oregon lumber town rides digital wave...
- US home prices rise at steady pace as sales...
- Stocks waver as investors monitor company...
- Why workers aren't taking vacation time, and...
- Provo airport expansion could provide up to...
- Get all your outdoors work done by... 13
- Provo airport expansion could provide... 5
- Verizon buys Yahoo for $4.83B, marking... 3
- Historic solar flight marks first... 1
- Why workers aren't taking vacation... 1
- 5 things to do when quitting your job 1
- Panel recommends options to reduce... 1
- Judge set to decide on $15B Volkswagen... 0


text:
Britannica Web sites
Articles from Britannica encyclopedias for elementary and high school students.
- Callimachus - Student Encyclopedia (Ages 11 and up)
(5th century BC). The Greek sculptor Callimachus is believed to have invented the Corinthian capital (one of the three major styles of columns in Greek architecture) after noticing acanthus leaves growing around a basket placed upon a young girl's tomb. In addition, he is often credited as the originator of the running drill for boring in marble.


text:
ModernTribe Atlanta is Now Closed.
June 19, 2015 | Posted by Jennifer rivlin
Welcome to ModernTribe Jewish Gifts' Blog about modern Jewish design and lifestyle. Find profiles of modern Judaica artists and Israeli design. You'll find the occasional post about trials, tribulations, and heaps of darn good fun we have bringing you the best Jewish gifts.
June 19, 2015 | Posted by Jennifer rivlin
August 26, 2014 | Posted by Jennifer rivlin
Come to ModernTribe's Atlanta shop and make a Fancy Caramel Apple as a sweet treat for Rosh Hashanah!
Tickets are $8 per person. Ticket includes making one Fancy Caramel Apple to bring home with you. (Additional apples are $5.)
September 02, 2010 | Posted by Admin
Jewish Woman Magazine's website is brand new and improved. American Jewish women need a virtual place to congregate. It is my hope that all of us can build this space for Jewish women everywhere to read and share information relevant to today's Jewish living. I'm very proud to present my first contribution: an entry to a column I'll be writing called:Identity by DesignExpressing Jewish Identity Through JudaicaBy Jennie Rivlin RobertsThough many of us talk about developing a Jewish identity, what does that really mean? To me, Jewish identity includes what...
0 Comments | Post A Comment | Categories: aimee golant, jewish woman magazine, Judaica, jwi.org
November 23, 2009 | Posted by Admin
This week, we are featuring some exciting wood products for the design enthusiast who wants to bring nature into their home. These wonderful gifts are perfect for the neo-folk/urban primitive/Americana fan. Wooden Kiddush CupsOn a search for new kiddush cups we wondered, "could there be a kiddush cup that reflects an earthy, eco-centered approach while at the same time holding true to modern design aesthetic?" Well, we found it! These cups are hand carved with love and are perfect for someone who likes the neo-folk or craftsman style. Enjoy Shabbat...
November 13, 2009 | Posted by Admin
This week's Parshat is Chayei Sarah and talks about the importance of understanding "true" love (hint: love ain't a feeling). Also featured are a video on Tallit from a feminist perspective and a "plug" for the Atlanta Jewish Music Festival from our friend Russell. Shabbat Shalom and have a beautiful weekend!Parshat Chayei SarahTallitAtlanta Jewish Music Festival From ModernTribe Jewish Gifts
November 07, 2009 | Posted by Admin
ModernTribe is very excited to add Pop Judaica and its funny Jewish gifts to ModernTribe's offering of modern design for Jewish lives. We have many exciting plans for our combined stores. Early 2010 we will integrate the shopping carts so you can shop both stores and check out once.Now, you can shop in both stores and pay for ground shipping once. Shop ModernTribe within 24 hours of placing your Pop Judaica order and use code SHOPPEDPJ during checkout (in the shopping cart before you click "checkout"). We will ship all...
0 Comments | Post A Comment | Categories: e-commerce, Judaica, ModernTribe, popjudaica


text:
PO Box 930
Clovis, NM 88102
(575) 763.6600
Email us
Documentations can target medical binary options software bills of designer or instances. Differential making losses can protect customers international calling plans at Disneyland or how good.
Copyright © 2015 Clovis EDC
All Rights Reserved
Designed and Powered by RC Web Design


text:
In previous papers, I have considered various phases of neurologic surgery. In this paper I shall discuss my series of brain tumors, which number eighty-five. These unquestionably present the most difficult and varied problems which the neurologic surgeon encounters.
In a recent paper read before the Clinical Congress of Surgeons, Cushing said that the time had passed when there is any purpose in speaking of the mortality in brain tumor cases any more than there is in speaking of the mortality in abdominal tumor cases. That such a generalization leads nowhere is also apparent in a study of my cases, and would be most discouraging were we to content ourselves with the bald statement that of the eighty-five patients, we lost twenty-nine, or 35.5 per cent. But when we study these cases from other angles, we find that eighteen of these twenty-nine deaths were in patients suffering with glioma, and
Thank you for submitting a comment on this article. It will be reviewed by JAMA Surgery editors. You will be notified when your comment has been published. Comments should not exceed 500 words of text and 10 references.
Do not submit personal medical questions or information that could identify a specific patient, questions about a particular case, or general inquiries to an author. Only content that has not been published, posted, or submitted elsewhere should be submitted. By submitting this Comment, you and any coauthors transfer copyright to the journal if your Comment is posted.
* = Required Field
Disclosure of Any Conflicts of Interest*
Indicate all relevant conflicts of interest of each author below, including all relevant financial interests, activities, and relationships within the past 3 years including, but not limited to, employment, affiliation, grants or funding, consultancies, honoraria or payment, speakers' bureaus, stock ownership or options, expert testimony, royalties, donation of medical equipment, or patents planned, pending, or issued. If all authors have none, check "No potential conflicts or relevant financial interests" in the box below. Please also indicate any funding received in support of this work. The information will be posted with your response.
Some tools below are only available to our subscribers or users with an online account.
Download citation file:
Customize your page view by dragging & repositioning the boxes below.
More Listings atJAMACareerCenter.com >
Enter your username and email address. We'll send you a link to reset your password.
Enter your username and email address. We'll send instructions on how to reset your password to the email address we have on record.
Need assistance?
Athens and Shibboleth are access management services that provide single sign-on to protected resources. They replace the multiple user names and passwords necessary to access subscription-based content with a single user name and password that can be entered once per session. It operates independently of a user's location or IP address. If your institution uses Athens or Shibboleth authentication, please contact your site administrator to receive your user name and password.


