Differential regulation of TNF-alpha production by listeriolysin-producing versus nonproducing strains of Listeria monocytogenes.

Only Listeria monocytogenes that produce listeriolysin O (LLO) elicit protective immunity. Given the importance of tumor necrosis factor alpha (TNF-alpha) in anti-Listeria immunity, we have investigated TNF-alpha production by macrophages after they ingested live LLO-producing compared to LLO-non-producing bacteria. We used two genetically engineered strains of Listeria that differed only in their ability (Ly+) or inability (Ly-) to produce LLO. Ly+ and Ly- caused the same kinetics of increased mRNA abundance for TNF-alpha during the first 90 min after phagocytosis. However, only Ly+ caused sustained transcription of TNF-alpha mRNA, and this may account for the increased release of TNF-alpha. The transcriptional inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) prevented the sustained abundance of cytokine mRNA 20 h after ingestion of Ly+. In addition, nuclear run-on assays indicated sustained transcription of TNF-alpha genes only after ingestion of Ly+. LLO itself was not responsible for the ability of Ly+ to stimulate the sustained transcription of the TNF-alpha genes. Instead, LLO may allow Listeria to survive within macrophages so that other bacterial products cause sustained TNF-alpha gene transcription. Both Ly+ and Ly- produced molecules, isolated by 50% ammonium sulfate, that induced cytokine production. In conclusion, we now report that Ly+ causes sustained transcription of the TNF-alpha gene and production of TNF-alpha by macrophages in vitro. We speculate that the TNF-alpha may activate endothelium and thus allow the recruitment of T cells to sites of infection. This may contribute to the ability of only LLO-producing Listeria to induce protective immunity.

Docosahexaenoic acid, a major constituent of fetal serum and fish oil diets, inhibits IFN gamma-induced Ia-expression by murine macrophages in vitro.

Decreased Ia expression by macrophages may account for the increased susceptibility of fetuses and neonates to infection. We chose to investigate the role of docosahexaenoic acid (DHA), an omega-3 fatty acid, on Ia expression in vitro, because neonatal serum concentrations of DHA (100-150 microM) are approximately 50 times higher than in the adult. In addition, DHA is a major component of fish-oil diets that ameliorate some autoimmune diseases and prevent renal allograft rejection. DHA inhibited IFN-gamma-induced Ia expression with a half-maximal inhibitory concentration of 25 microM. The inhibition was not caused by nonspecific damage, because oxidative metabolism via the mitochondrial electron-transport chain was not inhibited. There were strict biochemical requirements for inhibition of Ia expression. Polyenoic fatty acids with 22 carbons were more inhibitory than those with 20 carbons. Among 22-carbon fatty acids, those with more double bonds, and, in particular, with a double bond in the omega-3 position, were more inhibitory. Although DHA is known to inhibit cyclooxygenase and thus the production of eicosanoids, indomethacin did not inhibit Ia expression. This indicated that inhibition of cyclooxygenase was not responsible for inhibition of Ia expression. We divided induction of Ia expression by IFN-gamma into four phases, with IFN-gamma being present only during the second phase. DHA was most inhibitory when given before or with the IFN-gamma. This indicated that DHA inhibited early steps in IFN-gamma-induced Ia expression. Consistent with this idea, we found that DHA inhibited the increase in mRNA transcripts for Ia beta b, as assayed by Northern blotting. In summary, we found that DHA, a major component of fetal and neonatal sera as well as fish-oil diets, inhibited IFN-gamma-induced macrophage Ia expression in vitro by preventing increases in Ia mRNA transcripts.

Radiographic appearance of titanium alloy prefabricated posts cemented with different luting materials.

Eight luting materials were compared to determine which of them obscured the radiographic image of a cemented titanium alloy prefabricated post. Eight extracted, endodontically treated, maxillary molar teeth were selected for the study. A post channel was prepared in the lingual root canal of each tooth and a titanium alloy prefabricated post was cemented with one of the selected luting materials. Preoperative, precementation, and postcementation radiographs were compared visually to determine the outline of the cemented titanium alloy post. Glass ionomer cements obscured the outline more than composite resin cements but less than zinc phosphate or polycarboxylate cements. Further study is indicated before recommendation of a luting material with titanium alloy prefabricated posts.

Effect of inhaled preservatives on asthmatic subjects. II. Benzalkonium chloride.

Benzalkonium chloride has been used as a preservative in some antiasthma respirator solutions and is known to cause bronchoconstriction in asthmatic subjects. To increase understanding of how it causes bronchoconstriction, the characteristics of airway response in 28 asthmatic subjects were documented. Subjects inhaled histamine, in doses ranging from 0.03 to 7.8 mumol, or benzalkonium in doses ranging from 0.04 to 5.33 mumol on separate days. The dose of histamine or benzalkonium that caused a 20% fall in the 1-s forced expiratory volume (PD20FEV1) was measured. All subjects responded to histamine, with PD20FEV1 values in the range of 0.14 to 7.8 mumol and 17 responded to benzalkonium, with PD20FEV1 values in the range 0.35 to 5.55 mumol. Subjects who responded to benzalkonium were more sensitive to histamine (mean PD20FEV1 0.44 mumol) than subjects who did not respond (mean PD20FEV1 1.84 mumol) and, among the benzalkonium responders, there was a significant correlation between PD20FEV1 values for histamine and benzalkonium (r = 0.5, p less than 0.05). Inhalation of benzalkonium enhanced subsequent responses to histamine, causing a decrease in mean PD20FEV1 from 0.51 to 0.18 mumol histamine (p less than 0.001), but did not alter subsequent responses to benzalkonium. The response to benzalkonium reached a maximum 1 min after inhalation and was slow to recover, taking up to 60 min to return to baseline values. Response to benzalkonium was inhibited by 8 mg cromolyn sodium but not by 160 micrograms ipratropium bromide. The characteristics of the response to benzalkonium suggest a mechanism of action via release of mediators.

Clinical observations of a titanium-alloy prefabricated post.

Clinical observations led to the development of this article in order that the dental profession may become cognizant of the ethereal radiographic appearance of titanium-alloy posts.

A mathematical method for calculating the compensating anterior stop pin setting of a semiadjustable arcon articulator.

The geometry of a semiadjustable arcon articulator and development of mathematical formulae to calculate the setting of the anterior stop pin that compensates for the thickness of the interocclusal centric relation record are discussed. Measurements of the facebow mounted maxillary cast and interocclusal centric relation record are used in these formulae. Testing on 30 diagnostic mountings indicated that the formulae can reliably predict the amount of anterior stop pin opening necessary to allow near parallelism of the maxillary and mandibular bows of the articulator when the interocclusal centric relation record is removed and the mounted casts are brought into occlusion. The vertical component of dental malocclusion is identified as a source of variability in the setting of the anterior stop pin and therefore in the geometry of the articulator; it is defined as a need for further study.