Analysis of the directed and nondirected movement of human granulocytes: influence of temperature and ECHO 9 virus on N-formylmethionylleucylphenylalanine-induced chemokinesis and chemotaxis.

The directed movement of human polymorphonuclear leukocytes (PMN) in a plane (Zigmond chamber assay) is described by a statistical model. We demonstrate that (a) the movement of a single cell is a superposition of a directed and a random movement, and (b) the degree of orientation, P1, of moving cells in a chemotactic gradient can be determined either by the time average of a single cell or by the average of movement of multiple cells at a fixed time (Ergoden hypothesis). However, an homogeneous cell population is a necessary condition. P1, which is identical with the McCutcheon index, is derived from the measured angular distribution function of moving cells. The statistical model allows one to distinguish between chemotaxis and chemokinesis. Applying this model to the temperature-dependent changes of cell movement, we found that P1 = 0.82 (37 degrees C) decreased to P1 = 0.4 (22 degrees C). The average speed of moving cells exhibits a very strong temperature-dependent variation from 30 microns/min (37 degrees C) to 5 microns/min (22 degrees C), indicating a different temperature dependence of chemotaxis and chemokinesis. At a fixed temperature (37 degrees C) the stability of the chemotactic gradient can also be checked by the angular distribution function. In addition, this model was applied to investigate the enteric cytopathogenic human orphan, strain 9 (ECHO 9) virus-induced disturbances of cell movement. We found: (a) The average speed of cell movement is not affected by the virus. (b) The degree of orientation is not affected for virus doses below a critical virus dose, ao (virus/PMN = 0.8:1). (c) The degree of orientation above this critical value exhibits a time- and virus-dose-dependence. (d) At a fixed viral dose, the time-dependent decrease of P1 is described by an exponential law (virus/PMN = 5:1, the characteristic time is 110 min). (e) This characteristic time investigated as a function of viral dose results in a logarithmic law analogous with the Weber-Fechner law. These findings indicate that only chemotactic and not chemokinetic response is disturbed by ECHO 9 virus.

[Molecular mechanisms and consequences of cardiac viral infections]. / Molekulare Mechanismen und Konsequenzen kardialer Virusinfektionen.

Molecular biological methods have confirmed the pathogenetic role of enteroviruses, primarily coxsackieviruses of group B (CVB), in the induction and maintenance of inflammatory cardiomyopathy. More recently, adenoviruses, various herpes viruses, and increasingly parvovirus B19 (B19) have been identified as potential cardiotropic agents. While cardiac myocytes are target cells for enterovirus and adenovirus infections with virus-induced cytolysis, B19-associated inflammatory cardiomyopathy is characterized by infection of intracardiac endothelial cells of small arterioles and veins, which may be associated with endothelial dysfunction, impairment of myocardial microcirculation, penetration of inflammatory cells, and secondary myocyte necrosis. Recent observations showed that B19 is involved in intracellular calcium regulation by the viral phospholipase. B19-induced caspase activation can lead to proinflammatory/proapoptotic processes through dysregulation of STAT signaling. These cellular interactions may contribute to mechanisms by which B19 establishes persistent infection in endothelial cells and play a critical role in viral pathogenesis of inflammatory cardiomyopathy.

Simplified detection of microsatellite instability in colorectal cancer without the need for corresponding germline DNA analysis.

A panel of five quasimonomorphic mononucleotide repeats that dispenses with the need to analyse corresponding germline DNA was proposed by Suraweera et al for the detection of high-frequency microsatellite instability (MSI) in colorectal cancer. Using this panel, a simplified and a more sensitive (compared with the original) algorithm (p<0.05) was developed to define the instability of each repeat by assessing the morphological shape of its plot and not its absolute length. 103 cases of colorectal tumours were investigated and the results compared with those obtained by the analysis of five consensus microsatellites (Bethesda reference panel). By the proposed method, a higher specificity, but no loss of sensitivity, was found. Thus, the use of the five mononucleotide repeats in combination with the modified assessment technique simplifies the assessment of MSI, while retaining the sensitivity of the Bethesda panel for the detection of high-frequency MSI.

Comparison of two mechanical ultrasound devices with and without echogenic contrast dye.