text:
Product Reviews
No reviews yet for this product. Review This Product!
Product Reviews(0 Review)Add Your Review
Fantastic StickReview by Callum posted on 11/17/2015I purchased the M4 Pure at the clearance price of 9900 with a HallNugent Hopkins curve and couldnt be happier with the performance of the stick especially at the price point After a couple of uses Ive had no issues with durability and its certainly a significant upgrade on my old 55S as regards puck feel and power in the wrist shotHighly recommended


text:
I have a low power server, with 4 ethernet sockets.
It was running Fedora 11, with 3 of the network sockets named eth0, eth1 and eth2 (the final network socket was not in use --- it was eth3 but don't know for sure that it actually worked).
I installed Ubuntu Server 10.04 on it this morning. While it was installing, it recognised eth0 and used it to access the internet.
Now... it doesn't. It recognises eth1 as being eth1, and claims there is an eth5 somewhere, but doesn't recognise any other interfaces from eth0-eth7.
I require 3 working ethernet sockets.
I would like my ethernet sockets named eth0-eth3, as they were under Fedora. How do I make that happen? Why have they been renamed in the first place?
Ethernet sockets have always popped up with the names I was expecting before, so I don't know where to start.
There is some inconsistency in the hardware addresses three of the ethernet ports claim to have. Sometimes they claim to be at
xx:xx:xx:xx:xx:xx, at other times they claim to be at
00:00:00:00:xx:xx. (As far as I can tell, they never change hardware address when the OS is active, only when the machine is switched off, or possibly when rebooting.)
I don't know where I go from here.
Each ethernet port consistently has one of two hardware addresses, they're just inconsistent about which one they're using at any given time.
I tried telling udev that both hardware addresses belong to the same ethernet port --- I expected that udev would complain at this, but it accepts it and uses the hardware address that is present for that port.
So my problem seems to be solved.


text:
How long have you been on DeviantArt?
Ten years! OMG.
What does your username mean?
Sometime during the early 2000s, I joined a character illustration contest hosted by Animax Asia. I gave her the name 'Saber' because she's a swordsperson, and 'kite' was the name for the hoverboard she uses as transportation. It also is the reverse of my initials.
Describe yourself in three words.
Cautious, crazy, talented.
Are you left or right handed?
What was your first deviation?
What is your favourite type of art to create?
Photography, illustrations and drawings.
If you could instantly master a different art style, what would it be?
Realistic art using traditional mediums like watercolour, acrylic, oils, etc.
What was your first favourite?
What type of art do you tend to favourite the most?
The kind that's similar to the style I like to make, or want to learn.
Who is your all-time favourite deviant artist?
- this is only a partial list. Practically all the people I follow are my favourites!
If you could meet anyone on DeviantArt in person, who would it be?
Michael Zancan: zancan.deviantart.com/
How has a fellow deviant impacted your life?
They make me want to be a better artist.
What are your preferred tools to create art?
Pen and paper, traditional watercolour, Photoshop and my graphics tablet.
What is the most inspirational place for you to create art?
Anywhere I'm comfortable. Home remains the best place.
What is your favourite DeviantArt memory?
Whenever any one of my favourite artists would reply back to my messages or chat with me.


text:
Can a customer belong to more than one Membership Level?
I have a few PDF reports I want to sell as 10 day membership levels. Allowing customers access to a download page for 10 days. When I do a test signing up for a second membership level the existing membership levels is erased.
How do I sell more than one membership level to the same person?


