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Im Jahr 2019 wurden in Deutschland über 44.000 koronare Bypassoperationen und über 36.000 Operationen auf Grund von Defekten der Herzklappen durchgeführt. Gerade im Bereich der Bypassoperationen gibt es immer wieder Patienten, welche aus verschiedenen Gründen keine geeigneten autologen Gefäße besitzen. Nach derzeitigem Standard wird in einem solchen Fall auf die perkutane Koronarintervention (PCI) zurückgegriffen, obwohl insbesondere bei koronarer Dreigefäßerkrankung die Bypassoperation gegenüber der PCI im Vorteil ist. Um diesen Patienten trotzdem eine Bypassoperation zu ermöglichen, wird im Rahmen des Tissue Engineerings daran geforscht, einen Gefäßersatz zu schaffen, welcher die guten hämodynamischen Eigenschaften autologer Gefäße aufweist, ein möglichst geringes Thrombosierungsrisiko besitzt und nicht-immunogen ist. Zudem befasst man sich im Rahmen des Tissue Engineerings auch im Bereich des Herzklappenersatzes. Für beide Schwerpunkte gibt es Ansätze, bei welchen ein Gerüst, sei es biologisch oder synthetisch, mit vaskulären Zellen des späteren Empfängers besiedelt wird. In der vorliegenden Arbeit wurde sich im Bezug darauf mit der Isolierung und Differenzierung vaskulärer Zellen befasst. Diese Zellen wurden zunächst aus Bypassresten isoliert und die Unterscheidung erfolgte mittels verschiedener Antikörper für Immunfluoreszenzfärbung. Das Hauptaugenmerkt bestand vor allem bei den Antikörpern gegen Fibroblasten (FB), da die meisten für FBs aus anderen Geweben entwickelt wurden. Zudem können sich FBs in Kultur an Glattmuskelzellen (SMC) annähern. Zur Differenzierung der drei vaskulären Zelltypen wurden folgende Antikörper getestet: Cluster of Differentiation 90 (CD90, ein polyklonaler und ein monoklonaler AS02), Fibronektin, Fibroblast Surface Protein (FSP), Anti-Fibroblast TE7, alpha Glattmuskelzellaktin (αSMA), Desmin, Vimentin, Cluster of Differentiation 31 (CD31) und von Willebrand Faktor (vWF). Diese Antikörper wurden an FBs (dermal, selbstisoliert und kommerziell erworben), SMCs (selbstisoliert und kommerziell erworben) und selbstisolierten Endothelzellen (EC) nach Aceton-, Ethanol- und Formolfixierung getestet. Es wurden insgesamt drei Färbedurchläufe durchgeführt. Die Bewertung des Fluoreszenzsignals erfolgte im Hinblick auf die Intensität, die Lokalisation und das Färbemuster. Es zeigte sich, dass Fibronektin und Vimentin für alle Zelltypen stark positiv waren. Die Antikörper gegen FSP, Desmin und der polyklonale Antikörper gegen CD90 zeigten ein variables Signal ohne eine spezifische Tendenz. Der TE7 färbte sowohl die FBs als auch die SMCs schwach mit nukleärem und perinukleärem Signal an und die ECs zeigten kein Signal. Der Antikörper gegen vWF zeigte sowohl bei SMCs als auch bei FBs ein extrazelluläres, schlierenartiges Signal. Das Signal der kultivierten ECs war tupfen förmig innerhalb der Zellgrenzen gelegen. Eindeutiger war das Signal des CD31-Antikörpers. Er schien spezifisch die ECs zu färben. Der Antikörper gegen CD90 AS02 färbte ECs nicht an. Er war durch ein gleichmäßiges, flächiges Signal innerhalb der Zellgrenzen der FBs zu erkennen. Das Signal der selbstisolierten FBs war eher wolkig und glich dem der SMCs. Ein streifiges Signal zeigten die SMCs nach der Färbung mit αSMA, allerdings unterlag das Signal bei einigen SMCs stärkeren Schwankungen. Das Signal der FBs war punktförmig im Nukleus und perinukleär gelegen. Dieses entsprach dem Muster der schwach angefärbten SMCs oder dem der ECs, welche ein noch schwächeres Signal zeigten. Doch in der Literatur wurde bereits beschrieben, dass FBs αSMA ausbilden können und sich den SMCs im Sinne von Myofibroblasten annähern. Des Weiteren wurde auch von αSMA negative SMCs in anderen Arbeiten geschrieben. Es wird davon ausgegangen, dass diese Zellen im proliferativen Zyklus sind und es eher von Zellen ausgebildet wird, die sich im Wachstumsstillstand befinden. Die Fixierung mit Formol barg einige Schwierigkeiten, insbesondere in Bezug auf die Epitopdemaskierung. Zudem war die Signalintensität bei den anderen Fixierungsmethoden höher. Obwohl die Signalintensität nach Acetonfixierung stärker war als nach Ethanolfixierung, waren bei dieser die Unterschiede der einzelnen Zelltypen deutlicher und sie ist somit für eine Differenzierung besser geeignet. Zusammenfassend ist zu sagen, dass eine Differenzierung der Zellen am ehesten mit einer Antikörperkombination aus CD90 AS02, αSMA und CD31 möglich war, wenn nicht nur die Signalintensität, sondern auch die Lokalisation und das Färbemuster betrachtet wurde. Um die Differenzierung weiter zu verbessern, sollte noch die Morphologie der Zellen miteinbezogen werden und noch weitere Antikörper getestet werden.

