[Conditioning of the immune system-Already clinically usable?] / Konditionierung des Immunsystems ­ Schon klinisch nutzbar?

The brain and the immune system permanently exchange information via various neuronal and humoral signaling pathways. This communication network forms the basis for controlling peripheral immune functions via associative learning or conditioning processes. Establishing a learned immune reaction, an immunomodulatory drug that represents the unconditioned stimulus (US) is paired with a new odor or taste stimulus. Re-presentating this previously neutral odor or taste stimulus, its now functions as a conditioned stimulus (CS) and triggers reactions in the immune system similar to those formerly induced by the drug used as US. Using different learning protocols, it was possible to condition immunopharmacological effects in animal disease models, such as lupus erythematosus, contact allergy or rheumatoid arthritis, thereby reducing disease symptoms. Preliminary experimental studies in healthy volunteers and patients confirmed a possible clinical use of learned immune responses with the aim of using associative learning protocols as complementary measures to pharmacological interventions in clinical practice in order to reduce drug doses and thus undesirable drug side effects while maintaining therapeutic efficacy. However, there is still a great need for further research to understand the mechanisms of learned immune responses in preclinical studies and to optimize the associative learning processes for using them in the clinical routine in studies with healthy volunteers and patients.

Insights from a Multicenter Study on Adult H3 K27M-Mutated Glioma: Surgical Resection's Limited Influence on Overall Survival, ATRX as Molecular Prognosticator.

BACKGROUND: H3 K27M-mutated gliomas were first described as a new grade 4 entity in the 2016 WHO classification. Current studies have focused on its typical appearance in children and young adults, increasing the need to better understand the prognostic factors and impact of surgery on adults. Here, we report a multicentric study of this entity in adults. METHODS: We included molecularly confirmed H3 K27M-mutated glioma cases in patients >18 years diagnosed between 2016 and 2022. Clinical, radiological, and surgical features were analyzed. Univariate and multivariate analyses were performed to identify prognostic factors. RESULTS: Among 70 patients with a mean age of 36.1 years, the median overall survival (OS) was 13.6 + 14 months. Gross-total resection was achieved in 14.3% of patients, whereas 30% had a subtotal resection and 54.3% a biopsy.Tumors located in telencephalon/diencephalon/myelencephalon were associated with a poorer OS, while a location in the mesencephalon/metencephalon showed a significantly longer OS (8.7 vs. 25.0 months, p=0.007). Preoperative Karnofsky Performance Score (KPS) < 80 showed a reduced OS (4.2 vs. 18 months, p=0.02). Furthermore, ATRX loss, found in 25.7%, was independently associated with an increased OS (31 vs. 8.3 months, p=0.0029). Notably, patients undergoing resection showed no survival benefit over biopsy (12 vs. 11 months, p=0.4006). CONCLUSION: The present study describes surgical features of H3 K27M-mutated glioma in adulthood in a large multicentric study. Our data reveal that ATRX status, location and KPS significantly impact OS in H3 K27M-mutated glioma. Importantly, our dataset indicates that resection does not offer a survival advantage over biopsy.

[Infectious bone diseases]. / Infektiöse Knochenerkrankungen.

Bacterial infection of the bone is a severe disease with complications, potentially including long-term physical disability. The diagnosis and therapy of osteomyelitis include several elements: histopathology, microbiology, radiologic imagining, as well as antibiotic and surgical therapy. Histopathologists differentiate between acute osteomyelitis (infiltration of cancellous bone with neutrophil granulocytes); specific osteomyelitis (epithelioid-like granulomatous inflammation, tuberculosis, mycotic infections); primary/secondary chronic osteomyelitis (lymphocytic infiltration); and special forms of chronic osteomyelitis (varying histomorphology, Brodie abscess, SAPHO syndrome). Another important task in the histopathological diagnosis of inflammatory bone diseases is to differentiate osteomyelitis from malignant entities (sarcoma, lymphoma). Therefore, biopsy samples should be of sufficient size for safe diagnosis. Clinical information and imaging as well as interdisciplinary teamwork between radiologists, microbiologists, orthopedic surgeons and pathologists is mandatory to verify these diagnoses.

