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Patent application title: CARBOXYLATED LATEX PARTICLES
Albert Geiger (Penzberg, DE)
Gernot Frackmann (Wald-Michelbach, DE)
Hans-Ulrich Moritz (Bendestorf, DE)
Peter Sluka (Weilheim, DE)
IPC8 Class: AG01N33546FI
USPC Class:
Class name: Carrier is organic carrier is synthetic resin carrier is water suspendible particles
Publication date: 2008-10-30
Patent application number: 20080268552
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Patent application title: CARBOXYLATED LATEX PARTICLES
Albert Geiger
Gernot Frackmann
Hans-Ulrich Moritz
Peter Sluka
Origin: INDIANAPOLIS, IN US
IPC8 Class: AG01N33546FI
USPC Class:
The present invention relates to processes for preparing carboxylated
latex particles based on the copolymerization of a monoalkenylaromatic
monomer (A) and an aliphatic unsaturated carboxylic acid (B), where the
copolymerization takes place at a pH in the range from pH >1.5 to pH
<4.5. The pH is typically monitored during the copolymerization
process and regulated by suitable compensatory titration. The invention
likewise relates to the carboxylated latex particles obtainable by one of
the processes according to the invention, and to the use of these
particles in an immunological test method.
1. A process for preparing carboxylated latex particles comprising the
steps of:forming a mixture comprising a monoalkenylaromatic monomer and
an aliphatic unsaturated carboxylic acid, whereby copolymerization and
carboxylation of the monomer takes place and wherein the mixture has a pH
between 1.5 and 4.5,monitoring the pH during copolymerization,
andregulating the pH by compensatory titration so that it remains
constant during copolymerization, thereby forming carboxylated latex
2. The process of claim 1 wherein the monoalkenylaromatic monomer is selected from the group consisting of styrene and alpha-methylstyrene.
3. The process of claim 1 wherein the aliphatic unsaturated carboxylic acid is selected from the group consisting of acrylic acid, methacrylic acid, fumaric acid, and itaconic acid.
4. The process of claim 1 wherein the mixture is an aqueous emulsion.
5. The process of claim 1 wherein the mixture further comprises a water-soluble peroxide.
6. The process of claim 1 wherein the mixture further comprises a seed latex.
7. The process of claim 1 wherein the aliphatic unsaturated carboxylic acid is dissolved in the monomer prior to forming the mixture.
8. The process of claim 1 wherein the pH of the copolymerization mixture deviates from a target pH by a maximum of .+-.0.3.
9. The process of claim 1 wherein the regulation step utilizes an aqueous solution of an alkali metal carbonate, bicarbonate, or hydroxide as a regulating reagent.
10. The process of claim 1 wherein the pH is greater than or equal to 2.0 and less than 4.0.
11. A carboxylated latex particle prepared by a process comprising the steps of:forming a mixture comprising a monoalkenylaromatic monomer and an aliphatic unsaturated carboxylic acid, whereby copolymerization and carboxylation of the monomer takes place and wherein the mixture has a pH between 1.5 and 4.5,monitoring the pH during copolymerization, andregulating the pH by compensatory titration so that it remains constant during copolymerization, thereby forming carboxylated latex particles.
12. A conjugate comprising a particle according to claim 11 and a biomolecule selected from the group consisting of nucleic acids, peptides, proteins, haptens, lipids, and polysaccharides.
13. An immunological test method to determine an antigen or antibody comprising the steps ofcombining a sample containing the antigen or antibody with a conjugate comprising a particle according to claim 11 and a binding partner for the antigen or antibody whereby the particle aggregates with the antigen or antibody anddetecting aggregation of the particle as a measure of the antigen or antibody in the sample.
[0001]This application is a continuation of PCT/EP2006/006287 filed Jun.
26, 2006 and claims priority to EP 05014340.3 filed Jul. 1, 2005.
[0002]The present invention relates to processes for preparing carboxylated latex particles based on the copolymerization of a monoalkenylaromatic monomer (A) and an aliphatic unsaturated carboxylic acid (B), where the copolymerization takes place at a pH in the range from pH >1.5 to pH <4.5. The pH is monitored during the copolymerization process and regulated by suitable compensatory titration. The invention likewise relates to the carboxylated latex particles obtainable by one of the processes according to the invention, and to the use of these particles in an immunological test method.
[0003]Surface-modified latices have been employed for more than 40 years in many biomedical areas of application, such as, for example, in diagnostic tests as support material, inter alia for enzyme immobilization. Singer, J. M. and Plotz, C. M. (American Journal of Medicine 21 (195.6) 8988-896) used latex particles as long ago as 1956 in the development of homogeneous immunoassays.
[0004]Besides aminated latex particles, the latices mainly employed as starting material in biomedical applications have been functionalized on the surface for example by carboxyl, epoxide or hydroxyl groups. Biopolymers such as, for example, specific antibodies are linked to these latex particles either by passive coating (adsorption) or by covalent bonding, directly or with the aid of activating reagents such as, for example, N-hydroxybenzotriazole.
[0005]Many processes for preparing latex particles are known in the art.
[0006]For example, Roncari, G., (U.S. Pat. No. 4,226,747) describes a process for preparing latices which consist predominantly of styrene and butadiene and which are distinguished by anilylsulpho groups on the latex, surface. The co-monomers employed are sodium methallyl-sulphonate, acrylic acid and itaconic acid. The resulting latex is reacted with sodium dihexyl sulphosuccinate in water, α,α'-azobisisobutyramidinium chloride in water and p,p'-dithiobisaniline in styrene.
[0007]The application of Yaacoub, E. J., (DE 102 04 234) relates to a polymer latex prepared using at least one carbohydrate or carbohydrate derivative which is substituted by a group capable of free-radical polymerization and has surfactant properties. The polymer latex prepared by this process typically comprises styrene and methyl methacrylate as comonomers. These latex-particles can be employed in cosmetic, clinical or diagnostic applications.
[0008]Bowell, S. T. et al., (U.S. Pat. No. 2,734,883) prevented the decrease in pH during the emulsion copolymerization of styrene and butadiene, despite the acid produced by decomposition of the peroxo initiator, by continuous or intermittent addition of alkali so that the pH is kept at or above 9.7.
[0009]In Gasche, H. E. et al., (US 2003/0153678), the pH is adjusted to a pH of 9-11 before addition of the initiator in the batch polymerization of a latex, and is monitored during the reaction so that it does not deviate by more than ±0.5.
[0010]In the polymerization process for a polybutadiene latex filed by Sturt, A. G. and Feast, A. A. J. (DE 2 103 610), the pH is monitored up to a conversion of 15% in order to ensure emulsion-stable emulsion conditions.
[0011]To prepare an impact-resistant nitrile polymer in a multistage process of Takahashi, A., et al., (JP 61166813), the pH is adjusted to below 5 in the first copolymerization which is carried out batchwise, and the pH is reduced to 3 before the second, semicontinuous, stage.
[0012]A further process for preparing a stable latex was described by Sun-Lin, C. et al. (EP 0 476 528). Polymerization of the latex is initially candied out in the presence of a surface-active phosphate ester at a pal of below 3.5. The latex particles are then adjusted to a pH of between 7 and 10.5 by adding ammonia solution.
[0013]In the patent application of Meiners, C. et al., (DE 102 36 395), an aqueous polymer dispersion is obtained in a batch emulsion polymerization and is preferably used as rust-preventing paint. The emulsion is in this case adjusted to a pH of greater than or equal to 4.5, preferably 5-7, with aqueous ammonium hydroxide solution.
[0014]Brewer, J. F., (U.S. Pat. No. 3,032,524) developed a process in order to adjust the pH of a latex accurately. In this case, the pH is adjusted exactly to the desired pH with a tolerance of ±0.1 pH units by means of carbon dioxide during the preparation process. For this purpose, carbon dioxide is fed into the reactor and finely distributed by means of an impeller agitator.
[0015]The use of latex particles, including the use of carboxylated latex particles, for diagnostic purposes is known in principle. Thus, Fisk, R. T. (U.S. Pat. No. 3,088,875) in 1959 described an immunological test intended to diagnose antigens or antibodies in body fluids. Pure polystyrene latices were employed as support material. These were known latices manufactured by Monsanto Chemical Company or Koppers Company Inc. with particle sizes of 150-200 nm. The latex particles were coated with 7-S γ-G globulin (now called class G immunoglobulin) in order to detect the so-called rheumatoid factor (RF), a 19-S γ-M protein (now called class M immunoglobulin) in human serum by aggregation of the latex particles. The RF binds to the coated immunoglobulin G. This results in aggregation of the latex particles.
[0016]A latex functionalized on the surface by epoxy groups was prepared by Batz, H. G. et al. (EP 0 054 685). This latex is employed as support material to which biological and/or immunologically active substances are covalently bonded directly or via a coupling agent. One or more of the monomers glycidyl methacrylate, glycidyl acrylate, glycidyl vinyl ether, glycidyl vinyl phthalate and 3,4-epoxybut(l)ene is/are used as monomer containing epoxide groups. This/these is/are copolymerized for example with styrene, dienes, acrylamide, methacrylamide, alkyl hydroxyalkyl and aminoalkyl acrylate or methacrylate, vinyl ether, vinyl ester or N-vinylpyrrolidone.