Atomoxetin ist das erste Nicht-Stimulanz, das zur Behandlung der Aufmerksamkeitsdefizit-Hyperaktivitätsstörung im Kindes-, Jugend- und Erwachsenenalter in Deutschland zugelassen ist. Atomoxetin gilt als selektiver Noradrenalin-Wiederaufnahmehemmer. Weitere zellbiologische Wirkmechanismen sind bisher unzureichend untersucht. In dieser in vitro-Studie an HEK-293- und HEK-hNET-Zellen und an sich entwickelnden hippocampalen und kortikalen neuronalen Rattenembryonenzellen wurde untersucht, ob Atomoxetin dosis- und entwicklungsabhängig zu Veränderungen der Viabilität, Neuritogenese und Synaptogenese führt. Atomoxetin hat in klinisch relevanten Plasmakonzentrationen (3-5 µM) keinen Einfluss auf die Viabilität von HEK-293- und HEK-hNET-Zellen oder auf die Viabilität von hippocampalen und kortikalen neuronalen Zellen. Ab einer Konzentration von 50 µM bzw. 100 µM bei den HEK-hNET-Zellen zeigt sich bei allen Zellarten eine signifikante Verminderung der Viabilität. Bei Gabe von höheren Atomoxetinkonzentrationen zeigten sich weniger apoptotische Prozesse bei HEK-hNET-Zellen im Vergleich zu HEK-293-Zellen. Bei Behandlung mit höheren Atomoxetinkonzentrationen ging die Viabilität in den kortikalen neuronalen Zellkulturen signifikant stärker zurück als in den hippocampalen neuronalen Zellkulturen. Bei den kortikalen neuronalen Zellen kam es an allen Entwicklungstagen ab 50 µM zu einer signifikanten Verminderung der Neuritenanzahl. An DIV 5 und DIV 10 deutete sich ein Trend an, dass es zum Teil schon bei Behandlung mit 5 µM, 10 µM, 15 µM Atomoxetin zu einer Verminderung der Tertiärneuriten kam. An allen Entwicklungstagen kam es ab 50 µM zu einer signifikanten Verminderung der Synapsendichte, an DIV 10 schon bei Behandlung mit 15 µM Atomoxetin. Atomoxetin hat ab 50 µM entwicklungsunabhängig einen Einfluss auf die Viabilität der Zelllinien und der neuronalen Zellen sowie auf die Neuritenanzahl und die Synapsendichte kortikaler neuronaler Zellen.

CD8+ tumor-infiltrating T cells (CD8-TILs) are found in many types of tumors including human renal cell carcinoma. However, tumor rejection rarely occurs, suggesting limited functional activity in the tumor microenvironment. In this study, we document that CD8-TILs are unresponsive to CD3 stimulation, showing neither lytic activity, nor lytic granule exocytosis, nor IFN-γ production. Mechanistically, no deficits in TCR proximal signaling molecules (lymphocyte-specific protein tyrosine kinase, phospholipase Cγ) were identified. In contrast, distal TCR signaling was suppressed, as T cells of TILs showed strongly reduced steady-state phosphorylation of the MAPK ERK and were unable to increase phosphorylation of ERK and JNK as well as AKT and AKT client proteins (IκB, GSK3) after stimulation. These deficits were tumor-specific as they were not observed in CD8+ T cells infiltrating non-tumor kidney areas (CD8+ non-tumor kidney-infiltrating lymphocytes; CD8-NILs). Diacylglycerol kinase-α (DGK-α) was more highly expressed in CD8-TILs compared with that in CD8-NILs, and its inhibition improved ERK phosphorylation and lytic granule exocytosis. Cultivation of TILs in low-dose IL-2 reduced DGK-α protein levels, increased steady-state phosphorylation of ERK, improved stimulation-induced phosphorylation of ERK and AKT, and allowed more CD8-TILs to degranulate and to produce IFN-γ. Additionally, the protein level of the AKT client molecule p27kip, an inhibitory cell cycle protein, was reduced, whereas cyclin E, which promotes G1–S phase transition, was increased. These results indicate that the tumor-inflicted deficits of TILs are reversible. DGK-α inhibition and provision of IL-2 signals could be strategies to recruit the natural CD8+ T cells to the anti-tumor response and may help prevent inactivation of adoptively transferred T cells thereby improving therapeutic efficacy.

