Analytical accuracy of determinations of aminoglycoside concentrations by enzyme multiplied immunoassay, fluorescence polarization immunoassay, and radioimmunoassay in the presence of heparin

The accuracy of gentamicin, netilmicin, and tobramycin concentration determinations by enzyme multiplied immunoassay technique (EMIT; Syva Corp., Palo Alto, Calif.), fluorescence polarization immunoassay (TDx; Abbott Diagnostics, Irving, Tex.), and radioimmunoassay were compared in the presence of 0 to 3,000 USP units of porcine heparin per ml. Gentamicin, netilmicin, and tobramycin concentrations determined by EMIT decreased by 10 and 50% in the presence of 75 and 1,000 USP units/ml, 2 and 5 USP units/ml, and 2 and 7.5 USP units/ml, respectively. Accuracy of the TDx and radioimmunoassay determinations, however, were not affected by the presence of heparin. Blood samples for the determination of gentamicin, netilmicin, and tobramycin by EMIT should not be collected in evacuated heparinized tubes.     

Antimicrobial activity of heparin

Single clinical isolates of eight species of microorganisms were incubated in solutions of heparin and brain heart infusion broth at various concentrations to determine the possible antibacterial effect of heparin. At heparin concentrations ranging from 12.5 to 400 U/ml, the effect on Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Staphylococcus aureus, and S. epidermidis varied with brain heart infusion broth concentrations of 1.2 to 10% and inocula of 10² to 10⁶ colony-forming units per ml; similar effects were not observed with Klebsiella pneumoniae, Enterobacter aerogenes, and Citrobacter spp. The minimal inhibitory concentrations of heparin for ten strains of each species were determined in 2.5% brain heart infusion broth with inocula of 10⁴ colony-forming units per ml. All 10 isolates of S. aureus and all 10 of S. epidermidis were inhibited by heparin concentrations of 125 to 500 U/ml. Three E. coli, four P. aeruginosa, and nine C. albicans strains were inhibited by ≤500 U of heparin per ml. None of the K. pneumoniae, E. aerogenes, Enterobacter cloacae, and Citrobacter spp. was inhibited by heparin at ≤500 U/ml. Heparin inhibition of S. aureus in 2.5% brain heart infusion broth-500 U of heparin per ml could be quantitatively overcome by addition of magnesium, calcium, or magnesium and calcium. These data suggest that the growth of microorganisms from heparin-containing material may be suppressed.     

Prolonged stability of stored vancomycin, gentamicin, and heparin for use in the antibiotic-lock technique

The antibiotic-lock technique has been effective in salvaging tunneled catheters in hemodialysis patients with bacteremia. However, a practical concern exists with respect to the stability of the antibiotics and heparin in normal saline, when stored for a prolonged period. Vancomycin, gentamicin, and heparin were diluted in normal saline to a final concentration of 100 microg/ml of each antibiotic and 5000 units/ml heparin. Fresh samples, and samples refrigerated at 4 degrees C for 48 hours, 1 week, 2 weeks, 3 weeks, and 4 weeks, were assayed (in triplicate) for gentamicin and vancomycin concentration and bactericidal activity (Schlichter test) using methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa. An anti-Xa activity assay was used for monitoring heparin anticoagulant activity of the fresh samples and samples refrigerated for 2 and 4 weeks. Mean (+/- SD) anti-Xa activity for heparin/vancomycin solution was 7900 +/- 173 u/ml, and for heparin/gentamicin solution was 7467 +/- 751u/ml; both were stable over a 4-week storage period. Mean bactericidal titer for vancomycin was 1:121 +/- 11, and for gentamicin was 1:242 +/- 22; both were stable over a 4-week storage period. Mean vancomycin concentration was 97 +/- 4 microg/ml, and gentamicin concentration was 86 +/- 3 microg/ml; both were stable over a 4-week storage period. Vancomycin and gentamicin in a heparin/saline solution can be stored at 4 degrees C for up to 4 weeks without adversely affecting antibiotic concentration, bactericidal activity, or heparin anticoagulant activity.     