text:
Skip to main content
Search the history of over 498 billion pages on the Internet.
Search the Wayback Machine
All Texts
This Just In
Smithsonian Libraries
Lincoln Collection
Additional Collections
eBooks & Texts
American Libraries
Canadian Libraries
Universal Library
Shareware CD-ROMs
Community Texts
Project Gutenberg
Biodiversity Heritage Library
Open Library
Children's Library
Kimberly Kubus
All Video
This Just In
Prelinger Archives
Democracy Now!
Occupy Wall Street
TV NSA Clip Library
TV News
Animation & Cartoons
Arts & Music
Community Video
Computers & Technology
Cultural & Academic Films
Ephemeral Films
Understanding 9/11
News & Public Affairs
Spirituality & Religion
Sports Videos
Videogame Videos
Youth Media
All Audio
This Just In
Grateful Dead
Old Time Radio
78 RPMs and Cylinder Recordings
Live Music Archive
Audio Books & Poetry
Community Audio
Computers & Technology
Music, Arts & Culture
News & Public Affairs
Non-English Audio
Librivox Free Audiobook
Radio Programs
Spirituality & Religion
All Software
This Just In
Old School Emulation
MS-DOS Games
Historical Software
Classic PC Games
Software Library
Internet Arcade
Community Software
Software Sites
Tucows Software Library
Vintage Software
Console Living Room
Atari 2600
Magnavox Odyssey 2
Bally Astrocade
Sega Genesis
Sega Game Gear
Atari 7800
Sega Master System
All Image
This Just In
Flickr Commons
Occupy Wall Street Flickr
Cover Art
USGS Maps
Metropolitan Museum
NASA Images
Solar System Collection
Ames Research Center
Brooklyn Museum
Toggle navigation
1:00 am
veus san francisco. >> a great saturday. >> the fight is on to the finish. both candidates out pushing their plans to rev up our economy and we find t it's still not in full recovery mode andore u.s. companies are announcing major firing, not hiring. the president has his pamphlet and the governor has his overall plan. and more on the pamphlet later. and is governor romney's call right or wrong? i'm brenda buttner, it's bulls and bears. we've got gary b smith. tobin smith. jonas max ferris along with tim la camp and steve murphy, welme, everybody. okay. the folks at forbes will tackle the pamphlet plan in one hour, fair and balanced.
1:01 am
what about the plan to overhaul the tax code. what is it going to mean for jobs. >> i think it's going to be great for jobs. ovrhauling the taxes is the thing that the governor and hopefully president romney can do. and incents, if that's a word, every individual and corporation out there. provides a path of what they're going to do, what they're ing to pay and if it's simplified and overhauled, it will get rid of hopefully a lot of the special interests, whic takes the government out of the intervention and provides a clear playing field for everyone. this could turn around our economy, if done soon and done right. >> brenda: jonas, we don't know exactly what kind of overhaul we would get and isn't that adding uncertainty? wouldn't that keep people from hiring? >> moreover, it doesn't really, neither plan really addresses the deficit and the need for revenues. let's leave that problem off the table and talk about the job situation. i like what romney wantso do, the code has a lot of
1:02 am
deductions that shouldn'te there, we should just lower rates and get at them. and in fixing that, if you pay 25%, and then you get deduct the mortgage or don't get to deduct the mortgage and pay 20%. if you're net-net not getting taxed any more, i don't see how that's actually going to lead to more spending and i tell you a little dangerous in the short run, the housing market is still-- and to pull out major deductions the home mortgage deduction being one of them. he didn't say he was going to do that, if it didn't do that, but makes the budget work. >> brenda: i know, the tax code is like war and peace. overhauling it, will that help get jocks. >> like you've read that. >> not immediately, but look, we've got the most sophisticated tax code in the entire world, makes us uncompetitive and the u.s. falls on a competitive basis and a lot of reason is the tax code. what it's done, it's made small businesses at a big disadvantage to big business.
1:03 am
because what happens is big crony businesses getting their loopholes and their deductions from washington in ehange for higher rates, and it becomes way too complicated for small businesses to have to hire lawyers, they have to hire accountants, just to figure it out and sven hawking doesn't figure this out. it's not going to fix our economy immediately, but remove the biggest obstacle to growth and that's a complicated tax code. i'll tell yu, focusing on little things, little pieces of this, like the mortgage deduction,ocusing on bits and pieces, and the oblem. the problem is it's just way too complicated and small businesses cannot figure it out. >> brenda: you know, it's interesting though, steve, the other steve not hawking, because a lot of the bigger businesses have been laying f this week, ge, huge numbers coming out and they depend on a lot of these deductions, actually. >> yeah, no i think what we have to-- jim is absolutely right. first of all, steven hawking, would have looked at all this have and call it an alternate universe.
1:04 am
it doesn't, you know, it defies the reality that we live in here today. look, our economy is transforming. the consumers, we're not going to prot them into picking up the negative savings rate and spending that we had before the wall street collapse in 2008. they know better now. they're not going to be burned again. and this is an opportunity for exports are picking up, manufacturing's picking up and we need to stop consuming more than we've produced and start exporting more. so, yes, making our businesses more competitive is critical to that, and look, i would just get rid of every single corporate deduction, and bring the rate dramatically down, and cut it in half. 15%, something like that and they have to pay it on domestic profits, they have to pay it on foreign profits, minus whatever they're paying locally and just, and there are several other policies we're not allowed to get into this now, not enough time, it ought tbe about making american business more competitive and exporting and increasing our advanced
1:05 am
manufacturing or exporting. >> brenda: toby, is this going to increase jobs in the short-term? we're all looking long-term, even getting this whole tax code simplified would take forever. >> well, it would if we did two things. remember, we have this bizarre tax system that we started in the 19th century that taxes me as a company on money that i've made overseas twice. that makes no sense. we're the only country in the modern world and the stone age that continues to do that. and we also happen to be the only country in the world where you can deduct mortgage interest from your home. the only country in the world. canada, a few paces away from here has never done it, they have a much healthier home system. and we also can't deduct-- excuse me, a business and a person who gets benefits, i don't get taxed on my benefits, that came from the 40's. we have to get into the 21s century and tha would get companies to unleash this 2 1/2 trillion dollars in cash, that's sitting on the sidelines. >> brenda: well, gary b, you know, toby was back in the 19th century and understands
1:06 am
all of this from personal-- >> and it was a fine time. (laughter) >> and the points, that romneys going to have a lot of problems, kindf jonas alludes to, getting rid of a lot of these deductions. there's a lot of special interest, but it's the special interest that are kind of mucking up the economy. the government should not be in the housing business or pushing people into the housing business. that's exactly what the mortgage interest deduction does. and every industry and every service rely on a level playing field and that will unleash the power of the market, not the government mucking up things with all of these deductions here and there where, as you point out. like war and peace, you can't figure it out. the only industry that will suffer from this is the c.p.a. industry. >> okay, now, jonas, you get rid of the mortgage interest deduction, what happens-- we're talking about jobs. what happens to the housing market and the real, the wealth effect of people? they're going to stop spending, what's going to happen to jobs then.
1:07 am
>> ideally we would have gotten rid of that deduction before the bubble. now that we've had the crash, a lot of people buy hes because they look at the after tax benefit and for home sales, at best phased out. do you it overnight you'll see home prices drop, 10, 20% and people might start to by in anticipation of it going away. the other one is health care, get that free, take that deduction away and all of a sudden youhealth care costs go up. you can't get out of these in a short period time, too disruptive to the health care industry and home building industry. >> and here is the problem with that deduction when they're looking to buy a house. >> they're looking, but the health care, you'r wrong. ear not going to come out and say, geez, i have a heart attack and oops, wait a minute it's going to be taxable income. that's not how it works. >> it would act on a tax increase on a lot of people. >> and you can't tax--
1:08 am
>> and you if, phase it >> hold on. >> a $300,000 home-- >> this benefit primarily, it starts to phase out. over like a million dollars all of a sudden you're paying the marginal rate and the deducon, even the mortgage deduction, 1 million dollars. the point is, the effect is t make the tax code lets progressive, ultimaty somebody making $100,000 a year with a house with health care plan paid by the company is going to pay a higher rate and making 2 million dollars, those deductions got phased out after 2, 300,000, to the higher and lower rates-- >> hold on, that's got to be the last word. i love you guys, you get so passionate about taxes, i am so lucky. all right, no within's been following what this election means to your money, your job, and your taxes, like who else, but neil cavuto and he's back at it next weekend. first on saturday for a special cost of freedom, your ballot, your bucks, starting at 10 a.m. easte and then back on sunday for a special your world at 4 p.m. for neil, it's always about your wallet. so don't miss it.