The existence of autoantibodies is characteristic of several rheumatic diseases and indicates impaired self-tolerance. The analysis and detection of these autoantibodies represent an important factor in the diagnosis of rheumatic diseases. Immunofluorescence is an important technique in basic science and also one of the most commonly used techniques for the characterization of autoantibodies in clinical medicine thus facilitating the diagnosis and evaluation of rheumatic diseases. Addition of serum samples from individual patients to an antigen substrate (present in fixed cells or tissue) results in a specific reaction between the autoantibodies and the respective antigens. Incubation with a second, fluorescence-linked, anti-human antibody which binds to the primary antigen-bound autoantibodies from patient serum results in fluorescence patterns which are characteristic of certain rheumatic diseases.

Glioblastoma (GBM) is the most aggressive primary brain tumor and is resistant to all therapeutic regimens. Relapse occurs regularly and might be caused by a poorly characterized tumor stem cell (TSC) subpopulation escaping therapy. We suggest aldehyde dehydrogenase 1 (ALDH1) as a novel stem cell marker in human GBM. Using the neurosphere formation assay as a functional method to identify brain TSCs, we show that high protein levels of ALDH1 facilitate neurosphere formation in established GBM cell lines. Even single ALDH1 positive cells give rise to colonies and neurospheres. Consequently, the inhibition of ALDH1 in vitro decreases both the number of neurospheres and their size. Cell lines without expression of ALDH1 do not form tumor spheroids under the same culturing conditions. High levels of ALDH1 seem to keep tumor cells in an undifferentiated, stem cell-like state indicated by the low expression of beta-III-tubulin. In contrast, ALDH1 inhibition induces premature cellular differentiation and reduces clonogenic capacity. Primary cell cultures obtained from fresh tumor samples approve the established GBM cell line results.

Inflammations of the conjunctiva represent some of the most frequent eye diseases in general and ophthalmologic practice. Etiologically multiple diseases can be identified. A group of infectious conjunctivites contrasts with a very hereogeneous non-infectious group. Neoplastic processes such as low grade epithelial dysplasias and lymphomas may mascarade conjunctivitis. The differentiation of the various clinical pictures may be challenging for the practitioner. This article reviews the methodology of patient history and clinical work up as well as the symptomatology and treatment options for the most important conjunctivites in clinical practice.

Patients with pneumonia resistant to treatment are a common problem in chest hospitals. Pneumocystis jiroveci (Pc) should always be considered as an opportunistic pathogen in case of potential, especially T cell-related immunodeficiency, even if AIDS is not obvious. We report on cases of Pneumocystis jiroveci pneumonia (PcP) (n = 1921) without associated AIDS in a chest hospital.We assessed 1921 inpatients presenting with pneumonia during January 1 (st) 1996 and September 30 (th) 2005 in our hospital. Identification of the cases with PcP was based on discharge diagnosis as well as on our microbiological database (detection of Pc by immunofluorescence-test (IFT) and/or PCR).The diagnosis of PcP was made in 14 patients (11 males, 3 females) without associated AIDS. All patients were treated with immunosuppressive medication before admission. CD4 cell-counts were substantially decreased (CD4 cells200/microl) in 11 patients. To establish the diagnosis of PcP a Pc-PCR (using broncho-alveolar lavage (BAL) as the best material) was necessary in 8 patients, since the Pc-IFT proved positive only in 6 patients. Severe hypoxemia had to be managed by noninvasive ventilation in 3 patients, while another 4 patients had to be ventilated invasively. The mortality rate was 2 out of 14 patients.PcP is an important disease in the differential diagnosis of pneumonia and at the same time constitutes a severe pulmonary complication in immunodeficient patients. If immunodeficiency is suspected, a CD4 cell-count as well as bronchoscopy with BAL should be performed to diagnose PcP. As expected, Pc-PCR was superior to Pc-IFT and should therefore be used on a routine basis for diagnosing PcP.

Bullous pemphigoid (BP) is a cutaneous autoimmune disease predominantly affecting older patients which can cause death either due to severe clinical manifestations or due to the side effects of systemic immunosuppressive treatment. Topical treatment with corticosteroids is an established alternative to systemic treatment. However, prolonged application is accompanied by side effects such as skin atrophy. Recently, the immunomodulatory calcineurin antagonists tacrolimus and pimecrolimus have been introduced for topical treatment of skin diseases. Tacrolimus has been reported to be effective in several inflammatory skin disorders such as atopic dermatitis, psoriasis, lichen planus, lupus erythematosus and pyoderma gangraenosum. Efficacy has also been described in the topical treatment of BP in some cases. Here we present the case of a 89 year old patient with BP. He was treated with 0.1% tacrolimus ointment, which was able to control the disease. We briefly review the literature and discuss the potential role of tacrolimus as a novel option for the topical treatment of BP.