[Joint endoprosthesis pathology. Histopathological diagnostics and classification]. / Gelenkendoprothesenpathologie. Histopathologische Diagnostik und Klassifikation.

Prosthesis durability has steadily increased with high 10-year rates of 88-95%. However, four pathogenetic groups of diseases can decrease prosthesis durability: (1) periprosthetic wear particle disease (aseptic loosening) (2) bacterial infection (septic loosening) (3) periprosthetic ossification, and (4) arthrofibrosis. The histopathological "extended consensus classification of periprosthetic membranes" includes four types of membranes, arthrofibrosis, and osseous diseases of endoprosthetics: The four types of neosynovia are: wear particle-induced type (type I), mean prosthesis durability (MPD) in years 12.0; infectious type (type II), MPD 2.5; combined type (type III) MPD 4.2; and indeterminate type (type IV), MPD 5.5. Arthrofibrosis can be determined in three grades: grade 1 needs clinical information to be differentiated from a type IV membrane, and grades 2 & 3 can be diagnosed histopathologically. Periprosthetic ossification, osteopenia-induced fractures, and aseptic osteonecrosis can be histopathologically diagnosed safely with clinical information. The extended consensus classification of periprosthetic membranes may be a diagnostic groundwork for a future national endoprosthesis register.

Symmetry breaking in the female germline cyst.

In mammals and flies, only one cell in a multicellular female germline cyst becomes an oocyte, but how symmetry is broken to select the oocyte is unknown. Here, we show that the microtubule (MT) minus end-stabilizing protein Patronin/CAMSAP marks the future Drosophila oocyte and is required for oocyte specification. The spectraplakin Shot recruits Patronin to the fusome, a branched structure extending into all cyst cells. Patronin stabilizes more MTs in the cell with the most fusome material. Our data suggest that this weak asymmetry is amplified by Dynein-dependent transport of Patronin-stabilized MTs. This forms a polarized MT network, along which Dynein transports oocyte determinants into the presumptive oocyte. Thus, Patronin amplifies a weak fusome anisotropy to break symmetry and select one cell to become the oocyte.

SMARCB1/INI1 maternal germ line mosaicism in schwannomatosis.

Schwannomatosis is characterized by the development of multiple schwannomas of the nervous system, but without the occurrence of vestibular schwannomas. Most cases of schwannomatosis are thought to be sporadic, representing the first case in a family due to a new mutation in the causative gene. We recently identified SMARCB1/INI1 as a schwannomatosis-predisposing gene. Here, we analyzed this gene in a schwannomatosis family with two affected children, but with clinically unaffected parents. Both affected individuals carried a constitutional SMARCB1 mutation, c.1118+ 1G>A, that changes the donor splice site sequence of intron 8, causing skipping of exon 8 and resulting in the in-frame deletion of 132 nucleotides in the transcript. The mutation was not evident in constitutional DNA of the parents. Haplotyping revealed that the chromosome 22 segment that carries the mutant SMARCB1 allele originated from the mother. She transferred the same chromosome 22 segment, however, with a wild-type SMARCB1 copy, to a third unaffected child. Our findings indicate that the mother is germ line mosaic for the SMARCB1 mutation. In conclusion, our study shows for the first time that germ line mosaicism may occur in schwannomatosis, which has implications for genetic counseling in this disease.

[Is bioptic assurance reasonable in patients with Sjögren's syndrome? From focus score to diagnosing vasculitides]. / Ist eine bioptische Sicherung beim Sjögren-Syndrom sinnvoll? Vom Focus-Score zur Vaskulitisdiagnostik.