1:09 am
but first, neil and his gang will tell you why this 15 trillion dollar debt number may be bad, but a new number on how much of it you owe to china and other foreign countries may be worse. that's at the bottom of the hour. up here first, forge the political storm, it's the other perfect storm that could be hitting us all at the pump no matter where you livivivivivv
1:10 am
1:11 am
1:12 am
1:13 am
central appalachians. >> you can keep track of the hurricane on our website,
1:14 am
foxnews.com, i'm anna kooiman. >> brenda: so, just as gas prices are starting to slide. fears growing, hurricane sandy could drive them right back . the jor refiners on the east coast could take a direct hit. jim, how bad could this get at the pump? >> well, if you ask me how bad. it could get really bad. easynswer to say look, it isn't going to be a big deal because usually these kinds of things aren't, but let's take a worse case scenario, if you had a category 1 storm hit the east coast we're completely unprepared for it, first of all. gasoline prices are too high, even though they've come down recently they're too high and we don't have any slack in this enomy so this thing would be kicking our compli eco hat a time we're not circling the drain, but on the edge of the toilet. i don't think that consumers can afford the increase in the gasoline prices, nor can they afford the economic hit that they would take in that region, rebuildingt a time, again, where things are
1:15 am
difficult in the first place. >> gary b, are you worried. >> i am not. jim had it right and then talking until he had it wrong. the east coast area that might be hit by the hurricane has about 6% of the refining capacity, that will put a little dent, sure, a spike in prices, but i think what jim was talking about, we've been through this before. multiple times, hurricane katrina, hugo, et cetera, et cetera. the snow-ma geddon to hit the east coast, each time people thought this is it. they spike for a little bit. 24, 48 hours and then come back to where they were before. >> brenda: jonas? do you have any concerns about this? hurricane isaac when it hit, caused the biggest jump in gas prices in 18 months. >> and in this case, no, in connecticut i couldn't drive because tre were trees down and your mobility goes dn to the point where fuel consumption wi dro as much
1:16 am
as refining capacity unless it freakingly hits the refinery because you can't get to work, major power outages, no power for a week and you're not consuming energy when you can't turn on the power and can't get to work in your car. so it's a good thing, but keep the gas price froms going bananas in the first place. >> brenda: the east coast provides 6 1/2% of the gas that the u.s. uses. that's kinds of a-- could be a hit. >> i don't thinkt's going to be a very big hit. look, the prices may well go up, even if just one light bulb at one refinery gets knocked out because they rise and fall based on expectation of demand and supply, not the reality, so, they could go up a little bit. we could have a big one-day jump. they'll corrects in a couple of weeks and people won't be driving in this mess, and they suldn't be, just stay home, be safe and don't worry about it. >> brenda: i don't know, tobin, i'm worried, what about you? >> well, i'm looking to the nasty ugly speculators,
1:17 am
there'a futures market in gasoline. if the people who know the business best thought that gasoline was going be to be knocked out, a huge super spike, you would he seen the futures on gasoline go up accordingly, we haven't seen it. not to say we can't have a disaster, i don't have a ouiji board. but it would have a disaster to have a lasting effect as my colleagues said. >> brenda: thanks, guys, that's the last word. and homes sales heading higher, is a a a a a@7
1:18 am
1:19 am
1:20 am
1:21 am
>> here is something we've all been waiting for. housing numbers with green rrows, oh, new home prices jumping double digits last night and sales leaping the most in two and a half years, but jim says the government is about to knock housing back on its back. how so, jim? >> well, look, they're talking about tightening mortgage standards again because of the standards put forth by dodd-frank. lending standards are already too tight even though the housing market is improving, you talk to mortgage companies and title companies and tell you the same thing over and over again. these deals are very difficult to close right now. even though they're closing them. and this is the reason the housing market is not, has not moved a needle on the economy, like the statistics would suggest. keep in mind 31% of home owners out there are still underwater anyway so the housing market is held back anyway, if you tighten standards now, it's going to have a deleterious effect and not only that, but the federal
1:22 am
reserve board is out there and 75% of all mortgages, the mortgage market is becoming the same monster it was under fannie and freddie mac. we don't need this, i needs to be settled by iividual banks not by the federal government. >> brenda: okay. steve isn't this kind of the price we have to pay to not have another bubble and another burst bubble? >> absolutely. mortgage lending standards should be tough. you should have to be fully capitalized to offer mortgages and standards for the borrowers should be high and standards on the derivatives should be tough so we don't have it happen again. and other one thing i disagree wit jim on, where is the money going to pay for mortgages? it's being saved right now. we have a positive savings rate, 4 or 5%, 3 trillion in savings leads to an export economy. >> we've got to move on here, toby, what's it going to do to the economy? >> if they come in, they're draconian and my point the
1:23 am
government should not be in the private mortge business, if there's money to be made lending to gary and me and our house that is we can't refinance because, you know, we have different incomes, then there should be a market developing, but the new legislation keeps the private market out. >> brenda: right. gary b, is the pendulum swinging too far the other way? >> absolutely, the ultimate irony, i love steve's remarks, he has such faith and confidence in the government. this is the same government in ka hoots with the federal reserve that put so much free, easy money out there, and caused the housing bubble. now, what they're-- when things are finally starting to look green and prosperous, they want to clamp down on it. get the government out of the housing busins. >> brenda: okay. jonas. >> and first of all, the government is still the loose ninja lender of last resort. called the fha program, and you can put ltle down, 2, 3%, 0 and ma market existses.
1:24 am
the government is throwing away money, 3% program and home path program. the lenders almost all went out of businessecause of their own lousy credit system they were using and over exemptions thought that homes couldn't fall and now-- >> didn't have anything to do with fannie mae or freddie mac or the community reinvestment act or ultra low interest rates, doesn't have anything-- >> do you remember thornburg, the first were to go the non-fannie companies failed first and-- >> it was a long series of steps. >> those are the ones looking at ten cents on the dollar and don't tell me it was all fannie stuff. >> brenda: guys, there you go. >> jonas, you're right. >> brenda: gary b, last word. >> i get the last word? hey!. >> brea: yes. (laughter) >> that's all we needed. all right. and thank you very much, steve for joining us. wel one of the 80 ceo's warning about the looming fiscal cliff just announcing something that could make you a 40% bump by the spring.
1:25 am
1:26 am
1:27 am
1:28 am
>> predictions? and jim, you're up first. >> sirius xm and global car fleets are selling and we've got an aging u.s. fleet that needs replacing and their margins are expanding and looks really good to me. >> brenda: jonas, do u like sirius, that radio. >> no my cool internet phone is taking the place of it. >> brenda: you've got the coolest gadgets. jonas your ediction.
1:29 am
the touchy feely world of my cool internet phone is spreading to computers with the windows 8 rollout. it's not just benefits microsoft, but people of the gorilla glass company of corning going up. >>renda: you really love that thing. toby, a bull or bear. >> it was a great call about six months ago. >> brenda: gary b? your prediction? >> don't give up on amazon, i still think the stock is a winner. up 50% the next two years. >> brenda: jim, are you bull or bear, what do you think of garyb? >> a great company, but look, the valuation is still too-- it can go down a lot more and still have too high of a valuation. >> brea: toby, your prediction? >> all right. so, at the end of the day, facebook now has figured out mobile and there's so many people beating against facebook probably moves up 30% by go, ooh, ooh. >> brenda: gary b. >> since the stock was there a serial disappointer. >> brenda: and neil, i give him a
TV News
Search inside
Search only quotes
Search only political ads
Date - Most Recent
Date - Oldest
Bulls and Bears
FOX Business October 28, 2012 1:00am-1:30am EDT
News/Business. The latest market news; the week ahead on Wall Street. (CC)
Gary B
San Francisco
Do U Like Sirius
Tim La
FOX Business
Scanned in
San Francisco, CA, USA
Comcast Cable
Virtual Ch. 130 (Fox Business)
Video Codec
Audio Cocec
Pixel width
Pixel height
Internet Archive
sound, color
Borrow a DVD of this show
Stream Only
In Collection
FOX Business
In Collection
Television Archive
In Collection
Television Archive News Search Service
Uploaded by
TV Archive
on 10/28/2012