Effect of heparin on incorporation of thymidine-3H into A-1 cells in continuous culture

The effect of heparin on incorporation of thymidine-³H into A-1 cells in continuous culture was studied. Incubation of the cells in medium No. 199 containing heparin (1–200 units/ml) did not lead to any decrease in the percentage of labeled nuclei. The duration of the C₂-and S-periods likewise was unchanged compared with the control. However, the intensity of incorporation of thymidine-³H (mean number of grains above one labeled nucleus) was significantly reduced. The results indicate that heparin affects the permeability of the plasmalemma but has no effect on DNA synthesis. The antimitotic action of heparin is probably connected with its effect on the cell surface.Laboratory of Pathomorphology, Institute of Transplantation of Organs and Tissues, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 9, pp. 1119–1121, September, 1976.     

The contact phase of coagulation in the presence of heparin

The effect of heparin on the contact phase of coagulation has been investigated by a technique utilizing a solution of toluidine blue in calcium chloride. Heparin in a concentration of 7 units/ml. of plasma does not inhibit contact activation by glass. It is suggested that heparin does not prevent the activation of factor XII (Hageman factor) by glass or the subsequent formation of active factor XI (plasma thromboplastin antecedent).     

Heparin uncouples the muscarinic receptors from GK protein in the atrial cell membrane of the guinea-pig heart

The effects of heparin on activation of the G protein-gated muscarinic K⁺ channel were examined in atrial cells of guinea-pig heart. The inside-out patch clamp technique was used. The pipette solution contained 1.1 M acetylcholine (ACh). In the inside-out patches, intracellular GTP activated the muscarinic K⁺ channel. When heparin (0.05–5 units/ml) was further added to the intracellular side of the patch membrane, the channel openings were depressed in a concentration-dependent fashion. The effects of heparin were reversible after wash-out. Heparin did not affect GTP-S-induced activation of the K⁺ channel. Therefore, it is suggested that heparin may uncouple the muscarinic receptors from G_K protein in the cardiac atrial cell membrane.     

Untersuchungen zur kontinuierlichen Heparindosierung bei intermittierenden Dauerdialysen

Zusammenfassung Bei 6 Patienten in der intermittierenden Dauerdialysebehandlung (Kiil-Dialysator) wurden quantitative Heparinbestimmungen im Blut bei verschiedenen Heparininfusionsgeschwindigkeiten durchgeführt. Es zeigte sich, daß mit einer Anfangsdosis von 50 mg Heparin und einer konstanten Zufuhr von 0,3 mg Heparin/min die Konzentration in den meisten Fällen im Bereich von 5–10 Heparin/ml Blut gehalten werden konnte. Mit häufigen Gerinnungskontrollen und entsprechenden Änderungen der Heparininfusionsgeschwindigkeit war der Heparinverbrauch nicht wesentlich geringer als mit konstanter Zufuhr ohne Gerinnungsbestimmungen.Der Abfall der Heparinkonzentration im Blut nach Abschluß der Dialyse erfolgte bei drei oligo-anurischen und zwei doppelseitig nephrektomierten Patienten in gleicher Weise wie bei Normalpersonen. Dies spricht für eine überwiegend extrarenale Elimination von Heparin.

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Summary Quantitative determinations of heparin in blood at different heparin infusion rates were done in 6 patients during regular hemodialysis treatment (Kiil dialyser). It was shown that in most cases the concentration of heparin was kept between 5 and 10 /ml by an initial dose of 50 mg of heparin and by a constant infusion rate of 0,3 mg of heparin/min. By frequently checking blood clotting times with corresponding adjustments of the heparin infusion rate consumption of heparin was not significantly less than using a constant infusion rate without checking blood clotting times.The decrease of the heparin concentration after the dialysis was the same in 3 oligo-anuric and 2 nephrectomised patients as in normal persons. This indicates a predominantly extrarenal elimination of heparin.

     

Priming of human neutrophil functions by tumor necrosis factor: Enhancement of superoxide anion generation,degranulation, and chemotaxis to chemoattractants C5a and f-Met-Leu-Phe

