text:
|Magnetic particulate for immobilization of biological protein and process of producing the same
|Ikeda et al.
|Magnetic particles for use in separations
|Whitehead et al.
|Magnetic iron-dextran microspheres
|Magnetic polymer particles
|Polyglutaraldehyde synthesis and protein bonding substrates
|Magnetic organo-iron compounds
|Gable et al.
This Application is a Continuation-in-Part of application Ser. No. 784,863 filed Oct. 4, 1985, abandoned, of common inventorship and assignment herewith, which is incorporated by reference.
(a) combining a first aqueous solution of at least two species of transition metal ions capable of reacting with each other to form a magnetic precipitate and a polymer having available coordination sites in proportions adapted to produce a resuspendable product;
(b) reacting said transition metal ions in the presence of said polymer to form a magnetic precipitate comprising magnetic-polymer particles;
(c) recovering said magnetic-polymer particles.
(d) resuspending said magnetic-polymer particles in a second aqueous solution.
(d) filtering the suspension through a filter having pores of at most 0.44 um diameter.
(d) reacting said magnetic-polymer particles with a bifunctional compound specific to said polymer.
(d) reacting said magnetic-polymer particles with a bifunctional compound adapted to form primarily intra-particulate bonds.
(d) reacting said magnetic-polymer particles with a bifunctional compound adapted to form primarily intra-particulate bonds, said bifunctional compound having terminal groups selected from the group consisting of: aryl nitrenes, imido-esters, N-hydroxysuccinimide-esters, 2-diazo-3,3,3-trifluoropropionate, maleimides, pyridyl-disulfides, halo-nitrobenzenes, isothiocyanates, halo-sulfonates, active halogens, and active aldehydes.
(d) reacting said magnetic-polymer particles with a bifunctional compound; and
(e) reacting the product particle of step (f) with a biofunctional ligand.
(d) reacting said magnetic-polymer particles with an activating agent; and
(e) reacting the product particles of step (f) with a biofunctional ligand.
(d) reacting said magnetic-polymer particles with both a bifunctional compound adapted to form primarily extra-particulate bonds, said bifunctional compound having terminal groups selected from the group consisting of: aryl nitrenes, imido-esters, N-hydroxysuccinimide-esters, 2-diazo-3,3,3-trifluoropropionate, maleimides, pyridyl-disulfides, halo-nitrobenzenes, isothiocyanates, halo-sulfonates, active halogens, and active aldehydes and a biofunctional ligand selected from the group consisting of antigens, antibodies, lectins, avidin, biotin, staphloccal Protein A (SPA), enzymes, serum proteins, Clq, complement proteins, and rheumatoid factor.
(d) reacting said magnetic-polymer particles with both an activating agent selected from the group consisting of water-soluble carbodiimides, glutaraldehyde, cyanogen halides, periodates, and tannic acid and a biofunctional ligand selected from the group consisting of antigens, antibodies, lectins, avidin, biotin, staphloccal Protein A (SPA), enzymes, serum proteins, Clq, complement proteins, and rheumatoid factor.
(e) adding a salt to said suspension to increase its ionic strength to a molarity adapted to cause agglomeration and precipitation of said magnetic-polymer particles;
(f) separating said agglomerated precipitated particles;
(g) resuspending said magnetic-polymer particles in an aqueous solution of low ionic strength adapted to allow the formation of a stable suspension.
(a) preparing an aqueous solution of iron capable of reacting to form a magnetic precipitate and a polymer having available coordination sites;
(b) adding a stoichiometric excess of a strong base;
(c) agitating said solution;
(d) allowing the magnetic-polymer particles to precipitate;
(e) recovering said magnetic-polymer particles.
(a) preparing an aqueous solution of transition metal ions capable of reacting to form a magnetic precipitate and a polymer having available coordinate sites in proportions adapted to produce resuspendable particles;
(b) reacting said metal ions in the presence of said polymer to form a magnetic precipitate comprising magnetic-polymer particles;
(c) recovering said magnetic-polymer particles;
(d) resuspending said magnetic-polymer particles in an aqueous solution;
(e) reacting said magnetic-polymer particles with both a bifunctional compound adapted to form primarily extra-particulate bonds, and a biofunctional ligand, said biofunctional ligand being specific in its ability to bond with said predetermined species;
(f) exposing a mixture containing an unknown quantity of said predetermined species to a suspension of said magnetic-polymer particles containing said biofunctional ligand, thereby causing said biofunctional ligand to bond to said predetermined species;
(g) passing said mixture through a magnetic filter having a magnetic field, said filter adapted to retain said magnetic-polymer particles;
(h) removing said magnetic field from said filter and eluting said retained magnetic-polymer particles;
(i) analyzing said eluted magnetic-polymer particles to provide desired data relative to said predetermined species.
step d includes resuspending said magnetic-polymer particles in an aqueous buffer solution having low ionic strength and filtering said suspension through a filter having pores of at most 0.44 um diameter so as to sterilize said suspension.
said bifunctional compound having terminal groups selected from the group consisting of: aryl nitrenes, imido-esters, N-hydroxysuccinimide-esters, 2-diazo-3,3,3trifluoropropionate, maleimides, pyridyl-disulfides, halo-nitrobenzenes, isothiocyanates, halo-sulfonates, active halogens, and active aldehydes.
said biofunctional ligand is selected from the group consisting of antigens, antibodies, lectins, avidin, biotin, staphloccal Protein A (SPA), enzymes, serum proteins, Clq, complement proteins, and rheumatoid factor.
said biofunctional ligand is selected to provide half of a binding pair selected from the group of: antigens and their antibodies, haptenes and their antibodies, hormones and their receptors, vitamins and their receptors, toxins and their receptors, drugs and their receptors, and enzymes and their cofactors.
(a) combining a first aqueous solution comprising at least two species of transition metal ions capable of reacting with each other to form a magnetic precipitate, and a polymer having biofunctional activity and having available coordination sites in proportions adapted to produce a resuspendable precipitate product having biofunctional activity;
(b) reacting said transition metal ions in the presence of said polymer to form a magnetic precipitate comprising a complex of said metal and said biofunctional polymer;
(c) recovering said magnetic precipitate from said first solution.
This invention pertains to particles comprised of certain polymers in combination with magnetic metals, to compositions including such particles, and to methods of making and using such particles and compositions. More particularly, this invention pertains to such particles in which the polymer has a biochemical function.
Biologically active magnetic particles may find use in a variety of preparative and diagnostic techniques. Among these is high gradient magnetic separation (HGMS) which uses a magnetic field to separate magnetic particles from suspension. In instances where these particles are attached to biological materials of interest (e.g. cells, drugs), the material of interest may thereby be separated from other materials not bound to magnetic particles.
Several properties are important if a magnetic particle is to be useful in biological and diagnostic systems. First, the particle must possess the necessary biological activity, affinity, or reactive nature, by which it will perform its function. Second, the particles must be suspendable in an aqueous medium for delivery to the biological or reaction system. It may also be desirable that the particle suspension be stable (i.e. not settle out or agglomerate). Finally, small particle size may be desirable so that a suspension of the magnetic particles may be filter sterilized by conventional techniques.
Many techniques have been suggested in the prior art for the preparation of magnetic particles or organomagnetic materials. U.S. Pat. No. 4,001,288 to Gable et al. discloses a preparation of magnetic organo-iron compounds which are both water soluble and strongly magnetic. In the preparation taught by Gable et al., ferrous iron in solution is treated with hydrogen peroxide and then with ammonium hydroxide in order to precipitate an iron oxide compound. This compound is subsequently oxidized with peroxide, treated with a hydroxy-carboxylic acid, and reacted with an alkaline material in order to form a soluble product.
Gable et al., also disclose the introduction of proteins or "protein degradation products" into the ferrous iron solution which is subsequently reacted to precipitate iron oxides. The only material for which examples are presented is the hydrolysis product of gelatin treated with hydrogen peroxide.
U.S. Pat. No. 4,452,773 to Molday discloses "colloidal sized" iron oxide particles coated with a polysaccharide, (exemplified in the patent by dextran and dextran derivatives). Although Molday's particles are prepared in a manner somewhat similar to that used in preparing particles in the present invention, they are believed to be substantially different from the particles of the present invention.
U.S. Pat. No. 4,454,234 to Czerlinski teaches the preparation (generally by suspension polymerization on a magnetic particle substrate) of coated magnetic particles which are reversibly suspendable in solution. The suspension parameters taught by Czerlinski involve the effect of the Curie temperature of his magnetic particles which may be used alternately to cause the particles to be magnetic and non-magnetic. When the particles are magnetic, they tend to agglomerate, but may be resuspended after heating the particles to a temperature above the Curie temperature of the magnetite contained therein.
U.S. Pat. No. 4,230,685 to Senyei, et al., discloses the preparation of microspheres containing magnetite, albumin, and protein A. The preparation taught by Senyei does not involve the precipitation of magnetite in the presence of these other constituents, but rather is a coating of preformed magnetite particles.
Other patents which may be considered to be of interest include U.S. Pat. Nos. 4,152,210 to Robinson, et al.; 4,335,094 to Mosbach; 4,018,886 to Giaever; and 4,070,246 to Kennedy, et al. While these patents all disclose the preparation or use of magnetic-biologic particles, none of these are thought to be similar to those of the present invention.
Active Halogen: A halogen atom bonded to a carbon atom which is either bonded to or adjacent to a carbon bonded to an electron-withdrawing group. (For example: iodo-acetamide, iodo-acetate)
Available Coordination Site: A coordination site which is not sterically hindered (can be freely approached) and is adapted to coordinate a metal atom in metal compounds (e.g. Fe3 O4).