[0017]A patent application by Seksui Chemical Co. Ltd. (JP 59179609) is concerned with the preparation of a latex for the diagnosis of antigens and antibodies, with the pH being varied during the polymerization. In this case, styrene and styrene sulphonate in water is mixed with an emulsifier and a persulphate as initiator and polymerized initially under alkaline and subsequently under acidic or neutral conditions.
[0018]The publication by Miraballes-Martinez, I. et al. (J. Biomater. Sci. Polymer Edn. 8 (1997) 765-777) describes the preparation of a latex which is functionalized by chloromethyl groups on the surface and is suitable for covalent coupling of immunoglobulin G (IgG). The chloromethyl-styrene latex was in this case prepared in a two-stage process in a batch reactor. The experiments carried out in this publication showed that the IgG-latex obtained in this way can be employed for applications in immunodiagnostic tests.
[0019]Functionalized latices, predominantly carboxylated latices, are employed in the process patented by Hager, H. (U.S. Pat. No. 3,857,931).
[0020]In DE 27 12 044, Beskid, G. and Savard, E. V. describe the coupling of antibodies, antigenic substances or other biological materials to latex particles for serological tests. The Streptococcus group A antigen is conjugated, using a water-soluble carbodiimides, to the purified latex particles from this process.
[0021]Focella, A., et al. (DE 27 23 449) describe an agglutination test based on carboxylated latices and able to detect barbiturates.
[0022]Fischer, E. A. (DE 28 40 767) deals with the types of carboxylated latex support particles to which immunologically active materials such as, for example, primary amines, amino acids, peptides, proteins, lipo- and glycoproteins, sterols, steroids, lipoids, nucleic acids, enzymes, hormones, vitamins, polysaccharides and alkaloids can be coupled. Typical suitable latices are according to this carboxylated styrene-butadienes, carboxylated polystyrenes, acrylic acid polymers, methacrylic acid polymers, acrylonitrile polymers, acrylonitrile-butadiene-styrenes, polyvinyl acetate-acrylates, polyvinylpyridines, vinyl chloride-acrylates and the like.
[0023]A further patent by Fischer, E. A. (U.S. Pat. No. 4,264,766) describes the development of an immunological test in which a carboxylated styrene-butadiene latex (Dow CL 241) is activated with 1-amino-2-hydroxypropyl-dextran and further processed for the test.
[0024]Gallati, H. (DE 27 49 956) used a carboxylated styrene-butadiene latex to which an immunological reagent, e.g. HCG (human chorionic gonadotropin), is bound.
[0025]As explained above, latex particles can in principle be employed in diagnostic test methods. However, very great demands are made on the characteristics of the latices for use in diagnosis. In particular, it is very important for sensitive immunodiagnostic methods that the latex particles employed can be well characterized in terms of their size and/or in terms of their surface properties, in particular in terms of the charge distribution and charge density, and can be prepared reproducibly.
[0026]The defined and reproducible preparation of carboxylated latex particles and thus also the availability of carboxylated latex particles with defined charge distribution on the surface is difficult with prior art processes.
[0027]It was therefore an object of the present invention to establish processes which help to eliminate the disadvantages known from the prior art and provide carboxylated latex particles which can be employed for example advantageously in immunological test methods.
[0028]It has surprisingly been found that a process for preparing carboxylated latex particles based on the copolymerization of a monoalkenylaromatic monomer (A) and an aliphatic unsaturated carboxylic acid (B), where the copolymerization takes place at a pH in the range from pH >1.5 to pH <4.5, where the pH is monitored during the copolymerization process and regulated by suitable compensatory titration, is advantageous. Carboxylated latex particles with the desired size and charge density can be prepared very reproducibly in this process, and the latex particles obtained by this process show particular advantages and properties.
[0029]In a first embodiment, the present application relates to a process for preparing carboxylated latex particles based on the copolymerization of a monoalkenylaromatic monomer (A) and an aliphatic unsaturated carboxylic acid (B), where the copolymerization takes place at a constant pH in the range from pH >1.5 to pH <4.5. The pH is monitored during the copolymerization process and regulated by suitable compensatory titration. It is preferred for the monitoring of the pH in the copolymerization mixture and the compensatory titration to take place continuously. It is further preferred for the pH regulation to take place continuously and automatically. It is possible under the strongly acidic pH conditions of the process according to the invention for the charge density of the carboxyl functions on the latex particles to be controlled reproducibly and in a targeted manner.
[0030]In a further embodiment, the copolymerization takes place at a pH of greater than or equal to pH 2.0 and less than pH 4.0.
[0031]In a "copolymerzation", two or more monomers are simultaneously polymerized together, the resulting copolymer being composed of the constituent monomers. The properties of the copolymers depend on the combination and reactivity of the monomers, and the proportions of monomers and the management of the reaction. Copolymerizations can in principle be carried out in heterogeneous and homogeneous systems in reactors operated continuously, semicontinuously and batchwise. Copolymers with a wide variety of properties can be produced thereby. Although copolymers composed of three or more different monomers (ternary or quaternary copolymers etc.) exist, typically not more than three different polymerizable monomers are employed.
[0032]Seed polymerization is a special emulsion polymerization process used in particular for preparing latices with a very narrow particle size distribution, and latices composed of more than one type of monomer and of defined microstructure within the individual latex particles. It is carried Out by adding the monomer to be polymerized to a dispersion of latex particles of a uniform particle size distribution, called the seed latex. The monomer diffuses uniformly into the seed latex particles and is % polymerized therein. In the specific case of seed polymerization, the concentration of the emulsifier present in the emulsion is kept within the critical micelle concentration (CMC). Otherwise, newly produced, much smaller latex particles would lead to a great broadening of the size distribution. Seed polymerization corresponds according to the model of Smith, W. V. and Ewart, R. H. (Journal of Chemical Physics 16 (1948) 592-599) to phases II and III of an emulsion polymerization. The particle formation phase (I) of an emulsion polymerization is redundant due to the presence of the seed latex particles.
[0033]Numerous publications have appeared since 1950 on the theory of particle growth in seed polymerization (inter alia Poehlein, G. W. and Vanderhoff; J. W., J. Polymer Sci. 11 (1973) 447-452; Feeney, P. J., Napper, D. H., Gilbert, R. G., J. Colloid Interface Sci. 118 (1987) 493-505). The theoretical descriptions of latex preparation, based on the Smith-Ewart theory, deal with the influence of various factors on the particle size distribution. Poehlein and Vanderhoff describe therein for example "competitive growth" leading to a narrower particle size distribution. In this connection, they assume that, owing to differences in the specific surface area, smaller particles of the seed latex grow faster than larger ones, because there is a greater probability of monomers diffusing into the smaller particles.
[0034]Monodisperse latices with larger particle diameters are usually prepared by carrying out stepwise polymerizations. This entails initially a first seed latex being prepared by suitable polymerization. These seed latex particles then serve as starting material in the subsequent (seed) polymerization. It is possible in this way to prepare latices with a very narrow particle size distribution. Such polymerizations are frequently carried out as batch polymerizations, with the rate at which the monomers enter the seed particles, the average number of free radicals in the latex particle and the volume growth rate being decisive for the influence on the particle size distribution. In the industrial preparation of latices with particular size distributions, such as, for example, in the styrene-butadiene rubber, seed polymerizations are frequently employed with the reaction being managed semicontinuously or semibatchwise.
[0035]Semibatch emulsion polymerization is typically employed industrially when the aim is to obtain aqueous dispersions. For this purpose, two different so-called feed methods, are used, the monomer feed and the emulsion feed.
[0036]In the monomer feed, the initial charge comprises water, emulsifier and initiator, and usually a small proportion of the monomer, and the remaining monomer and possibly also initiator are added during the polymerization.
[0037]In the emulsion feed, for example the initial charge comprises a portion of the emulsion and, after the reaction has started, the remaining monomer emulsion, which may differ in composition from the initially charged emulsion, is metered into the reactor.
[0038]The advantage of semibatchwise management of the reaction is that the polymerization rate and, the heat of evolution can be controlled via the feed rate, the concentration of unreacted monomer can be kept low, and the reaction mixture can be cooled with cold feed. In order to produce a uniform polymer in the copolymerization of two monomers differing in reactivity, it is possible either for the initial charge to comprise an excess of the monomer which polymerizes more slowly, and to feed, in the more reactive monomer, or for the monomer composition to be adjusted during the feeding (Chujo, K., et al., J. Polymer Sci. 27 (1969) 321-332; Snuparek, J. and Kr{hacek over (s)}ka, F., J. of Applied Polymer Science 20 (1976) 1753-1764). To control the copolymer composition, various monomer dispensing strategies have been developed and quantified in order to set the necessary, time-dependent metering rates (Hamielec, A. E., et al., Compr. Polym. Sci 3 (1989) 17-31). It is thus possible by suitable feed conditions to vary the degree of polymerization, the particle size and the particle size distribution without altering the overall composition of the latex. If the feed rate is not too high, according to Krackeler, J. J. and Naidus, H., J. Polymer Sci. 27 (1969) 207-235, a quasi steady-state is set up with monomers which comply which the Smith-L-wart mechanism, i.e. no formation of new particles takes place during the monomer metering, with the net reaction rate R and the feed rate F being approximately identical, and R depending only on the feed rate (Wessling, R. A., J. of Applied Polymer Science 12 (1968) 309-319). The monomer concentration [M] in this case assumes a value below the saturation concentration and changes only slightly during the feed period.
