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Im Hinblick auf die Anwendung komplexer Therapien bei an Multipler Sklerose (MS) erkrankten Patienten entwickelten wir aus dem in Deutschland am weitesten verbreiteten MS-Dokumentationssystem (MSDS) ein innovatives Patientenmanagement-System (MSDS 3D). Es integriert multidimensional und interaktiv Daten von Arzt, MS-Schwester und Patienten und vergleicht diese mit einem festgelegten internen Qualitatsstandard. Das System kann aber nicht nur genutzt werden, um Daten des Patienten zu erfassen und zu interpretieren, sondern als interaktives System auch Informationen an den Patienten vermitteln. Im Rahmen eines speziellen Moduls fur die Anwendung von Immunmodulatoren innerhalb von MSDS 3D werden in einem definierten klinischen Pfad die notwendigen Vor- und Begleituntersuchungen ebenso abgebildet wie Patientenbefragungen.

Das sich stetig vergrosernde Spektrum von Behandlungsoptionen der Multiplen Sklerose (MS) erfordert ein spezifisches Therapie- und Risikomanagement, um das individuelle Ansprechen, aber auch potenzielle Nebenwirkungen zu erfassen. Der vorliegende Artikel stellt das neuartige computergestutzte Patientenmanagementsystem Multiple-Sklerose-Dokumentationssystem 3D (MSDS 3D) vor. MSDS 3D integriert Daten von Patienten, betreuenden Arzten und MS-Schwestern (3 Dimensionen). Es erlaubt die Dokumentation von Vorstellungsterminen (Visiten) sowie Untersuchungsbefunden uber interaktive Bildschirmoberflachen (Touchscreens). Ein spezifisches, in die aktuelle Version von MSDS 3D integriertes Modul dient der monatlichen Evaluation von Patienten unter der Therapie mit Natalizumab. Die Entwicklung einer Checkliste fur Zeichen einer progressiven multifokalen Leukenzephalopathie (PML) sowie eines Natalizumab-Fragebogens konnten dazu beitragen, diese in Fruhstadien gut therapierbare Erkrankung zeitnah zu erkennen.

Vitamin B(12) (VitB(12), cobalamin) deficiency has been associated with various neuropsychiatric conditions, such as peripheral neuropathy, subacute combined degeneration, affective disorders, and cognitive impairment. Current assays analyze vitamin B(12), of which only a small percentage is metabolically active. Measurement of its active fraction, holotranscobalamin, might be of greater relevance, but data in populations with neuropsychiatric populations are lacking. In this study, in order to validate VitB(12) and holotranscobalamin (holoTC) serum levels for the detection of VitB(12) deficiency in neuropsychiatric conditions, we compared the validity of VitB(12) and holoTC in a patient cohort with neuropsychiatric conditions suspicious for VitB(12) deficiency. The cohort included all patients admitted to the Department of Neurology at our university between 2005 and 2009 with at least two parameters of the VitB(12) metabolism available (n = 1,279). We used elevated methylmalonic acid as the external validation criterion for VitB(12) deficiency and restricted our analyses to subjects with normal renal function. Among all normal renal function patients, 13.2% had VitB(12) deficiency. In receiver operating characteristic curve (ROC) analysis, correlation of VitB(12) and holoTC with vitamin B(12) deficiency was generally weak, and the areas under the curve (AUC) were not significantly different for holoTC compared to vitamin B(12) in all subjects (AUC: 0.66 [95%CI: 0.51-0.82]; p = 0.04 vs. 0.72 [0.65-0.78], p0.0001) and in subcohorts of patients with classical VitB(12) deficiency syndromes. The positive predictive values for holoTC and vitamin B(12) were low (14.7 vs. 21.0%) and both were associated with more false-positive than true-positive test results. holoTC does not show superior diagnostic accuracy compared to VitB(12) for the detection of VitB(12) deficiency in subjects with neuropsychiatric conditions. Neither test can be recommended to diagnose VitB(12) deficiency in subjects with neuropsychiatric disorders.