Recombinant human tumor necrosis factor- (rTNF) stimulated increased generation of Superoxide anion (O ₂ ^– ) by human neutrophils in a concentration-dependent fashion. Preincubation of human neutrophils with rTNF (2.2–2200 units/ml) for 10 min enhanced the subsequent generation of O ₂ ^– in response to C5a and f-MetLeu-Phe(FMLP). Recombinant TNF did not enhance O ₂ ^– generation by neutrophils stimulated with phorbol myristate acetate (PMA). Recombinant TNF alone failed to induce release of myeloperoxidase (MPO) and lysozyme by neutrophils. However, it did enhance the release of MPO and lysozyme by neutrophils stimulated with C5a and FMLP, but not with PMA. Although rTNF alone (0.001–50,000 units/ml) was not chemotactic for neutrophils, preincubation of neutrophils with rTNF (0.001–0.1 units/ml) enhanced the chemotactic activity of suboptimal concentrations of C5a (0.1 nM) and FMLP (5 nM). Neutrophils treated with high concentrations of rTNF (100–10,000 units/ml) showed inhibition of random movement and of chemotaxis induced by C5a or FMLP. We conclude from these studies that rTNF primes neutrophils for enhanced responses to subsequent stimuli and thus may augment the inflammatory response by increased oxidant production and lysosomal enzyme release and promote down-regulation of chemotactic movement.     

Activation of parathyroid hormone by heparin in vitro

Pieces of the parietal bone of inbred C57BL/Mib mice aged 3 days were grown in culture for 3 days. The following substances were added to the culture medium: 0.01 unit/ml (series I) and 0.1 unit/ml (series II) parathormone, 0.1 unit/ml (series III) and 1 unit/ml (series IV) heparin, and 0.01 unit/ml parathormone +0.1 unit/ml heparin (series V). Resorption of bone tissue of the explants was observed in the experiments of series II and V, but not in those of series I, III, and IV. Parathormone (0.01 unit/ml), combined with heparin (0.1 unit/ml), stimulated resorption of the bone tissue of the explant (series V), whereas if added separately in the same dose it had no such action.Laboratory of Experimental Genetics, Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 80, No. 9, pp. 97–99, September, 1975.     

A low molecular weight heparin in hemodialysis

Summary The purpose of our study was to check whether the dosage recommended for the low molecular weight heparin tested here, i.e., 50% of the corresponding unfractionated heparin dose, is adequate to prevent clot formation in the extracorporeal system. Sixteen dialysis treatments of 4–5 h were given to each of six chronic dialysis patients. In dialyses 1, 2, 15 and 16 unfractionated heparin (initial dose 35 IU/kg, continuous dose 20 IU/kg/h) was given, and in dialyses 3–14 low molecular weight heparin (initial dose 17.5 anti-Xa U/kg, continuous dose 10 anti-X U/kg/h). At these dose levels of low molecular weight heparin, clot formation occurred in the extracorporeal system in five of the six patients, despite the fact that the plasma anti-Xa level of 0.5 U/ml recommended by the manufacturer had been attained. For this reason the continuous dose of low molecular weight heparin had to be raised to approx. 80% of the corresponding continuous dose of unfractionated heparin. A plasma anti-Xa level of 0.7 U/ml is necessary to prevent extracorporeal clot formation.Abbreviations anti-Xa U Anti-factor Xa unit - aPTT Activated partial thromboplastin time - AT III Antithrombin III - IU International unit - LMWH Low molecular weight heparin - UFH Unfractionated heparin     

Long term catheterization of the portal vein in cats

Summary A method of chronic catheterization of the major blood vessels, including the portal vein, may be successfully used for studying the interstitial metabolism and hemodynamics. For catheterization of the portal vein in cats in conditions of chronic experiment we used catheters made of polyethylene, 1–1.5 mm in diameter and 120–180 mm in length with a dilatation at the peripheral end. The catheter was introduced surgically through one of the branches of the mesenteric vein with the aid of a special guide. To avoid thrombosis during the postoperative period the catheters were washed once in 2–3 days with heparin solution (400–500 units/ml).(Presented by Academician V. N. Chernigovskii) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 55, No. 1, pp. 122–123, January 1963     

Testing small insulin concentrations by means of isolated epididymal rat fat

Summary The study of glucose uptake by rat epididymal fat has demonstrated that the effect occurs at concentrations above 10 units per ml; it then increases progressively up to a concentration of 100,000 units per ml. Within a range of concentration from 50 to 100,000 units per ml, there is a linear relationship between the logarithm of the insulin concentrations in the medium and its effect; the correlation co-efficient is +0.889. A regression equation was calculated which gave a mean error for the determination of the insulin concentration of 1.78. Thus, this method is not only simple, but is also sufficiently precise for the determination of insulin concentrations in an incubation medium.(Presented by Active Member AMN SSSR V. G. Baranov) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 52, No. 7, pp. 121–124, July, 1961     