To evaluate the impact and limitations of intracoronary ultrasound, 64 segments of 13 isolated coronary arteries were examined and 2 mechanical devices (device A, 30 MHz 5Fr [CVIS] and device B, 20 MHz 4.8Fr [Diasonics/Boston Scientific] were compared with the corresponding histologic specimens. Luminal dimensions were assessed with and without echogenic contrast dye (Laevovist, Schering AG, Germany). After application of contrast dye, correlation of luminal area between histology and ultrasound was improved from r = 0.62 to r = 0.77 (device B; p < 0.05) and from r = 0.82 to r = 0.88 (device A; p = NS). Low accuracy of lumen measurements in segments < 2.5 mm could be improved by application of contrast dye. The number of quadrants in which wall thickness measurements were impossible was significantly higher for device B (n = 56; 22%) than for device A (n = 28; 11%); p < 0.01. This may be due to the different ringdown diameters of both systems (B, 2.6 mm; A, 2.0 mm; p < 0.0001). In assessing wall thickness only in segments of > 2.5 mm, a reliable correlation between ultrasound and histology was found (A, r = 0.80; B, r = 0.60). Sensitivity of plaque (n = 51) detection was lower for device B (63%) than for device A (82%, p < 0.05), and measurements correlated with histology only for device A. There are considerable differences in the accuracy of ultrasound measurements between mechanical systems. Nevertheless, additional application of contrast dye can improve accuracy of luminal measurements, especially in smaller vessels.

Investigation of bone marrow lymphocyte subsets in normal, reactive, and neoplastic states using paraffin-embedded biopsy specimens.

Bone marrow lymphocyte subsets in normal and reactive states and in neoplastic diseases involving the marrow were investigated with a select panel of monoclonal antibodies reactive on routinely processed, paraffin-embedded trephine biopsy material. In all cases, the antibodies beta F1 and UCHL1 (CD45RO) stained virtually equal numbers of T cells (reactive and neoplastic), whereas antibody OPD4 stained only about one half of this number of T cells. Antibody L26 (CD20) stained B cells (reactive and neoplastic) in all specimens. The T-cell to B-cell ratio in the normal marrow was between 4:1 and 5:1, and a significant increase in T-cell numbers was observed in reactive and myelodysplastic states. A significant increase in B-cell numbers, however, was seen only in marrow infiltrated by B-cell lymphoma. Bone marrow exhibiting infiltrates of B-cell lymphoma, acute leukemia, or myeloproliferative disorders showed normal or decreased numbers of T cells. These findings show that antibodies UCHL1, beta F1, and L26 can be used to determine the numbers of B and T lymphocytes in paraffin-embedded, formalin-fixed bone marrow specimens and thus may help to distinguish reactive T lymphocytosis from B-cell lymphoma.

Ectopic prostatic tissue in the spleen.

Ectopic prostatic tissue was found in the spleen in a 49-year-old white man who died of wide-spread malignant mesothelioma. The prostatic origin of the tissue was affirmed by positive immunohistochemical staining for prostatic specific antigen and prostatic acid phosphatase.

Extensive pectoral muscle necrosis after defibrillation: nonthermal skeletal muscle damage caused by electroporation.

A 37-year-old Italian male developed a myocardial infarct with subsequent ventricular fibrillation. He was defibrillated seven times with up to 360 Joules. Thirteen days later the patient died of recurrent myocardial infarct due to thrombotic occlusion of the left circumflex coronary artery. At autopsy, necrosis of the right pectoralis muscle was observed. Electroporation is the pathogenetic mechanism of skeletal muscle damage due to multiple defibrillations with high energy levels.

Demonstration of Epstein-Barr viral DNA in paraffin-embedded specimens of lymphoproliferative syndrome. Evidence for a productive infection comparable to lymphoblastoid cell lines.

B-cell lymphoproliferative syndromes (LPS) occurring in immunodeficient subjects are frequently associated with EBV infections. Histology as well as EBV-related serology are not diagnostic, but demonstration of EBV DNA in LPS suspected lesions might be useful for diagnosis. We studied four autopsied cases of LPS that developed in the setting of bone marrow transplantation, with proliferations ranging from poly- to monoclonal. Our protocol of DNA extraction allowed detection of EBV DNA in formalin-fixed, paraffin-embedded tissue specimens of all four cases. In dot blot hybridization the sensitivity in these specimens was 10% as compared to fresh frozen material, but still sufficient for a biotinylated probe. Southern blotting with the former DNA was not successful due to extensive degradation. In situ hybridization resulted in positive signals in all cases, using either 35S or 3H labeled probes. The labeling pattern suggested virus replication in B cells of LPS. By this, LPS resembles productively infected lymphoblastoid cell lines rather than latently EBV-infected Burkitt's lymphoma (BL). These findings strengthen the concept of LPS as a distinct clinicopathologic entity, differing from monoclonal, latently EBV-infected BL, as well as from polyclonal infectious mononucleosis, and the more common EBV-negative Non-Hodgkin lymphomas of the immunocompetent host.

Selective inhibition of human neutrophilic chemotaxis by echo virus, type 9. Virus-induced changes in membrane fluidity.

By means of the excimer-forming lipid technique changes of membrane fluidity of human PMNs exposed to a chemoattractant (FMLP) and/or Echo virus, type 9, strain A. Barty has been investigated. It was shown that preincubation of PMNs with Echo 9 virus results in a viral dose-dependent decrease of membrane fluidity of granulocytes exposed to FMLP. The virus-induced rigidity of cell membrane causes obviously a disturbance in the aggregation of membrane particles to form receptors, and thus can lead to a failure of regular processing of chemotactic signals.