In the summer of 2001, a nationwide epidemiological multiple sclerosis (MS) register was initiated under the auspices of the German MS Society (DMSG). This project aimed at collecting epidemiological data on the number of patients with MS, course of the disease, and their social situation in Germany. During the 2-year pilot phase, five MS centers with various regional differences and treatment methods participated, leading to a representative selection of patients. In December 2003, standardised data sets of 3,458 MS patients were available for evaluation. After examining the quality of the data, 3,223 sets remained for further analysis. The demographics were similar to those obtained from other epidemiological studies: 72% of the patients were female, mean age was 42.9+/-11.2 years, mean disease duration 12.6+/-8.7 years, and 64% suffered from the relapsing-remitting form of the disease. The median EDSS was 3.0, and 69% of patients had an EDSS

Das Multiple-Sklerose-Dokumentationssystem (MSDS) wurde wahrend der vergangenen 4 Jahre an der Neurologischen Universitatsklinik Dresden entwickelt. Die erste Version der Datenbankapplikation befindet sich seit 10/2000 im praktischen Einsatz. MSDS verwaltet Patientenstammdaten, Arztestammdaten, anamnestische Angaben, korperliche Untersuchungsbefunde, Ergebnisse relevanter Zusatzdiagnostik, MS-relevante Scores sowie Bioproben. Grundsatzlich erlaubt MSDS die “Online”-Dateneingabe und generiert halbautomatisch Arztberichte an alle betreuenden Haus- bzw. Facharzte. Patientenaufklarungsblatter wie auch Informationen fur Arzte sind Teil des Systems. Im Rahmen einer 3-monatigen Evaluierung wurde die erste Version von MSDS an 9 Kliniken (8 Universitatskliniken, ein Landeskrankenhaus) getestet und positiv bewertet. Anderungswunsche und Verbesserungsvorschlage nebst den eigenen praktischen Erfahrungen fanden bei der Entwicklung von MSDS 2.0 Berucksichtigung. Diese Version wird bis Ende 2001 verfugbar sein.

Through extensive serologic, chemical, and molecular studies involving monoclonal Ig proteins and B cell-related populations, we provide definitive evidence that the V lambda IV subgroup of human light (L) chains is separate and distinct from V lambda III and all other known V lambda gene families. lambda IV and lambda III L chains were differentiated immunochemically using well-characterized polyclonal and monoclonal anti-V lambda subgroup-specific Abs. Prototypic L chains, originally classified as lambda IV on the basis of distinctive framework region 1 residues, were distinguished serologically from lambda IIIa, lambda IIIb, and lambda IIIc proteins and also from lambda I, lambda II, lambda VI, and lambda VIII L chains. Furthermore, by using anti-lambda IV reagents, we identified eight additional monoclonal V lambda IV-related populations, including three IgM rheumatoid factor-producing cell lines. The percentage of homology among the lambda IV proteins ranged from 83 to 100, vs 53 to 72 when compared with lambda III components. Moreover, lambda IV proteins shared particular subgroup-associated FR and complementarity-determining region residues. At the molecular level, the nucleotide sequences encoding two of the IgM lambda IV rheumatoid factors were identical to that found for the genomic counterpart, as well as to the previously reported IGLV3S1 and Humlv418 germ-line genes. Two other lambda IV cDNAs contained additional non-germ-line-encoded nucleotides at the V-J joint. The single or pauci-gene nature of the V lambda IV family was evidenced from Southern blotting and from the extensive sequence homology among lambda IV components. Our studies have provided further evidence for the prevalence of lambda IV L chains among Ig lambda autoantibodies, thus implying a functional significance for the V lambda IV subgroup.

The increasing therapeutic options for relapsing-remitting multiple sclerosis require a specific treatment and risk management to recognize the individual response as well as potential side effects. To switch from pure MS documentation to MS management by implementing a new multiple sclerosis management and documentation tool may be of importance.This article presents the novel computer-based patient management system "multiple sclerosis management system 3D" (MSDS 3D).MSDS 3D allows documentation and visualization of visit schedules and mandatory examinations via defined study modules by integration of data input from patients, attending physicians and MS nurses. It provides forms for the documentation of patient visits as well as clinical and diagnostic findings. Information is collected via interactive touch screens. A specific module which is part of MSDS 3D's current version allows the monthly monitoring of patients under treatment with natalizumab. A checklist covering clinical signs of progressive multifocal leukoencephalopathy (PML) and a detailed questionnaire about the handling of natalizumab in practice have additionally been added.The MS patient management system MSDS 3D has successfully been implemented and is currently being evaluated in a multi-centre setting. Advanced assessment of patient data may allow improvements in clinical practice and research work. The addition of a checklist and a questionnaire into the natalizumab module may support the recognition of PML during its early, treatable course.