Charge-related alterations of the cerebral endothelium

In a short-term rat brain perfusion model, the luminal surface of cerebral endothelium was exposed to the following solutions: (a) the polycation protamine sulfate (PS) in a dose of 50, 100, and 500 micrograms/ml for 1 or 2 minutes; (b) PS in a dose of 100 micrograms/ml (or 500 micrograms/ml) for 1 or 2 minutes followed by the polyanion heparin in an equivalent dose of 12 units/ml (or 60 units/ml) for 1 or 2 minutes; (c) heparin alone for 1 or 2 minutes, and (d) Krebs-Ringer-bicarbonate solution as control for 1, 2, or 4 minutes. We studied in the cerebral endothelium: (a) structural alterations by electron microscopy, (b) permeability changes to horseradish peroxidase (HRP) by light and electron microscopy, and (c) charge alterations of luminal surface visualized with colloidal iron at pH 1.8 by electron microscopy. We found that: (a) PS resulted in extravasation of HRP throughout the perfused hemispheres in a time- and dose-dependent fashion. In this experimental group, colloidal iron binding decreased on the luminal surface of the cerebral blood vessels; (b) heparin perfusion following PS reversed the colloidal iron staining but failed to prevent the blood barrier opening to HRP; (c) heparin perfused alone also induced extravasation of HRP in the treated brain hemispheres; (d) in Krebs-Ringer-bicarbonate-perfused control brains extravasation of HRP was encountered only in occasional vascular segments. In all brain hemispheres showing tracer extravasation, electron microscopy revealed HRP reaction product in compartments of endothelial tight junctions suggesting opening of interendothelial routes as the structural basis of blood-brain barrier opening. Endothelial cell death reflected by swelling and influx of HRP into endothelial cytoplasm in PS- and/or heparin-perfused hemispheres was probably an additional mechanism explaining tracer extravasation into the neuropil. Our results indicate a correlation between the effect of polycation PS and a decrease in the anionic sites of cerebral endothelium. The relationship between charge alteration and barrier opening in the short-term rat brain perfusion model is not clear.     

Effect of prostacyclin on the circulatory histamine during cardiopulmonary bypass

Changes in plasma histamien levels were studied in sixteen dogs under cardiopulmonary bypass (CPB) or bypass with the addition of prostacyclin (PGI₂) infusion. In both groups, plasma histamine rose immediately after anaesthetic induction (median above pre-induction: 0.4 ng/ml) and following heparin infusion (median above pre-induction: 0.7 ng/ml). During CPB, plasma histamine levels were elevated throughout the 90 minute period of perfusion (median above pre-induction: 0.5–1.2 ng/ml). PGI₂ infusion reduced the elevation in plasma histamine levels (median above pre-induction: 0–0.7 ng/ml). These data support the hypothesis that histamine release occurs during CPB. Platelet aggregation in the extracorporeal circuit may be contributory since prostacyclin premedication reduces histamine release.     

Relationship Between the Concentration of Bacteria in Saliva and the Colonization of Teeth in Humans

The relationship between the salivary concentration of bacteria and their number that can be recovered from tooth surfaces has been studied in 12 human subjects. The mean salivary concentration of naturally occurring Steptococcus mutans and lactobacilli, determined on selective media, was 3.7 × 10⁵ and 3.8 × 10⁵ colony-forming units (CFU) per ml, respectively. In subjects with salivary concentrations of S. mutans of about 10⁴ CFU/ml or less or about 10⁵ CFU/ml or less of lactobacilli, these organisms could not be isolated from cleaned teeth after 2 to 3 h of oral exposure. In experiments with streptomycin-labeled S. sanguis cells held in the mouth for 15 min, the minimal salivary concentration required for their recovery from the teeth was about 10³ CFU/ml. Both S. mutans and lactobacilli were found to be highly localized on teeth. This evidence suggests that the concentrations of S. mutans and lactobacilli generally present in saliva are insufficient for the initiation of their firm attachment to relatively nonretentive tooth surfaces. The low efficiency of their intraoral spread, as suggested by their highly localized distribution on teeth, or of their transmission between subjects may be essentially due to the interrelated factors bacterial affinity and number of colony-forming units available for attachment.     