The specific structure of a dental implant's surface is often used as an argument for a better overall performance and durability. Nonetheless, it is not yet fully clear to what extent an implant's material or surface structure improves its performance. For a better understanding of the early integration-related processes, growth and initiation of mineralization of human MG-63 osteoblasts grown on glass slides or on polished (MS: machine-surfaced) or structured (SB: deep-structured) titanium surfaces were monitored. Cells were cultured under non-confluent conditions in absence and presence of osteogenic medium (OS). The 3D-architecture of growing cells was documented by confocal laser scanning microscopy (CLSM) upon fluorescently labeling for collagen I. Collagen I distribution was found comparable under all conditions. In control cultures on glass, MS and SB, cells had grown as multilayered but not yet confluent networks, which additionally comprised dome-like structures on the two Ti-surfaces. While on glass and SB cells essentially maintained this architecture also in presence of OS, three-dimensional structures on MS were--if at all--only barely visible. This was reflected also in the protein content of the cultures that was significantly lower under OS in the case of MS only. The activity of the early mineralization marker alkaline phosphatase (ALP) was higher in the controls on glass than on MS or SB. In presence of OS it was significantly higher on MS and SB than in the controls, yet not on glass. Obviously both, the surface material and the surface structure contribute--possibly synergistically--to early processes of implant integration, such as matrix formation and mineralization. The impact of such effects is discussed, in relation to implant loading and implant durability.

Inflammatory articular cartilage diseases such as arthritis and osteoarthritis are characterized by a loss of articular cartilage due to an imbalance between synthesis and degradation of the extracellular cartilage matrix. These diseases are accompanied by an increased induction of cytokines such as interleukin 1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha). The increased release of cytokines leads to an enhanced production of matrix-degrading enzymes e.g. the matrix metalloproteinases (MMPs). In this study the direct antirheumatic effects of an extract of the secondary root of the African devil's claw (Harpagophytum procumbens DC) on the production of MMPs in IL-1beta-stimulated human chondrocytes were examined. A detailed evaluation by immunomorphological methods and Western blot analysis showed that the extracts of Harpagophytum decreased significantly the production of MMPs (MMP-1, MMP-3, MMP-9) in chondrocytes. The IL-1beta-induced production of MMPs was also significantly reduced by both a JM-extract (Jucurba) containing 210 mg dry extract and JF-extract (Jucurba forte) containing 480 mg dry extract. After all it could be shown that the effect of JF-extract on the MMP-synthesis was more pronounced in untreated and cytokine-stimulated chondrocytes when compared with the effect of the JM-extract. The capability of the JM-extract to suppress the MMP-production via the inhibition of the synthesis of inflammatory cytokines could explain its therapeutic effect in arthritic inflammations. In these in vitro experiments the JF-extract showed a higher efficacy than the JM-extract.

Dermatology and internal medicine have in common that many systemic diseases manifest with skin symptoms that are easily accessible for both diagnostic and investigative procedures. Even if modern practice of medicine tends towards organ specific particularity of specialists, dermatology traditionally strives for interdisciplinary communication. Cutaneous manifestations of internal diseases present in various forms. In a syndrome, a number of symptoms present in an identical pattern, and pattern recognition is diagnostic. Vice versa, a distinct skin sign can be a clue to various internal disorders, that have to be considered in the differential diagnosis. Purpura and livedo due to either inflammatory (immune complex mediated or associated with ANCA) or vasoocclusive microthrombotic vascular disease are presented as examples. Finally, subtile variations in key symptoms of the skin, such as of pyoderma gangrenosum, may give a clue to a specific underlying disease, such as rheumatoid arthritis, inflammatory bowel disease, monoclonal gammopathy, or hematologic disease, as exemplified by the variants of pyoderma gangrenosum.

Lipoprotein lipase (LPL) is the central enzyme in plasma triglyceride hydrolysis. Various genetically modified mouse lines expressing either native or mutated LPL have been established, and the function of LPL in the metabolism and during lipoprotein transport has been characterized. Some of these animals showed an LPL-mediated myopathy or cardiomyopathy. Comparing the histological changes with metabolic data provides new insights in the function of LPL in metabolism and for atherogenesis.Histological and electron microscopy specimens of muscle and heart from different LPL transgenic mouse lines have been evaluated and compared to metabolic data.Presence of LPL causes a myopathy and cardiomyopathy primarily by augmenting nonenzymatic selective cholesterol ester uptake. LPL-mediated fatty acid and lipoprotein uptake is not sufficient to harm muscle tissue.It is suggested, that LPL derived from endothelial cells and macrophages--besides other atherogenic mechanisms--can mediate cholesterol ester influx into the vessel wall and thereby promote proatherogenic modifications. Although due to its hydrolytic function, LPL is generally considered an antiatherogenic enzyme, endothelial LPL can locally facilitate atherogenesis.

