Blockade of Ca2+‐activated K+ channels in T cells: an option for the treatment of multiple sclerosis?

Voltage- and Ca(2+)-dependent K(+) channels in the membrane of both T and B lymphocytes are important for the cellular immune response. In the current issue of the European Journal of Immunology, Reich et al. demonstrate that selective blockade of the intermediate-conductance Ca(2+)-activated K(+) channel (the IK channel encoded by the KCNN4 gene) prevents cytokine production in the spinal chord and ameliorates the development of EAE caused by injection of myelin oligodendrocyte glycoprotein (MOG)(35-55) in mice. These data renew the focus on the IK channel as a potential target for the development of new immune-suppressant drugs for the treatment of autoimmune diseases.     

Pore size in implanted polypropylene filters is critical for tissue organization

Widely different implant materials induce surprisingly similar tissue reactions in vivo in contrast to their in vitro responses. Increasing attention has recently been given to the surface texture of the material. When both the material composition and the surface topography are varied, the surface topography seems to be the predominant factor for the induced tissue response. The present study addresses differences in the tissue response to commercially available Millipore mesh filters of polypropylene with pore sizes of 0.6, 10.0 or 30.0 microm. The Millipore filters with adjacent tissue were directly sectioned in a cryostat and evaluated via an immunofluorescence technique with double and triple staining, allowing simultaneous analysis of different antigens in tissue sections. These results show that macrophages, total cells, necrotic cells, nitric oxygen distribution, early angiogenesis, and capsule thickness were influenced by the surface structure. Implants with pore sizes of 0.6 microm, where entrance of inflammatory cells was inhibited, induce the most pronounced foreign body capsule formation. The 10- and 30-microm filters, in contrast, had large amounts of macrophages inside the filter structure, although very few inflammatory cells were found outside the filters. The inflammatory cells within the filters appeared not to influence the foreign body capsule induction. The critical factor for the formation of a foreign body capsule seems to be the localization of implant-close macrophages. Whether this is due to differences in cell activation or in signal transduction to collagen-synthesizing fibroblasts remains an open question.     

Increased phagocytic capacity of the blood,but decreased phagocytic activity per individual circulating neutrophil after an ultradistance run

The effect of a long strenuous endurance exercise on the phagocytic function of neutrophils was examined. 9 athletes [7 males, 2 females, age: 36–68 years, body mass: 64 (SD 10) kg, height: 175 (SD 10) cm] completed a competetive 100 km run in 8:07 (median value; range: 7:29–9:50 hours). In a whole blood assay the phagocytosis of opsonized E. coli, the receptor density of the Fc receptor 3 (CD16) and the complement receptor 3 (CD11b, direct immunofluorescence) of neutrophils were measured on a per cell basis by flow cytometry before and up to 3 hours after the race. The phagocytic rate (percentage of neutrophils incorporating bacteria) was unchanged after exercise, whereas the phagocytic activity (number of incorporated bacteria per cell) was significantly reduced by –34 (SD 8) % (Wilcoxon test, P<0.001). The total phagocytic capacity of the blood increased 2-3fold post exercise. The surface antigen expressions of CD11b and CD16 were unaffected by the ultradistance run. The results indicate either a reduced phagocytic function of neutrophils on a single cell basis or the mobilization of neutrophils of the marginal pool with a lower phagocytic activity. However, after a long endurance exercise the phagocytotic capacity of the blood was enhanced due to increased cell concentrations.     

Cellular and developmental patterns of expression of Ret and glial cell line-derived neurotrophic factor receptor alpha mRNAs

 Glial cell line-derived neurotrophic factor (GDNF) has recently been shown to signal by binding to GDNF receptor-alpha (GDNFR-α), after which the GDNF-GDNFR-α associates with and activates the tyrosine kinase receptor Ret. We have localized Ret messenger RNA (mRNA) in the developing and adult rodent and compared with to the expression of GDNF and GDNFR-α mRNA. Ret mRNA is strongly expressed in dopamine neurons and α-motorneurons as well as in thalamus, ruber and occlumotor nuclei, the habenular complex, septum, cerebellum, and brain stem nuclei. Ret mRNA was also found in several sensory systems, in ganglia, and in nonneuronal tissues such as teeth and vibrissae. Very strong Ret mRNA signals are present in kidney and the gastrointestinal tract, where Ret and GDNF mRNA expression patterns are precisely complementary. The presence of Ret protein was confirmed in adult dopamine neurons using immunohistochemistry. GDNFR-α mRNA was strongly expressed in the developing and adult dopamine neurons. It was also found in neurons in deep layers of cortex cerebri, in hippocampus, septum, the dentate gyrus, tectum, and the developing spinal cord. In the kidney and the gastrointestinal tract, GDNFR-α mRNA and Ret mRNA distribution overlapped. Dorsal root ganglia, cranial ganglia, and developing peripheral nerves were also positive. GDNFR-α was additionally found in sensory areas and in developing teeth. Sensory areas included inner ear, eye, olfactory epithelium, and the vomeronasal organ, as well as developing tongue papillae. The temporospatial pattern of expression of GDNFR-α mRNA did not always match that of Ret mRNA. For instance, GDNFR-α mRNA was also found in the developing ventral striatum, including the olfactory tubercle, and in hippocampus. These areas seemed devoid of Ret mRNA, suggesting that GDNFR-α might also have functions unrelated to Ret. Received: 2 January 1997 / Accepted: 26 February 1997     