Sjögren's syndrome is an autoimmune disease which targets the salivary and lacrimal glands in particular, causing sicca syndrome. Extraglandular manifestations are often seen. Chronic sialadenitis of the parotid gland is the most common symptom to be assessed for differential diagnosis. Common HE and Giemsa slices are histopathologically examined and graduated for lymphocyte infiltration (focus): grade 0: absent, grade 1: slight, grade 2: moderate non-focal infiltration, grade 3: 1 focus (> or =50 lymphocytes) per 4 mm2, grade 4: >1 focus. Grade 3 infiltrates correspond to a focus score of 1, which is one of four disease-classifying criteria acknowledged for diagnosis. Bioptic examination is also performed to rule out different (non-) immunologic sialadenitises, such as the necrotizing or epithelioid-like form (in sarcoidosis), and the extranodal marginal-zone lymphoma. Extraglandular manifestations of Sjögren's syndrome can also be safely diagnosed by histopathological examination. Emphases lie on vasculitides and myositides. Bioptic work-up, therefore, is not only reasonable but also an essential tool for diagnostics in Sjögren's syndrome.

[Meniscal degeneration score and NITEGE expression : immunohistochemical detection of NITEGE in advanced meniscal degeneration]. / Meniskusdegenerationsscore und NITEGE-Expression : Immunhistochemischer NITEGE-Nachweis in der schwergradigen Meniskusdegeneration.

BACKGROUND: Meniscal degeneration (MD) is a structural change of fibrous cartilage that is common in orthopaedic diagnostics and relevant for health insurance matters. So far, there has been neither a standardised scoring system nor an immunohistochemical marker for MD. MATERIAL AND METHOD: In this retrospective trial, the meniscal tissue of 60 patients was assessed immunohistochemically for NITEGE (G1 fragment of the proteoglycan aggrecan) expression. NITEGE expression was correlated with defined grades of MD: little (grade 0/1), medium (grade 2), or severe (grade 3). RESULTS: Detection of extracellular NITEGE deposits in grade 2 or 3 MD had a positive predictive value and specificity of 100%, whereas no deposits were found in grade 0/1 MD. Sensitivity in advanced MD was 55%. Detection of extracellular NITEGE correlated positively with the grade of degeneration, as did patient age and the grade of degeneration. The patient age of those with grade 0/1 MD was significantly lower than for grade 3 (p<0.0001). CONCLUSION: The thoroughly defined degeneration score (grade 1 - grade 3 MD) is suitable to assess the severity of degeneration. Extracellular NITEGE deposits can be regarded as an immunohistochemical marker for advanced (grades 2 and 3) MD.

[Histopathological degeneration score of fibrous cartilage. Low- and high-grade meniscal degeneration]. / Histopathologischer Degenerations-Score des Faserknorpels. Low-grade- und High-grade-Meniskusdegeneration.

Although histopathology of meniscal degeneration plays an important role, no criteria to assess severity of the degeneration are available to date. Our aim was to create a histopathological scoring system for meniscal degeneration with good interobserver variability, taking matrix degradation and cellularity in meniscal tissue into consideration. Degeneration is classified as follows: grade 1 (low), grade 2 (intermediate), grade 3 (high). The pattern of NITEGE deposits (G1 fragment of aggrecan) was assessed immunohistochemically (n=38) and compared with the grades of degeneration. In 48% of the patients with grade 2 or 3 degeneration extracellular NITEGE deposits (specificity 100%) were found, whereas grade 1 patients showed no deposits. Extracellular NITEGE deposits correlated positively with the grade of degeneration. In all, 30 cases (10 per grade) were assessed by three pathologists (A, B, C). Grading conformity was 70% for grade 1, 66% for grade 2 and 100% for grade 3. Cohen's Kappa coefficient was 0.6--0.7 between pairs of observers. Combining grade 1 and 2 to low-grade degeneration, compared to a grade-3 high-grade degeneration achieved Kappa coefficients of between 0.93 and 1.0. This reproducible degeneration score for fibrous cartilage could form the basis for the standardized assessment of meniscal degeneration.

[Value of histological work-up for synovial diseases]. / Stellenwert der histologischen Diagnostik der Synovialkrankheiten.