Elevated free fatty acid (FFA) levels are believed to be one of the major contributing factors in the pathogenesis of type 2 diabetes. FFAs enhance peripheral insulin resistance, promote beta-cell dysfunction, and trigger beta-cell death. The purpose of this study was to investigate whether the lipoapoptotic effect of FFAs is determined by the degree of saturation and the chain length, and which pathways might be involved.After incubation of RIN1046-38 rat insulinoma cells and primary human islet cells with different FFAs, apoptosis was assessed by flow cytometry (sub-G1-DNA formation) and TUNEL assay, respectively. Intracellular distribution of protein kinase C-delta (PKC-delta) was determined by confocal laser microscopy.In RIN1046-38 cells, the sub-G1 fraction increased 40-fold over basal (0.5% to approximately 20%) after treatment with the saturated FFAs palmitate and stearate (1 mM, 24 h), while the monounsaturated FFAs palmitoleate and oleate as well as the polyunsaturated FFA linoleate had no proapoptotic effect. Unsaturated FFAs even more prevented palmitate- and stearate-induced apoptosis completely. Furthermore, these results could be confirmed in primary beta-cells of human islets. Investigating potential underlying signaling pathways, it was found that only the saturated FFAs were able to induce nuclear translocation of PKC-delta.FFAs exert different effects on beta-cell survival. Saturated FFAs turned out to be pro-apoptotic, whereas unsaturated FFAs protected from saturated FFA-induced apoptosis. Furthermore, our data suggest an involvement of PKC-delta in apoptosis induced by saturated FFAs.

Hypersensitivity vasculitis (leukocytoclastic vasculitis) is defined as small-vessel vasculitis mediated by deposition of immune complexes (Arthus reaction) after exposition to various agents, such as drugs, toxins and infections. Due to a wide spectrum of precipitating agents and symptoms, classification systems and synonyms, understanding of the disease, its diagnosis and therapeutic strategy are difficult.Based on an extensive analysis of the literature, causal agents, etiopathogenesis as well as symptoms and their frequencies are summarized. The widespread differential diagnoses and the development of a diagnostic strategy for practical management of the disease are discussed.The course of disease is generally benign, and spontaneous remission is often observed. Severe organ manifestations and chronic courses have, however, been described. Clinical diagnosis must be confirmed histopathologically. Once the diagnosis is made, search for etiologic agents (successful in only 50% of the cases) and an intensive organ diagnostics should be performed. Subsequently, treatment with steroids, antihistamines and slow-acting antirheumatic drugs such as cytotoxic substances and immunosuppressive drugs is planned according to severity and prognosis.

Circulating autoantibodies are diagnostic markers for a variety of autoimmune diseases including rheumatoid arthritis, scleroderma, Sjögren's syndrome, systemic lupus erythematosus, and autoimmune hepatitis. Since only view studies exist on individuals without known diseases, we analyzed the prevalence of frequently determined autoantibodies in "healthy" individuals.111 individuals (43 female, 68 male; mean age 58 +/- 13 years, median 58, range 22-89), in whom either known or actual clinical evidence for autoimmune or internal disease was found, were included. Antinuclear and anti-smooth muscle antibodies (ANA and ASMA, respectively) were detected by immunofluorescence on rat organ sections and Hep 2 cells. Antibodies to liver-kidney-microsomes-1 (anti-LKM-1) and antimitochondrial antibodies (AMA) were detected and semiquantified by immunofluorescence. Additionally, anti-LKM-1 and AMA were determined by ELISA and Western blot. Antibodies against soluble liver antigens (anti-SLA) were quantified by ELISA. Sera with a titer of 1 : 40 or higher were classified as positive.Sera of "healthy" adults displayed high frequencies of ANA and ASMA (28/111, 25%, and 48/111, 43%, respectively). Although no sex differences were found for ASMA, sera of healthy women tested more often positive for ANA (p0.01). Since at least one in three or four healthy individuals tested positive for ANA or ASMA, the positive predictive value of these autoantibodies is low, and clinical interpretation should include additional information.

Anti-Neutrophil Cytoplasmic Antibodies (ANCA) are a heterogenous group of autoantibodies with a broad spectrum of clinically associated diseases. The diagnostic value is established of Proteinase 3 (PR3)-ANCA for Wegener's granulomatosis (WG) as well as Myeloperoxidase (MPO)-ANCA for microscopic polyangiitis (MPA). Within the last 20 years these antibodies were subject of intensive studies and a growing body of evidence arose for a distinct role of ANCA in the pathogenesis of the ANCA associated vasculitides WG and MPA. Our current concept of whether ANCA directly or indirectly contribute to vascular damage (ANCA-Cytokine-Sequence-Theory) was mainly developed from in vitro studies. It is plausible and it is supported by data from clinical investigations as well as animal models. Nevertheless our knowledge of the etiological and pathogenetic pathways remains incomplete.