[Multiple cavernous haemangiomas of the colon (author's transl)]. / Multiple kavernöse Hämangiome des Kolons.

Extensive involvement of the colon with cavernous haemangioma is described in a 26-year old man. Clinically the disease manifests itself by recurrent rectal bleeding and abdominal distension, leading to his admission. Double contrast examination of the colon showed involvement of the ascending, transverse and descending colon, the sigmoid and the rectum. Arteriography demonstrated vascular tumours in the territory of the jejunal artery, the left colic artery and the superior rectal artery. This is a rare condition which has been described only 53 times in the literature (1, 3, 5, 7). The diagnostic value of double contrast examination of the colon and of selective angiography for differential diagnosis is stressed.

Fibromuscular dysplasia of the renal artery responsible for renovascular hypertension: a histological presentation based on a series of 102 patients.

BACKGROUND: Fibromuscular dysplasia (FMD) is a rare non-atherosclerotic and non-inflammatory disease in the arterial system. The purpose of the study was a retrospective analysis of FMD in the renal artery. PATIENTS AND METHODS: A total number of 102 patients (mean age: 36.9 years) who suffered from renovascular hypertension underwent a surgical therapy. The operative specimens of the renal arteries were analysed with the lightmicroscop using histological and immunohistochemical methods. RESULTS: 101 patients (99.02%) presented a medial FMD (extensive-medial subtype in 56 patients, 54.9%, subadventitial subtype in 29 patients, 28.4% and combined subtype in 16 patients, 15.7%). In 1 patient (0.98%) an adventitial FMD was found. We observed the following complications: true and dissecting aneurysms (75 patients, 74.5%), arterio-venous fistulae (2 patients, 1.96%) and chronic thrombosis (10 patients, 9.8%). CONCLUSIONS: With the progress in angioplasty, not all patients suffering from FMD undergo a primary surgical therapy and therefore this lesion is less seen in the daily work of the histopathologist.

[Traumatic cysts of the spleen]. / Traumatische Milzcysten

During a routine clinical examination of a 7-year-old girl and a 17-year-old man a large tumor was discovered in the left upper quadrant of the abdomen. Large splenic cyst were found in the spleen at splenectomy. The traumatic etiology of the cysts can be assumed on the basis of histologic results, as well as of a significant history of previous trauma. The etiology, differential diagnosis, therapy, and classification of splenic cysts are discussed.

[Pathogenesis, diagnosis and therapy of heart disease caused by enteroviruses (molecular biological study)]. / K patogenezu, diagnostike i terapii porazheniia serdtsa, vyzvannogo énterovirusami (molekuliarno-biologicheskoe issledovanie).

Molecular hybridization studies have demonstrated that human enteroviruses, including group B coxsackieviruses (CVB), are detectable in myocardial tissue of patients with acute and chronic myocarditis. As well, such infections are observed in some patients with end-stage dilated cardiomyopathy indicating the possibility of persistent heart muscle infection. Enterovirus persistence in the human heart is supported by the discovery in various murine models of chronic myocarditis, demonstrating that coxsackievirus B3 (CVB3), typically a cytolytic virus, is capable of evading immunological surveillance in a host-dependent manner. Currently attention is focused on the analysis of molecular mechanisms of virus persistence, the characterization of viral and host factors and their impact in determining the natural course of myocardial enterovirus infections. The evidence for a causal linkage of enterovirus infection with heart muscle diseases has emerged therapeutic implications. From the view of a virologist, immunosuppressive treatment of patients revealing enterovirus infection in the myocardium with steroids is clearly contraindicated. The evaluation of potent antiviral agents, such as interferons, in established in vitro and in vivo model systems of enterovirus infection is expected to contribute significantly to new therapeutic strategies in human enteroviral heart disease.

Molecular and clinicopathological findings in a tonsillar synovial sarcoma. A case study and review of the literature.

Synovial sarcoma (SS), 3-5% of which occurs in the head and neck region, has generally been regarded as high grade sarcoma. Recent analysis of clinical, morphological, and molecular characteristics of SS, however, identified low and high risk group of patients, resulting in important implications for the treatment of patients diagnosed with SS. We describe the case of a 31-year-old male who presented with biphasic SS with poorly differentiated areas (clinical stage IIA) in a palatine tonsil, an extremely rare site of SS. Molecular analyses revealed typical t(X;18) translocation of the SYT gene and a SYT/SSX1 fusion type. The tumor was surgically resected with free margins. Adjuvant radiotherapy or chemotherapy was not considered indicated. To date, the patient has remained free of tumor for 4 years after surgery. Literature review reveals that primary tonsillar HNSS has previously been documented only in three patients. In all of these patients the tumor was histologically biphasic; however only one published case and the case presented here showed areas of poor differentiation. We discuss the relevance of the presented findings with regard to prognostic and therapeutic considerations in SS in the head and neck region.