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  1. December 18, 2018: DNA-Based Facial Reconstruction – A Multidisciplinary Discussion
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Verstraete et al. The instantaneous, pure substitutive treatments allow these deficient or nonexistent coagulation factor by intravenous infusion to bring into the bloodstream in order to re-establish a normal hemostasis. If this complication occurs, the Substitionsbehandlung becomes ineffective by supplying the loss-FVIII, and one has to resort to other methods to treat the thus immunized hemophiliacs.

One of the possible means immunized hemophiliacs and subjects who develop autoantibodies anti-FVIII antibodies particularly associated with autoimmune diseases , to help is to purify the blood plasma of these antibodies.

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This plasma cleaning of anti-FVIII antibodies is generally done by running to be purified plasma through an extracorporeal circulation through a column containing an immunoadsorbent. So wurde beispielsweise bereits vorgeschlagen, insbesondere von Regnault V. Thus, for example, been proposed, particularly Regnault V. Protein A reacts. Hjelm H. Its properties make this protein at a good ligand for the purification of total IgG, and therefore it is used in the purification of various antibodies for a long time.

Moreover, this carrier has insufficient stability, which can lead to a gradual desorption of protein A in its use. This is then carried along in the blood stream of the patient, wherein the plasma cleaning is carried out, resulting in a harmful activation of the complement system. This method is extremely costly in terms of the manufacturing price of this column type and does not protect against a possible viral contamination by protein A. Recently, in particular in patent application FR-A-2,,, proposed a method for the separation of anti-FVIII antibodies by adsorption columns, consisting of carriers based on polymers cellulose polyesters, Cellulosepolyethern, polystyrene PS , polyisoprene or polybutadiene , which are functionalised with sulphonate and Chlorsulfonylfunktionen that carry amino acid residues such as glutamic acid, hydroxyproline, threonine, alanine or phenylalanine.

In dem Artikel von C. Boisson-Vidal et al. In the article by C. However, such carriers are considering their insufficient adsorption specificity to anti-FVIII antibodies and their poor cleaning degree of anti-FVIII antibody does not always satisfactory. Der Artikel von L. Dahri et al. The article by L. Moreover, the presence of impurities chlorine was detected. To eliminate these problems, the inventors have surprisingly developed the subject of the invention. These resins are able to specifically adsorb anti-FVIII antibodies, allowing their use as immunoadsorbent for the production of columns for extracorporeal purification in individuals who have developed these antibodies.

According to a preferred embodiment of the invention, the polystyrene is cross-linked by divinylbenzene. The use of polystyrene with a degree of crosslinking than 5 leads to difficult to be functionalized resins. The resins of the formula I are generally in the form of beads, which have in the dry state has a diameter of preferably about 35 to The resins of the formula I can be used for the preparation of chromatographic columns, particularly for the production of columns for plasma and blood purification.

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Accordingly, the invention also provides purification columns that contain at least one resin of the above defined formula I , which is equilibrated in buffer having a physiological pH of generally between about 7. These columns may be part of a device for the purification of anti-FVIII antibodies, consisting of a closed loop with continuous circulation, in which one or more defined above purification columns may be incorporated, these columns are arranged in series or parallel to thereby increasing the degree of purification of anti-FVIII antibodies and, if appropriate, to reduce the time for cleaning the same time.

This cleaning cycles are preferably provided with an injection valve and a pump which provides for circulation.

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When this cleaning device is used for the treatment of blood drawn, it may also be equipped with a cell separator, which ensures the separation of the plasma from the blood cell components making up before it is run by the inventive column. The present invention therefore comprise one or more resins of the above defined formula I for the preparation of columns for the purification of liquids, the anti-FVIII antibody to the use.

The liquids to be treated are preferably plasma or whole blood, and most preferably plasma or whole blood of patients suffering from hemophilia A. In fact, the inventors have shown that the purification of plasma from patients with haemophilia A using plasma cleaning devices comprising at least one column containing at least one inventive resin of the formula I , allows the largest part of the anti-FVIII to remove antibodies in this plasma.

After this cleaning, the content of residual total plasma IgG and of coagulation factors also in very narrow limits, which shows also the very high specificity of the resins of formula I for the adsorption of anti-FVIII antibodies. Another advantage of this plasma cleaning apparatus is that the throughput of the plasma does not perform the purification column according to the invention to a significant Dekomplementierung the plasma by adsorption of proteins of the complement system.

According to the inventive use, and when the cleaning is finished, can be defined as above purification columns, comprising at least one resin of formula I can be regenerated by desorption of the adsorbed on the resin plasma fraction by a salt solution, generally a 3 M sodium chloride solution, running through the columns. The resin is then equilibrated with a buffer solution at physiological pH between about 7. According to a particular embodiment of the invention, and when the cleaning circuit is provided with at least two parallel columns according to the invention, is regenerated one of them while the other is used for adsorption of the anti-FVIII antibodies.

