Ouabain Enhancement of Compound 48/80 Induced Histamine Secretion from Rat Peritoneal Mast Cells: Dependence on Extracellular Sodium

Abstract: Purified populations of rat peritoneal mast cells were used to study the effect of ouabain on compound 48/80-induced histamine secretion and on ⁸⁶Rb⁺ uptake. ⁸⁶Rb⁺ was used as a tracer for extracellular K⁺. The calculated value of the ouabain-sensitive uptake of K⁺ and ⁸⁶Rb⁺ was considered a measure of the Na⁺–K⁺ pump activity of the cells. Ouabain caused an immediate inhibition of the pump activity and a time-dependent increase in histamine secretion in the absence of extracellular calcium. No effect on the secretion was observed in the presence of calcium. The effect of ouabain on the secretion occurs in the presence of sodium but not when sodium was replaced by lithium. Preservation by ouabain of a high intracellular sodium content in sodium-loaded cells was associated with preservation of the secretory response in a calcium-free medium. In the presence of lanthanum in a calcium-free medium, the pump activity was inhibited and the enhancement by ouabain of the secretion of histamine was blocked. A less marked inhibition of the pump was found in a calcium-free medium containing magnesium. The inhibition exerted by magnesium was concentration-dependent (0–5 mM) as was the counteraction of magnesium of the enhancement of ouabain of the secretion of histamine. These observations indicate that the enhancement by ouabain of the secretory response of mast cells preincubated in a calcium-free medium is associated with accumulation of sodium inside the cell. In addition to a decreased rate of sodium-calcium exchange caused by a decreased inward directed sodium gradient, the mechanism by which ouabain enhances the secretory response is likely to involve an increased binding of calcium to membrane binding sites.     

Association between alcohol consumption and aeroallergen sensitization in Danish adults

BACKGROUND: It has been proposed that alcohol consumption may be one of the lifestyle factors associated with a westernized, urban, and affluent lifestyle contributing to the rise in atopic disease. OBJECTIVE: The aim was to investigate the association between alcohol consumption and atopy (aeroallergen sensitization). METHODS: In 1982, a population-based cross-sectional study of 3608 Danes (79% of the invited), aged 30, 40, 50, and 60 years, was carried out. Information on alcohol consumption was obtained by a questionnaire. Aeroallergen sensitization was defined as a positive test for the detection of specific IgE against a panel of 19 common inhalant allergens in stored serum samples. A total of 3317 subjects with complete information on all variables were included in the analyses. RESULTS: We found a statistically significant association between alcohol consumption and aeroallergen sensitization (independent of the type of alcoholic drink consumed). This association appeared to relate only to those who consumed more than 8 drinks/week. After adjustment for confounders this association was only statistically significant for those who consumed 15-21 drinks/week (adjusted odds ratio 1.8, 95% confidence interval 1.2-2.8). CONCLUSION: In this adult general population, self-reported alcohol consumption was positively associated with aeroallergen sensitization.     

Duplex ultrasonography of the portal vein

Although the ability of ultrasonography to provide anatomic detail and physiologic information about the arterial system is well established, its applicability for the venous system and the splanchnic circulation has only recently been recognized. We have found duplex scanning, which is non-invasive, rapid, inexpensive, and reproducible, to be highly accurate in (1) establishing or confirming the diagnosis of portal hypertension, (2) demonstrating portal and splenic vein patency and direction of flow, (3) assessing portosystemic shunt patency, and (4) providing novel anatomic and physiologic information regarding the normal and diseased splanchnic venous system. These ultrasonographic techniques also have a significant role to play in the surveillance of patients who have undergone liver transplantation or massive liver resection. To a great extent, ultrasonography may supplant the invasiveness, discomfort, and expense of contrast angiography in the evaluation of many patients with advanced liver disease.     

Loss of somatal neuropeptide y immunoreactivity in the rat hippocampus following transient cerebral ischemia

Adult male Wistar rats were subjected to 20 min of global cerebral ischemia and allowed to survive for 1, 2, 4, or 21 days. The brains were processed for immunocytochemistry and the hippocampal neuropeptide Y (NPY)-immunoreactive (-i) neurons were counted and compared to control values. In order to map out the subregional distribution of ischemic cell loss in the hippocampus, cells were also counted in hematoxylin-eosin (HE)-stained brain sections processed from additional ischemic rats after 21 days survival. Cell counts demonstrated a significant loss of hippocampal NPY-i somata 1-21 days after ischemia. The ischemic loss of somatal NPY-i was in the CAI stratum oriens, the CA1 stratum radiatum, and the CA3(ab) subfield not correlated to hippocampal cell loss. NPY-i fibers were found in all subfields of the hippocampus 1-21 days after ischemia. It is known that the majority (>50%) of hippocampal somatostatin-i (SS) neurons also costore NPY-i and the SS-i neurons in the CA1 and CA3(ab) regions of the hippocampus are preserved following an ischemic insult. The present results showed a 90% ischemic loss of CA1 and CA3 NPY-i somata. Based on these findings, it is concluded that ischemia selectively damaged NPY-i and not SS-i within some surviving hippocampal neurons that co-localized both peptides prior to the ischemia.     