Blood transfusion requirements in coronary artery surgery with and without the activated clotting time (ACT) technique

Summary Control of anticoagulation during cardiopulmonary bypass (CPB) with the automated activated whole blood clotting time (ACT) and reversal of heparin after CPB using a computerized ACT dose-response curve method resulted in significant reductions of blood transfusion requirements, surgical time, and protamine doses in 150 patients undergoing coronary artery bypass grafting procedures (ACT group) as compared to 200 patients for whom a standard fixed dose protocol for heparin and protamine was used (control patients). Mean transfusion requirements were 1,938±60 SEM ml whole blood and 853±48.3 SEM ml red blood cells for control patients and 1,397±59 SEM ml whole blood (P<0.001) and 695±34 SEM ml red blood cells (P<0.01) in the ACT group. ACT group patients also required less protamine with 26.2±0.60 SEM ml Protamine 1,000 (Roche) as compared to 33.9±0.49 SEM ml for control patients (P<0.001) but more heparin with 31,440±783 SEM I.U. versus 26,760±263 SEM I.U. (P<0.001). Surgical time decreased from 321±5.5 SEM min for control patients to 289±5.4 SEM min for ACT group patients (P<0.001).Abbreviations AB autologous blood - ACD right coronary artery - ACT activated clotting time - ACT_o ACT — before heparin administration - ACT₃₆₀ ACT — 5 min. after 360 I.U. heparin/kg body wt. - CPB cardiopulmonary bypass - Cx circumflex branch of the left coronary artery - DIAG diagonal branch of the left coronary artery - ECC extracorporeal circulation - FB fresh blood - FFP fresh frozen plasma - POD postoperative day - RBC red blood cells - RIA descending branch of the left coronary artery - RIP posterior descending branch of the right coronary artery - WB whole blood     

Immune Responses to Vi Capsular Polysaccharide Typhoid Vaccine in Children 2 to 16 Years Old in Karachi,Pakistan, and Kolkata,India

The geometric mean concentration (GMC) and the proportion maintaining a protective level (150 enzyme-linked immunosorbent assay (ELISA) units [ELU]/ml) 2 years following a single dose of 25 μg of injectable Vi capsular polysaccharide typhoid vaccine was measured against that of the control hepatitis A vaccine in children 2 to 16 years old in cluster randomized trials in Karachi and Kolkata. The GMC for the Vi group (1,428 ELU/ml) was statistically significantly different from the GMC of the control hepatitis A vaccine group (86 ELU/ml) after 6 weeks. A total of 117 children (95.1%) in the Vi group and 9 (7.5%) in the hepatitis A group showed a 4-fold rise in Vi IgG antibody concentrations at 6 weeks (P < 0.01). Protective antibody levels remained significantly different between the two groups at 2 years (38% in the Vi vaccine groups and 6% in the hepatitis A group [P < 0.01]). A very small proportion of younger children (2 to 5 years old) maintained protective Vi IgG antibody levels at 2 years, a result that was not statistically significantly different compared to that for the hepatitis A group (38.1% versus 10.5%). The GMCs of the Vi IgG antibody after 2 years were 133 ELU/ml for children 2 to <5 years old and 349 ELU/ml for children 5 to 16 years old. In conclusion, Vi capsular polysaccharide typhoid vaccine is immunogenic in children in settings of South Asia where typhoid is highly endemic. The antibody levels in children who received this vaccine remained higher than those in children who received the control vaccine but were significantly reduced at 2 years of follow-up.     

Colonization of Teeth in Humans by Streptococcus mutans as Related to Its Concentration in Saliva and Host Age

The relationship of the salivary concentration of Streptococcus mutans and host age to the colonization of this organism on erupting teeth was studied in humans. Plaques were obtained from fissures and buccal surfaces of erupting permanent first and second molars of children 6 to 7 and 11 to 14 years old, respectively. In subjects of both age groups with salivary S. mutans concentrations below 5 × 10² colony-forming units per ml, the organism was detected on only a few of the tooth surfaces; at concentrations of 5 × 10² to 4.9 × 10⁴ colony-forming units per ml more than half of the surfaces and at concentrations of 5 × 10⁴ colony-forming units per ml or higher most of the surfaces were colonized by S. mutans. The frequency of detection and concentration of S. mutans in plaque as well as its concentration in saliva were higher in the case of the older children. However, when younger and older children with similar salivary S. mutans concentrations were compared, S. mutans was more frequently isolated from plaque from older children only in the case of children with below 5 × 10² colony-forming units per ml of saliva. Eleven of the 64 children studied had low or undetectable S. mutans levels in plaque and saliva. The salivary S. mutans levels of the parents of these children were lower than those of parents of a group of children with normal oral S. mutans levels.     