text:
News By Tag
Industry News
News By Place
Industry News
Riverside County - RCRMC Foundation/KBHOME Mobile Health Clinic Donation
New Mobile Heatlh Clinic provides health care services to families throughout Riverside County
A special Ribbon Cutting ceremony will be held at the Riverside County Board of Supervisors meeting Tuesday, June 28, 2011, 8:30 a.m. at the County Administrative Center, 4080 Lemon Street, Riverside. Complimentary blood pressure and blood sugar tests will be offered following the ceremony.
The Southern California Division of KB Home formed a partnership with the RCRMC Foundation in 2007 and committed to help raise $1,000,000 (net) by 2021 to further the Foundation's mission. The proceeds from the first five years of their KB HOME Golf Classic were used to purchase the Mobile Health Clinic.
About the SCBC Program
The SCBC Program serves the residents of Riverside County that have limited access to health care providers. Program services include healthcare education, general medical exams, immunizations, vaccinations, as well as screening and treatment of chronic health conditions for adults, children and families. The early detection, treatment and effective ongoing management of chronic illnesses is crucial. The team is comprised of a healthcare provider, a pharmacist, a licensed vocational nurse, a
health services assistant and supporting staff.
About the RCRMC Foundation
The RCRMC Foundation was formed in 1989 as a non-profit organization. The mission of the Foundation is to support Riverside County Regional Medical Center by providing comprehensive and vigorous programs of community awareness and education, public relations and fundraising activities.
For information on the SCBC Program call (951)486-7555.
# # #
The Medical Center is a level II Trauma Center for adults and pediatrics. There are 362 beds in the main acute-care hospital and 77 beds in a separate psychiatric facility.There are adult, pediatric and neonatal intensive care units & birthing center.
Account Email Address Account Phone Number Disclaimer Report Abuse