[0039]In this case, the reaction rate R [mol/s] and the monomer metering rate F [mol/s] can be described by the following equation of the Formula I:
[0040]In this, the constant G depends on the average number of free radicals per particle, the number of particles, the growth rate constants, the reactivity coefficients and the partition coefficients of the monomers between the latex and the aqueous phase.
[0041]The time-conversion plot for semibatchwise polymerizations is linear over a wide range, and the molar mass distribution or the composition of the polymer becomes homogeneous because a substantially constant polymerization rate is maintained over the reaction time (Snuparek, J. and Kr{hacek over (s)}ka F., J. of Applied Polymer Science 20 (1976) 1753-1764).
[0042]It is important and preferred for the copolymerization to proceed in a controlled manner under so-called "starved conditions".
[0043]It has proved, to be particularly advantageous to construct the carboxylated latex particles according to the invention according to the core-shell principle. In this process, the initial charge comprises a core or seed latex, and the shell with the desired properties is prepared by copolymerization on this seed latex. Suitable, in principle as seed latex are all latex particles. It is preferred for the seed latex to be able to enter into covalent connections with the reagents employed to construct the carboxylated shell. The seed latex typically employed is a polystyrene mixed latex or a pure polystyrene latex. The skilled person is able to select suitable seed latices without an inventive step. The skilled person is able to calculate from the size and the concentration of seed latex-particles the amounts of monoalkenylaromatic monomer which are required to form the desired shell. The amount of aliphatically unsaturated carboxylic acid can be ignored in the calculation and adjustment of the particle size, because the proportionate amount thereof is negligible in this context.
[0044]The final size or target size of the particles depends substantially on the number and size of the seed latex particles and on the amount of monoalkenylaromatic monomer present. The relation between particle size, target molecule, weight of target latex and weight of seed latex is represented by Formula II:
where DT=average diameter of the target latex, DS=average diameter of the seed latex (average), wtot=wseed+wmonomer in g, wseed=weight of seed latex employed in g, and monomer=weight of monomer employed in g.
[0045]The number (amount) and size of the seed particles thus specifies the amount of monomers necessary to achieve a particular target particle size of the final product.
[0046]The mixture for a stepwise polymerization is normally chosen so that the target latex results as an approximately 3 to 30% strength suspension. The conditions are typically chosen so that the target latex is in the form of a 5 to 20% strength suspension.
[0047]The monoalkenylaromatic monomer (A) will typically be a styrene, an alpha-methylstyrene or a vinyltoluene. Preferred monoalkenylaromatic monomers are styrene and alpha-methylstyrene. If necessary, it is also possible to employ a mixture of two or more monoalkenylaromatic monomers in the process according to the invention. However, typically not more than two of these monomers are mixed. Styrene is typically used as monoalkenylaromatic monomer.
[0048]The aliphatically unsaturated carboxylic acid (B) will typically be acrylic acid, methacrylic acid, fumaric acid, itaconic, acid, maleic acid or maleic anhydride. If necessary, it is also possible to employ a mixture of two or more aliphatically unsaturated carboxylic acids in the process according to the invention. However, typically not more than two of these aliphatically unsaturated carboxylic acids will be employed. It is further preferred for the aliphatically unsaturated carboxylic acid to be selected from the group consisting of acrylic acid, methacrylic acid, fumaric acid and itaconic acid. Acrylic acid is the preferred aliphatic unsaturated carboxylic acid.
[0049]It is necessary in many processes for preparing latex particles to add the various monomers employed for the copolymerization separately to the reaction mixture. A further advantage of the process according to the invention is that a monoalkenylaromatic monomer (A) and the aliphatic unsaturated carboxylic acid (B) can be matched with one another in such a way that (B) dissolves in (A). The aliphatic unsaturated carboxylic acid (B) is therefore typically dissolved in the monoalkenylaromatic monomer (A) and, in the process according to the invention, the mixture of (A) and (B) is fed into the reaction mixture.
[0050]As mentioned, it is particularly advantageous that the process according to the invention for preparing carboxylated latex particles proceeds at a strongly acidic pH and that this pH is continuously measured and kept constant during the reaction. The desired pH for the reaction conditions is also referred to, as target pH. A constant pH for the chosen reaction conditions in the context of the invention described herein is present when the deviation of the pH in the copolymerization mixture, from the desired target pH amounts to a pH of 0.3 or less. It has further proved advantageous for the process according to, the invention that the pH in the copolymerization mixture does not deviate by more than pH 0.3 from the target pH. The process according to the invention is accordingly typically characterized in that the pH in the copolymerization mixture deviates by a maximum of ±pH 0.3 from the target pH. The process according to the invention is further typically characterized in that the pH in the copolymerization mixture deviates by a maximum of ±pH 0.2 or likewise typically by a maximum of ±pH 0.1 from the target pH.
[0051]It is possible in principle to use any suitable alkaline solution to adjust the pH. The pH-regulating reagent typically employed is an aqueous solution of an alkali metal carbonate, bicarbonate or hydroxide.
[0052]As appreciated in the prior art, there are various possibilities for dealing with the problem of the poor solubility of (forming) latex particles. It has proved appropriate in the process according to the present invention for the copolymerization to be carried out in aqueous emulsion. In a further preferred embodiment, the process according to the invention is thus characterized in that the copolymerization takes place in aqueous emulsion. This copolymerization can moreover-typically take place in the absence of emulsifiers.
[0053]The (co)polymerization reaction as is characteristic for the production of latex particles can be initiated by various agents. Known in the art are in particular gamma rays and reagents which release free radicals. The use of reagents which form free radicals under suitable conditions is preferred according to the invention. Peroxide-based free-radical formers are typically employed. Preference is given to water-soluble peroxides, in particular persulphates and peroxides, such as, for example, sodium or ammonium persulphate and hydrogen peroxide.
[0054]The Carboxylated latex particles which can be prepared by the process according to the invention are particularly distinguished also by the fact that the charge-density on the surface of these particles can be predetermined and reproduced in a targeted manner. The charge density is ascertained by means of atomic force microscopy (AFM) (Tan, S., et al., Langmuir 21 (2005) 43-49). The so-called parking area is stated as a measure of the charge density. The parking area indicates, for example for the carboxylated latex particles according to the invention, the average area occupied by a carboxyl, group on the latex surface.
[0055]It is possible in the process according to the invention, as shown in Example 3, to adjust a particular, desired charge density by suitable choice of the reaction parameters. The carboxylated latex particles typically prepared by the process, according to the invention have an average parking area of from 10 to 160 (10-10 m)2, likewise typically from 20 too 130 (10-10 m)2, or likewise typically from 30 to 120 (10-10 m)2.
[0056]It is possible for the parking area not only to be adjusted empirically but also to be calculated in advance by Formula III:
[0057]The y axis of the function describes the parking area in (1010 m)2, and the x axis represents the acrylic acid concentration, set in the formulation in 10-1 mol/l. This value corresponds to the total concentration of acrylic acid in the emulsion achieved during the metering. When the pH=2, then y=15.99x-0.9805, when the pH=3, then y=41.065x-0.4706, and when the pH=4, then y=51.061x-0.4922.
[0058]The values for a and b in Formula III can be ascertained empirically for each polymerization mixture and at each pH. The charge density is typically set in accordance with Formula III in the process according to the invention.
[0059]Since the carboxylated latex particles according to the invention have an exactly defined charge density, they are very particularly suitable for use in diagnostic methods. The defined charge density has the effect that the carboxylated latex particles show a uniform and reproducible behaviour.
[0060]The present invention thus includes in a preferred embodiment also the carboxylated latex particles prepared according to the invention.
[0061]The carboxylated latex particles prepared by the process according to the invention are outstandingly suitable for covalent attachment of biochemical molecules.
[0062]Carboxylated latices are activated for example by using carbodiimides as coupling reagents. Biomolecules can be linked to the latex via amide groups directly or via an active ester. The coupling reagents used in this connection are typically water-soluble carbodiimides (WSC). The following commercially available, water-soluble carbodiimides are particularly suitable for activation and coupling of biomolecules to carboxylated latex particles; 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimides metho-p-toluenesulphonate and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride. These coupling agents can be employed for direct linkage of biomolecules via amide groups. However, it is likewise possible for free carboxyl functions initially to be converted into an active ester in order then to undertake the coupling via this active ester (e.g. an O-acylurea intermediate). Both procedures are depicted diagrammatically below.
Reaction for Direct Coupling to Carboxylated Latex:
[0063]Activation of Carboxylated Latices Via an Active Ester:
[0065]One advantage of the coupling via an active ester is that the carbodiimides can be removed before the antigen or antibody is coupled to the latex, and thus has no influence on the immunological test.
[0066]Suitable biomolecules are all molecules employed in conventional diagnostic test methods. They particularly typically include nucleic acids, peptides and proteins. However, it is also possible advantageously to employ other biomolecules such as, for example haptens, lipids or polysaccharides.
[0067]Preferred biomolecules are from the group of proteins, especially those proteins which represent good binding partners for further biomolecules, with very particular preference in this connection for lectins, avidin, streptavidin and antibodies.