Biofunctional Ligand: A molecule having a biological activity or a particular affinity in a biological system which can be linked to the ferromagnetic-polymer particles of this invention so as to impart particular biological properties to those particles.
Coordination Sites: An atom in a molecular structure which has a "free" electron pair capable of forming a coordinate bond with a transition metal atom.
Denatured Proteins: Proteins which have lost a specific biofunctional activity (e.g. enzymatic, antigenic, antibody, etc.) through a chemical or structural alteration. As used by Gable et al: Protein residues remaining following cleavage of some amide bonds and having molecular weights <10,000.
Extra-Particulate Bonds: Linking of one functional group of a bifunctional compound to a site on a ferromagnetic-polymer particle while the other functional group is linked to a site on a different molecule or (typically a "ligand").
Ferromagnetic: Permanently magnetic iron compositions (having a net magnetic moment).
Intra-Particulate Bonds: Linking of both functional groups of a bifunctional compound to different sites on the same magnetic-polymer particle.
Low Ionic Strength: An aqueous solution of near-neutral pH and having a total concentration of cations (usually from a buffer) of <40 mM.
Protein: Amino acid polymers linked with amide (peptide) bonds and having molecular weights >10,000.
Resuspendable: A material which once agglomerated (e.g. by centrifugation, HGMS, or flocculation) is capable of being redispersed to yield a stable suspension.
Sonification: Exposure to high intensity ultrasound.
Stable Suspension: A suspension which does not settle or otherwise agglomerate if left quiescent at standard temperature and pressure for 2 days.
The present invention comprises a process for the preparation of suspendable and resuspendable magnetic-polymer particles and the particles produced thereby. Such particles exhibit useful properties, particularly in immunoassays wherein the particles are prepared with a particular biofunctional ligand and are subsequently separated by high gradient magnetic separation techniques.
The process of the invention includes the coprecipitation of metal ions (eg. [Fe(II+Fe(III)] or [Fe(II)+Cr(III)] as magnetic compounds in the presence of a polymer having available coordination sites, reaction of the polymer and the metal to form a precipitate and recovery of the magnetic-polymer particles. Additionally, various biofunctional groups may be incorporated into the particles in order to yield an effective biofunctional reagent for use in immunoassay, cell capture, enzyme immobilization reactors, NMR imaging, and other diagnostic and analytical techniques.
According to one embodiment of the present invention, a solution containing Fe(II) and Fe(III) (typically FeCl2 and FeCl3), and a polymer (e.g. a protein) having available coordination sites is treated (by titration or otherwise) with a strong base such as ammonium hydroxide (NH4 OH) in order to precipitate magnetic iron oxides such as magnetite (Fe3 O4) in a form which is intimately combined with the polymer. The precipitation is typically carried out with rapid stirring and optional agitation by sonication, in order to produce resuspendable magnetic-polymer particles.
After precipitation, the particles are washed and subsequently resuspended in a buffer solution at approximately neutral pH.
Other embodiments of the present invention involve the use of metals other than iron in the coprecipitation reaction. In particular, Fe(III) may be replaced by any of a wide range of transition metal ions. In some cases, iron may be completely supplanted by appropriately selected transition metal iron. In many cases, the use of metals other than ion produces colored particles ranging from white to dark brown.
Magnetic-polymer particles prepared according to the present invention exhibit many useful properties. These particles are magnetic due to the inclusion of a form of magnetic metal compound (eg. iron similar in form to magnetite, or a similar compound). The particles may be formulated to be resuspendable after aggregation and to produce relatively stable suspensions which do not settle even after several days of quiescent storage. Furthermore, particles produced according to the present invention may be relatively small (approximately 0.01-0.2 micron) and therefore filter sterilizable. Finally, particles produced according to the present invention can be tailor-made to include specific biofunctional ligands useful in various analytical, diagnostic, and other biological/medical applications.
Subsequent to precipitation and resuspension of the magnetic-polymer particles, they may be treated with a bifunctional reagent in order to cross-link reactive sites present on the polymer. This cross-linking may be effective as either an intra-particulate cross-linking in which reactive sites are bound on the same particle, or may be a reaction of an extra-particulate ligand which is then cross-linked to the polymer on a given particle. In the second case, a bifunctional reagent having a relatively short distance between its two functional groupings is desirable to promote linkage between the particle polymer and the extra-particulate species. Conversely, intra-particulate cross-linking is promoted by the use of a bifunctional reagent which is longer and is not sterically hindered from bending so that two reactive sites on a single particle may be linked by a single bifunctional molecule.
As an alternative to the use of sonication during either the precipitation or resuspension steps outlined above, another type of agitation (such as mechanical stirring) may be employed.
Resuspension of the magnetic-polymer particles of the invention is typically carried out in a low ionic strength buffer system (e.g. 40 mM phosphate). The buffer system enables resuspension of particles which are not resuspendable in non-ionic solutions. In addition to phosphate buffers, borate and sulfate systems may also be used.
The intimate association of polymer and metal in the present invention is thought to result from coordination of metal present during coprecipitation by coordination sites on the polymer. It is hypothesize that certain coordination sites are more "available" than others, based on both the strength of the coordinate bond which may be formed by the particular atom, and the spatial hindrances imposed by surrounding atoms. It is known, for instance, that oxygen atoms having a "free" electron pair complex iron more strongly than amine nitrogen atoms and, to an even greater degree, a hydroxyl oxygen atom. Thus, a polymer bearing oxy-acid functional groups should provide better product particles than an amine-substituted polymer. Similarly, coordination sites which may be freely approached to close distances should yield better performance than sites which are hindered in either a path of approach or in approach distance.
The above-described trends are qualitatively observable in various experiments performed by the present inventors. The presence of "available coordination sites" appears necessary to the production of the resuspendable magnetic-polymer particles of the present invention. For example, such diverse polymers as natural proteins, synthetic proteins, poly-amino acids, carboxy-poly-alkyls, alkoxy-poly-alkyls, amino-poly-alkyls, hydroxy-poly-alkyls, and various copolymers of these have all been demonstrated to produce the particles of the present invention. In addition, other polymers such as sulfoxy-poly-alkyls, poly-acrylamines, poly-acrylic acid, and substituted poly-alkylenes produce the particles of the present invention.
In selecting the transition metals to be employed in the coprecipitation reaction, several criteria appear to be important. First, the final compound must have one or more unpaired electrons in its structure. Second, one of the metals must possess an available coordination site for bonding to a polymer. Third, the coprecipitate must be capable of forming a cubic close-packed or hexagonal close-packed (eg. for cubic: spinel or inverse spinel) crystalline structure. This last requirement is thought to result from the need for a very close packing in order for a compound to be magnetic.
Finally, polymers useful in preparing the particles of the present invention may be "tailor-made" to include monomers which exhibit a specific biofunctional activity. Using such a polymer permits direct precipitation of a biofunctional magnetic-polymer particle which requires little or no further treatment in order to be useful in assays which rely on the particular biofunctional activity of the polymer.
In certain applications, larger, less stable particles are useful. The particles of the present invention may be made to agglomerate while still retaining both their biofunctional and magnetic characteristics. Agglomeration of the particles may be accomplished by treatment of a suspension with a predetermined amount of, for example, barium chloride solution. This treatment may be designed to cause the particles to settle out of suspension in a predetermined period of time in order to allow the performance of further procedures, or to allow the larger particles to be easily attracted by relatively small magnets.
The following examples serve to illustrate various parameters of magnetic-polymer particle preparation and use:
The starting aqueous solutions for a typical (micro) preparation are as follows:
1 mg./ml solution of a given protein
500 mg./ml FeCl3
200 mg./ml FeCl2
20 mM phosphate buffer (approximately pH 7)
7.5% or 15% ammonium hydroxide solution (NH4 OH)
The preparation begins by diluting 2.5 ml of the protein solution to a volume of 20 ml with distilled water. 175 ul of the FeCl2 and 140 ul of the FeCl3 is added. Finally, about 200 ul of ammonium hydroxide solution is added.
It should be noted that the amount of base added is usually pre-calculated to allow for the gross amount of protein or polymer in the reaction mixture and the buffering ability of that particular protein or polymer. The objective is to raise the pH to a sufficient degree to permit precipitation of the iron oxides, but to retain the essential activity of the protein or polymer by not exceeding the pH at which the material denatures.
These materials are all added to the reaction vessel with continuous agitation in order to promote a homogeneous reaction mixture. Immediately upon addition of the base (NH4 OH), black particles precipitate. The reaction mixture is centrifuged, typically at 3,000 rpm for 15 minutes and the supernatant is discarded. The pellet is then broken up and the material washed in 20 ml of phosphate buffer, typically for three wash cycles, with centrifugation. After the final wash, the pellet is again broken up and the material resuspended in buffer, under the influence of sonification. The final solution is a transparent amber or brown solution which is a stable suspension of ferromagnetic-polymer particles. Throughout the procedure, no residual aggregate of iron is ever observed.
Subsequent treatment of the particles may proceed by any of the procedures detailed below, including intra-particulate cross-linking, or extra-particulate cross-linking to biofunctional reagents.
As described above, metals other than iron may be incorporated into the magnetic-polymer particles of the present invention. Presented below is a (non-exhaustive) table of those ions which may be used in preparation of magnetic-polymer particles:
In addition to the list above, Fe(II) may be used in combination with selected transition metal ions whose electromotive potential is insufficient to oxidize the Fe(II) to Fe(III). Of the above listed metals, only V(III) is capable of oxidizing the Fe(II), and is therefor unsuitable.