Novel mouse model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis

Pseudomonas aeruginosa causes a chronic infection in the lungs of cystic fibrosis (CF) patients by establishing an alginate-containing biofilm. The infection has been studied in several animal models; however, most of the models required artificial embedding of the bacteria. We present here a new pulmonary mouse model without artificial embedding. The model is based on a stable mucoid CF sputum isolate (NH57388A) with hyperproduction of alginate due to a deletion in mucA and functional N-acylhomoserine lactone (AHL)-based quorum-sensing systems. Chronic lung infection could be established in both CF mice (Cftr(tmlUnc-/-)) and BALB/c mice, as reflected by the detection of a high number of P. aeruginosa organisms in the lung homogenates at 7 days postinfection and alginate biofilms, surrounded by polymorphonuclear leukocytes in the alveoli. In comparison, both an AHL-producing nonmucoid revertant (NH57388C) from the mucoid isolate (NH57388A) and a nonmucoid isolate (NH57388B) deficient in AHL were almost cleared from the lungs of the mice. This model, in which P. aeruginosa is protected against the defense system of the lung by alginate, is similar to the clinical situation. Therefore, the mouse model provides an improved method for evaluating the interaction between mucoid P. aeruginosa, the host, and antibacterial therapy.     

Pseudomonas aeruginosa hemolytic phospholipase C suppresses neutrophil respiratory burst activity

Pseudomonas aeruginosa is a persistent pathogen in the airways of patients with cystic fibrosis or bronchiectasis from other causes and appears to have evolved strategies to survive the inflammatory response of the host. We hypothesized that the secreted hemolytic phospholipase C (PLC) of P. aeruginosa (PlcHR) would decrease neutrophil respiratory burst activity. We found that while intact wild-type P. aeruginosa cells stimulated moderate respiratory burst activity from human neutrophils, an isogenic mutant pseudomonas (DeltaHR strain) containing a targeted deletion of the plcHR operon induced a much more robust oxidative burst from neutrophils. In contrast, a second pseudomonas mutant (DeltaN) containing a disruption in the gene encoding the nonhemolytic PLC (PlcN) was not different from the wild type in stimulating neutrophil O2.- production. Readdition of purified PlcHR to the DeltaHR strain suppressed neutrophil O2.- production to levels stimulated by wild-type bacteria. Interestingly, purified PlcHR decreased phorbol myristate acetate (PMA)- but not formyl methionyl-leucyl-proline (fMLP)-induced respiratory burst activity, suggesting interference by PlcHR with a protein kinase C (PKC)-specific signaling pathway. Accordingly, the PKC inhibitor bisindolylmaleimide inhibited the oxidative burst induced by either PMA or intact pseudomonas, but not by fMLP, whereas the p38 kinase inhibitor SB-203580 fully inhibited the respiratory burst induced by fMLP or the PlcHR-replete wild-type bacteria, but not PMA or the PlcHR-deficient DeltaHR bacterial mutant. We conclude that expression of PlcHR by P. aeruginosa suppresses bacterium-induced neutrophil respiratory burst by interfering with a PKC-dependent, non-p38 kinase-dependent pathway.     

Carcinoembryonic antigen like antigen in granular cell myoblastomas

Summary A series of granular cell myoblastomas (GCM) and other benign and malignant tumours of soft tissue were examined for cytoplasmic content of carcinoembryonic antigen (CEA) by the two-layer conjugated immunoperoxidase technique. Using a commercial rabbit anti-CEA serum only granular cell myoblastomas showed positive cytoplasmic reaction. Pretreatment with periodic acid made this reaction less intense, but when the commercial rabbit anti-CEA serum was absorbed with tissue powder from normal human spleen the positive reaction was totally abolished. It is concluded that the positivity of GCM for CEA using commercial rabbit anti-CEA serum is due to the content of non-specific cross-reacting antigen (NCA) and maybe other cross-reacting glycoproteins in this tumour, and not to CEA as claimed in a previous study.