Human lambda L chains of a major V lambda subgroup, V lambda III, have been differentiated serologically and chemically into three V lambda III sub-subgroups designated V lambda IIIa, V lambda IIIb, and V lambda IIIc. Antisera prepared against lambda III Bence Jones proteins were obtained that recognized distinctive V lambda III-related epitopes expressed by monoclonal lambda III L chains. After appropriate absorption, these reagents were rendered specific for three distinct populations of lambda III proteins--lambda IIIa, lambda IIIb, and lambda IIIc. The antisera were used in comparative immunodiffusion analyses of 28 monoclonal lambda III L chains, 10 of which were classified as lambda IIIa, 4 as lambda IIIb, and 14 as lambda IIIc. The isotypic nature of the three lambda III sub-subgroups was demonstrated serologically through analyses of lambda-chains derived from the serum IgG molecules of normal individuals. The amino acid sequences of five serologically classified lambda III chains, which included members of the three V lambda III sub-subgroups, had been previously determined. This information, in addition to our establishment of the complete (or virtually complete) V region sequence of 15 and the partial sequence of eight other lambda IIIa, lambda IIIb, and lambda IIIc proteins, made it possible to correlate chemical data with serologic classification. Proteins within each of the three serologically-classified lambda III sub-subgroups typically possessed a high degree (approximately 83%) of intra-sub-subgroup sequence homology that included both framework and complementarity determining region residues. Furthermore, within the framework and complementarity determining regions, sub-subgroup-specific residues were identified. Taken together, these data reveal that the human V lambda III genome consists of (at least) three distinct V lambda IIIa, V lambda IIIb, and V lambda IIIc germline genes that encode for lambda IIIa, lambda IIIb, and lambda IIIc L chains, respectively.

In the summer of 2001, a nationwide epidemiological multiple sclerosis (MS) register was initiated under the auspices of the German MS Society (DMSG). This project aimed at collecting epidemiological data on the number of patients with MS, course of the disease, and their social situation in Germany. During the 2-year pilot phase, five MS centers with various regional differences and treatment methods participated, leading to a representative selection of patients. In December 2003, standardised data sets of 3,458 MS patients were available for evaluation. After examining the quality of the data, 3,223 sets remained for further analysis. The demographics were similar to those obtained from other epidemiological studies: 72% of the patients were female, mean age was 42.9+/-11.2 years, mean disease duration 12.6+/-8.7 years, and 64% suffered from the relapsing-remitting form of the disease. The median EDSS was 3.0, and 69% of patients had an EDSS/=4.0. The great effect of this disorder was underscored by the fact that one third of the patients had prematurely retired due to MS. After successful completion of the pilot phase, the MS register will provide reliable data and thus serve as an important tool to improve the overall situation of MS patients in Germany.

The MSDS (multiple sclerosis documentation system) has been developed at the Department of Neurology, Technical University of Dresden, Germany, during the last 4 years. The first version of this database application has been in use since October 2000. The MSDS manages information on MS patients, their treating physicians, patient history (symptoms, other diseases, biographical history, family history, habits, medication), clinical signs, results of laboratory examinations (blood chemistry, autoantibodies, borrelia serology, evoked potentials, cranial and spinal cord magnetic resonance imaging), clinical scores relevant for MS, and biosamples. In principle, MSDS allows online data input and semiautomatically generates reports to all general practitioners and neurologists treating the respective patient. Patient information sheets and internal treatment guidelines are part of the system. During a 3-month evaluation, the first version of MSDS was tested at eight university multiple sclerosis ambulatory care units and one general neurology hospital. The overall judgement was favorable. Suggestions for changes and improvements, as well as practical experiences, were considered when developing MSDS 2.0, which will be available by the end of 2001.