Two solutions (each 40 ml) were mixed rapidly in an ultrasonic bath to promote mixing. Both contained 1.5 mg/ml bovine serum albumin (BSA). One contained ammonium hydroxide (8 ml of 30%, final concentration=74 mM). The other contained 140 mg of Fe(II) Chloride and 280 mg of Fe(III) Chloride (total iron concentration, 1 mg/ml after mixing). A black precipitate immediately formed. The mixture was neutralized by addition of 6 ml of glacial acetic acid while stirring. The sample was divided into 4 tubes and the precipitate was washed by centrifugation (3000 rpm, 15 min) and the small amount of iron remaining in the gold-colored and slightly cloudy supernatent was discarded. The pellet was resuspended in 20 ml of 20 mM phosphate buffer at neutral pH. Each of the 4 tubes containing 5 ml of particles was sonicated for 5 min in a Branson Sonifier with a cup horn attachment.
The available amino groups on the particles were reacted with succinimidyl-propiono-dithiopyridine (SPDP) to prepare a portion of them for later coupling to antibody. This reaction (20 ml particles+5 mg SPDP) proceeded for 1 hour in the cold with stirring. Then 25 mg of the amino-reactive cross-linking reagent ethylene-glycol-disuccinimide-ester (EGS) was added to further stabilize the particles. The mixture was reacted for a further hour with stirring. The EGS cross-linked any remaining amino residues which were in close enough proximity to be coupled. Other remaining amino groups would have been bound by one of the ends of the EGS molecule while the other would eventually hydrolyze to a carboxylic group. This results in a net conversion of positively charged to negatively charged groups on the particle surface which is thought to promote stability of the colloidal suspension.
At the end of the reaction period, the preparation was transferred to a 50 ml tube and 10 ml of 3-molar NaCl solution in water was added to "salt out" the particles. After allowing 10 minutes at room temperature for aggregation, the particles were centrifuged at 1500 rpm for 10 minutes. The clear colorless supernatant was discarded. The pellet was resuspended and centrifiged two more times before receiving a final suspension in 20 ml of 20 mM phosphate buffer (no sonication was usually necessary at this point).
Particles were coupled to goat antiserum specific for horseradish peroxidase, as a conveniently assayed antigen. Antiserum was activated by reaction with SPDP. For 30 minutes at room temperature, 0.128 ml of antiserum, containing 1.28 mg total protein, was reacted with 12.8 micrograms of SPDP, diluted into 0.512 ml phosphate buffer. After 30 minutes 3.1 mg dithiothreitol (DTT) was added to convert the SPDP to its free sulfhydryl form. The reacted Ab was separated (desalted) on a small gel filtration column.
Thiolated antiserum and SPDP-activated particles were reacted by the addition of 6.4 ml containing the antibody (0.64 mg of antiserum protein if there was 100% recovery) to particles containing 14 mg of iron. Concentrations in the reaction mixture were 1.0 mg/ml for antiserum protein and 2.2 mg/ml for particle iron, in a total volume of 6.4 ml. After an hour at 4° C., the particles were salted out and washed. The pellet was resuspended in 3.2 ml phosphate buffer, with sonication, for 1 minute.
The antigen-binding activity of the magnetite-bound anti-peroxidase was assayed by incubating an aliquot of particles with free horseradish peroxidase (HRP), salting out the particles, washing them and assaying the resuspended particles for enzyme activity in a colorgenic assay. The antibody-coupled particles took up more than ten times as much enzyme as a control particle preparation which was coupled to a similar antibody that was specific for an irrelevant antigen.
Particles were formed by the method described in Example 2, with the exceptions that a small amount of radio-iodinated (125 I) BSA was added to the reaction as a tracer for the protein component, and that four preparations were made which varied the amounts of iron and BSA in the precipitation reaction as follows:
A. 1.25 mg Fe/ml and 1.5 L mg BSA/ml (usual concentrations)
B. 3.75 mg Fe/ml and 1.5 mg BSA/ml (higher iron concentration)
C. 1.25 mg Fe/ml and 5.0 mg BSA/ml (higher BSA concentration)
D. 3.75 mg Fe/ml and 5.0 mg BSA/ml (higher iron and BSA concentrations)
Immediately after precipitation but before any washing procedures, a sample was counted to quantitate the amount of radioactive label in the mixture. Then the particles were centrifuged, washed, resuspended, and counted. The results showed complete utilization of the BSA (incorporation into the magnetite pellet) for mixtures A and B, only 60% incorporation in the high-BSA sample (C), and a return to complete incorporation when the amount of iron was elevated to return to approximately the original ratio of iron to protein (sample D). Assuming that the iron was completely converted to magnetite (Fe3 O4), the compositions of the four preparations were:
A. 46% protein, 54% magnetite
B. 22% protein, 78% magnetite
C. 63% protein, 37% magnetite
D. 49% protein, 51% magnetite
The stability of the protein-magnetite particles against loss of BSA during sedimentation and resuspension with sonciation was tested. After resuspension, particles were again "salted out" and radioactive BSA left behind in the supernatent was counted. An average of 40% (st. dev.=11%) of the counts which had been incorporated into the particles was lost. When the process was repeated, the losses in the next wash were less (14%, st. dev.=4%).
Particles were precipitated and coupled to human IgM with SPDP, as described in Example 2 above, to make a magnetic antigen. The particle-IgM magnetic antigen bound commercial antibody against human IgM to which was coupled alkaline phosphatase enzyme (Ab-AP). A 1:500 dilution of Ab-AP was incubated with approximately 250 micrograms of IgM-magnetite in 100 microliters of phosphate containing 1% BSA. The amount of Ab-AP which bound to the IgM-magnetite was quantitated by passing the incubation mixture through a small magnetic filter. The filter bed was then washed with excess buffer, and loaded with buffer containing enzyme substrate. After 15 minutes incubation, the buffer was eluted and the amount of reaction product generated by enzyme captured on the filter bed was determined by measuring the optical density.
The above procedure formed the basis of a competitive immunoassay for human IgM. When small amounts of free IgM were added to the incubation mixture, the uptake of enzyme by the magnetite-antigen was specifically inhibited. The decrease in enzyme activity on the filter as a function of IgM in the incubation mixture was graphed to produce a calibration curve which allowed the process to be used as a competitive immunoassay to measure unknown amounts of IgM. The sensitivity was approximately 0.15 mg/ml (the concentration of IgM which resulted in 50% reduction in the specific capture of enzyme activity on the filter).
Through Preparation
Seven precipitation reactions were performed using mixtures of BSA and goat antibody (IgG fraction of goat anti-rabbit-immunoglobulins). The total amount of antibody in each was either 0.75 or 0.375 mg/ml. To this was added zero BSA, or enough BSA to bring the total protein up to 0.375, 0.75, 1.5 or 3.0 mg/ml. To each 2-ml sample was added 3.5 mg FeCl2 and 7.0 mg FeCl3. Particles were precipitated by the addition of 20 microliters of 30% NH4 OH, to bring the pH to 9.4. The preparations were then neutralized with acetic acid and spun down in a centrifuge and washed. The pelleted particles were resuspended with 2 minutes of sonification each. Half of each preparation was treated with the bifunctional reagent EGS (0.31 mg/ml) for several hours. After EGS treatment, particles were salted out with 1.5M NaCl, washed and resuspended with 1 minute sonication each.
The particles which were made with IgG (no BSA), and not EGS-treated, spontaneously settled out of suspension within about 24 hours. This was true of both the 0.75 mg/ml and the 0.375 mg/ml preparations. In both cases, however, the half of the prep which received the EGS treatment was stable in suspension and could be used.
Twelve samples were tested for goat-anti-rabbit-Ig activity by hemagglutination. A series of microtiter wells was set up containing sheep red blood cells (SRBC) and a sub-agglutinating concentration of rabbit antibody against SRBC. In each series, particles were added in concentrations which decreased two-fold in each successive well in the series. After several hours, the maximum well in which agglutination could be seen was read for each series. More active particles agglutinated at lower concentrations (larger well numbers). A table of results for the 12 preparations tested is given below.
|Hemagglutination Results Goat-anti-Rabbit Particles 0.375 mg/ml Ab 0.75 mg/ml Ab Total Protein in Precipitation in Precipitation in Precipitation No EGS +EGS No EGS +EGS
0.375 mg/ml
0.75 mg/ml 8.5 8.5 <S> 6.5
1.5 mg/ml 8.5 8.0 9.5 10.5
3.0 mg/ml 7.5 7.5 9.0 9.5
<S> = settled and so not tested in hemagglutination Note: Fractional wells refer to pattern in which last well was not definitively agglutinated (Eg 9.5 = 9 wells agglutinated and tenth one partially so).
After two weeks storage at 4° C., hemagglutination was repeated from the samples which were made with 1.5 mg/ml total protein. Activity was essentially unchanged. Specificity of binding was demonstrated by the fact that no hemagglutination was seen in a control series of wells in which the anti-SRBC antibody was omitted.
(i) Antigen binding activity was hardly affected by EGS. In particles made with the same total protein, more activity was seen in preparations which contained greater amounts of Ab.
(ii) In particles which contained either amount of Ab, but increasing amounts of BSA, improvements were seen up to 1.5 mg/ml total protein, and a slight drop-off in activity above that point.
Experiments were performed on magnetic particles containing 125 I-labelled protein (HSA) in order to assess the stability of the particles with respect to protein composition.
Particles were prepared using 125 I-labelled HSA that was iodinated by the ICl method. Three preparations were made containing total protein concentration of 0.05, 0.5 and 1.0 mg/ml of HSA. Each of these samples was prepared using the same amount of Fe. The relative proportions of "hot" and "cold" protein were adjusted such that the specific activity (4,000 c.p.m./ug) was the same in each case.
In each case the amount of radio-label present in the supernatant immediately after particle formation was assessed. The amount of radio-label lost in the subsequent washing procedure was also determined. The final sonicated particle suspension was assayed for radio-label and Fe content and a protein/Fe ratio thus determined. The results are summarized in Table IV.