The regeneration of these purification columns does not lead to degradation of the resin of formula I , as a result, and the advantage that does not change its specificity for anti-FVIII antibodies. The invention further provides a process for the production of resins from functionalized polystyrene of the formula I. The preparation of the resins of formula I is performed in two main steps: Chlorosulfonation, which permits the insertion of Chlorsulfonylfunktionen to the aromatic rings of the polystyrene, and then binding of the L-Tyrosinmethylesters on the chlorosulfonyl groups.

This method of synthesis can be represented in simplified form by the following general scheme A, in which the index d has the same meaning as that indicated above for the compounds of formula I. According to a preferred embodiment of this manufacturing process, the molar ratio R is equal to 0. The beads used in the first step of cross-linked, non-functionalized polystyrene are preferably cross-linked with Devinylbenzol polystyrene beads.

They are preferably previously swollen in a solvent liquid for the polystyrene left as dichloromethane, nitromethane, perchlorethylene, dimethylformamide, methylene chloride or tetrahydrofuran. The source allowing the polystyrene beads in the solvent liquid makes it possible to increase the accessible surface so that the reagent can fully penetrate the beads, so as to obtain a good yield. The chlorosulfonation reaction of the first step is preferably carried out according to the previous sources of PS in dichloromethane in two sub-steps: dichlorosulfonic acid reacts at the non-functionalized polystyrene and leads to the formation of an intermediate polymer, poly sulfone styrene, and then the formation of poly parachlorsulfonyl styrene ,.

At the end of the first step, the PSSO2Cl-resin is preferably washed one hand with dichloromethane to remove excess acid, and secondly with acetone to remove all traces of residual acid in the medium and to facilitate drying of the resin. Before carrying out the second step, the PSSO are preferably dried 2 Cl beads under vacuum and then in a solvating medium such as those used in the first step and are as defined above, is allowed to swell, so that the reagent for the employed in the second step accessible surface is increased.

In this second step, the binding of the L-Tyrosinmethylesters is carried out by condensation of the hydrochloride form of the latter to the PSSO2Cl in an anhydrous solvent in the presence of a tertiary amine such as triethylamine NEt 3. The tertiary amine converted to the hydrochloride in free amine, which then reacts with the Chlorsulfonpolystyrol. In a second step, the amino acid solution is contacted with the PSSO2Cl resin which was allowed to swell preferably previously, in contact.

The hydrochloric acid formed during the reaction is neutralized by adding a tertiary amine such as NEt 3 to the reaction medium. The determination of the input amount of reactant tertiary amine and hydrochloride of L-Tyrosinmethylesters depends on the amount of SO 2 from Cl-functions, which has been bound in the first step, which is argentometrically and determined by elemental analysis.

Hierzu wird das Harz vorzugsweise mit einem Alkalisierungsmittel wie Natriumhydroxid gewaschen, das in der Lage ist, die restlichen para-Chlorsulfonylfunktionen zu hydrolysieren, ohne die Methylesterfunktionen des L-Tyrosins zu verseifen. For this purpose, the resin is preferably washed with an alkalizing agent such as sodium hydroxide , which is able to hydrolyze the remaining para-Chlorsulfonylfunktionen without saponify the Methylesterfunktionen of L-tyrosine.

After about ten washes the residual chlorine content is negligible. Prior to its use, the resin is preferably conditioned by a sequence of washing and centrifugation to remove any trace of impurities which could interact with the plasma proteins. For this purpose carried out a number of washing steps of the resin, various types are used by solutions: sodium chloride, sodium citrate, Michaelis buffer.

Between each of these washes, the resins are preferably washed thoroughly with double distilled water. After each step of the synthesis of a number of physicochemical characterizations resins of the invention of formula I are preferably subjected. In particular, it allows before microscopic examination of the resins, information about the shape and size distribution of the particles and obtain after each synthesis step.

The determination of the degree of substitution of each resin can be performed by elemental analysis of the resin constituting elements, nitrogen, sulfur, chlorine and sodium. The chlorine content of the resins can be determined by argentometric analysis of the released after hydrolysis of a sample of the resin in suspension in an aqueous sodium hydroxide chlorine ions. At the end of synthesis, the fine particles which originate from a possible breakage of the polystyrene beads in the various synthesis steps, are preferably removed.

This removal can be done by a series of consecutive washing and centrifugation of the resins in buffer solution, preferably at a pH between 7.

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These washing steps are preferably carried out until the supernatant is perfectly clear. The units focus primarily on oral skills. You learn the language by speaking and always according to your individual needs. I support you in all areas of learning German - from A1 to B2, language training for physicians for the German Why choose Stevan D.

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This trial is an affront to democracy. On Sunday a leftist German blog wrote to ask if we could explain Brexit. As far as they could see, it was a simply a question of the British trying to gain trade advantages and […]. That the elections were suddenly brought forward by more than a year was a surprise in itself — but the standout surprise […]. Lars P. Article Chris Bambery. EU politics.