Somatic deletion mutations in the cell adhesion molecule E-cadherin are present in almost 50% of diffuse type gastric cancer. We recently generated monoclonal antibodies against an in-frame deletion of exon 9. The aim of this study was to generate and characterize monoclonal antibodies against the second mutational hot spot, in-frame deletions of exon 8. Lou/C rats were immunized using a KLH-coupled peptide that represents a unique sequence generated by fusion of exon 7 and exon 9 from an E-cadherin deletion mutation lacking exon 8. Hybridoma supernatants were tested in a solid-phase immunoassay using BSA-coupled peptide. Positive reacting hybridomas were confirmed by Western Blots, FACS analysis, and immunohistochemistry of E-cadherin negative carcinoma cells that had been transfected with mutant and wild-type E-cadherin cDNA, respectively. In addition, routine formalin fixed and paraffin embedded tissues from gastric cancer patients were analyzed using both mutation-specific and commercial monoclonal antibodies against E-cadherin, including HECD-1 and AEC. Two hybridoma supernatants, termed E-cad delta 8-1, were selected that reacted with the mutant peptide used for immunization and gave strong signals in Western Blot and FACS analysis with cells expressing mutant E-cadherin lacking exon 8. Wild-type protein expressing cells only reacted with the commercial antibodies but not with the two selected hybridoma supernatants. In contrast to AEC, monoclonal antibody HECD-1 did not react with exon 8 deleted E-cadherin, suggesting that the previously unknown epitope for this often used monoclonal antibody is located at least in part within exon 8. Four gastric cancer specimens known to express mutated E-cadherin mRNA strongly reacted with both mutation-specific supernatants and with AEC monoclonal antibody but not with HECD-1. Taken together, we succeeded in generating monoclonal antibodies reacting with mutant E-cadherin protein lacking exon 8. Furthermore, using both HECD-1 and the new mutation-specific antibodies E-cadherin immunoreactivity can for the first time be evaluated in an allele-specific manner in archival tissues.

The NF-kappaB transcription factor is activated by a wide variety of stimuli, including phorbol esters such as 12-O-tetradecanoylphorbol-13-acetate. In its inactive state, NF-kappaB is sequestered in the cytoplasm tethered to an inhibitor protein, IkappaB. Activation comprises the rapid phosphorylation of IkappaB-alpha at N-terminal sites, which presumably marks IkappaB-alpha for proteolytic degradation and leads to release of NF-kappaB into the nucleus. In addition, IkappaB-alpha is constitutively phosphorylated at the C terminus, which may be a prerequisite for proper IkappaB function. Protein kinase C (PKC) is activated by 12-O-tetradecanoylphorbol-13-acetate and has been previously reported to phosphorylate IkappaB-alpha in vitro. As PKC has turned out to constitute a multigene family encoding isozymes with different biological functions, we have reinvestigated IkappaB-alpha phosphorylation by PKC using recombinant PKC isozymes expressed in insect cells. While crude PKC preparations were efficient IkappaB-alpha kinases, highly purified PKC isozymes completely failed to phosphorylate IkappaB-alpha. Biochemical separation of porcine spleen yielded at least two fractions with IkappaB-alpha kinase activity, both of which were devoid of detectable PKC isozymes. One peak contained both Raf-1 and casein kinase II (CKII). Purified Raf-1 does not phosphorylate IkappaB-alpha directly, but associates with CKII, which efficiently phosphorylates the C terminus of IkappaB-alpha. Two-dimensional phosphopeptide mapping and high pressure liquid chromatography-mass spectroscopy analysis showed that all IkappaB-alpha kinases induced phosphorylation at the same prominent sites in the C terminus. Our results clearly indicate that PKC isozymes alpha, beta, gamma, delta, epsilon, eta, and zeta as well as Raf-1 are not IkappaB-alpha kinases. They furthermore demonstrate that IkappaB-alpha is targeted by several kinases, one of which appears to be CKII.