Simulating activations with cytoarchitecture

Cytoarchitectonic delineation of areas in post-mortem human brains provides the precise location of these areas. It has been possible to study the size and location of areas between post-mortem brains with multi-subject cytoarchitectonic data. If the structure–function relationship is assumed to be a one-to-one mapping for the purposes of inter-subject variability, then functional areas in the cortex will also adhere to the structure, and therefore, the location and size of cytoarchitectonic areas in the brain. Thus, it is possible to use the cytoarchitectonic data as being representative of the size and location of functional activations. Under this assumption, we simulated activations in cytoarchitectonic areas from ten post-mortem brains in this study. We then treated these data as we would a normal PET experiment. The purpose of this study is to demonstrate a standard PET image analysis on a simulated ten-subject PET study using cytoarchitecture to localize the activations. By doing so, we simulate activations with real inter-subject variability with the size and location of each area. Significant activations were obtained for activations simulated in areas 3a and 3b. A voxel-wise conjunction between simulated data and experimental data was made to better determine the underlying areas activated by the experimental tasks. This study presents a novel technique for demonstrating the effect of standard image analysis on the location and size of simulated activations as determined by cytoarchitectonic data from multiple subjects. Furthermore, this technique has been applied to better determine the underlying areas activated in an experiment.     

Comparison of anatomical location of squamous cell carcinoma within the oral cavity and oropharynx with the incidence of in vitro hyperdiploidy

The anatomical location of the squamous cell carcinoma (SCCA) within the oral cavity and oropharynx influenced the association of SCCA with the biomarker in vitro hyperdiploidy in human dermal fibroblast cultures (IVH). There was a strong association of IVH with the occurrence of SCCA in the anterior 2/3 of the tongue, floor of the mouth and lower alveolar ridge of the oral cavity and in the base of the tongue and pharyngeal wall of the oropharynx. There was a lower association of SCCA with IVH in the tonsillar region of the oropharynx. IVH showed no association with SCCA located in other anatomical parts of the oral region. The patient group whose diagnosis of SCCA in the anterior 2/3 of the tongue occurred prior to the age of 50 years were invariably IVH-, whereas those diagnosed after the age of 50 years were IVH+, providing evidence for heterogeneity. There was no such correlation of biomarker subgrouping with age of diagnosis demonstrated for SCCA at any other anatomical location within the oral cavity or oropharynx.     

Prostaglandin content in blood and lung tissue during alveolar hypoxia

The aim of the present work was to investigate whether prostaglandins (PGs) are synthetized and released from isolated blood-perfused rat and cat lungs secondary to vasoconstriction induced by alveolar hypoxia. The lungs were perfused with autologous blood with constant volume inflow via the pulmonary artery in a recirculating system. They were ventilated with constant volume positive pressure, and acute alveolar hypoxia was induced by ventilation with a gas containing 2% O₂. A superfusion bioassay technique was used to measure PG-like activity in the perfusate from the lungs, the blood being re-oxygenated before reaching the assay tissues. The oxygenator prevented the perfusate hypoxia induced by ventilation hypoxia to affect the bioassay tissues. The assay tissues were rat stomach strip, rat colon and chick rectum. They were sensitive to calibrating doses of 0.5–1 ng/ml PGE₂ and 1–2 ng/ml PGF_2α. In another series of experiments PGs of the F-series were measured in lung tissue from normoxic and hypoxic lungs with radioimmunoassay technique. No increase in PG-like activity could be detected in the venous effluent by means of bioassay during hypoxia, nor was the lung tissue content of immunoactive PGF increased by hypoxia. The present findings indicate that alveolar hypoxia does not stimulate PG-synthesis in lungs, refuting that PGs are important mediators of the pulmonary vasoconstrictor response to alveolar hypoxia. It is concluded that PGs play no significant role in producing the pressor response to alveolar hypoxia.     