At times clinicians and pathologists underestimate the value and significance of histopathological diagnostics for synovial tissue. However, the most common diseases of the synovium show specific morphological hallmarks that allow an exact diagnosis. Using the synovitis score allows one to distinguish between degenerative (low-grade synovitis) and inflammatory rheumatic (high-grade synovitis) diseases. Synovial biopsies are not only especially indicated when there are atypical patterns of arthritis, clinical options have been exhausted or monarthritis of unknown origin occurs, but also in patients with known rheumatoid arthritis. Joint infections, crystal-induced arthritis or pigmented villonodular synovitis can also be diagnosed as secondary synovial diseases. Providing clinical information when submitting biopsies/tissue specimens is essential to classify even unspecific morphological changes. Immunohistochemical staining, polarization microscopy or molecular biology techniques (PCR) may be used to ensure diagnoses.

[Scleroderma and fibrosing diseases]. / Sklerodermie und fibrosierende Erkrankungen.

Fibroblasts and myofibroblasts play an important role in the pathogenesis of systemic sclerosis, fibromatoses, arthrofibrosis, and Ormond's disease. These conditions are characterized by an excessive fibroblast proliferation and partly accompanied by inflammation. Scleroderma is either localized or systemic, and features additional vasculopathy. Scleroderma-like skin lesions can be found in graft-versus-host disease following allogeneic hematopoietic stem cell transplantation, complicated malignoma or can represent an adverse drug reaction. The fibromatoses are found in superficial, or as semi-malignant desmoids in deep body compartments. Ormond's disease is a chronic periaortitis of unknown origin which extends into the retroperitoneal space. The diagnostic relevance of a histopathological diagnosis of fibrosing diseases varies and ranges from a disease-supporting to a disease-defining value.

[MMP- and FAP-mediated non-inflammation-related destruction of cartilage and bone in rheumatoid arthritis]. / MMP- und FAP-vermittelte inflammationsunabhängige Destruktion von Knochen und Knorpel in der rheumatoiden Arthritis.

INTRODUCTION: Due to morphological similarities of high-grade synovitis in rheumatoid Arthritis (RA) and mesenchymal, semimalignant tumors and the hypothesis that RA progression is not only inflammation-related, but also determined by tumor-like mechanisms, a comparison was made between expression profiles of RA, giant cell tumor of bone (GCT) and normal synovium (ND). METHODS: Array data of selected genes were validated through immunohistochemical staining of paraffin-embedded and deep frozen tissue samples of GCT, RA and normal synovium. RESULTS: With microarray analysis, CCR1, CCR5, MMP-1, MMP-2, MMP-3, MMP-9, MMP-14 and FAP were found to be significantly upregulated in RA and GCT compared to ND. A significant upregulation in RA and GCT compared to ND could be validated by immunohistochemistry for MMP-1, MMP-9, MMP-14 and FAP. DISCUSSION: For MMPs, and MMP-9 in particular, an important role in early cartilage destruction of RA was suggested. The presence of FAP in RA and in stroma of a semimalignant tumor indicates tumor-like tissue destruction in chronic synovitis associated with RA.

[Histopathologic diagnostics in endoprosthetics: periprosthetic neosynovialitis, hypersensitivity reaction, and arthrofibrosis]. / Histopathologische Diagnostik in der Endoprothetik: Periprothetische Neosynovialitis, Hypersensitivitätsreaktion und Arthrofibrose.

The durability of endoprosthetic implants of the large joints has increased over the last decades. North American studies have shown a 10-year durability of 94% for prosthetic hip implants, and European studies have shown 10-year durabilities of 88-95%. Pathologists differentiate three etiological disease patterns for the"pathology of endoprosthetics" that lead to reduction of implant durability: 1) periprosthetic particle disease (aseptic loosening), 2) infection, and 3) arthrofibrosis. Four types of neosynovitis/periprosthetic membrane have been determined in a consensus classification: particle-induced type (type I), with a mean prosthesis durability (MPD) of 12 years; infectious type (type II), MPD 2.5 years; combined type (type III), MPD 4.2 years; and indeterminate type (type IV), MPD 5.5 years. There are three histopathologic degrees of arthrofibrosis; grade 1 always needs clinical information for diagnosis, whereas grades 2 and 3 are distinct histopathologic entities.