[0068]The term "antibody" means, besides the intact immunoglobulins, also all antibody fragments. These include for example Fab, Fab' or F(ab')2 fragments. The term antibody without the addition of "monoclonal" or "polyclonal" always includes both types of antibodies, plus chimeric constructs and all fragments listed above.
[0069]After attachment of the biomolecules to the carboxylated latex particles according to the invention, the skilled person refers to biomolecule-latex conjugates or latex conjugates for short. The latex conjugates based on carboxylated latex particles according to this invention likewise represent a preferred embodiment of the invention.
[0070]The latex conjugates based on carboxylated latex particles according to this invention are particularly advantageously employed in immunodiagnostic test methods. The design and procedure for such test methods are familiar to the skilled person.
[0071]The following examples further illustrate the invention characterized in the appended claims.
Preparation of Seed Latex Particles (Seed Latex)
[0072]A seed latex consisting of completely polymerized and emulsifier-stabilized latex particles is prepared by an emulsion polymerization. The procedure for this follows for example the experimental method of van den Brink (M. v. d. Brink: "On-line monitoring of polymerization reactions by Raman spectroscopy, application to control of emulsion copolymerizations and copolymerization kinetics", Technische Universiteit Eindhoven, 2000).
[0073]The stated amount of sodium dodecyl sulphate (SS) and NaHCO3 is weighed into a 1000 ml glass beaker and dissolved in the stated amount of water by stirring. The solution is cautiously flushed with nitrogen. The solution is transferred together with the styrene into a 1150 ml laboratory reactor with jacket and anchor stirrer. After the reaction mixture has reached a constant temperature, the prepared initiator solution is added.
[0074]The seed is prepared at a reaction temperature of 80° C. with a stirring speed of 300 rpm.
TABLE-US-00001 TABLE 1 Exemplary formulation for a seed preparation according to v.d.Brink in the emulsion polymerization of styrene Seed preparation according to v.d. Brink Component Content in % by mass Styrene 27.06 Emulsifier (sodium dodecyl sulphate) 2.88 Buffer (sodium bicarbonate) 0.09 Initiator (sodium peroxodisulphate) 0.19 Continuous phase (deionized water) 69.78
[0075]In the emulsion copolymerization, in each case one part of the monoalkenylaromatic monomer (e.g. styrene) is replaced by the seed polymer latex of Example 1. The amount of monomer necessary to achieve a particular target particle size can be determined according to Formula I with the aid of the particle concentration and size present in the seed.
pH-Controlled Copolymerization Under Strongly Acidic Conditions
[0076]The advantage of this method is that the surface charge can be adjusted to the particular requirements by targeted setting of the pH and appropriate choice of the acrylic acid concentration.
General Procedure for Carrying the pH-Controlled Copolymerization
[0077]The styrene employed is destabilized by distillation before use, and the demineralized water is degassed with a stream of nitrogen. In the examples specifically detailed hereinafter, the styrene was divided into 2 portions, and the amounts listed in the table were (a) added to the seed latex (monomer feed), or (b) metered continuously into the reaction mixture (emulsion feed). The further components were employed directly without any processing.
[0078]The stated amount of NaHCO3 is weighed into a 1000 ml glass beaker and dissolved in the stated amount of water by stirring. The solution is cautiously flushed with nitrogen. The solution is transferred with the seed emulsion and with the weighed amount of stryene (a) into a 1150 ml laboratory reactor. After the reaction mixture has reached a constant temperature of 72° C., the prepared initiator solution is added. The styrene (b) and the acrylic acid are added by means of metering pumps (piston diaphragm pump, Prominent Gamma 4 type). The sodium bicarbonate solution used to adjust the pH is metered in with a perfusor or likewise with a metering pump.
[0079]The stated amount of styrene (b) is metered in together with the acrylic acid dissolved therein over a period of 240 min. A latex is produced with a core of polystyrene and a shell of styrene-acrylic acid copolymer with an average particle size of about 125-135 nm diameter.
[0080]The latex was prepared at a reaction temperature of 72° C. with a stirring speed of 180 rpm.
[0081]In the copolymerizations carried out, a sodium bicarbonate solution was added through a second metering line to set and regulate the stated pH values.
[0082]Table 2 shows the initial weights in the exemplary formulations for pH-controlled copolymerization of the carboxylated latices:
TABLE-US-00002 TABLE 2 Exemplary formulations for pH-controlled preparation of carboxylated latices Initial Initial Component weight [g] weight [wt %] Polystyrene seed 26.9% (39 nm) 9.57 1.15-1.165 Initial amount of styrene (a) 2.57 0.31-0.313 Sodium bicarbonate (initial amount) 0.01-0.05 1.2-1.6 10-3 Initiator (sodium peroxodisulphate 1 0.12 Continuous phase (water) 740.0 89.39-90.082 Styrene (b) (metered over 4 h) 68.03 8.218-8.281 Acrylic acid (metered over 4 h) 0.706-7.06 0.853-0.086 Sodium peroxodisulphate (metered 1 0.12 over 4 h) Sodium bicarbonate to adjust the pH 0.24-1.14 0.029-0.14 during the reaction
Preparation of Various Carboxylated Latices
[0083]In the semibatchwise reactions in Table 3, the pH was kept constant during the metering time at the stated pH values of respectively pH=2, pH=3 and pH=4, and the influence of the pH during the reaction on the parking area was investigated.
[0084]The pH was adjusted with a sodium bicarbonate solution which was continuously metered in by means of a perfusor during the monomer metering. The concentration of the NaHCO3 solution depends on the amount of acrylic acid to be metered and is between 0.25 and 1.15 mol/l, and the total amount of metered sodium bicarbonate under the described experimental conditions is 2.9-13.5 mmol, depending on the amount of acrylic acid and pH.
[0085]At pH 2, sodium peroxodisulphate was additionally metered over a period of 240 min, because the acid-catalyzed hydrolysis of the initiator is no longer negligible at this pH.
[0086]The initial weights of the polystyrene seed and of the initial charge of styrene and metered styrene, of the continuous phase (demineralized water) and of the initiator employed are described in Table 2. Table 3 below shows besides the exact amount of acrylic acid employed, broken down according to the particular pH set, also the resulting parking area of the latex particles. The particle size of the target latex particles was found to be 125-135 nm.
TABLE-US-00003 TABLE 3 Resulting parking area in the pH-controlled preparation of carboxylated latices Acrylic acid concentra- parking area Batch Mass of acrylic acid [g] tion [10-1 mol/l] [10-10 m]2 pH = 2 1 0.706 0.132 107.6 2 0.884 0.166 100.8 3 1.009 0.189 78.1 4 1.765 0.331 51.1 5 7.06 1.32 11.8 pH = 3 6 0.706 0.132 130.5 7 1.009 0.189 83.5 8 1.07 0.201 76.8 9 1.765 0.331 65.2 10 7.06 1.32 38.2 pH = 4 11 0.706 0.132 151.5 12 1.009 0.189 116.5 13 1.479 0.277 99.1 14 1.765 0.331 72.5 15 7.06 1.32 47.6
[0087]As the amount of acrylic acid falls there is an exponential increase in the parking area, while there is a linear decrease in the mass of metered acrylic acid relative to styrene. In the experiments in which the pH was kept constant at pH=4, less acrylic acid is incorporated on the latex surface by contrast with a constant pH of pH=3. More acrylic acid is present in protonated form in the acidic-range, and is thus more hydrophobic and copolymerizes more readily with the styrene. In this case, more acrylic acid is incorporated in the copolymerization than dissolves in the styrene.
[0088]As is evident from Table 3, carboxylated latex particles can be produced with a desired, predetermined charge density in a very targeted manner with the aid of the present process.
Reproduction Batch
[0089]It was attempted in this experiment to reproduce the particles from batch 2 in Example 3.
TABLE-US-00004 TABLE 4 Parking area on repetition of batch 2 (at pH 2) Mass of acrylic Acrylic acid concentration Parking area Batch acid [g] [10-1 mol/l] (10-10 m)2 16 0.884 0.166 98.5
[0090]As is evident from Table 4, virtually identical reproduction of batch 2 was possible in new batch 16. The described method of pH-controlled emulsion copolymerization to prepare carboxylated latex particles is thus also distinguished by optimal reproducibility.
Preparation of an Anti-CRP-Latex Conjugate
[0091]15 mg of latex were activated in 0.75 ml of 20 mM MES (2-(N-morpholino)ethanesulphonic acid) buffer of pH 6.1 in the presence of 4 mM sulpho-NHS(N-hydroxysuccinimide) and 4 mM EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimides hydrochloride) and incubated in a roller incubator at room temperature for 1 h. Then 705 μl of a MAb<CRP> solution (C=0.96 mg of MAb/ml) (Mab<CRP>=monoclonal antibody against C-reactive protein (CRP)) in 20 mM MES buffer of pH 6.1 were added and incubation was continued for 20 min. Subsequently, 45 μl of a 2% strength Synperonic solution in MES of pH 6.1 were added. The reaction mixture was incubated for a further 100 min and the reaction was stopped by adding 30 μl of a 2M glycine HCl solution (pH 11). The conjugate mixture was centrifuged, and the supernatant was removed and redispersed in buffer (50 mM glycine HCl, pH 8.0 with 0.03% Synperonic and 0.05% sodium azide).