Under the conditions employed, final Fe content was approximately 1-1.56 mg/ml. Almost all of the labelled protein was incorporated into the initial precipitate in the case of preparations containing protein concentrations of 0.05 and 0.5 mg/ml. However, when protein was used at a concentration of 1 mg/ml a substantial proportion (approximately 30%) of the radio-label remained in the supernatant. This indicated that under these conditions, particle formation was efficient with regard to protein incorporation up to protein concentrations of 0.5 mg/ml. Higher protein concentration appeared to result in a saturation effect whereby excess protein remained in solution. This conclusion was further substantiated by the fact that the protein Fe ratio rose substantially in going from 0.05 to 0.5 mg/ml of protein, but no further increase was observed at 1.0 mg/ml.
The particles appeared to be relatively stable with regard to protein content throughout the washing procedure. (Significantly, the particles formed at high HSA concentration, lost more radio-label during the subsequent washing of the particles). It should also be noted that some of the loss of 125 I-label may be accounted for by the loss of small but intact protein/magnetite particles.
Experiments were also performed in which the presence of free protein in these particle preparations was assessed by gel filtration. Preliminary experiments indicated that very little of the protein appeared to become "particle-free" after sonication. Also, no further protein appeared to "leach-off" when preparations were stored for up to 7 days. Furthermore, a second sonication did not appear to increase the proportion of free protein.
|Original Protein 0.05 0.5 1.0 Concentration (mg/ml) Original 125 I-HSA 1.106 10.106 20.106 (c.p.m.) 1st Supernatant (%) 2.2 7.2 28 (5 mls) Precipitated Particles* (%) ##STR1## ##STR2## ##STR3## 1st Wash (%) (20 ml) 3.5 3.5 2.7 2nd Wash (%) (20 mls) 7.5 8.2 5.6 3rd Wash (%) (20 mls) 4.5 8.3 21 Sonicated Particle 83 78 70 Suspension (%) Fe Content (mg/ml) 1.1 1.2 1.6 Protein: Fe ratio 1:30 1:3 1:3
*Obtained by difference
By the above technique, a polymer having reactive groups may be employed to provide a photoactivatable particle which may then be coupled to specific biofunctional compounds.
In an attempt to replicate the preparative procedures of Molday and compare these to the procedures of the present invention, two series of reaction mixtures were prepared. The first series of mixtures began with the dextran and iron concentrations as taught by Molday and serially decreased dextran concentrations by powers of 10 in order to assess dextran concentration effects upon final particle characteristics. The second series of reaction mixtures began at dextran and iron concentrations in accordance with the present invention and serially increased dextran concentrations by powers of 10. The results of these series of preparations are summarized in Table III.
|Results Fe (III) Fe (II) Dextran After Base After Heating Procedure (mg/ml) (mg/ml) (mg/ml) Addition (Molday Only) After Sonication
75 32 250 Stable Susp.
Stable Suspension
Same 25 Magnetic Ppt.
No change
Stable Magnetic Susp.
Same 2.5 Same
Same 0.25 Same Non-Resuspendable
Example 1
4 1.6 0.25 Magnetic Ppt.
== Stable Magnetic Susp.
Same 0.5 Amber Solut'n.
Same 1.0 Same == NA
Same 256 Same == NA
Example 1
4 1.6 0.0025
Magnetic Ppt.
== Stable Magnetic Susp.
Same 2.5 Amber Solut'n.
Notes: NA = Not Attempted == = Not Applicable to procedure used.
As is apparent from the comparative data, preparations in accordance with procedures taught by Molday fails to yield resuspendable particles. As concentrations of dextran are decreased toward the range employed in the present invention, the quality of product particle is degraded and the suspension of these particles becomes unstable. In contrast, concentrations of dextran and iron as taught by the present invention yield marginally acceptable stable suspensions of weakly magnetic particles. As dextran concentrations are increased toward that taught by Molday, however, a failure to precipitate results and no particles are produced. This divergence in conditions, as well as certain qualitative differences in particle size and iron to polymer ratio, are indicative of the extreme differences between the process and product taught by Molday and that of the present invention.
From the foregoing examples, the following conclusions have been drawn by the present inventors:
(i) The particles of the present invention are
(a) Magnetic
(b) Stable in Aqueous Suspension
(c) Resuspendable
(d) Small (and thus filter sterilizable)
(e) Easily produced to include a specific biological activity
(f) Clear in Suspension
(ii) Particles may be produced and subsequently bonded to specific biofunctional ligands using convention bifunctional reagents.
(iii) Specific biofunctional ligands may also serve as the polymer during coprecipitation without a significant loss of biological activity.
(iv) Treatment of particles with bifunctional reagents may improve overall stability, both of the particles and of suspensions, without excessively impacting on particle size.
(v) The particles, when coupled to specific biofunctional ligands, may be used in magnetic immunoassays for biological materials of interest.
(vi) The particles decrease the NMR relaxation time of neighboring protons in solutions or tissues. This decrease is most pronounced for the T2 (spin-spin) relaxation.
While this invention has been described with reference to specific examples, it will nonetheless be understood by those skilled in the art that other variations of process conditions and parameters may be employed without departing from the true spirit of the invention. It is intended that the claims which follow should be construed to encompass all such variations.
The present invention comprises novel magnetic-polymer particles and methods for making them. These particles are useful in a variety of biological/medical fields including cell capture, use as a contrast reagent for NMR imaging, immobilized enzyme reactors, immunoassay, and other analytical and diagnostic techniques.


text:
Welcome to NCM!
How many hours do your employees work on average? Is it too much? Too little? Are they even working while they're at work? Expert Robin Cunningham shares his observations about the ways successful dealerships are scheduling their employees.
Have another question for Robin or the other #AskNCM experts? Email us at email@example.com!

subset: commoncrawl


text:
The New York Islanders have seven collegiate prospects – three in the WCHA, three in the ECACHL and one in the CCHA. There are four forwards, two defensemen and one goaltender. The future of these Islanders prospects is bright as they are all enjoying successful seasons with their teams, having garnered five "Player of the Week" awards collectively.
Kyle Okposo, RW
University of Minnesota Golden Gophers (19-1-3)
2006 first-round draft pick (7th overall)
pan style="font-size: 10pt; font-family: Arial;">Freshman forward Kyle Okposo has all the makings of a star. As charismatic as he is gifted with handling the puck, he has already become one of the more popular players on the University of Minnesota Golden Gophers. He scored a highlight reel goal by putting the stick between his legs and collected his first hat trick. On Oct. 31, Okposo was named WCHA Rookie of the Week following three goals in a two-game sweep of Colorado College. Dec. 5 was a tremendous day for Okposo, who was again named WCHA Rookie of the Week after a three-point weekend against rival Minnesota State, which helped continue his team's 16-game unbeaten streak. On that same day he was also named to the U.S. National Junior Team, where he played on the team's top two lines and was instrumental in capturing the bronze medal at the recent World Junior Championships.
The University of Minnesota hockey environment is conducive to Okposo's development as a player. He has an excellent support system both on the roster and in the coaching staff. The team holds the No. 1 ranking in the nation. Currently, Okposo leads all rookies on the team in points, with 16 goals and nine assists for 25 points to go along with a +17 rating and 26 penalty minutes in 23 games played.
Rhett Rakhshani, RW
Denver University Pioneers (14-6-2)
2006 fourth-round draft pick (100th overall)
In his freshman year for Denver University, Rhett Rakhshani is off to a strong start, averaging close to a point per game. The Pioneers have enjoyed success as well, going undefeated in the month of November with a 5-0-1 record. A rookie from California, Rakhshani has shown no shortcomings in his game so far and has lived up to the hype in being considered one of the nation's top freshmen forwards. On Nov. 14th, Rakhshani was honored with the WCHA Rookie of the Week award for stats that included one game-winning goal (with :15 seconds left), 11 total shots on net, a +2 rating, four hits and two blocked shots, in the two-game sweep of Wisconsin. He was again named Rookie of the Week on Jan. 2 for his performance in the Wells Fargo Denver Cup in which he was named Most Valuable Player after collecting three assists in the tournament.
Rakhshani has proven to be the very definition of a well-rounded player. He leads all rookies on his team and ranks third overall in points. He has shown no hesitance in going after the puck along the boards, in the corners, or in the neutral zone. This aggressive style of hockey and playmaker's touch make Rakhshani a definite prospect to keep an eye on. To date, he has put in five goals, added 20 assists, for a total of 25 points, a -1 rating and 26 penalty minutes in 28 games played for DU.
St. Cloud State Huskies (14-3-3)
2006 fourth-round draft pick (108th overall)
For both freshman goaltender Jase Weslosky and the St. Cloud State hockey team, this will be a learning season. Weslosky is learning the collegiate game as backup to returning All-American Bobby Goepfert. The Huskies, on the other hand, are coming to understand that they have something special with Weslosky. While he's played in only five games this season, he's certainly racked up the highlights. In his first start, Weslosky earned a victory with a 28-save performance. In his second game, he shut out the Clarkson Golden Knights, including two saves against fellow Islanders prospect Shea Guthrie. For his 24-save effort in that game, Weslosky was named the first star.
St. Cloud State has used Weslosky sparingly thus far in the season but each time he's been put in, he's proven to be smart, patient and quick in the crease. His numbers are very telling of his ability and when Goephert finishes his senior year this season, the Huskies will inherit a quality starting goaltender for the next three seasons. To date, Weslosky has posted a 5-0-0 overall record, with nine goals allowed for a 1.80 goals-allowed average, 116 saves, and a .928 save percentage.