The putative precursor molecule of a human AL type amyloid fibril protein was isolated from an ultrafiltrate after hemofiltration. Subsequent separation of this protein was achieved by high performance liquid chromatography (HPLC) after reduction and carboxymethylation of the disulfide bonds. The protein was separated into several fractions which were further analyzed by automatic amino acid sequence determination. It was deduced from the sequence data that the precursor molecule is an immunoglobulin L-chain of the lambda-type. The V-region of this protein is most closely related to the proteins of subgroup II. Internal splits occurred in the molecule after lysine residues in positions 110, 129 and 179. The predominant fragment commences with either serine or alanine in position 9 and extends to a serine in position 65 of the V-region. Tryptic peptides generated from the fragments cover nearly the entire V- and C-region of the L-chain, with the exception of positions 1-8, from which no peptide has been isolated.

The Fc-receptor of normal human leukocytes, of CLL-cells, and of hematopoietic cell lines was demonstrated with soluble peroxidase-anti-peroxidase (PAP) complexes. In about 9% of normal lymphocytes an almost continuous, strong labeling of the cell membrane was established. Some of these lymphocytes were characterized by a peculiar uniform fine structure. The percentage of PAP-labeled monocytes was in the range of 25%, neutrophils nearly 100%, eosinophils 0%, CLL-cells 10%. Labeled portions of the membrane were interiorized from monocytes. The lymphoid cell-line Daudi established from a Burkitt's lymphoma appeared almost negative, the cell line K 562 established from a myeloid leukemia in 75% of the cells strongly positive. PAP-labeling was not influenced by preincubation with trypsine or with neuraminidase; it was negative when PAP-F(ab)2 was used. Results of PAP-labeling were not always in agreement with EA-rosettes or with agg-Ig.

Anti-T cell globulin (ATCG) prepared from antihuman thymocyte serum by absorption with kidney, cells from patients with chronic lymphatic leukemias, and several lymphoblastoid cell lines was shown to react specifically with human thymus-derived lymphocytes. While high activity against thymocytes and a T-lymphoblastoid cell line could be demonstrated, ATCG remained negative against several chronic lymphatic leukemias and B-lymphoblastoid cell lines. The ATCG was used in the cytotoxic test, electronmicroscopy, and immunoautoradiography for identification of T cells in thymus, tonsils, spleen, blood, bone marrow, lymphatic leukemias, and lymphoblastoid cell lines. A comparison of these results with the ability to form spontaneous SRBC-rosettes revealed remarkable deviations between both markers in leukemias. Absorption with human brain failed to remove specific activity of ATCG. Labeling experiments by immunoautoradiography and investigations by complement fixation permitted quantitation of relative T-cell antigen concentration on different cell populations. As further evidence for specificity it could be shown that ATCG was no longer toxic for hemopoietic progenitors, whereas unabsorbed globulin reduced the number of colonyforming cells considerably.

The unlabeled antibody enzyme method was used to delineate the anatomical distribution of lymphocytes positive with a-MBΘ and a-MIg in tissue sections of thymus, spleen, lymph nodes and Peyer's patches of mice. The known T- and B-cell areas were established. Moreover, individual B-cells were detected in T-cell regions, especially in the thymus medulla, and in the periarteriolar section of spleen white pulp. Similarly, individual T-cells occur in B-cell areas, namely in the marginal zone of spleen white pulp, in the medullar cords of lymph nodes, and in germinal centers.

The antigen specificity of hybridoma antibodies with activity against H-2 antigens was analyzed by testing lymphocytes of inbred lines and their recombinants, as well as B10.W lines and wild mice in relation to the reactivity of alloantisera. Within the limited test panel of 11 inbred strains and their recombinant derivatives, the hybridoma antibodies reacted concordantly to alloantisera with public (H-2.5, H-2.11, and Ia. 13) and private (H-2.23) antigens. However, when tested on cells of B10.W lines or wild mice discordant reactions were observed, indicating that the antigenic determinants recognized by the hybridoma antibodies are included in the antigens defined by alloantisera within the inbred test panel. In wild mouse populations, these determinants do also occur independently of those which comprise the alloantisera-defined specificities. The results point to an even higher complexity of MHC antigens than envisaged so far, and it appears that a certain allele might be defined only by a certain set of antigenic determinants. Because of the discrepant reactions of alloantisera and hybridoma antibodies, it has been proposed that antigenic determinants detected by hybridoma antibodies receive a numerical designation according to their description but independent of antigens defined by alloantisera, even if they match the reactivity pattern of an alloantiserum-defined antigen specificity within the inbred panel.