Common missense mutation G1528C in long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. Characterization and expression of the mutant protein, mutation analysis on genomic DNA and chromosomal localization of the mitochondrial trifunctional protein alpha subunit gene.

Mitochondrial trifunctional protein (MTP) is a recently identified enzyme involved in mitochondrial beta-oxidation, harboring long-chain enoyl-CoA hydratase, long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) and long-chain 3-ketothiolase activity. A deficiency of this protein is associated with impaired oxidation of long-chain fatty acids which can lead to sudden infant death. Furthermore, it is clear that this inborn error of fatty acid oxidation is very frequent, second to medium chain acyl-CoA dehydrogenase deficiency. In most patients only the LCHAD activity of MTP is deficient with near normal activity of the two other enzyme activities of the complex. We recently described the occurrence of a frequent G1528C mutation in the cDNA coding for the a subunit of MTP. Using S. cerevisiae for expression of wild type and mutant protein we show that the G1528C mutation is directly responsible for the loss of LCHAD activity. Furthermore, we describe a newly developed method allowing identification of the G1528C mutation in genomic DNA. The finding of an 87% allele frequency of the G1528C mutation in 34 LCHAD deficient patients makes this a valuable test for prenatal diagnosis. Finally, we show that the gene encoding the alpha subunit of MTP is located on chromosome 2p24.1-23.3.

Method to obtain absolute impurity density profiles combining charge exchange and beam emission spectroscopy without absolute intensity calibration.

Investigation of impurity transport properties in tokamak plasmas is essential and a diagnostic that can provide information on the impurity content is required. Combining charge exchange recombination spectroscopy (CXRS) and beam emission spectroscopy (BES), absolute radial profiles of impurity densities can be obtained from the CXRS and BES intensities, electron density and CXRS and BES emission rates, without requiring any absolute calibration of the spectra. The technique is demonstrated here with absolute impurity density radial profiles obtained in TEXTOR plasmas, using a high efficiency charge exchange spectrometer with high etendue, that measures the CXRS and BES spectra along the same lines-of-sight, offering an additional advantage for the determination of absolute impurity densities.

Tumor regression by means of magnetic drug targeting.

The expression 'magnetic drug targeting' is understood as meaning the targeted administration of a drug, for example, a cytostatic, with the intention of optimizing the local therapeutic effect. A magnetic field strength of 0.6 T is applied externally to the body. Iron oxides are administered intravasally into a vein. Cytostatics are bonded to the iron oxides. This form of administration, also known as sluicing, is particularly suitable for cytostatics, since the intention is to achieve a high concentration of the cytostatic at the target site (site of the tumor), but to minimize the harmful effect in the rest of the tissue. A reduction in tumor volume under the magnetic field and in the liver of 45-90% has been detected by MRI.

[Synovitis score: value of histopathological diagnostics in unclear arthritis. Case reports from rheumatological pathological practice]. / Synovialitis-Score: Wertigkeit histopathologischer Diagnostik bei unklaren Arthritiden. Fallberichte aus der rheumatologisch-pathologischen Praxis.

Histopathological assessment of synovial biopsies has an established value. The value for inflammatory joint diseases without standardized rating mechanisms was, however, unknown until recently. The exemplary use of the synovitis score in four cases all including recurrent bruises of the knee joint portrays its value for diagnosis and therapy. Usage of the score includes assessing the enlargement of the lining layer, cellular density of synovial stroma and leucocyte infiltration by giving each a score of 0-3 points and adding them. Presence of high-grade synovitis (>or=4 points) in all cases displayed the reason for the joint bruises within a primarily inflammatory, rheumatoid circle. In this report we show the broad variety of uses for the synovitis score dealing with cases of Lyme arthritis, rheumatoid arthritis, seronegative monarthritis and HLA-B27-positive peripheral arthritis.