Shea Guthrie, W
Clarkson Golden Knights (13-5-1)
2005 third-round draft pick (76th overall)
Coming in with the expectations of exceeding last season's All-Star statistics for the Clarkson Golden Knight hockey team, second-year forward Shea Guthrie had difficulty finding the net at the beginning. However, while he may not have been contributing in that particular statistical category, he was busy setting up plays and recording assists, leading to a six-game point scoring streak from Oct. 14th to Nov. 11th, including a two-game stretch in which he recorded five points in two games. In December, Clarkson competed in the Badger Showdown. The Golden Knights won the championship, beating Lake Superior State 3-2, with Guthrie having scored the game-winning goal. Recently, he totaled three assists in a surprising victory over the reigning NCAA champion Wisconsin.
Guthrie has since risen to be among the offensive leaders for the Golden Knights and has begun scoring with more consistency. For the most part, he seems to be getting back into the groove of things and the second half of the season will be telling of how he's maturing as a hockey player. To date, Guthrie has recorded five goals, 12 assists for 17 points and 20 penalty minutes in 20 games this season.
Doug Rogers, C
Harvard Crimson (7-10-0)
2006 fourth-round draft pick (119th overall)
A big and mobile centerman, Doug Rogers is playing well for the Harvard Crimson. The freshman is patient with the puck and sets up plays very well. In only his second collegiate game of his career, Rogers notched his first goal when the Crimson lost to fellow Islanders prospects Guthrie and Tyrell Mason of the Clarkson Golden Knights. For Rogers personally, November was a good month for him, one that saw him tally 9 points in only 11 games played. The team, however, did not fare that well, dropping out of the top-20 national ranking and slipping to 3-9-0.
Rogers has leadership experience, having captained St. Sebastian's School. He is currently second on the team in scoring and has done well on the power play, recording three of his points with the man advantage. He's got three more seasons to go and is certainly a prospect to watch. To date, Rogers has recorded four goals, nine assists for 13 points and 10 penalty minutes in 17 games played.
Tyrell Mason, D
Clarkson Golden Knights (13-5-1)
2005 sixth-round draft pick (180th overall)
More along the lines of the defensive-minded defenseman, Mason has improved upon his game with the Clarkson Golden Knights. Last season, he recorded six points total and had a team-leading +10 rating. In the first six games of the season alone, Mason recorded three assists and tallied up a +7 rating. Continuing his stellar play into the New Year, he has already passed his statistical achievements of last season with seven assists to date.
To go along with his teammate Guthrie's efforts against Wisconsin, Mason also registered three assists in the impressive victory. Also, following Clarkson's championship game win at the Badger Showdown tournament, Mason was named to the Badger Hockey Showdown All-Tournament team. Truth be told, he isn't the type of player who will consistently put up big numbers game in and game out for his team. He is, however, the player you want guarding your team's blue line. He makes smart decisions with the puck and knows how to turn the play around, as best evidenced by his plus/minus rating. To date, Mason has yet to register a goal, recorded eight assists for eight points and 38 penalty minutes in 23 games played.
Wes O'Neill, D
Notre Dame Fighting Irish (17-4-1)
2004 fourth-round draft pick (115th overall)
Wes O'Neill passed on the opportunity to join the Islanders organization this season in favor of graduating with a degree and to also trying to help the Notre Dame Fighting Irish into the NCAA playoffs. At this point in the season, O'Neill has already seen tremendous success. On Oct. 20, there was the headline grabbing 7-1 defeat of the previously No.1 ranked Boston College. Shortly thereafter, on Oct. 23, O'Neill was named CCHA Defensive Player of the Week for his efforts in the Boston College game (two assists) and for his play against Providence (helped to kill 15 of 16 penalties). The alternate captain and his mates also won the school's first hockey tournament since 1981, the Lightning College Hockey Classic. The team's wins in the tournament included impressive shutouts of both Army and Air Force.
The Irish rode a nine-game unbeaten streak into the middle of November and saw their team's ranking rise from 10th to fourth in the nation. They continued their strong play into December, winning six of their games, moving their overall ranking up to No. 3. The head coach of the Fighting Irish is, in fact, former Islanders assistant coach Jeff Jackson. To date, O'Neill has yet to find the net, registering no goals, 12 assists for 12 points, to go along with a +9 rating and 24 penalty minutes in 26 games played.
Copyright 2007 Hockey's Future. Do not reprint or otherwise duplicate without permission of the editorial staff.


text:
Vintage Birds Quilt
Beautiful vintage quilt in peach and cream colors with embroidered bird designs. Just a few stains, but overall great condition! Really amazing addition to a child's room. Measures approx. 66" x 68".
Vintage pieces are beautiful. Each item is unique and different. We source only the best curated items, but please expect some wear. Email us prior to purchase if you would like to see additional images and a more detailed description of the item.


text:
River Ecclesbourne
This article needs additional citations for verification. (January 2013) (Learn how and when to remove this template message)
|River Ecclesbourne
|Physical characteristics
|• location
|Wirksworth, Derbyshire
|Confluence with the Derwent
• location
|Duffield, Derbyshire
• coordinates
|52°59′5″N 1°28′48″W / 52.98472°N 1.48000°WCoordinates: 52°59′5″N 1°28′48″W / 52.98472°N 1.48000°W
|15 km (9.3 mi)
|Basin size
|60 km2 (23 sq mi)
|• location
|Duffield [1]
|• average
|0.63 m3/s (22 cu ft/s)
|Basin features
|• left
|Franker Brook
|• right
|Alton Brook, Holm Brook, Sherbourne Brook
|Progression : Ecclesbourne—Derwent—Trent—Humber—North Sea
The River Ecclesbourne is a small river in Derbyshire, England, which starts in the upper part of the town of Wirksworth, flows for 9 miles to Duffield,[2] and then enters the River Derwent just outside the town.
The River Ecclesbourne's source is in the town of Wirksworth. This is also where the Ecclesbourne Valley Railway, which is named after the river and follows it for its entire length, begins. The start of the river is located between Steeple Grange and Sough Lane. The river's two main headwaters come from Steeple Grange, below Bolehill, and the fields below the National Stone Centre, Ravenstor. The main source is about 100 metres down the railway tracks from Ravenstor railway station, where the Meerbrook Sough Mine area lies. The Meerbrook Sough drained water from the nearby quarries and led them in a tunnel to Whatstandwell where the excess water from the Sough joined the River Derwent towards Ambergate. The source of the Ecclesbourne contributed to the water powering the Meerbrook Sough. The Ecclesbourne flows mostly underground through the area of the Sough, but momentarily appears above ground in several places, near Ecclesbourne Cottages at the bottom of Sough Lane, in a shallow ditch next to the railway (now piped) and also near the bottom of Fanny Shaws Park, again near the main railway track. Part of the main headwaters of the Ecclesbourne flowed past the rectangular Sough Reservoir, which transported water up to the quarries around Wirksworth. This reservoir was located at the bottom left area of Fanny Shaws Park. The Meerbrook Sough Engine House was built in the middle of the Sough area and powered the waterworks of the Sough and made materials for the quarries. Below Fanny Shaws Park and skatepark the Ecclesbourne leaves the Sough area and goes underground and reappears in the Hannages further down the railway tracks, where it powered the mills at the bottom of Wirksworth. The housing estate of Spring Close has recently been built over the area of the Meerbrook Sough; the Engine House still remains in its original place, but has recently been renovated into a house.
After leaving Wirksworth the River Ecclesbourne is now a fairly large river. It flows slowly through the fields, collecting smaller streams as it goes (among them the Alton Brook and Holm Brook), to its next port of call, the village of Idridgehay. Further down, the Ecclesbourne flows through Turnditch. After leaving Turnditch the Ecclesbourne increases in volume and speed. In some areas the river is piped underground to reduce the risk of flooding, but this rarely happens any more.
A few miles beyond Turnditch the river enters the town of Duffield. The Ecclesbourne School, the town's secondary school, is built on its banks in the town centre. The Ecclesbourne then flows out of Duffield and leaves the railway after flowing underneath its tracks. It reappears and flows through a few more fields before joining the River Derwent, which has just come down from Milford and Belper. At the confluence where the Ecclesbourne ends it is about 10 metres wide. The area around the confluence is popular with visitors and a good area for angling. After the river has met the Derwent, the railway line joins the main line to Derby.
Wildlife and conservation[edit]
The River Ecclesbourne is a popular fishing river, known for producing good-quality fish, especially in its lower reaches. It supports a population of brown trout.
Species of conservation interest found along the Ecclesbourne include white-clawed crayfish, kingfisher, otter and water vole. Threats include pollution, weirs that restrict fish migration, and invasive Himalayan balsam. The river is one of 25 river catchments chosen by Defra for a Pilot Catchment Project, with the aim of encouraging co-operation between various agencies and interested parties to improve water quality, as part of the UK's commitment to the EU Water Framework Directive.[2]
- ^ "28055 - Ecclesbourne at Duffield". The National River Flow Archive. Centre for Ecology & Hydrology. Retrieved 10 February 2013.
- ^ a b Turley, Heather (2012). "The Ecclesbourne: Going with the flow". Wild Derbyshire. Derbyshire Wildlife Trust. Winter 2012/Spring 2013: 14.