Production of prostaglandin E2 in monocytes stimulated in vitro by Chlamydia trachomatis, Chlamydophila pneumoniae, and Mycoplasma fermentans

Chlamydia trachomatis (CT) as well as Chlamydophila pneumoniae (CP) cause chronic inflammatory diseases in humans. Persistently infected monocytes are involved in the pathogenesis by inducing mediators of inflammation. An in vitro system of chlamydial persistence in human peripheral blood monocytes (HPBM) was used to investigate prostaglandin E(2) (PGE(2)) production and the expression of the key enzyme for prostaglandin production, cyclooxygenase-2 (COX-2). PGE(2) production was determined by PGE(2)-ELISA of HPBM-culture supernatants. Cox-2 mRNA expression was measured by real-time RT-PCR of total RNA isolated from HPBM. Both, CT and CP, stimulated PGE(2) production of HPBM in vitro. Equivalent numbers of CT per host cell induced a higher PGE(2)-response compared to CP. The amount of synthesized PGE(2) depended on the chlamydial multiplicity of infection (MOI). Even at an MOI of 10 the amount of CT- and CP-induced prostaglandin, respectively, was lower than the amount of prostaglandin induced by E. coli lipopolysaccharide (LPS) at a concentration of 10microg/ml. In contrast to stimulation with LPS, Chlamydia-induced PGE(2) production as well as cox-2 mRNA decreased after day 1 post infection (p.i.). These data indicate that Chlamydia stimulate PGE(2) production in human monocytes. Since Chlamydia are often contaminated by mycoplasma, the influence of mycoplasma on the prostaglandin production was investigated additionally. Mycoplasma fermentans (MF) also stimulated PGE(2) production. The co-infection of mycoplasma and Chlamydia resulted in an additive effect in the production of PGE(2). Thus it is important to use host cells and Chlamydia free of mycoplasma contamination for the analysis of Chlamydia-induced prostaglandin production.     

The use of blood components in surgical transfusion therapy

Blood components can be prepared either by separation of ordinary whole blood units or, selectively, by apheresis techniques. In recent years, new methods for improvement of quality and length of storage have been developed. The additive solution approach is now being applied increasingly. Its advantages and the difference between some available systems are described. Blood component therapy must be integrated into the patients' water/electrolyte balance and nutrition schedule. An outline is given of the role of blood components in the treatment of shock. The question of excessive bleeding, the possibilities of making the diagnosis of its cause(s) in the individual case, and the use of blood components is described. Coronary pulmonary bypass is used as an example of a complicated situation that can be handled effectively by a limited number of diagnostic and therapeutic tools.

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Resumen Los componentes sanguíneos pueden ser preparados bien por separación de unidades ordinarias de sangre total o, selectivamente, por técnicas de aferesis. Estas últimas tienen la ventaja de permitir la selección de donantes particularmente apropiados en relación a compatibilidad inmunológica o a la ausencia de agentes infecciosos transmisibles, así como a la posibilidad de que un mismo donante pueda ser usado en forma repetida dentro de un período de tiempo corto. Nuevos métodos de mejoramiento y prolongación del período de almacenamiento han sido desarrollados. El enfoque de las soluciones aditivas es empleado con creciente frecuencia; se describen sus ventajas y las diferencias con otros métodos disponibles en la actualidad.La terapia con componentes sanguíneos debe ser parte integral del manejo del equilibrio de agua y electrolitos y del programa de soporte nutricional. Se provee una guía sobre el uso de componentes sanguíneos en el tratamiento del shock.Se describe el problema del sangrado excesivo, la posibilidad de establecer el diagnóstico de su causa en cada caso individual, y el uso de los componentes sanguíneos. El bypass coronario es presentado como ejemplo de una situación compleja que puede ser manejada en forma efectiva por medio de un número limitado de métodos de diagnóstico y tratamiento.

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Résumé Les constituants du sang peuvent être isolés soit par séparation des unités de sang total ordinaire, soit sélectivement par techniques d'apharèse. Au cours des récentes années des méthodes pour améliorer la qualité et la durée du stockage ont été mises au point. Pour ce faire, l'emploi d'additifs est largement répandu. Les avantages et les différences entre quelques méthodes disponibles sont décrits par les auteurs. Le traitement par constituants du sang doit être associé à l'équilibre hydroélectrolytique et nutritif. Le rôle des différentes parties constituantes du sang dans le traitement du choc est souligné. La question de l'hémorragie excessive, les possibilités de faire le diagnostic de ses causes dans les cas individuels et l'emploi des parties constituantes adéquates du sang sont décrits. A titre d'exemple le By pass pulmo-coronarien, situation particulièrement délicate, peut être contrôlé efficacement par un nombre limité de méthodes diagnostiques et thérapeutiques.