The diagnosis of herpes zoster is mostly based on clinical findings. In atypical cases, rapid and reliable virological diagnostics is necessary because effective antiviral chemotherapy is available.The meaningfulness and practicability of conventional diagnostic methods mostly used such as antigen staining, virus isolation and detection of virus-specific antibodies were compared with polymerase chain reaction (PCR) in 100 patients with zoster.PCR using oligonucleotides selected from the open reading frames 28 and 29 of the varicella-zoster virus (VZV) genome showed the highest sensitivity of 94% and a specificity of 100%. Direct immunofluorescent antigen staining carried out within few hours had a sensitivity of 82% and a specificity of 70%. However, results could only be obtained in 71% of the cases. On the basis of serologic parameters, an active VZV infection was detected in 61% of the patients. Virus culture frequently regarded as "gold standard" was successful in 20%. There was a strong dependence on the time of collecting specimens and the immunocompetence of the patients.The study shows that the PCR has to be considered the method of choice for the diagnostics of zoster with an atypical course. Serological methods can only be recommended to confirm the diagnosis retrospectively.

The aim of this study was to evaluate whether the presence of SLA/LP-autoantibodies in PBC-patients gives evidence for a secondary AIH, also called AIH/PBC-overlap-syndrome.Out of 233 consecutive patients with PBC who had been followed between October 1980 and April 2000, we evaluated the data of anti-SLA/LP-positive patients and compared them to patients with an anti-SLA/LP-negative AIH/PBC overlap syndrome as well as to patients with a classical course of AIH and PBC.In total we could identify nine PBC patients with anti-SLA/LP antibodies (six women/three men) or 3.9% of the study population, Anti-SLA/LP-positive PBC patients were slightly younger at diagnosis in comparison to anti-SLA/LP-negative PBC-patients (49.9 vs. 53.2 years). Transaminases and gamma-globulins were significantly higher in anti-SLA/LP-positive PBC-patients in comparison to anti-SLA/LP-negative PBC-patients (mean: 235 vs. 55 IU/l and 27.6 vs. 19.5 g/l). Anti-SLA/LP-positive patients significantly more frequently had an HLA-type that is characteristic for AIH (B8; DR3; DR4). Immunosuppressive therapy reduced inflammatory activity and cholestasis significantly. Relapses were frequent after reduction or discontinuation of immunosuppressive therapy.The presence of SLA/LP autoantibodies in PBC patients has a high specificity for a secondary AIH (AIH/PBC overlap syndrome). These patients have a good response to immunosuppressive therapy. The autoantibody profile and immunogenetics may help in future to identify PBC patients that benefit most from immunosuppressive therapy.

We compared a universal fungal PCR assay with fluorescence microscopy for the diagnosis of Pneumocystis carinii pneumonia. 82 bronchoalveolar lavages (BALs) of 64 immunocompromised patients with atypical pneumonia and 50 BALs of 50 immunocompetent adults without lung disease were examined. 10 immunocompromised patients were clinically and/or histologically proven to suffer from PCP. For fluorescence microscopy, sensitivity and specificity in detecting P. carinii were 80.0% and 98.1%, for the PCR assay 100.0% and 96.2%, respectively. The PCR assay is a useful method for the diagnosis of PCP and is recommended as an additional test to microscopical methods.

Several studies have demonstrated an association Chlamydia pneumoniae with coronary artery disease, suggesting that infection with C. pneumoniae increases the risk for coronary artery disease by factor 2 or more. Since atherosclerosis is considered to be a chronic inflammatory process, these data would fit into the response-to-injury hypothesis of atherosclerosis rather than representing a completely novel concept. Several pathomechanisms as increased cytokine synthesis, proliferative and proaggregatory effects could transmit the effects of chronic C. pneumoniae infection. Animal models and first clinical trials using antibiotic therapy seem to support an etiological role of C. pneumoniae in coronary artery disease. In this paper the current knowledge of the role of C. pneumoniae in coronary artery disease is reviewed and possible pathomechanisms are discussed.

A 40-year-old man with histologically proven sarcoidosis, known for 15 years, which had involved the myocardium was hospitalized because of intractable heart failure (NYHA class IV). An implantation of an intracardiac defibrillator for ventricular arrhythmias (Lown type IVa) had preceded. On physical examination severe dyspnea at rest cough and fever were noted.The erythrocyte sedimentation rate (88/104 mm), C-reactive protein (250 mg/l) and white cell count (20/nl) were markedly increased. Serum sodium (113 mmol/l), potassium (3.0 mmol/l) and chloride (64 mmol/l) were markedly reduced, while creatinine (2.5 mg/dl) and urea (82 mg/dl) were elevated due to renal failure. The chest radiogram demonstrated central venous congestion, cardiomegaly and right pericardial infiltration. There were no obvious changes due to sarcoidosis and computed tomography did not indicate pulmonary involvement by sarcoidosis. The echocardiogram revealed severe impairment of left ventricular function with an ejection fraction of ca. 14%.Heart failure (NYHA class IV), caused by a dilated cardiomyopathy of sarcoidosis, was accompanied by pneumonia which responded to antibiotics. But the chronic heart failure failed to improve on drug treatment and cardiac transplantation was undertaken. The explanted myocardium was examined histologically, immunologically and virologically. Antibodies were demonstrated against vascular endothelium, sarcolemma and endocardium (IgG and IgA), but not by PCR against cytomegalovirus, enterovirus and adenovirus. The transplantation was without complication and, under azathioprine and methylprednisolone, one rejection had occurred until now. The patient has been working full-time since 2 years in his previous occupation of lorry driver. There has been no evidence of renewed sarcoidosis activity.Sarcoidosis may involve the myocardium in up to 25% of cases. Clinically relevant symptoms are even more rare. Sarcoidosis should be included in the differential diagnosis of unexplained serious arrhythmias or cardiomyopathy, particularly in young persons. Cardiac transplantation may have to be contemplated if drug or pacemaker treatment fails to control heart failure.