text:
When Things Don't Go As Planned, Here's What To Do
When things don't go as planned, we beat ourselves up. Read on to learn how to move forward with your plans and dreams even when things don't go as planned.Read More
Communication In Relationships For Times Of Transition
Communication in relationships is key. Read on to learn why communication is so crucial right now and what you can say when you're at a loss for words.Read More
Calendarize Your Tasks To Make Sure They Get Done
Want to know my secret to never missing events, appointments, payments, birthdays, and other important dates? It's really simple: I calendarize my tasks.Read More
What You Need When Embarking On A New Adventure
Throughout your life, you'll embark on new adventures. Here's what you need to carry to make your transition less rocky and make the most of your journey.Read More
Dealing With Opposition From Every Which Way
No matter what you do in life, somebody will always oppose it. When they challenge your decisions, it's up to you to be strong and keep your commitment.Read More
Impressing Your Date With Charm And Authenticity
Surface-level won't cut it when you're searching for your soulmate. I'll guide you through the mental and physical prep work (by just being wonderful you).Read More
Dealing With Insecurities Before They Swallow You Whole
Insecurities take the fun out of everything. They can swallow us whole, if we let them. Here's how to deal with your insecurities next time they pop up.Read More
Your Plan To Take An Easy, Inspiring Road Trip
A road trip can be an amazing way to explore the world and discover something new about yourself. Follow my Guide to prepare for your trip and get going.Read More
Uplift Yourself, Be Confident, And Rise Above Self-Deprecation
I learned a life lesson during a job interview, and I have grown from it since. The gist of it is this: don't ever think of yourself as "just" anything.Read More
Four Life Lessons I Learned From Taking An Interviewing Class In College
I learned interviews aren't just for jobs. Many common interactions you experience with coworkers, family, and friends are considered interview situations.Read More


text:
Klaus Kumrow
Developments within an artist's oeuvre lead to new definitions of the ideas and structures that form it. At a certain moment, all considerations focus on the new form without actually touching the original form, like larvae which must take on another form in order to remain themselves. Klaus Kumrow has taken a similar step: from his photographic sculptures (folded photographs presented as sculpture) he has now developed large-format, flat photographs. The effect of this reduction, from the three-dimensional, to the two-dimensional, gives the works more power. Instead of seeming flat, they open themselves to more dimensions, and thus the concept of sculpture has expanded even as it technically disintegrates.
Three of a total of four photosculptures were shown. Ohne Aussicht (Without a view, 1992) is based on many photographs of rail-road tracks that Kumrow folded to look like toys. These
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text:
Respiratory Muscle Training
The benefits of using exercises for breathing muscles
An online course with the Physioplus Team
Trusted by over 100,000 physiotherapists
Improve your clinical knowledge and skills today!
Learn anytime, anywhere, on any device
Are you aware of the implications of the fact that the muscles involved in respiration are also involved in core stability as well as other upper body movements and exercises. Research has demonstrated that exercising to increase the strength and endurance of the respiratory muscles can benefit back pain, balance and falls, artistic and athletic performance as well as address breathing difficulties caused by chronic conditions such as COPD and ageing.
- Section One: What is respiratory muscle training
- Section Two: Practical Application
- Section Three: Learn from the expert – Interview with Alison McConnell
Target audience
This course is aimed at Physiotherapy and Physical Therapy professionals, clinicians and students.
This course is divided into 3 sections and it is suggested (but not required) that each section be completed over the duration of approximately a week. Please note that no deadlines are applied and this course can be started and completed according to your own schedule. We expect the required elements of each section/week to take around 4-5 hours depending on your schedule and learning style. Additionally there are many optional resources provided and if you choose to review these the course could take longer to complete.
It's not going to be easy, we'll expect you to work hard for your completion certificate and PP+ points! You won't be sitting back and watching webinars, we'll expect you to reading tasks, complete quizzes, perform literature searches and other learning activities. You'll need to reflect on your own experiences and contribute them within the discussion forum, this is where we can learn from each other's experiences and knowledge from all around the world! At the end of the course, when you have completed all of the required elements, you will be able to download a certificate of completion and 5 PP+ points will be added to your personalised learning dashboard.
Requirements to complete this course
In order to complete this course and receive a course completion certificate you will need to:
- abide by the Physiopedia Plus Community Culture
- log as completed all the required learning activities
- actively participate in the course forum discussions
- pass a final quiz with a score of 80% or more
- complete a course evaluation form


text:
Face Off Max 184.108.40.206 Full Version With PATCH
Face Off Max - Make fun photo by putting your face on another body using any photo or template. Haven't you ever wondered how you would look if you had another body? Wouldn't it be great to see your face in a celebrity group or on a magazine cover? Now Face Off Max enables you to embarrass your coworkers, make yourself look cool and make your friends look like dweebs all with a few clicks of the mouse. Our amazingly simple and easy interface, step-by-step instruction and hundreds of fantastic templates allow anyone to be a professional artist.
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DONASI VIA PAYPAL Bantu berikan donasi jika artikelnya dirasa bermanfaat. Donasi Anda membantu Admin untuk lebih giat lagi dalam membagikan template blog yang berkualitas. Terima kasih.
Newer Posts Newer Posts Older Posts Older Posts


text:
Pajama Day is basically every high schooler's dream; arriving to your classes in footie pajamas, slippers, robes, or whatever else your preferred form of bedtime comfort may be makes the burden of high school much easier.
For sophomore Jillian Lenicka, her festive pajamas of choice was a Dr. Seuss inspired 'Thing 1," onesie.
"My sister wore the same one for pajama day during her junior year in 2013," Lenicka said.
She said that her experience walking around the school in a bright red onesie with cartoon heads on the feet was both embarrassing and fun.
"People liked my school spirit, and I made friends with people who also wore onesies," Lenicka said.
Whether they were pajamas for sheer comfort or to show off outrageous novelty PJs, many students embraced their sassy sleepwear with pride on Monday.


text:
Superintendent gets extension
Staff writer
Peabody-Burns superintendent Ron Traxson got a vote of approval from school board members who voted to extend his contract Jan. 10.
After an executive session including Traxson to discuss his evaluation and contract, the board voted to extend his contract another year.
The board also met in executive session to discuss personnel matters related to certified staff, but no action was taken.
Business Professionals of America sponsor Mary Miller presented information about upcoming state and national conventions for middle and high school members. The board voted to table a decision on the national conference until February.
Last modified Jan. 17, 2018
© 2020 HOCH PUBLISHING | Marion County Record | Hillsboro Star-Journal | Peabody Gazette-Bulletin | Checkerboard


text:
London Dance Parties
Dance parties are great fun for girls and boys of all ages and the perfect stress-free birthday party idea for parents.
At Let's All Dance, we understand that every child is different, which is why we pride ourselves on creating totally unique birthday dance parties for every child. We're more than happy to work with your choice of music and dance style, or you can leave it to our professional team of dancers to prepare an age-appropriate playlist and choreograph a fun routine for your child and their friends.
On the big day we will liaise with you carefully to work around the games and birthday tea you've prepared, or you can sit back and relax, leaving us to manage the party. So whether you want a dancing Teddy Bear's Picnic for your 4 year old or a funky Street Dance party for your sweet sixteen, we'll create the perfect birthday dance party that your child and their friends will remember for years!
- Warm up
- Children learn a specially choreographed routine designed for your child
- Rehearsal
- Party games (if required)
- Tea (provided and served by you, with dancer helping)
- Mini Makeovers (if required and with parents' permission)*
- Rehearsal
- Performance for parents
Our team of professional dancers have teacher training and are highly skilled in a wide variety of dance styles. We can lead a party with any dance style, any music and you can even add a theme such as princesses or pirates.
Your child can choose from:
- Disco
- Street dance
- Ballet
- Contemporary
- Musical theatre
- Hip-hop
- Tap dancing
- Bollywood
- and many more
We travel to most parts of the UK.
Please contact us for more information and to check our availbility:
020 8265 4634
4 year old ballet party, Clapham
Our team of professional dancers have training and specialise in a wide variety of different dance styles. We'll take into account any specific dance styles your child wants to try, any particular bands or musicians they love or we can work with the theme you've chosen for their birthday party, such as princesses or pirates.
Our dancers specialise in a range of different dance styles, including but not limited to:
- Disco
- Street dance
- Ballet
- Contemporary
- Musical theatre
- Hip-hop
- Tap dancing
- Bollywood
Please contact us if you want more information on the varieties of dance we offer at our children's birthday parties. Whilst based in London, at Lets All Dance, we regularly provide birthday dancing parties throughout England.
If you're interested in our children's dance parties, please get in touch with us or call 020 8265 4634 for more information.
- 1 hour
1 dancer- £140, 2 dancers – £230, 3 dancers – £320
- 1.5 hours
1 dancer- £165, 2 dancers – £255, 3 dancers – £345
- 2 hours
1 dancer- £190, 2 dancers – £280, 3 dancers – £370
- 3 hours
1 dancer- £240, 2 dancers – £330, 3 dancers – £420
All prices are + VAT
*Makeovers cost £1 extra per child and include face glitter and lip gloss for girls/hair gel and temporary tattoos for boys.


text:
Parsing with Perl 6 Regexes and Grammars
This book is 65% complete
Last updated on 2018-12-29
About the Book
As humans, we are incredibly good at finding patterns. So we assume that as programmers, we must be good at using patterns to parse text. But it's a skill that needs to be learned. This book aims to teach you how to write good regexes and parsers with Perl 6.
It starts from the very basics of regular expressions, and then explores how they integrate with regular Perl 6 code. The result of a successful regex match is a Match object, which contains all the useful information for extracting data, so that is the next topic.
Then we discuss how regexes work under the hood, and how Perl 6 uses a mixture of finite state machines and backtracking to do its magic. With this understanding, we can explore common techniques for constructing regexes and exploring the data under scrutiny.
So far, the regexes can only match relatively simple formats. But with reusable named regexes and grammars, the sky is the limit. Figuratively, of course. We discuss techniques for code reuse in grammars, and how to write parser for more involved data formats.
One of my favorite topics is the generation of good error messages for when the input can't be parsed by a grammar, so there will be a separate chapter on that.