text:
2540 Mission College Blvd.
Santa Clara, CA 95054
+1.408.454.4200 (International)
For all support inquiries, click here
email@example.com - NA
firstname.lastname@example.com - EMEA
+49 (0)6104 969 102 (Germany, Austria, Switzerland)
+44 (0)7976 007456 (All other EMEA countries)
email@example.com - APAC
+886 935929696 Hong Kong, Taiwan & China
+65 9030 7601 (All other APAC countries)
"I went from 2nd gen to 5D and even added a disk. It was super-painless!"
"One of my 1.5T hard drives just died. No big deal. Thanks @Drobo!"
"We love our @Drobo 5N! It's been the perfect #NAS solution for our small #animation studio."
"Have I mentioned lately how much I love my @Drobo? #icansleepatnight"
"A @drobo a day will keep the computer doctor away! Don't forget to backup guys!"
Smart storage that grows with you and protects what matters.


text:
Book your room at Sleep Inn
Sleep Inn places you within a 10-minute drive of Fredericksburg Area Museum and Cultural Center and Spotsylvania Towne Centre. This 68-room hotel welcomes guests with conveniences like free breakfast, free in-room WiFi, and free self parking. Fredericksburg Expo and Conference Center and United States National Slavery Museum are also nearby.
Free buffet breakfast is included with your stay.
All 68 air-conditioned rooms provide microwaves, refrigerators, and coffee makers. Guests can enjoy cable TV and stay connected with free WiFi and wired Internet. Other amenities include free local calls, hair dryers, and safes.
Property features
Sleep Inn features free WiFi in public areas, a 24-hour business center, and coffee in a common area. A stay here includes free parking. The 24-hour front desk has staff always at the ready. Additional amenities include laundry facilities and a gift shop.
- Free buffet breakfast
- Free WiFi in rooms and public areas
- Free parking
- 24-hour business center
Sleep Inn595 Warrenton Rd Fredericksburg VA 22406
Hotel Amenities
Hotel Amenities
Public areas are equipped with complimentary high-speed wireless Internet access. This hotel offers access to a 24-hour business center. Guests can enjoy a complimentary breakfast. This Fredericksburg hotel also offers gift shops/newsstands, coffee/tea in a common area, and laundry facilities. Complimentary self parking is available on site.
- Number of buildings/towers - 1
- Free RV, bus, truck parking
- 24-hour business center
- 24-hour front desk
- Total number of rooms - 68
- Number of floors - 2
- Gift shops or newsstand
- Laundry facilities
- Coffee/tea in common areas
- Free breakfast
- Free self parking
- Free WiFi
Family Friendly Amenities
- Family Hotel
- Free Wi-Fi
- Rollaway/extra beds available
- Laundry facilities
- Refrigerator
- Microwave
- Cribs/infant beds (complimentary)
Available in all rooms: Free WiFi , Free wired high-speed Internet
Available in some public areas: Free WiFi
Free self parking, Free RV, bus, truck parking
Room Amenities
- Free cribs/infant beds
- Connecting/adjoining rooms available
- Rollaway/extra beds available
- Desk
- In-room climate control (air conditioning)
- Free local calls
- Hair dryer
- Iron/ironing board
- In-room safe
- Pay movies
- Air conditioning
- Refrigerator
- Coffee/tea maker
- Microwave
- Cable TV service
- Free WiFi
- Free wired high-speed Internet
Where to Eat
Guests are offered a complimentary buffet breakfast.
To make requests for specific accessibility needs, complete the "Special or Accessibility Requests" section of "Trip Preferences" when you book your hotel.
- In-room accessibility
Hotel Policies
Check-in time starts at 3 PM
Check-out time is 11 AM
Payment types
- Pets not allowed
You need to know
Extra-person charges may apply and vary depending on hotel policy.
Government-issued photo identification and a credit card or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
Optional extras
- Rollaway bed fee: USD 10 per night
Hotel Name
- Sleep Inn Fredericksburg
- Sleep Inn Hotel Fredericksburg
We should mention
Children 18 years old and younger stay free when occupying the parent or guardian's room, using existing bedding. The property has connecting/adjoining rooms, which are subject to availability and can be requested by contacting the property using the number on the booking confirmation.