A 57-year-old patient presented with malaise and severe persistent arthralgia of the left shoulder. He reported an acute illness with fever, generalized myalgia and arthralgias of the large joints which had started one month earlier during his flight back to Germany after a two weeks trip to the South Pacific. Physical examination showed extensive pain on palpation of the glenohumeral and acromioclavicular joints with impairment of active and passive mobility. Investigation of the cervical spine was normal.Apart from elevated C-reactive protein and erythrocyte sedimentation rate levels, routine laboratory investigations were normal including negative immunodiagnostic tests for autoantibodies and various global infections that may be associated with arthritis. Immunofluorescence tests showed significant levels of specific IgM- and IgG-antibodies against Ross River virus (RRV) but not against other arboviruses endemic in the South Pacific and Australia (Dengue, West Nile, Chikungunya, Sindbis, Barmah Forest). This was confirmed by a positive RRV neutralisation test. Attempts at virus isolation and detection of viral RNA by PCR were not successful.Symptomatic treatment with high doses of diclofenac quickly led to pain relief, and arthralgias receded within 10 days after begin of treatment. However, several bouts of arthralgia of the left shoulder and left knee occurred during a period of 4 months.Because of the current epidemiological situation in the South Pacific and Australia, infections by arboviruses like RRV should be considered in travellers returning from these areas with severe and persistent arthralgia of unknown origin, even in the absence of fever and other symptoms of acute infection.

In amphibians, Chlamydia psittaci infections are known since 1982 as a fatal disease. Only documented in two amphibian species, the pathogen seems to be widespread in these coldblooded vertebrates. The detection of intracytoplasmatic inclusion bodies and following examinations (electronmicroscopy) were necessary for the postmortal diagnosis. In 26 amphibians (3 species) a specific immunological testkit (immunofluerescence test) have been used. In 18 cases a positive result was found. This is the first report about using an IFT for the detection of Chlamydia psittaci in amphibians. Two new host species for this pathogen were found, one bufonid anuran (Bufo maculatum) and the new Pachytriton sp.

The simultaneous occurrence of lupus erythematosus (LE)- and lichen ruber (LP)-like symptoms is called LE/LP-overlap syndrome (LE/LP-OS). It is defined by concomitant clinical, histologic and immunhistologic features of both diseases. To date, 47 cases of this rare dermatosis have been reported with marked differences in the skin lesions: They are either of intermediate appearance between LE and LP (type I = intermediate type), or show a coexistence of LE- and LP-specific lesions (type II = polar type). To determine the frequency and characteristics of the LE/LP-OS we reviewed our LE-patients from 1984-1995. 5 cases were diagnosed. The frequency of LE/LP-OS in our patients is higher than generally assumed. Due to its variable clinical, histological and immunhistological appearance and the lack of unequivocal pathognomonic signs, this overlap-dermatosis may be underdiagnosed. Since therapeutic consequences result from the diagnosis, criteria are suggested to facilitate the recognition of the LE/LP-OS.

In up to 73% of ophthalmia neonatorum, Chlamydia trachomatis is the causative agents. Untreated sequelae to the eyes and organs may be the result. The aim of this study was to determine the bacterial spectrum of ophthalmia neonatorum with special regard to chlamydia and their diagnostic tests.We compared the results of 15 newborn with ophthalmia neonatorum. For the diagnosis we used a rapid diagnostic test, Immunofluorescent Antibody Staining and Culture on McCoy cells. Bacteria that were cultured on culture media were also identified.In 66% the newborn showed a positive rapid diagnostic test result that was confirmed by Immunofluorescent Antibody Staining. In 5 patients all tests were negative.In this study C. trachomatis was the most frequent pathogen. In the culture media we isolated mostly gram-positive cocci but not Neisseria gonorrhoeae. We point out the value of an exact rapid diagnosis and specific treatment to avoid sequelae to the eye and organs.