[0092]The prepared conjugates were assessed using the Roche Diagnostics Cobas Mira system. The performance of the conjugates based on the latices prepared according to the invention complied with the requirements for use in a homogeneous immunoassay for detecting CRP.
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Company / App Name: Software Proposal Portal Inc
Twitter – @SoftProposal
What does it do?
Swproposal is a user friendly web based software proposal portal to create elegant and attractive software project proposal for software projects in less time span .
Why do we need it?
Swproposal is designed with the purpose of automating the tedious routine business tasks by creating software project proposal attached with CRM features for tracking sales activities and Online Scheduler for time management.
Who is it for?
Swproposal is a web based user friendly application designed for freelancers , entrepreneurs and software development companies to ease the routine business tasks .
What makes it stand out from the crowd?
The two distinctive features which make Software Proposal Portal unique are :
Primarily , CRM Features to automate sales activities and manage negotiation process .
Secondarily , Online Scheduler to manage your time and project work load.
What's next?
We are trying to make the Software Proposal Portal much more robust as per the reviews of the people and their suggestions.

subset: commoncrawl


text:
from Wiktionary, Creative Commons Attribution/Share-Alike License
- n. Plural form of integration.
Sorry, no etymologies found.
To me this is just a clear admission of the shortcomings of longer term integrations and of the serious parameterization and other flaws in current climate models.
Long-term integrations predict that some moons could collide in less than one million years.
For example, even supercomputers are inadequate to allow long-term integrations of the relevant equations at adequate spatial resolutions.
Think Progress » An Inconvenient Truth and An Intolerable Summer
Let's call the integrations build as hourly build, assuming it will try to integrate a couple of smaller continuous builds every hour.
The only limits to the integrations are your imagination and needs.
Many partners have also developed KnowledgeTree plugins such as integrations with popular business platform SugarCRM (news, site), and document management tools such as Kofax (news, site) Capture,
The company will also build "integrations" into MySQL allowing it to work more productively with other key Oracle software, as well as making it easier to deploy.
Indieclick is doing some very cool "integrations" of brands into established music and related communities they represent.
"integrations" (the two different values of local time at the end of the experiment) are supposed to be the same, and that the difference shows that this "vector field" is not conservative, and that that is a counterexample to relativity.
Quickly after proclaiming Remember the Titans -- a movie starring Denzel Washington about the integrations of a Virginia high school played on the backdrop of a memorable football season -- the best movie she had ever seen, she proudly proclaimed: "My sons don't even know what segregation is."
Domonique Foxworth: We Must Accept and Learn From Our Past -- Not Drown in It


text:
Browse Books
Open Access
Classical Music
Russian Music at Home and Abroad
New Essays
Richard Taruskin
Available worldwide
Roth Family Foundation Imprint in Music
Read Chapter 1 (PDF)
Unjacketed Hardcover, 560 pages
September 2016
$85.00, £71.00
Other Formats Available:
E-Book Version
Request a Desk or
Examination Copy
Foreign & Other Subsidiary Rights
Author Bio
This new collection views Russian music through the Greek triad of "the Good, the True, and the Beautiful" to investigate how the idea of "nation" embeds itself in the public discourse about music and other arts with results at times invigorating, at times corrupting. In our divided, post–Cold War, and now post–9/11 world, Russian music, formerly a quiet corner on the margins of musicology, has become a site of noisy contention. Richard Taruskin assesses the political and cultural stakes that attach to it in the era of Pussy Riot and renewed international tensions, before turning to individual cases from the nineteenth century to the present. Much of the volume is devoted to the resolutely cosmopolitan but inveterately Russian Igor Stravinsky, one of the major forces in the music of the twentieth century and subject of particular interest to composers and music theorists all over the world. Taruskin here revisits him for the first time since the 1990s, when everything changed for Russia and its cultural products. Other essays are devoted to the cultural and social policies of the Soviet Union and their effect on the music produced there as those policies swung away from Communist internationalism to traditional Russian nationalism; to the musicians of the Russian postrevolutionary diaspora; and to the tension between the compelling artistic quality of works such as Stravinsky's
Sacre du Printemps
or Prokofieff's
and the antihumanistic or totalitarian messages they convey.
Russian Music at Home and Abroad
addresses these concerns in a personal and critical way, characteristically demonstrating Taruskin's authority and ability to bring living history out of the shadows.
Introduction: My Wonderful World; or, Dismembering the Triad
1. Non-Nationalists, and Other Nationalists
2. Revenants
3. Crowd, Mob, and Nation in Boris Godunov: What Did Musorgsky Think, and Does It Matter?
4. Catching Up with Rimsky-Korsakov
5. Not Modern and Loving It
6. Written for Elephants: Notes on Rach 3
7. Is There a "Russia Abroad" in Music?
8. Turania Revisited, with Lourié My Guide
9. The Ghetto and the Imperium
10. Two Serendipities: Keynoting a Conference, "Music and Power"
11. What's an Awful Song Like You Doing in a Nice Piece Like This? The Finale in Prokofieff's Symphony-Concerto, Op. 125
12. The Birth of Contemporary Russia out of the Spirit of Music (Not)
13. Just How Russian Was Stravinsky?
14. How The Rite Became Possible
15. Diaghilev without Stravinsky? Stravinsky without Diaghilev?
16. Resisting The Rite
17. Stravinsky's Poetics and Russian Music
18. Did He Mean It?
19. In Stravinsky's Songs, the True Man, No Ghostwriters
20. "Un Cadeau Très Macabre"
Richard Taruskin
is the Class of 1955 Professor of Music emeritus at the University of California, Berkeley, where he taught from 1987 to 2014, after twenty-six years at Columbia University (man and boy). He is the author of
Stravinsky and the Russian Traditions
On Russian Music
Defining Russia Musically
, and the six-volume
Oxford History of Western Music
"In surveying the continent of Russian music, Richard Taruskin has breathtakingly altered its scholarly appearance, displaying its arc in space as if through a telescope and its textures as if through a microscope. His new book casts a resolute and penetrating eye on contemporary Russia and the processes now underway there, which are shaping a new awareness of music within the cultural traditions that are at the heart of Russian spiritual life."—Liudmila Koynatskaya, Saint Petersburg Conservatory
"Taruskin's book does not read like a book by 'the other' on 'our' music, although his view from the outside often radically alters our perspective. Nor is it just a matter of the author's sincere sympathy, which shows through even the most ironic passages, but rather that Taruskin is waging an all-out fight for an approach to Russian music that no longer defines it primarily in terms of 'alterity' or 'exceptionalism,' but seeks to consider it, both from within and from without, as part of a single Western cultural sphere."—Olga Manulkina, Saint Petersburg Conservatory
"In the preface to his new book, Professor Taruskin promises the reader a responsibly academic rather than a loose journalistic performance. And yet the author, having long since made a home for himself amid the vast expanse of Russian music, is innately incapable of speaking or writing about it as a dispassionate observer. This 'wonderful world,' as he calls it, is his own. Taruskin treats octatonicism and other specialized matters with the same intensity as he does the vicissitudes of Chaikovsky's private life. His eye is keen and ardently engaged, and the reader is constantly reminded of the Russian criticism of the nineteenth century, and especially of Vladimir Vasil'yevich Stasov, the godfather of the Mighty Kuchka, a fiery tribune, a polemical journalist, and an academic scholar all in one."—Svetlana Savenko, Moscow State Tchaikovsky Conservatory
"Professor Taruskin regards the phenomenon of Russian music the way Joseph Brodsky regarded Pushkin, as 'a sort of lens into which has gone the past and out of which has come the future.' Armed with penetrating words, having set his historical sights with care and X-rayed the major players of the nineteenth and twentieth centuries, the author offers us a fresh look at our national art and the behavior of the key figures in the history of Russian music."—Lidia Ader, Senior Researcher, Saint Petersburg State Museum of Theatre and Music
Related Titles
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Music and the Elusive Revolution
The Castrato
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Arnold Schoenberg's A Survivor from Warsaw in Postwar Europe
Art of Suppression
The Magna Carta Manifesto
Europe's Promise
Stalinism for All Seasons
Smyrna's Ashes
Making New Music in Cold War Poland
Music of the Renaissance
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text:
Districts working on Pre-K plan
Updated 10:52 pm, Wednesday, October 17, 2012
San Antonio school superintendents have been meeting to fine-tune a legal "memorandum of understanding" that would govern the relationship between districts and the city if voters approve the Pre-K 4 SA initiative Nov. 6 — and if the districts decide to participate.
The 1/8-cent sales tax increase would set up city-run centers offering full-day pre-K to thousands of 4-year-olds and provide teacher training and grants for school districts to eventually expand and improve their own programs.
Virtually all local superintendents have attended the private monthly huddles, discussing the draft document and homing in on issues such as transportation, special education and liability.
The negotiations are part of the process of developing the program, not debating its pros and cons or selling it to the public, city officials said.
In recent interviews, superintendents said the talks can't get too far ahead of the voters' decision on Pre-K 4 SA — or those of their own school boards — and they can't finalize anything before the election.
"They're anticipating, of course, that it's going to pass and they're trying to get ready for it," said Ronnie Beard, executive director of the Texas Education Agency's Region 20 service center, where the superintendents have gathered.