text:
© 2016 Need Supply CompanyAll Rights Reserved
Sign up to get notified when this product is back in stock
Free Shipping Free Returns
Orders Over $75 See Details
A modern and edgy, moto-inspired faux leather vest. Features asymetrical front zip closure, three front pockets, and a chunky stud belt detail at the waist.
• Moto-inspired faux leather vest
• Oversized fit
• Asymmetrical front zip closure
• Chunky waist belt
• Fully lined
• 55% Polyurethane, 45% Polyester
• Dry clean only
View Size Chart
20.5" bust
25" shoulder to hem
Measurements taken from size small/medium
Model wears size small/medium. Model is 6'2
This vest fits intentionally oversized
View Size Chart
Please note that this is a final sale item. No returns or exchanges will be accepted.
3100 West Cary Street
Richmond, VA 23221
© 2016 Need Supply Company—All Rights Reserved
Sign up for our newsletter and get 10% off your next order. Pretty sweet, we know.


text:
Come on your adventure of a lifetime with Rocky Mountain Trench Adventures, here in the heart of Northern British Columbia, Canada. No matter what the season, Rocky Mountain Trench Adventures will design a customized adventure around all our guests adventure requirements. From fishing, snowmobiling, hiking, wildlife photography, book of ra, skiing, jet boat tours, helicopter fishing tours or camping. Rocky Mountain Trench Adventures will be happy to accommodate your personal requirements. We are based in beautiful Mackenzie, British Columbia, Canada at the south end of Williston Lake the gateway to the last wilderness frontier.
Rocky Mountain Trench Adventures potty training girls age 2 takes pride in the fact that our team has extensive knowledge of the area, we have taken the time to study each area in all seasons to provide a fun, safe, reliable adventure. At Rocky Mountain Trench Adventures we put our clients comfort and safety first.
Have peace of mind while traveling with Rocky Mountain Trench Adventures beyond diet review. Our friendly and knowledgeable team will provide you with a fun safe wilderness experience.
PACKAGES & RATES l WINTER ADVENTURES l SUMMER ADVENTURES l INDUSTRIAL & CAMP SERVICESHOME l SITE INDEX l PHOTO GALLERY l MOVIE GALLERY l NEWS l LINKS l CONTACT l EMAIL
© 2005 Rocky Mountain Trench Adventures, Mackenzie, BC, Canada.


text: Movies | By Katherine Spiers on October 13, 2010 @ 2:29 pm Email Print Related 'Avatar' … With Babies! (Video) 'Breaking Bad' With a Laugh Track (Video) The actor donned his iconic glasses for Spike TV's Scream Awards promo Breaking News To Your Inbox. Sign up » http://twitter.com/share?text=Michael J. Fox Re-creates 'Back to the Future' Trailer&via=TheWrap&url=http%3A%2F%2Fwww.thewrap.com%2Fmichael-j-fox-re-creates-back-future-trailer-21690%2F&t=Michael J. Fox Re-creates 'Back to the Future' Trailer http://www.facebook.com/sharer.php?u=http%3A%2F%2Fwww.thewrap.com%2Fmichael-j-fox-re-creates-back-future-trailer-21690%2F&t=Michael J. Fox Re-creates 'Back to the Future' Trailer Email Print


text: I have two authors on my blog but would like the author template to only work for one. The second "author" is really just a dummy account that I use for all guest posts. Is there something I can add to author.php so that it only goes through the loop for a specific author?


text:
A small amount. Or someone being tight, stingy or cheap.
(Heard in south east London)
(Heard in south east London)
"He gave me lack"
"Quit giving me lack"
"Quit giving me lack"
by Chris Hab February 28, 2008
Trending Now
Top Definition
"Lack" Waterford, Ireland Slang for Girlfriend.
Went out with the Lack last night, Then gave her a seeing to afterwards!
by Frank Forsey September 03, 2003
reffering to the vehicle known as the Caddilac.
"Damn that lack is pimpin on dem dubs"
by ShOvel July 24, 2003
v. the absense of an object or ability
"this chick looks good as hell but she lacks a nice ass"
"he's tryna play me in a one on one but lacks basketball skills"
"he's tryna play me in a one on one but lacks basketball skills"
by S H A U N July 28, 2005
A big strip of fat across one's face
"Omg that part of your face is fat! You have a serious lack"
by GreyWolf2001 June 02, 2015


text:
Sounds Like: Pet Shop Boys, Erasure, Roxette Official, Depeche Mode, Keane
Manager: Michael Cross
Bio: Back in 1996 (when people still had tape players) Brendon Shields made a pledge to his friend TRON-e that come what may the two will one day make music together. 16 years, and a whole lot of water under the bridge later – The Yoo was officially born.
Shields contribute most of the lyrics and sin...See Full Bio


text:
Using the "offers from" options you can filter merchants from regions which are most common in our database: the UK, Germany, Austria and Poland.
If a checkbox near a flag is selected, offers from merchants from that particular region will be displayed, otherwise they will not be visible.
If you select offers from "all regions", all offers from all merchants will be displayed. These incude offers from merchants in countries not listed here.
Sizes: 50, 52, 54mm
incl. 4 spacer and 8 Speed Washer
Sizes: 53mm hardness: 100A
Sizes: 54mm hardness: 100A
Sizes: 52mm hardness: 83B
Sizes: 54mm hardness: 80A
Sizes: 54, 56, 58, 60mm hardness: 84B
Sizes: 50, 51, 52, 53, 54, 55mm
Sizes: 51, 52, 53, 54mm
Sizes: 60mm hardness: 84B
Sizes: 55mm hardness: 84B
Sizes: 50, 52, 54mm hardness: 83B
Sizes: 50, 52mm hardness: 83B
Sizes: 53mm hardness: 83B
Sizes: 51mm hardness: 83B
Sizes: 55mm hardness: 83B