Table of Contents
- 1. This Book Will Be Published By Apress
2. What are Regexes and Grammars?
- 2.1 Use Cases
- 2.2 Regexes or Regular Expressions?
- 2.3 What's So Special about Perl 6 Regexes?
3. Getting Started with Perl 6
- 3.1 Installing Rakudo Perl 6
- 3.2 Using Rakudo Perl 6
- 3.3 Summary
4. Building Blocks of Regexes
- 4.1 Literals
- 4.2 Meta Characters vs. Literals
- 4.3 Anchors
- 4.4 Pre-Defined Character Classes
- 4.5 Quantifiers
- 4.6 Disjunction
- 4.7 Conjunction
- 4.8 Zero-Width Assertions
- 4.9 Summary
5. Regexes and Perl 6 Code
- 5.1 Smart-Matching
- 5.2 Modifiers and Quote Forms
- 5.3 Comb and Split
- 5.4 Substitution
- 5.5 Crossing the Code and Regex Boundary
- 5.6 Summary
6. Extracting Data from Regex Matches
- 6.1 Positional Captures
- 6.2 The Match Object
- 6.3 Named Captures
- 6.4 Backreferences
- 6.5 Match Objects Revisited
- 6.6 Summary
7. Regex Mechanics
- 7.1 Matching with State Machines
- 7.2 Regex Control Flow
- 7.3 Backtracking
- 7.4 Why Would You Want to Avoid Backtracking?
- 7.5 Frugal Quantifiers and Backtracking
- 7.6 Longest Token Matching
- 7.7 Summary
8. Regex Techniques
- 8.1 Know your Data Format
- 8.2 Think about Invalid Inputs
- 8.3 Use Anchors
- 8.4 Matching Quoted Strings
- 8.5 Testing Regexes
- 8.6 Summary
9. Reusing and Composing Regexes
- 9.1 Named Regexes
- 9.2 Whitespace
- 9.3 Grammars
- 9.4 Code Reuse with Grammars
- 9.5 Proto Regexes
- 9.6 Summary
10. Parsing With Grammars
- 10.1 Understanding Grammars
- 10.2 Starting Simple
- 10.3 Assembling Complete Grammars
- 10.4 Parsing Whitespace and Comments
- 10.5 Keeping State
- 10.6 Summary
11. Extracting Data From Matches
- 11.1 Action Objects
- 11.2 Building ASTs with Action Objects
- 11.3 Keeping State in Action Objects
- 11.4 Summary
- 12. Generating Good Error Messages
- 13. Acknowledgements
- Notes
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My favorite drinks in summer are green herbal or sweet fruity lemonades – home made of course.
If you happen to have a similar blessing of mint like me, cut it to encourage even better growth and make this refreshing green lemonade:
Place 10 freshly cut mint stalks with the leaves in the blender, add 500 ml still water, 1 cup of crushed ice, the juice of 1 organic lemon and approximately 2 tablespoons of agave syrup or acacia honey (take more if you love the lemonade sweeter).
Blend the ingredients well for around 30 seconds, then strain through a fine sieve.
Enjoy the green drink immediately after preparation to avoid discoloring and loss of freshness.
This drink has a convincing strong green and minty flavor, is full of chlorophyll, menthol and Vitamin C and is my secret weapon against the summer heat and thirst. The fresh breeze you feel on your taste buds when you take the first sip is something you won't want to miss anymore once you have tried it.
Stay healthy and enjoy the summer!
Another fruity and mild lemonade recipe will be featured soon on this blog.


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Issue 89
November 2010
Local Transport Plan Consultation
The final draft long term transport strategy and implementation plan for Leicestershire is now available for comment. The strategy looks at the challenges that Leicestershire may face to 2026 and how transport policy could help to address these.
The Plan consists of a long-term strategy and shorter-term implementation plan. The County Council would like as much feedback as possible and you have until 26 November to lodge your comments. A summary of the proposed plan can be viewed at: www.leics.gov.uk/current_LTP3_consultation
While cycling features, much is made of the concerns of the public about safety. Perhaps this is one area where the County could devote some resource to overcoming the false perception that cycling is particularly dangerous.
Also there appears to be no mention of working with the NHS with whom they should have a symbiotic relationship in that more cycling improves health and reduces the number of cars on the road, the need for new roads (plus wear and tear on existing ones) and atmospheric pollution together with CO2 emissions.
East Midlands Cyclists' Forum meeting
At the most recent meeting, hosted in Nottingham on Saturday 2nd October by Pedals, it was decided that the EMCF should continue through the current local government turmoil, both to share ideas and experience, and to make sure that we are all fully briefed so as to know how best to take advantage of the various opportunities afforded when new agencies (transport, health, etc.) do emerge, even if they have much more limited budgets for cycling. Some, such as the new business-led Local Enterprise Partnerships will only deal with parts of the present region, covering new boundaries that include some areas outside the present East Midlands.
There were 26 at the meeting and just over half stayed on for the afternoon ride, on The Big Track route, in lovely autumn sunshine, with plenty of refreshment and opportunities for further informal networking. Photos and the slides from the two main presentations can be found on the Pedals website. A third, shorter, presentation, was given by Joanna Ward, Project Director for Sustrans / Cycling England about the 'UCycle' Higher Education Project in Nottingham.
Andy Salkeld, Cycling Co-ordinator for Leicester City Council, has offered to host the next meeting in Leicester with a provisional date of Saturday 12 March 2011.
East Midland to set limit of 2 bicycles per train
It is reported (but not yet confirmed) that East Midlands Trains are planning on making it compulsory for cyclists to reserve their place on the train, ban bikes from the vestibule and (strictly) allow only two cycles per train.
Dave Holladay of Transport Management Solutions (who works with CTC on public transport issues) reacted with these comments:
How the apparent idea that bikes cannot be placed in any other location than the van space suggests that those making this proposal have failed to actually observe how the service has to operate at Loughborough, Long Eaton and I think a few other places.
In my book, bike spaces are like sleeper berths as a special facility - if a sleeper berth is available at the time of departure the train staff are aware that it is available for the remaining part of the route (no booking), and you can get a place on a first come first served basis. The same should apply to bike spaces.
For Mr Souter's other rail operation he sees the clear economic and self-interest gain in getting passengers with cycles using the trains. By converting both peak hour commuters and off-peak travellers to cyclists he overcomes a major problem and potentially massive costs, and revenue loss to the company, for the provision of car parks. On his other rail operation the fact that car parks were being filled by 07.30 meant that fewer people were coming in to use off-peak services and fill the empty seats because there is no parking.
Patrick Lingwood's review of Bedford Station is telling in this respect - just 19% of the passengers are delivered through the car parking facility but this requires a totally disproportionate claim on resources (money and space) to supply and maintain those parking spaces. Weighing up in land terms alone the picture emerged of being able to 'deliver 7 passengers by bike for every one delivered through car parking. With a multi-level parking space costing around £12,000-£15,000 to build and approaching £1000/year in running costs (lighting, CCTV, management) it becomes rather difficult to see how a car park actually manages to make a serious commercial proposition. NB the car parking capital costs are paid for either by the TOC (funded by the subsidy or reduced payments to DfT- ie public money) or Network Rail (public money) or local/national government direct funding (public money) Does anyone actually look at this as a value for (public) money exercise?
A refurbishment of the Meridian trains should :
- clear out the bay with the daft pigeon-holes over the bike space, remove the locked door and bulkhead and install the German-style Gamma securing units, which can be accessed directly through the single outer door, as dedicated and permanent bike space for bikes;
- install a bay of tip-up longitudinal seating in the opposite end to the toilets in each coach, with a wider access to the door vestibule to take the surge of passengers at station stops, store additional bikes/wheelchairs/luggage outside peak commuter times, and provide a bay for the catering service to operate from when the trolley cannot get through the train. This could provide a pilot/demonstration for flexible internal space design for the many routes where a long distance train has to deliver a commuter service at certain times of day.
Editor's note: the current storage system and lack of luggage capacity on trains serving 2 major airports means that passageways and vestibules are often blocked, and these would be the main escape routes in an emergency. Dave's suggestion would address this safety issue.
Sustrans Interactive Cycle Maps
Sustrans' online mapping service aims to prove the information you need to help you travel on foot or by bicycle. It helps you find walking and cycling routes and information on public transport, as well as local schools, shops, sports facilities and other destinations.
The mapping service can also be used to find all routes of the National Cycle Network.
Who Cycles?
A report for market researchers Mintel "Bicycles in the UK 2010" reveals that regular cyclists – those who cycle at least once a week – are also disproportionately likely to read broadsheet newspapers, be well educated, have a household income of at least £50,000 per year and shop at Waitrose. They are also twice as likely to be men as women.
Bicycle sales are being driven by 35- to 45-year-old family men. Where this age group might once have treated themselves to a sports car – in an attempt to hang on to their youth – they now invest in a luxury bike instead. Alan Sugar is a case in point, showing off the full-carbon Pinarello machines he has bought for his many residences at a cost of many thousands of pounds each. Halfords has confirmed the trend - premium sales as a whole are up by around 54% in the past two years. However the Association of Cycle Traders says the "cycling boom" is confined to a few places, mainly London.
Just 12% of adults questioned said they cycled regularly, while 65% said they never rode a bike. One in seven (15%) said they were "lapsed cyclists" who had a bicycle which they no longer rode. The main reason given for not cycling was safety, with 39% of respondents saying it is too dangerous to ride a bicycle on the road. Ten percent viewed cyclists as "a nuisance". That increased to 14% among those who regarded cycling as too dangerous.
Roger Geffen for the CTC has pointed out that where cycling is perceived to be safe, as in the Netherlands and Denmark, where far more people cycle, 55% of all bike trips are made by women.