They and city officials both want to head off any "surprises" that may arise, he said.
Mayor Julián Castro's signature education plan would require cooperation from the school districts, some of whose boards already have registered either support or skepticism.
The districts are being asked to give up a stream of pre-K students and almost all the existing state and federal funding that comes with them.
In exchange, the districts' teachers and principals who deal with students up to the third grade would receive access to professional development.
More importantly, school districts would get back students better prepared to start kindergarten, city officials have said. The program would aim to close achievement gaps measured by third-grade standardized tests.
Changes made
Northside Independent School District superintendent Brian Woods said school officials have requested one round of changes so far, amending a section about transportation that opponents had criticized as vague.
Woods said city officials have been receptive to the edits.
"We asked the city to put in some language that school districts weren't required to provide transportation and they did that," Woods said. "We asked them to put in some language that if (school districts) decided to provide transportation, it would be cost neutral."
Other requests include one that would allow school districts to keep some of the average daily attendance funding that flows from the state for each child in pre-K classes. Because school districts will be required to continue tracking students who attend the city centers, districts have asked that 3 percent of that funding stay with the district to cover administrative costs — a percentage that still might be negotiated higher after the North East Independent School District board asked for that possibility.
"We're hitting those (issues) head on," Assistant City Manager Peter Zanoni said.
David Thompson, a prominent education lawyer who's been advising city staff and working with school officials on the memo's wording, said this week that so far, the city has made all changes the districts have requested.
Zanoni said other questions, such as who would be responsible if a child at the center was injured or needs special education, still are being worked out. City officials are waiting for the superintendents to review the latest draft with their staffs to decide on a new round of proposed changes, he said.
The sheer number of school districts involved makes it a slow process, said Sylvester Perez, the San Antonio Independent School District's interim superintendent.
"One of the things that I have asked for is for a smaller number of us getting together ... because it's just real simple: The more legal eagles that get involved, the more potentially complicated that it can get," Perez said.
Neutral posture
School boards will be asked to approve the partnership if voters pass Pre-K 4 SA. Some opponents have said voters should know ahead of time if school boards will say yes or no.
Early in the city's campaign, the SAISD board adopted a resolution supporting the initiative.
Other school board endorsements may follow. Sources involved in the city's campaign have said the Harlandale and Southwest ISD superintendents have shown early support for it, though they and other superintendents publicly have maintained a neutral posture on the proposal — especially in districts on the North Side where opposition appears strongest.
"This school district will not make a decision about whether or not we'll participate in this program until after the election," NEISD Superintendent Brian Gottardy said.
A spokesman for the Alamo Heights Independent School District said it was premature to comment. Fort Sam Houston ISD Superintendent Gail Siller also declined to indicate where the district stood on the proposal.
Because eligibility is determined by state requirements that include the children of military personnel, all preschoolers in military districts within the city limits would be eligible for the program.
Districts outside the San Antonio city limits — such as Somerset and Randolph ISD — won't be able to participate at all. School officials at Southside ISD, which only has one elementary school in the city limits, said earlier this month that city officials had not contacted them or shown them a draft of the legal memo.
Carl Scarbrough, director of curriculum and instruction at Southside ISD, said he figured the city wasn't interested in partnering after the district was left out of the city's plans to apply for a competitive Head Start grant, a federal pre-K program.
"We would love for them to include us," Scarborough said, adding that the district has seen the benefits of its own pre-K classes.
Although Gottardy has said he "feels better" about transportation after city officials promised it would be "cost neutral" to districts, the concern has become a sticking point for detractors of the plan who are trying to present it as half-baked.
"Many of these elections come down to a trust factor and how much capacity you have in terms of credibility," said Perez at SAISD. "The city is going to have to react to those criticisms" that the plan lacks detail.
Woods, who was critical of the plan early on, said he agrees with the argument of city staff that certain specifics can't be delineated until after the vote.
"Some of the issues that we have questions about can't reasonably be nailed down until some implementation like, for instance, transportation," Woods said. "I think the city staff and the school districts agree that unless you know how many kids and where they live, it's really hard to nail down transportation."
Twitter: @mlcesar


text:
I address you tonight not as the President of the United States, not as a leader of a country, but as a citizen of humanity. We are faced with the very gravest of challenges. The Bible calls this day 'armageddon' - the end of all things. And yet, for the first time in the history of the planet, a species has the technology to prevent its own extinction. All of you praying with us tonight need to know that everything that can be done to prevent this disaster is being called into service. The human thirst for excellence, knowledge; every step up the ladder of science; every adventurous reach into space; all of our combined technologies and imaginations; even the wars that we've fought have provided us the tools to wage this terrible battle. Through all of the chaos that is our history; through all of the wrong and the discord; through all of the pain and he suffering; through all of our times, there is one thing that has nourished our souls, and elevated our species above its origins, and that is our courage. The dreams of an entire planet are focused tonight on those fourteen brave souls traveling into the heavens. And may we, citizens the world over, see these events through. God speed, and good luck to you.
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text:
An initiative of the University of Texas Departments of History and of Radio/Television (of which KLRU was part), the Texas Educational Media Program initiative brought to Austin forty-one of the foremost American historians and taped them in two or three 30-minute lectures. Part of a course entitled "The History of American Civilization - By Its Interpreters," the lectures were intended to be a component upper undergraduate courses that "should be the liveliest, most-talked-about in a department's offerings." Originally headed by Walter Prescott Webb, Joe Frantz took over after Webb's unexpected death and completed the project in 1963.
Consisting of 94 two-inch quadruplex videotapes the KLRU-TEMP Video Collection, 1961-1963, documents American historiography and historians in post-World War II America. Richard Hofstadter, Samuel Eliot Morison, Merle Curti, Arthur Schlesinger Jr., John Hope Franklin and other lecturers discuss a wide range of topics relating to American history, including "The Mormons," "American Foreign Policy," "20th Century Collective Security," "Latin America Viewpoint," "The Intellectual and the Spirit," "The Rise of the City," "American Social History," "Historical Writing and Research," and the "Southern Historian and his Subject," to name a few.
This collection is open for research use.
This collection is stored remotely. Advance notice required for retrieval. Contact repository for retrieval.
KLRU-TEMP Video Collection, 1961-1963, Dolph Briscoe Center for American History, The University of Texas at Austin.
Basic processing and cataloging of this collection was supported with funds from the National Historical Publications and Records Commission (NHPRC) for the Briscoe Center's


text:
Dear Prospective Applicant:
Thank you for your interest in the Endocrinology & Reproductive Physiology graduate program at the University of Wisconsin-Madison. Our program focus is in the areas of endocrinology and reproductive physiology, and as such a background in the biological sciences is a priority. While we do not place specific requirements on entering students due to the breadth of our program faculties endocrine studies from whole animal physiology through molecular and cellular signaling, applicants should be aware our core course requirements include intermediate and advance biochemistry and cell biology in addition to endocrine and reproductive physiology. A copy of our first year PhD course requirements is available as a pdf file. Applicants with strengths in these areas will be more likely to both be admitted and match to an advisor. Further admission requirements are listed below. We also urge you to look at our current students biosketches listed on this site and view our annual Symposium Booklet on this site to get a feel for the kind of studies you are likely to undertake. Please note, our application deadline for Fall 2018 is December 1, 2017.
How to Apply:
The Endocrinology & Reproductive Physiology program is a member of the Biomedical Sciences Consortium. Our admissions process is outlined on the consortium's admissions website.
NIH Required Data:
As a National Institutes of Health training grant funded PhD program, we are required to provide all applicants with information on our acceptance rates and time to degree completion. This information is available here.
Information and Advising for Applicants with an Interest in Medical School


text:
D-Tox Rockfest With Lamb of God, Hatebreed, NOFX — Live Review
Montebello, Quebec, is a small place with a country feel. About an hour away from Ottawa, Ontario, its population is slightly over 1,000. From what I saw, many of those "slightly over 1,000″ were themselves slightly over 60. In short, Montebello's got a cottage town feel; the coffee houses all have homemade jam for sale and the people who live there are wont to sit on their front porches and, well, sit on their front porches.
It is not, in any way, the kind of place where most people would think to situate a two-day music festival featuring some of the biggest and best names in punk rock, hardcore and metal. Yet that's exactly what happened there last weekend (June 17-18). Though its scheduling overlapped somewhat with the juggernaut that is Toronto's NXNE, D-Tox Rockfest was, by all measurements, a rousing success.
Headlined by Richmond, Virginia heavyweights Lamb of God (who were making their only 2011 appearance and, judging by their set, wanted everyone to be aware of it) as well as pioneers NOFX, the festival attracted some of the biggest names in the business. Scene stalwarts like Pennywise, the Bouncing Souls, Hatebreed, A Wilhelm Scream, Cancer Bats and Underoath were complimented by reunited legends like Hot Water Music and the Descendents. Hell, even CJ Ramone played a set.
The fest was heavy on quality CanCon as well, boasting the likes of the Flatliners, the Sainte Catherines, Slaves on Dope and Montreal psychobilly stalwarts the Brains (in addition to the aforementioned Cancer Bats).
"It's been awesome," said organizer Alex Martel of his festival a few hours before it came to a close. "I'm crazy tired and I've slept about two hours in the past two weeks — in the end, it's worth it. There's tons of people and it's really been a success."