Clostridial collagenase

Leukocyte chemoattractants markedly alter the morphology and membrane functions of leukocytes. Bacterial collagenase causes a change in cell shape similar to that seen with the leukocyte Chemoattractant, f-Met-Leu-Phe, and also promotes capping of concanavalin A. Human neutrophils in suspension or adherent to cover glasses were exposed to clostridial collagenase (10–250 units/ml) for up to 30 min at 37°C and then fixed. Collagenase (125 units/ml) caused more than 85% of PMNs to assume an asymmetric or motile morphology even in the presence of 1% gelatin or 10 mg/ml bovine serum albumin. Trypsin alone (0.01–1%) did not induce a shape change. A similar morphology was seen in some untreated PMNs (less than 5 % of all cells) and is characteristic of f-Met-Leu-Phe-treated cells (more than 90%). Collagenase inhibitors (i.e., reduced glutathione, cysteine, and acid-soluble collagen), however, prevented the shape change induced by collagenase but not by f-Met-Leu-Phe. At 4°C, fluorescein-Con A (20 g/ml) bound uniformly to both untreated and collagenase-treated cells. Upon further incubation at 37°C, Con A was internalized over the entire cell periphery of the rounded, untreated cells but on collagenase-treated PMNs was rapidly gathered into a cap overlying the uropod or protuberant region of cytoplasm where it was subsequently internalized. Checkerboard Boyden chamber assays showed clostridial collagenase to be chemokinetic and chemotactic for human PMNs. In receptor binding experiments, the clostridial collagenase preparation competed poorly with [¹²⁵I]formylhexapeptide for binding to PMN formylpeptide receptors (less than 15% reduction in binding at 200 units/ ml collagenase). Thus, collagenase does not seem to interact strongly with the neutrophil formylpeptide receptor and may stimulate PMN motility by interacting at an altogether different site.     

Ureaplasma diversum infection in vitro alters prostaglandin E2 and prostaglandin F2a production by bovine endometrial cells without affecting cell viability.

Bovine epithelial and stromal cells of the endometrium were inoculated with Ureaplasma diversum, pathogenic strain 2312, at 10(6) or 10(3) color-changing units (ccu)/ml in the presence of 1% fetal bovine serum (depleted of steroids by dextran-charcoal treatment) to assess the effect of infection on prostaglandin biosynthesis. When the inoculum of U. diversum was 10(6) ccu/ml, the concentration of U. diversum in the culture medium decreased with time. U. diversum was found on the epithelial and stromal cell monolayers, increasing in titer 100-fold, indicating that attachment and eventually growth occurred. When the inoculum was 10(3) ccu/ml, the titer of U. diversum remained the same or increased in the supernatant and increased on epithelial and stromal cells. The effect of infection was evaluated by measurement of the primary prostaglandin produced by each cell type, prostaglandin F2a for epithelial cells and prostaglandin E2 for stromal cells. Infection with U. diversum significantly decreased prostaglandin F2a accumulation, by 44.7% +/- 6.0% at 10(6) ccu/ml (P < or = 0.005) and 15.8% +/- 5.3% at 10(3) ccu/ml (P < or = 0.05) in epithelial cells. Prostaglandin E2 accumulation by stromal cells was decreased by 34.0% +/- 4.0% at 10(6) ccu/ml (P < or = 0.001) and by 13.5% +/- 2.7% at 10(3) ccu/ml (P < or = 0.005). Infection with 10(6) ccu/ml did not alter endometrial cell viability, as shown by protein measurement, trypan blue dye exclusion, and cell plating efficiency tests. Thus, alterations in prostaglandin production were not due to cell deterioration. These observations suggest that U. diversum can alter prostaglandin E2 and prostaglandin F2a patterns in primary cultures of bovine endometrial cells without affecting cell viability.