Bronchoalveolar lavage was performed in 128 pigs from five fattening units showing acute pneumonia (48 animals), subclinical purulent pneumonia (17 animals), and chronic purulent pneumonia (63 animals). These samples were investigated for bacteria. Additionally immunofluorescence microscopy as well as serological investigations were performed to detect antibodies against several bacteria and viruses. Pasteurella multocida could be detected in more than a half of the samples of pigs with acute pneumonia. Bordetella bronchiseptica and mycoplasmas were isolated in a lower amount. Probably these bacteria infected the pigs of at least one herd after a primary infection with influenza virus because (i) influenza virus could be detected in three of four animals investigated for influenza virus by culture methods, (ii) the virus could be detected in one third of the animals investigated for by immunofluorescence microscopy, and (iii) antibodies against influenza virus could be detected in almost all animals. From pigs with subclinical purulent pneumonia Bordetella bronchiseptica as the only bacterial lung pathogen could be isolated exclusively from nearly each sample. From the samples of pig suffering from chronic purulent pneumonia first of all Bordetella bronchiseptica, Pasteurella multocida and different mycoplasma species could be detected. Using cultural methods Actinobacillus pleuropneumoniae could be isolated from six samples only, in contrast to frequent positive reactions against Actinobacillus pleuropneumoniae antigens obtained by immunofluorescence microscopy and CFT.

To investigate molecular aspects of the mechanism(s) involved in orthodontic tooth movement, periodontal ligament fibroblasts (PDL) were isolated and grown on culture dishes with a flexible bottom. The cells were stimulated by stretching the bottom of the culture dishes and whole cell extracts were prepared and subjected to sodium dodecyl sulfate polyacrylamide (SDS-PAGE) electrophoresis, electrotransferred to nitrocellulose membrane and immunoprobed with antibodies specific for vimentin and alpha- and beta-tubulin. No apparent alterations in the expression profile of the above mentioned major cytoskeletal elements were evident after mechanical stretching. Moreover, immunofluorescence against the same proteins revealed no changes in their topographical organisation between stretched and unstretched cell cultures.

In contrast to the cartilaginous preformed bones of the skeleton, ossification of human thyroid cartilage begins at the end of the second decade of life. The process continues until an advanced age and shows sex differences. An in-growth of blood vessels by cartilage canals is a prerequisite for ossification. The aim of the study was to investigate the temporal and spatial appearance of cartilage canals in thyroid cartilage. Extracellular matrices of cells in cartilage canals were characterized immunohistochemically. Corrosion casts indicated that the cricothyroid branch of the superior thyroid artery was involved in vascularization of the thyroid cartilage. The first cartilage canals were detectable at the end of puberty, always lying at certain distances to almost ossified areas of the thyroid cartilage. The extracellular matrix localized at the border of the cartilage canals reacted with antibodies against type I collagen and type III procollagen, as well as with antibodies against type II collagen. This finding suggests that these cells are intermediate between fibroblasts and chondrocytes.

We report on a 2-year-old girl with chronic bullous disease of childhood (CBDC). Concomitantly with a feverish gastrointestinal infection caused by Salmonella enteritidis, the child presented with lesions resembling impetigo contagiosa on the legs and face. Following antibiotic treatment with cephalosporins p.o. and flucloxacillin i.v. she developed typical symptoms of CBDC, i.e. tense blisters on erythematous skin in the perineal area, on the flexor aspects of the thighs and upper arms and on the face. Direct immunofluorescence revealed a linear IgA deposition along the basement membrane zone, confirming the diagnosis of CBDC. These lesions cleared rapidly after treatment with dapsone p.o. This case prompted us to consider new aspects of the pathogenesis, clinical entity and treatment of this rare bullous disease of childhood.

Human herpesvirus 7 (HHV-7) has been recently isolated from CD4-positive peripheral blood lymphocytes of a healthy person. The present study was performed to find the antibody prevalence of this virus in the healthy Mexican population. Two hundred blood samples from candidates for blood donation at the General Hospital of Mexico were studied with the indirect immunofluorescence test (IFA) in HHV-7 infected SupT1 cells. 83.5% were male persons and 16.5% female, the mean age for the male group was 28.8 years and for the female group 31.5. The donors came from 12 different states in Mexico, predominantly from the city (60.8%), and had different occupations. Almost all samples (98.5%) were positive to HHV-7. Other studies done revealed 1% positive to brucella, 1% positive to Hepatitis B, 2% positive to syphilis, hepatitis C and HIV test were negative in the whole group studied. There was a high incidence of HHV-7 in the group studied: more than 50% of the subject had high titers. This results should be further studied determine titers indicative of an active infection and to search for any association with illnesses.

We report on a patient with multicentric juvenile xanthogranuloma, whose unusual clinical and histological pattern initially obscured the diagnosis. Spontaneous remission of the tumour, which has now been maintained for over 2 years, the change from the primary monomorphological fibrocytic appearance to a polymorphological histiocytic picture with multiple fat-storing giant cells, and the lacking dermonstration of specific cell organelles finally allowed the nature of the disease to the definitely diagnosed.