Asked about the atypical location for his festival, Martel explained that as a local, it was something that he'd been thinking about for years before starting Rockfest in 2005.
"I am from Montebello, so I went to the University of Ottawa and [traveled] pretty much all over the place, but I always came back here and [to] this idea to have this big fest in this particular venue. I just thought that the site was awesome, [and] that it'd be pretty cool to have more of a rock-oriented festival in this tiny place in the middle of nowhere."
For the purposes of the Rockfest, the venue — an open area halfway between the Auberge Montebello resort and the waterfront/marina — was turned into a little city. There were food and drink kiosks as well as miniature markets that supplemented the various bands' merch booths. The two main stages featured alternating performances which rarely interfered with the third stage, which featured younger, up-and-coming bands (including an act featuring one of the more fear-inducing female vocalists you're apt to run across).
At various locations nearby, festival-goers were permitted to camp out. Many patrons took full advantage of this; the area directly adjacent to the venue dubbed "tent city." Some of the campfires burned all night.
While there were a few alcohol-related dust-ups, the main focus for the weekend was almost always where it should have been: on the music. That said, the Auberge, which housed most of the musicians, was an interesting home base, featuring a rooftop sauna and hot tub which, word has it, has been the site of some debauchery in years past. The hotel also offered a trampoline, which proved to be irresistable for Lamb of God vocalist Randy Blythe, who was seen happily bouncing around, smoking a cigarette while tooling around on his phone.
Unfortunately, Blythe was far less interested in doing press interviews. Fortunately, this wasn't really a major concern since the majority of us journalists were far more interested in the naked man in the Hatebreed pit and another in a wheelchair who insisted on crowd-surfing, tilting, falling out of his chair, getting put back into his chair, and getting back where he wanted to be — only to be mocked, good-naturedly by NOFX's Fat Mike.
Additional highlights of the festival: the unicyclist in a mosh pit, countless flashed breasts, kids so amped up on energy drinks they talked like auctioneers, various witnessed "I don't speak French" mishaps, some truly terrible French-Canadian accents (mine, most prominently), some truly excellent poutine, and children who may or may not have been conceived in or around the hot tub.
Occasionally, we even talked to and watched bands.
The unofficial festival standouts included Lamb of God, the Descendents and the Sainte Catherines, who earned high praise from everyone who saw them. Despite being long in the tooth, the Descendents were riveting from start to finish, with frontman Milo Aukerman seemingly unaware he was covered in Mayflies, which were gross and absolutely everywhere. He rambled and thrashed through his set like he was a man twenty years younger.
NOFX were entertaining as always. Fat Mike earned a lot of laughs when explaining towards the end of the band's set that they couldn't go over their allotted time because Alcoholica, the famous Metallica cover band, were closing the show. Mike was his usual acerbic and sarcastic self; the highlight of his weekend was apparently the round of golf he snuck in before the show.
When asked about the festival moving forward, Alex Martel was succinct. "I have a good idea of what I want to do next year," he said. "I want to get bigger bands and try to expand the festival."
Truer and more anti-climactic words were never spoken, but if Martel and Co. can top Rockfest 2011, it'll be a sight to see.
Watch Cancer Bats Perform at D-Tox Rockfest 2011


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Happy Birthday, my dearest friend! I wish you AAALLL the best!
Just stopping by to say Hello Richelle. Hope all is well with you & yours =) Your Angel Friend Mindi~~
Happy Belated Birthday Chelle, hope your special day was delightful =) Your Angel Friend Mindi~~
Dear Richelle.A red rose in the early morning hours with dew drops still on the petals smelling of morning freshness is yours .As I can`t forward or indeed send you anything,but god wishes. Know that I will always stand by you no matter what. the rose is a queen among flowers the colour red symbolises love the feeling that i must share with you my dear friend.I hope you are all well.Lots of love and light from Lisbeth and the lads.
Dear Richelle.Thank you for the bow of friendship.If the messaging service had worked I would have sent it to you.Instead a few words of friendship must surfice.How are you and yours?My lads are doing well and I am studying for my exams.Wich will be in eight weeks time.There is so much to get through.However I am enjoying every minute of it.Connor was attacked and bitten by a stray dog the other day.He diden´tgrowl or bite back.He sat and looked at the other dog as if to ask what did you do that for? Needless to say Mike my whippet and me chased it off.This was not the dogs fault but its careless owner.She knows that her dog is agressive and should kept him on a leash.Connor did not recieve any serious bites thank the gods for that.Lots of love hugs and cuddles from Lisbeth and the lads.
Just dropping by to say Hi Richelle, and wishing you a fantastic week =)


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Testing and Training for Physical Fitness (not offered in 2014)
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Beauty News: Fashion Week Backstage Must-Haves
Behind all the glitz and glamour of Fashion Week is the hard work of the designers, hair stylists and makeup artists. They create the hottest, trend-setting looks for the rest of the year.
Want to know what the must-have makeup artist essentials are for this Fashion Week? Here is a sneak peek of everything they'll need to create the incredible and flawless looks seen on the runways this season!
Koh Gen Do Cleansing Spa Water, $39– As models bounce from show to show, it's essential to cleanse and prep for the next look. Koh Gen Do Cleansing Spa Water removes dirt, oil, and makeup skin impurities in a single sweep with while nourishing the skin.
Koh Gen Do Makeup Sponges, $18– These unique sponges are made of dense elastic in a convenient palm-size shape, which allows for seamless application and a perfect, show-stopping finish.
Dermablend Professional Brush Line, $22-40– Perfect tools are a must. These brushes are necessary for every makeup step—from under eyes and face to neck and even body.
La Roche-Posay Thermal Spring Water, $11.95– To help set makeup even under the runway lights, spritzing Thermal Spring Water over makeup prevents drying for a fresh finish.
Nature's Gate Glow Lotion, $12.99- Don't forget about the arms and legs! Nature's Gate Glow Lotion provides the perfect and quick solution to even, glowing skin. This moisturizing body lotion enhances skin tone and won't run off on clothes.
Goody "Start.Style.Finish" Gel Purse Brush, $4.99- To keep hair cat-walk ready, stylists use this brush backstage to keep models' locks shiny and frizz-free. This brush is also perfect for show-goers to keep in their bags for touch-ups between shows.
Shop the essentials:
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Pregnancy Facts: Interesting Things About Conceiving
Many couples today put off conceiving until they are established in their careers, have their own homes, and feel financially and emotionally ready to take on the responsibilities of rearing a child.
While this is a wise decision, the one thing most couples overlook is the fact that getting pregnant may not be as easy as it seems. While some couples seem to be able to conceive within a month or two of trying, others may have a difficult time getting pregnant, and still, others may find getting pregnant impossible.
If you are planning on becoming pregnant in the future, here are some pregnancy facts that you need to be aware of.
The 20s is the best age to get pregnant
The mid-20s is the most fertile time for both men and women. Women are the most fertile in the early to mid-20s. As a woman ages, her eggs age along with her. This eventually causes deterioration of the eggs quality and likelihood of being properly fertilized.
During your mid-20s you have a 20%-25% chance of conceiving during each reproduction cycle.
These odds are fantastic for anyone who wants to conceive quickly in their 20s.
There are other great benefits to getting pregnant in your 20s. People in their 20s are able to get up early and stay up late without much consequence or toll on their bodies.
Because of their youthful energy, they are easily able to get up to tend to the baby's needs more so than someone in their 30s or 40s. That isn't to say it can't be done by women later in life, but it's certainly easier.
There are some downsides to getting pregnant in your 20s. Many couples put off getting pregnant because they're still working on their careers.
Several young people get Master's degrees or extra schooling to advance in the workplace. These lifestyle choices can affect when a couple will want to have their child. Make sure you're at the right point in your life to try for a baby.
The 30s is still a great time to get pregnant
Women in their 30s have a 15% chance of conceiving during each reproduction cycle. Although this is slightly lower than when a woman is in their 20s, they can still conceive and have the baby they've always wanted.
When a woman reaches 30, often times she's in a great place in her career. She and her partner have been working near a decade advancing in their careers. By this point, they are financially and emotionally ready to begin a family.
Although it might be scary thinking about how your schedules will change and how you'll make time for a 3rd member of the family, the joy that comes with pregnancy will outweigh those feelings. Once you have a baby, you'll learn how to handle a healthy balance of time for the baby and time for you.
By age 30, the miscarriage rate slightly increases. The odds of having a baby with disabilities also increases as you age through your 30s. Although there are negative effects as you age, this doesn't mean you should rush into anything now.
You may feel the need to rework your life so you can have a baby sooner rather than later. It's important to discuss with your partner to see when it's realistic to bring a new life into this world!
It takes time to conceive
15% of couples who are trying to get pregnant do not conceive within the first year. However, about 95% of couples do manage to become pregnant within 2 years.
With chances of conceiving in a single month being so low, it's not easy to get pregnant in a single month. There are a lot of factors when trying to get pregnant and even if you time it correctly, there's still a large chance you won't conceive.
Not to worry though. If you can't get pregnant right away, it doesn't mean that you or your partner has fertility problems. Factors like your age, health, and reproductive system can make chances of getting pregnant greater or smaller.