Tea and Cakes at Swithland in 2011
St. Leonard's Church Swithland will be open for "Tea and Cakes" on the second Saturday & Sunday of each month commencing in April 2011 through until September 2011 from 2pm until 4.30pm
Loughborough CTC Mince Pie Run
Loughborough CTC will be holding their traditional Mince Pie Run at Belton Village Hall on Sunday 19th December from 10am to 1pm. All cyclists welcome.


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I am trying to upgrade a SQL server with Analysis Services to SQL 2017 cu5, 14.0.3023.8, (I tried CU4 before this) and I'm running into a problem with an SSIS package that processes my cubes, but only when run as a SQL Agent job.
The package is very simply, 1 Analysis Services Processing Task that connects to AS on the same server and tries to process the cubes. This worked fine in previous versions of SQL, the latest we have is SQL 2014. I've tried creating a new package from scratch and get the same error when running the package as a job.
The error is:
Executed as user: <myUser>. Microsoft (R) SQL Server Execute Package Utility Version 14.0.3023.8 for 64-bit Copyright (C) 2017 Microsoft. All rights reserved. Started: 9:22:29 AM Error: 2018-04-10 09:22:29.59 Code: 0x00000000 Source: Analysis Services Processing Task Analysis Services Processing Task Description: Could not load file or assembly 'Microsoft.AnalysisServices.AdomdClientUI, Version=22.214.171.124, Culture=neutral, PublicKeyToken=89845dcd8080cc91' or one of its dependencies. The system cannot find the file specified. End Error DTExec: The package execution returned DTSER_FAILURE (1). Started: 9:22:29 AM Finished: 9:22:29 AM Elapsed: 0.531 seconds. The package execution failed. The step failed.
The package runs fine in VS2017 on the server and from DTExecUI.exe, but not when run as a SQL Job. I just installed, and reinstalled, SQL 2017 developer edition on a new Windows 2016 server. I've tried this on 2 different server and received the same results.
I tried installing the Analysis server Feature pack related items (AS AMO, AS OLE, AS ADOMD) along with copying some AS DLLs into the GAC, but nothing has worked.
The connection string to the cubes should be fine since it works in VS, but its:
Data Source=Server1\Instance1;Initial Catalog=MyCubes;Provider=MSOLAP.8;Integrated Security=SSPI;
The TargetServerVersion is SQL 2017.
I tried configuring the job step to "Use 32 bit runtime", no luck.
Has anyone else experienced this? Does anyone know of a fix for this?
Brian S.
Brian Shaw - Tuesday, April 10, 2018 7:27 AM
A couple of things to check (which, I agree, do not seem to bear any relevance to the error message!):
If the answer to your question can be found with a brief Google search, please perform the search yourself, rather than expecting one of the SSC members to do it for you.
See https://www.sqlservercentral.com/articles/forum-etiquette-how-to-post-datacode-on-a-forum-to-get-the-best-help/ for details of how to post T-SQL code-related questions.
Phil Parkin - Tuesday, April 10, 2018 7:40 AM
Thanks for the reply Phil.
All data sources are pointing to the correct DB instance. Since it processes fine in VS I know those connections are good.
The data source impersonation was set to "Default", I changed it to "ImpersonateServiceAccount", but still received the error. The SSAS service account does have access to the datasource.
Brian Shaw - Tuesday, April 10, 2018 7:53 AM
The annoying thing is that I had this error not so long ago ... and, you know how it is, I can't remember how I solved it.
I'll post back if my memory manages to dredge up anything useful.
If the answer to your question can be found with a brief Google search, please perform the search yourself, rather than expecting one of the SSC members to do it for you.
See https://www.sqlservercentral.com/articles/forum-etiquette-how-to-post-datacode-on-a-forum-to-get-the-best-help/ for details of how to post T-SQL code-related questions.
SSC Guru
Points: 110551
Brian Shaw - Tuesday, April 10, 2018 7:27 AM
This seems likely to be either a permissions issue, or a file path issue that wouldn't be a problem within VS, but is when run as a SQL Agent Job. The two execution contexts are likely rather different. I'd start looking at permissions to the file locations of the assembly being run, and see if the Agent Service account has execute permissions or even read permissions to that folder. It may not. Your error effectively says "file not found", which usually means you "just can't see it". That can be anything from a path not being specified correctly, to a folder permissions issue. I'd see if I could get that path into a package variable and somehow log it before that part of the package executes. That might be rather revealing...
Steve?(aka sgmunson)?:) 🙂 :)?
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Thanks Steve.
The SQL agent is running with a service account in the administrators group so its not a permission thing, but that got me thinking.
I tried running the package as the service account with DTExec.exe (32 and 64 bit) and received the same error with both, so its not related to SQL agent.
I fired up Process Monitor and discovered it was looking for Microsoft.AnalysisServices.AdomdClientUI.dll in a directory where it didn't exist. I found this DLL, copied to where it wanted it and it works now. It was looking for it in several places, I copied the file from D:\Program Files (x86)\Microsoft SQL Server\140\DTS\Binn to D:\Program Files\Microsoft SQL Server\140\DTS\Binn. I know one directory is for 32 bit DLLs and the other is for 64 bit DLLs, but I tried it and it worked.
As far as I am concerned this is a bug in the installation process or DTExec.exe.
Brian Shaw - Tuesday, April 10, 2018 12:29 PM
Well done on tracking that down.
If the answer to your question can be found with a brief Google search, please perform the search yourself, rather than expecting one of the SSC members to do it for you.
See https://www.sqlservercentral.com/articles/forum-etiquette-how-to-post-datacode-on-a-forum-to-get-the-best-help/ for details of how to post T-SQL code-related questions.
SSC Guru
Points: 110551
Brian Shaw - Tuesday, April 10, 2018 12:29 PM
Excellent that you were able to track it down, but I have to wonder if the PATH environment variable might be where the problem started. The order in which the path elements occur can sometimes be critical to finding files, and while copying a file to where it's "expected" can solve the problem, it may be that the PATH just isn't set right, or failed to be updated to the new file location by the most recent product upgrade installation or other similar install. I'd check your current PATH environment variable to see if that file would be found in any of the folders listed. Alternatively, there may just be a registry entry that just points to the wrong folder, very possibly for the same reasons I suggested were possible causes of PATH problems. Admittedly, the registry is a dangerous place to go, and one would want to verify the correctness (or incorrectness) of what you have before making a change, as well as have a backup of same before making a change.
Steve?(aka sgmunson)?:) 🙂 :)?
Health & Nutrition
Make Guaranteed Income
SSC Enthusiast
Points: 147
Hi guys,
I got the same issue here after installing the CU5.
found the DLL in the Visual studio folder of my dev computer.
once copied in the BINN folder of SQL Server everything starts working.
is microsoft aware about this problem?
its certainly a dev guy which references this DLL by mistake.
and pass all the Microsoft unit tests.... which mean... they dont test all the SSIS tasks before releasing a CU.
this is a huge problem.
Valued Member
Points: 53
SQL Server Version: 2017 CU 5
I had the same problem as described here. I tried to modify the PATH variable to remove all old instances of SQL Server. However, no change in behavior. I then attempted to find the DLL for the ADOMD client, but none existed on the server. After installing the 64-bit ADOMD client from https://docs.microsoft.com/en-us/azure/analysis-services/analysis-services-data-providers, I was able to copy Microsoft.AnalysisServices.AdomdClientUI.dll from C:\Program Files (x86)\Microsoft SQL Server\140\Tools\Binn\ManagementStudio to both C:\Program Files (x86)\Microsoft SQL Server\140\DTS\Binn and C:\Program Files\Microsoft SQL Server\140\DTS\Binn. This resolved the issue for me.
Also, another symptom I was experiencing is when deploying my Tabular model using the deployment wizard, it would fail and lock my domain account even though it only appears to attempt the deployment one time. Super frustrating!! I'm not sure if this was related or if I was doing something else wrong, but thought I would mention it.
Valued Member
Points: 57
I had the same problem after installing SQL SERVER 2016 SP2.
I can't find the 130 version of the Microsoft.AnalysisServices.AdomdClientUI.dll file.
I tested with the 140 version but I have an error that the assembly version la ont compatible.
Any idea ?
Many thanks
Hall of Fame
Points: 3661
micheal.staples - Wednesday, April 18, 2018 9:34 AM
Thanks Guys. I had the same issue. It saved my day. I have copied dll in "C:\Program Files\Microsoft SQL Server\140\DTS\Binn" and it worked.
Old Hand
Points: 302
We recently upgraded from SQL Server 2016 CU8 to SQL Server 2016 SP2. After upgrading, SSIS packages which perform cube processing functions throw the following error:
Error Code: 0 Description: Could not load file or assembly 'Microsoft.AnalysisServices.AdomdClientUI, Version=126.96.36.199, Culture=neutral, PublicKeyToken=89845dcd8080cc91' or one of its dependencies. The system cannot find the file specified.
This has happened on two servers we upgraded in our lower environments. Anyone heard anything from MS? There was a posting on the community PowerBI board about this issue last week, but it seems to have been pulled.
SSC Enthusiast
Points: 147
SQL 2017 suffered the same issue.
you'll found the missing DLL from a PC where VS is installed. this missing DLL is not available on the server itself.
Copy it on the BINN folder of your server.
SSC Enthusiast
Points: 104
After installation of SQL Server 2016 SP2, was also getting this error (for Version 188.8.131.52), I was getting this error. I tried copying from a bunch of different ones, and also had to add to the GAC, but can only get it to where it now returns "Error: Method not found: 'Boolean Microsoft.AnalysisServices.AdomdClientUI.AdomdConnectionStringBuilder.IsAzure()'
Viewing 15 posts - 1 through 15 (of 28 total)