A simple method has been established for tritium labeling immunoglobulins. This label avoids certain disadvantages of 125 iodine such as high radiation hazard and requirement for frequent labeling owing to the short half-life of this isotope. High specific activities (range 1.8–9.0 cpm/μg protein) were obtained with no loss of functional activity as demonstrated by passive hemagglutination. The labeled conjugate is especially suitable for detection of activity in hybridoma supernatants by indirect binding assay. Adapted to microtiter plates, the method is suitable for large numbers of samples, e.g. up to 800 supernatants may be examined daily. The utility and advantages of the binding assay with respect to reproductibility, sensitivity, binding kinetics, easy and rapid performance are described.

Iodine 125, an emitter of beta-radiation with an energy lying between that of tritium and carbon-14, is investigated for its applicability in quantitative autoradiography. Absorption and geometric factors of radiation are elucidated. From this, appropriate measuring conditions are derived. The simultaneous exposure of radioactive standard sources permits the evaluation of absolute amounts of radioactivity. Standard cells with labelled membranes are a suitable source of reference taking into account the physical properties of the isotope. Sheep red blood cells are examined for their suitability as standard cells after enzymatic radioiodination. The absolute number of antigenic substances on the surface of single cells is obtained by determining the specific activity of the labelled antibody molecules, and by measuring the silver grain densities of the cells under investigation and of the standard cells. The radioactivity per standard cell can be assessed by conventional procedures. The new method is applied to the quantification of membrane-bound immunoglobulin molecules of the IgG-type on single human lymphocytes. The determination of an immunologic saturation of the labelled antibody is essential for this purpose. On the lymphocytes of a normal person and of a patient with chronic lymphatic leukaemia quite different amounts of immunoglobulins have been evaluated.

Antisera against murine thymus-derived lymphocytes were raised in rabbits injected with mouse brain. It was possible to remove contaminating antibodies which cross-reacted with B cells and hemopoietic stem cells by absorption with liver and plasmacytoma followed by purification and fractionation over DEAE-cellulose. T cell specificity of the absorbed anti-mouse brain serum (AMBS) and its purified globulin fraction (AMBG) was demonstrated by cytotoxicity, complement fixation and unaltered numbers of plaque-forming cells and colony-forming units. In vivo, the effect of absorbed AMBG on the graft-versus-host reaction was investigated in an H-2-incompatible parent-to-F1 hybrid donor-recipient combination. (C57BL/6 × CBA)F1 mice irradiated with 900 rad and transfused with C57BL/6 spleen cells died from acute secondary disease within 20 days. They survived as complete chimaeras beyond day 300 without symptoms of secondary disease when the donor cells had been preincubated with absorbed AMBG. The implications of the suppression of secondary disease in this model are discussed.

Klinik und Verlauf eines IgG-Plasmocytoms mit ossarer und tumorartiger extraossarer Ausbreitung bei einer 64jahrigen Patientin werden beschrieben. Pleurabefall mit Exsudatbildung des Plasmocytoms bestimmten Verlauf und therapeutische Bemuhungen wesentlich. Immunelektrophoretische Befunde sowie lichtmikroskopische Besonderheiten dieses Plasmocytoms in Sternalpunktat, Pleuraexsudat, Myelotomiepraparat und autoptisch gewonnenen Geweben werden dargestellt. Von besonderem Interesse sind die feinstrukturellen Befunde an den Plasmocytomzellen aus Sternalmark, peripherem Blut und postmortal gewonnenem Gewebe, vor allem die reichliche Ausbildung fibrillarer Strukturen in Nachbarschaft der gelappten und segmentierten Kerne. Die Problematik der genauen Einordnung derartiger Krankheitsbilder und der Rolle des Elektronenmikroskops hierbei werden diskutiert.

Im heterologen Antimause- bzw. Antirattenlymphocytenserum vom Kaninchen liesen sich die folgenden Antikorpertypen unterscheiden: 1. Anti-Erythrocyten/Leukocyten/Thrombocyten/Leber/Nieren-Antikorper; 2. Anti-Leukocyten /Thrombocyten/Leber/Nieren-Antikorper; 3. Antileukocytenantikorper; 4. Antierythrocytenantikorper; 5. Antiserumproteinantikorper.