It's best to understand the facts about how long it takes to get pregnant. Understand that it's normal to not get pregnant right away. It's recommended that you try for a full year before seeking any help.
Trying for a second pregnancy can be tough
It comes as a surprise to many couples that the second time around can be more difficult to conceive. Once you have a baby, you may have an idea of the perfect timeframe in which to space your children. You might be in for a surprise when you get back to trying to conceive.
5-6% of couples have already conceived one child and then have difficulty conceiving a second child. This is called secondary infertility. Because it's not as well-known, many people assume that if you gave birth to one baby, you can easily get pregnant with another.
Well-meaning close family or friends might ask you why you haven't had another child yet and this can take a toll on you emotionally. It's hard for people to grasp that there might be more wrong than just 'not trying' enough. It's easy to let stress build up since you have a first pregnancy to compare this situation with.
The most important thing to realize is that you cannot compare this second attempt at having a child with your first successful attempt. It's likely been a few years since your first pregnancy. Factors like age, development of eggs, and health issues may be impacting you or your partner. If you aren't able to conceive within 6 months, it's a great idea to visit your doctor and find out what you can do to remedy this issue.
Infertility is not just for women
Infertility can be related to problems with the female, male or both. A couple is considered be infertile or have fertility problems if the woman is over 35 and has not conceived within 6 months of having unprotected sex. Or if she is under 35 and has not become pregnant within a year.
It's easy to assume that the woman has an issue that needs to be resolved. But actually, there is a whole side of infertility that many people forget about. Male factor infertility is something that is often overlooked.
About 20% of infertile couples have issues with male fertility. It's not as uncommon as you think, although it's often not discussed. In about 50% of the cases, doctors are able to determine why a man is infertile. In many cases, there are remedies that can assist him in regaining the fertility he needs to help his partner conceive.
People try to get pregnant in all different walks of life. Some younger, others older. Some are first timers and others have conceived successfully in the past. Hopefully these facts enlighten you on your own journey towards pregnancy!


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So we now know that there was a plan to wreck the coal-mining economy of the UK in 1984 and put nothing significant in its place in industrial Yorkshire.
We also know that the planning for that confrontation dated back a year and was not the subject of debate in Ministries, before Parliamentary Committees or on the floor of the Commons.
We now know that the Miners Union was right when they said that 70 pits would quickly close and that such a move should be resisted. We also know that when McGregor, the man in charge, sent letters to miners saying that he knew nothing of this, he was lying.
The 1984/5 coal strike taught us all a number of lessons. One was that working men, and the women who supported them, get prison records for relatively minor misdemeanors when they seek to protect their livelihood but bankers who manipulate bank rates to their own advantage and cripple economies are reprimanded, apologise and are not imprisoned.
We also know that if you believe that it is better to buy in Russian coal than produce it, you are as foolish as those who say that 'we are all in this together'.
Also if you sell the the family silver of transport, civic housing stock and essential social services onto the businessmen the social profits drop like a stone but bonus culture flourishes.
Brian Lewis
Linden Terrace


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As with humans, cats that have been diagnosed with diabetes have special dietary requirements and while you may believe that all cat foods are nutritionally balanced and ideal for all cats, this is not always the case. Indeed, there is a fair range of specifically formulated diabetic cat foods on the market, even these will not always be appropriate for every diabetic cat. Your veterinarian should be your first source of information about what to feed your diabetic cat as nutrition has an immediate impact on the progression and symptomology of diabetes.
Since diabetes is often related to other health concerns like obesity, kidney and pancreas problems, each diabetic cat will likely have very specific nutritional needs. However, in the broad sense, cats with diabetes should be fed a diet that is low in carbohydrates and high in protein. Naturally, cats are carnivores so it makes sense that they have evolved to eat a protein rich diet. In fact, traditionally cats would have had very little access to carbohydrates yet many inexpensive dry cat foods are rich in cheaper carbohydrate-based additives that are filling like proteins. Canned cat foods are often a good alternative to dry varieties as they are usually high in protein. Still, many canned cat foods can be unnecessarily high in sugars so be selective and read labels.
In some cases, veterinarians will recommend a prescription diabetic cat food that provides your cat with highly specified nutrition. The downside to these diabetic cat food prescriptions is that they can be very expensive and unaffordable for many households. Ultimately, you should discuss your budget and your cat's needs with your veterinarian to find an effective, yet affordable diabetic cat food. Also remember that many cats are picky with their food choices so you may want to have a few reasonable options available to help you find something to your cat's taste.


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Allison Mouland Gets Opportunity to Shine with Expansion Lady OutlawzMay 14, 2013
The multi-talented Allison Mouland has a great love of sport.
Competing for the expansion Okotoks Lady Outlawz, Mouland is getting the opportunity to accomplish a lifelong dream of playing tackle football. Competing on offense as a quarterback and on defense as a cornerback, Mouland brings enthusiasm to the growing Western Women's Canadian Football League.
"I can't really remember a time when I didn't like football, because I always liked all sports. I think I started developing a real interest in it around grade nine. My younger brother was playing flag football, and he played tackle when he was older, so I would watch him and it looked like a lot of fun. My parents were always encouraging when it came to staying active and so they always had me involved in some kind of sport.
"I played soccer and did swimming lessons when I was young. I had really wanted to play hockey and finally got the opportunity when I was fifteen, and I've been playing that ever since. In grade 10, I made new friends who were also football fanatics, and we started playing pick-up football games at lunchtime and after school nearly every day."
Joining the Lady Outlawz would reunite her with a former educator and mentor. Rick Crawford has known Mouland since high school and provided her with direction in pursuing her love of the game. Having competed at the touch-football level, contributed as a trainer and assisted in coaching, Mouland has paid her dues.
"Rick Crawford, the defensive coordinator for the Lady Outlawz, was actually my grade 10 social studies teacher and the senior football coach. My love for football was obvious to him, and he set me up with a ladies touch-football team. I have not stopped playing football since. He also drafted me to his arena football intramural team in high school.
"I took sports medicine in high school and did many of my training hours with his team, and he was always encouraging and let me be involved in the drills, which was great. It helped my understanding of the game. Last year, I ran into Rick at the university. After I explained that I was finishing my kinesiology degree and moving into education, he offered me the opportunity to learn to coach with his senior high-school football team."
Contributing on Crawford's coaching staff helped bring Mouland a new perspective on the game. The valuable learning experience is one that has paid dividends for her. Employing great acumen on the gridiron, Mouland is quick to credit Crawford as a positive influence.
"This was an excellent opportunity, and it allowed me to gain coaching experience while learning more about the game. He is also the one who recruited me to check out the Lady Outlawz. He has been extremely influential in my life and he has encouraged me and provided me with many football opportunities, and I am grateful for that."
A proud member of the Lady Outlawz, Mouland understands the responsibility that her status as an athlete brings. The thought of possibly being a role model for young girls is one that she would gladly welcome. While the league is still growing and the level of parity has not yet been reached, Mouland hopes that today's competitors can lay the foundation for a strong tomorrow.
"I would love to be a role model to young girls. I know there has been a lot of change in recent years when it comes to gender equality and the opportunities available to girls, and I know that my generation is very fortunate."
Having had the opportunity to compete in the first game in Okotoks Lady Outlawz franchise history on May 4, 2013 was one that brought great happiness for her. After years of aspiring to compete, and having spent several seasons serving in coaching or trainer's roles, the opportunity to don the helmet and step on to the field was the culmination of a dream.
"It was so exciting to hit the field for the first time. I have to admit, I was extremely nervous and had butterflies, but the experience was amazing. I love this sport, and now that I have a taste for playing the tackle version of it, I don't ever want to stop. I'm so grateful that this opportunity finally exists for women."
The first game also brought with it humility. In playing the defending Western Conference champion Lethbridge Steel, an overwhelmed Outlawz team was defeated by a 74-0 score. While the loss was difficult, it provided Mouland and her teammates with an invaluable learning experience. Despite the heartbreak of losing their first game on home soil, Mouland noted the remarkable team spirit that existed.
"It was a very tough loss and a rude awakening against Lethbridge. What I found very encouraging was how we managed to not get down on one another. There was no blame being thrown around. Compared to other teams that I have played on, I was really impressed by the lack of negativity among the team, both during and after the game. It was tough, but we took it as a learning experience, and we are using it to inspire us for move forward and get better."
"All quotes obtained first hand unless otherwise indicated"


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A Passion for Blue & White (Broadway Books) Interior designer Carolyne Roehm examines her fascination with the color scheme that has served as a constant inspiration throughout her 35 years in fashion and decorating.
Defining Luxury (Rizzoli) Exploring what it means to live in luxury, renowned interior designer Jeffrey Bilhuber takes a look at all the elements of haute living in the American home.
Dreaming Green (Random House) Husband-and-wife team Paul Gleicher and Lisa Sharkey share their secrets on how to live in an eco-conscious home without sacrificing quality or style in both urban and rural settings.
Tiffany Style (Abrams) A...


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|Stickers at KRLX. Photo: J. Waits
Students at Carleton prevailed, however, launching a campus-only carrier current station in 1948, which morphed into FCC-licensed FM station KRLX in 1974. Read my full report on Radio Survivor.
|Stickers at KRLX. Photo: J. Waits
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