Human salivary lactoperoxidase (HS-LP) is synthesized and secreted by the salivary glands, whereas myeloperoxidase (MPO) is found in PMN leukocytes, which migrate into the oral cavity at gingival crevices. HS-LP levels vary with changes in salivary gland function, but increased numbers of MPO-containing leukocytes indicate infection or inflammation of oral tissues. To determine the contribution of each enzyme to the peroxidase activity of mixed-saliva samples, activity was assayed at pH 5.4 with tetramethylbenzidine as the substrate, with and without the inhibitor dapsone (4,4'-diaminodiphenylsulfone). Dapsone blocked the activity of HS-LP but not MPO. The enzymes were also separated and partially purified from the soluble portion of saliva samples and from detergent extracts of the saliva sediment. Chromatographic properties of the proteins were similar to those of LP from bovine milk (BM-LP) and MPO from human leukocytes. The identity and amounts of the enzymes were confirmed by the absorption spectra and by immunoblotting with antibodies to BM-LP and human MPO. Eosinophil peroxidase (EPO), a distinct enzyme found in eosinophilic leukocytes, was not detected by chromatography or with antibodies to human EPO. On average, 75% of the activity in samples from normal donors was due to MPO and 25% to HS-LP. When corrected for the lower specific activity of HS-LP in this assay, the average amount of MPO (3.6 micrograms/mL) was twice the amount of HS-LP (1.9 micrograms/mL). The amount of MPO corresponded to 1 x 10(6) PMN leukocytes/mL of saliva. The enzymes were distributed differently: Eighty-nine percent of the HS-LP was in the soluble portion of saliva, and 78% of the MPO was in the sediment, which contained 51% of the total activity. In contrast to results obtained with PMN leukocytes from blood, detergent was not required for MPO activity to be measured in saliva, indicating that the enzyme was accessible to peroxidase substrates. The results indicate that MPO is responsible for a large portion of peroxidase-catalyzed reactions in mixed saliva. The unique function of HS-LP may be carried out within the salivary glands, prior to secretion into the oral cavity.

In secreted fluids, the enzyme lactoperoxidase (LP) catalyzes the oxidation of thiocyanate ion (SCN-) by hydrogen peroxide (H202), producing the weak oxidizing agent hypothiocyanite (OSCN-), which has bacteriostatic activity. However, H202 has antibacterial activity in the absence of LP and thiocyanate (SCN-). Therefore, LP may increase antibacterial activity by using H202 to produce a more effective inhibitor of bacterial metabolism and growth, or LP may protect bacteria against the toxicity of H202 by converting H202 to a less-potent oxidizing agent. To clarify the role of LP, the antibacterial activities of H202 and the LP-H202-SCNsystem were compared by measuring loss of viability and inhibition of bacterial metabolism and growth. The relative toxicity of H202 and the LP system to oral streptococci was found to depend on the length of time that the bacteria were exposed to the agents. During incubations of up to 4 h, the LP system was from 10 to 500 times more effective than H202 as an inhibitor of glucose metabolism, lactic acid production, and growth. However, if no more H202 was added, the concentration of the inhibitor OSCNfell because of slow decomposition of OSCN-, and when OSCNfell below 0.01 mM, the bacteria resumed metabolism and growth. In contrast, the activity of H202 increased with time. H202 persisted in the medium for long periods of time because H202 reacted slowly with the bacteria and streptococci lack the enzyme catalase, which converts H202 to oxygen and water. After 24 h of exposure, H202 was as effective as the LP system as an inhibitor of metabolism. H202 also caused a time-dependent loss of viability, whereas the LP system had little bactericidal activity. The concentration of H202 required to kill half the bacteria within 15 s was 1.8 M (6%) but fell to 0.3 M (1%) at 2 min, to 10 mM (0.03%) at 1 h, and to 0.2 mM (0.0007%) with a 24-h exposure. The results indicate that if high levels of H202 can be sustained for long periods of time, H202 is an effective bactericidal agent, and the presence of LP and SCNprotects streptococci against killing by H202. Nevertheless, the combination of LP, H202, and SCNis much more effective than H202 alone as an inhibitor of bacterial metabolism and growth.

A model baiting system suitable for the delivery of an oral rabies vaccine to freeranging raccoons (Procyon lotor) was developed and tested on barrier islands in South Carolina (USA). Features of barrier island physiography and ecology were studied relative to selective bait deployment and site biosecurity. Capture-mark-recapture data were obtained from 228 raccoons. 93 to 100% of placebo baits were consistently disturbed by 7 days post-bait deployment, and bait acceptance rates by raccoons ranged from 49 to 85%, by using a modular systems approach to select the optimum combination of bait attractant, biomarker, matrix, density, and distribution. These results suggest that a large proportion (up to 85%) of a free-ranging island raccoon population can be selectively and safely targeted, marked and monitored utilizing a proposed oral bait delivery system for recombinant or other rabies vaccines.

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The present study was conducted to determine the prevalence and significance of Pneumocystis carinii antigenemia in patients with acquired immunodeficiency syndrome (AIDS) and clinically or invasively diagnosed P. carinii pneumonitis. Single serum specimens from 20 AIDS patients invasively examined for P. carinii organisms and 106 AIDS patients with a clinical diagnosis only of P. carinii pneumonitis were blindly tested for P. carinii antigenemia by a counterimmunoelectrophoresis assay. In the 20 specimen-documented cases, the antigen test demonstrated a sensitivity of 75% and a specificity of 90%. The positive predictive value of the test was 90%, while the negative predictive value was 70%. In AIDS patients with specimen-documented P. carinii pneumonitis, the prevalence of P. carinii antigenemia coincided almost exactly with the prevalence of positive invasively obtained specimens (60 and 59%, respectively). In patients with a clinical diagnosis only of P. carinii pneumonitis, half as many (30%) were found to exhibit antigenemia. Sequential P. carinii antigen titers determined by a new latex agglutination technique on three AIDS patients with specimen-documented P. carinii pneumonitis demonstrated the influence of specific therapy upon P. carinii antigenemia and its potential prognostic application.