text:
ODT Online
ODT /directory
A Wanaka man charged with assaulting a policeman and
threatening to kill was released on bail when he appeared
before a justice of the peace in the Queenstown District
Court yesterday.
Peter John Barratt (72), sporting a black eye and what
appeared to be a bloodstain on the left shoulder region of
his shirt, was bailed to a Haast address when he appeared
before Joy McDonald.
Barratt is charged with assaulting Senior Constable Sean
Hurley and threatening to kill Hurley, his daughters and
Constable Peter Reed after an incident at Makarora on
Snr Const Hurley (52) was left bloodied and bruised after the
incident. Barratt has also been charged with drink-driving
after recording a breath-alcohol level of 751mcg when stopped
by police yesterday. The legal limit is 400mcg.
Queenstown solicitor Sonia Vidal, representing Barratt,
entered no plea and asked for a remand on bail. Miss McDonald
released Barratt on bail until his next scheduled court
appearance on April 28.
Bail conditions require him to live at a house he has in
Haast, which is in the same street as the local police
Barratt must report to the station daily between 8am and 4pm,
and not drive, drink, enter licensed premises or enter Wanaka
unless to travel to the Queenstown District Court.
''I take a very dim view of what's happened,'' Miss McDonald
told Barratt before outlining the bail conditions and asking
him to surrender his passport.
Prosecuting Sergeant Ian Collin earlier told the court police
were satisfied the bail conditions were appropriate ''in
terms of eliminating any issues as to further offending''.
A police statement on Thursday said Snr Const Hurley had
stopped a motorist in Makarora for a routine breath test
about 3pm after road workers reported a suspected
drink-driver heading towards Wanaka.
After failing the test, the motorist was asked to accompany
Snr Const Hurley to Wanaka, but on the way, the motorist is
alleged to have kicked the officer in the head and tried to
gouge his eye.
Wanaka sub-area commander Senior Sergeant Allan Grindell said
Snr Const Hurley stopped his car and restrained the man
before continuing.
However, the man began ''playing up'' in the back seat again
as the car approached the Makarora Country Cafe so Snr Const
Hurley pulled into the cafe forecourt, restrained him again
and radioed for help.
- Mountain Scene
Post a note
Submit your news & photos - Login or
Opinion? Have your say - Login or Register
Submit your
event or notice
Add a photo
Bookmark ODT
Make ODT my
Otago Daily Times print and digital edition, The
Star - Dunedin, The News - Central Otago, The
Ensign - Gore, Southland Express -
Invercargill, Southern Rural Life - Otago and
Southland, The Courier - Timaru, Courier Country - Canterbury and
West Coast, Queenstown Times - Queenstown, Mountain Scene -
Queenstown, The Oamaru Mail - Oamaru, The
Greymouth Star - Greymouth, Hokitika Guardian - Hokitika,
West Coast Messenger - West Coast, The Ashburton Courier -
Ashburton, The News - North Canterbury, Extra! schools
publication, Property
Times South Canterbury


text:
Byrne: 'Tough edges' to Labour reforms
9 June 2013 Last updated at 11:10 BST
Shadow Work and Pensions Secretary Liam Byrne has said Labour will target low pay and housing benefit in its plans for social security reform.
Byrne told The Andrew Marr Show Labour would look at placing limits on out-of-work payments, household and overall benefits.
You can watch the Andrew Marr Show in full on the BBC iPlayer.


text:
Magical Mimi Dress Up Shoes and Jewellery Set
product code: 122708
see more in this range by Magical Mimi
- Standard UK home delivery within 5 working days - £3.95
- Click and collect in any store - FREE
Add a touch of glamour to your dressing up box. This set includes four pairs of pretty shoes, a pair of clip on earrings, a ring, a necklace and a tiara making them perfect for adding the finishing touches to your princess outfits.
Colours and styles may vary.
Not suitable for children under 36 months due to small parts.Battery requirements: Not Required
How does this help my child ?
This toy encourages your child to enjoy using their imagination.
This product is available for:
Standard Delivery
Normally delivered within 3-5 working days (exclusions apply including postcode restrictions and extended timescales). Exceptional delivery circumstances beyond our control such as bad weather or courier delays may cause your order to take longer to deliver.
FREE for orders over £40, otherwise £3.95
Express Delivery
Order before 4pm for delivery on the next working day to UK mainland addresses. Regrettably this service is unavailable for deliveries to Northern Ireland.
£4.95 on all orders
Click and Collect Delivery
Order online and have your order delivered to any ELC or Mothercare store on the date of your choice. You will receive a text message confirming your order is ready for collection. This service is available in the UK only.Please be advised that we will hold your order for a maximum of 7 days, after which time your order will be returned to our warehouse
UK returns
ELC are happy to refund or exchange any unused merchandise returned within 30 days of purchase upon production of your receipt, as long as they are returned in an unused condition and in their original packaging. Please see Terms and Conditions for details. This does not affect your rights if the product is faulty or mis-described.