[Kinetical studies on phytochrome-induced protein synthesis]. / Kinetische Studien zur Phytochrominduzierten Proteinsynthese.

Phytochrome control of protein metabolism has been analyzed in the cotyledons of the mustard seedling (Sinapis alba L.) which can be regarded as being representative of dicotyledonous seedlings. A mustard seedling is a closed system for nitrogen under our experimental conditions. Fig. 2 shows the kinetics of protein content in the cotyledons of the mustard seedling in darkness and under the influence of continuous far-red irradiation, i.e. under the influence of a low but virtually constant concentration of phytochrome 730. -In order to understand these two curves it must be realized that the cotyledons of the mustard seedling contain much storage protein which will be degraded during the development of the seedling. The resulting amino acids are either translocated to other parts of the seedling or used within the cotyledons for the synthesis of enzymes and structural protein. -Histochemical studies have shown (HäCKER, 1966) that under the influence of far-red light the degradation of storage protein in the cotyledons is enhanced; at the same time, however, phytochrome 730 stimulates a strong de novo synthesis of structural and enzyme protein in the cotyledons. -This interpretation of the data on protein content is supported by evidence of the phytochrome-enhanced incorporation of (14)C into the protein fraction after an application of (14)C-Leucine (U) (Figs. 5, 6). - All present findings on phytochrome-controlled protein synthesis agree with the hypothesis that phytochrome 730 exerts its function through a differential gene activation (MOHR, 1966b).

[Computer-assisted cognitive training of schizophrenic patients. Use of evaluation outcome for developing an individualized training method]. / Computergestütztes kognitives Training bei schizophrenen Patienten. Die Nutzung von Evaluationsergebnissen für die Entwicklung eines individualisierten Trainingsverfahrens.

The paper deals with the evaluation of a computer-based cognitive group training programme for schizophrenic patients. The evaluation of the training results of 100 patients who participated in an earlier training scheme showed ceiling effects in up to 46% of the cases. In the remaining cases the improvements varied in a wide range between 1.6 and 20%. These results implied that the training had to be adapted more closely to the specific needs of the individual patient. To this end we developed criteria for each task to decide whether training in this task is necessary or the performance is sufficient. This approach limits training to those tasks in which patients display deficits. Satisfactory improvement terminating training for the specific task, is defined as an increase in correct responses of at least 10%. For those tasks in which reaction times are used as performance measure, the criterion is a 10% increase in percentile rank. An analysis of the new training programme with the data of 20 patients revealed that 27% of the tasks showed ceiling performance with no further training need. In the remaining tasks the patients' performance attained the training criterion in 68%, at instances mostly even after one additional session. These preliminary results indicate that the new training scheme is able to provide more flexibility, individualization and an enhanced focus on the patients' deficits.

Slit-scanning technique using standard cell sorter instruments for analyzing and sorting nonacrocentric human chromosomes, including small ones.

We have investigated the performance of two types of standard flow cell sorter instruments, a System 50 Cytofluorograph and a FACSTar PLUS cell sorter, for the on-line centromeric index (CI) analysis of human chromosomes. To optimize the results, we improved the detection efficiency for centromeres in two ways. A higher efficiency was obtained first by elongation of the chromosomes and second by introducing a high resolution lens system for laser beam focusing. In the two-parameter flow karyotype of CI and DNA content of human chromosomes, distinct peaks are produced not only by the larger chromosomes 1-8 and X, but by the smaller nonacrocentric chromosomes 9-12 and 16-20 as well. As the chromosomes 9-12 cannot be distinguished by other flow karyotyping methods, we discriminated and sorted chromosomes 12 and 10 from 9 and 11 to investigate the capacity for the separation of chromosomes in this group. A purity of at least 90% was achieved; in the isolated population the fraction chromosomes 12 was 55%; the remaining 45% were chromosomes 10 (40%) and unidentifiable chromosomes (5%).