Incomplete Freund's adjuvant enhances locomotor performance following spinal cord injury.

Following spinal cord injury (SCI), the pathological sequelae which ensue through the secondary mechanisms of degeneration produce myelin deposits which are potent inhibitors of endogenous neuroregeneration. We have enhanced the immune-mediated response following a hemisection lesion by immunizing adult C57Bl/6 female mice against the inhibitor of neurite outgrowth Nogo-A(623-640) peptide. Moderate anti-Nogo-A(623-640) antibody titre levels were obtained by using Montanide as the adjuvant. However, this antibody response was not obtained using incomplete Freund's adjuvant (IFA). Significant benefit in locomotor performance was demonstrated only in animals which were vaccinated with IFA and not with Montanide. No further benefit could be demonstrated with the Nogo-A(623-640) peptide beyond that seen for adjuvant alone. These data imply that generating antibodies against Nogo-A(623-640) in vivo alone is not sufficient to enhance locomotor recovery and that subcutaneous injection of IFA prior to SCI can enhance locomotor performance.

The hemostatic effects of desmopressin on patients who had total joint arthroplasty. A double-blind randomized trial.

The effects of desmopressin on postoperative bleeding and postoperative transfusion requirements were studied in ninety-two hemostatically normal patients who had had an elective primary total hip or total knee arthroplasty. The patients were randomized into either a placebo or a desmopressin group in a double-blind prospective clinical trial. During closure of the wound, desmopressin (0.03 microgram per kilogram of body mass) or the placebo was infused into a peripheral vein over a twenty-minute period. Compared with the placebo, desmopressin did not significantly decrease blood loss or transfusion requirements, and it did not affect the postoperative platelet or fibrinogen levels or the bleeding time. The results were no different even when the treatment and control groups were matched according to surgeon, use of cement for the femoral and knee components, preoperative use of non-steroidal anti-inflammatory agents, or performance of a lateral release for total knee arthroplasty. We concluded that desmopressin does not reduce blood loss or transfusion requirements after total joint arthroplasty.

Intraocular pressure increases with fenoldopam, but not nitroprusside, in hypertensive humans.

Fenoldopam mesylate stimulates adenyl cyclase in porcine ocular trabecular meshwork and raises intraocular pressure in humans. To clarify whether this results from direct activation of the dopamine-1 receptor or indirectly from baroreflex sympathetic stimulation after blood pressure reduction, intraocular pressure was measured in 14 patients with accelerated/malignant hypertension, randomized between intravenous fenoldopam or sodium nitroprusside. Intraocular pressure was measured with a Perkins tonometer, before and at the twentieth minute of each dose increment. In seven patients with a mean blood pressure of 232/131 mm Hg treated with fenoldopam, intraocular pressure increased in a dose-dependent fashion, from 16 +/- 1 to 20 +/- 2 mm Hg (p less than 0.005). In contrast, seven patients with a mean blood pressure of 225/134 mm Hg treated with sodium nitroprusside exhibited no change in intraocular pressure (15 +/- 1 versus 14 +/- 1 mm Hg) despite similar blood pressure reduction. Increases in heart rate were not significantly different. Rates of urinary excretion of norepinephrine plus epinephrine increased significantly relative to baseline (p less than 0.05) but were not different between groups. These data suggest that the increase in intraocular pressure with fenoldopam results from specific activation of the dopamine-1 receptor and is not caused by baroreflex sympathetic stimulation. Because dopamine-1 receptors may modulate intraocular pressure, dopamine-1 receptor blockers might be useful therapy for glaucoma.

Effects of dopamine receptor activation on the level of cyclic AMP in the trabecular meshwork.

We examined the effects of dopamine and of a selective DA1 agonist, fenoldopam, on the levels of cyclic AMP in the trabecular meshwork, freshly excised from porcine and canine eyes. As measured by radioimmunoassay, fenoldopam at a concentration of 10(-5) M caused a 4-fold increase in the cyclic AMP content, from a basal level of 24.4 +/- 1.9 to 101.8 +/- 6.3 pmol/mg protein, of the trabecular meshwork samples from porcine eyes. In tissue samples from canine eyes, fenoldopam at a concentration of 10(-4) M increased the endogenous cyclic AMP level from a basal value of 26.0 +/- 4.6 to 64.2 +/- 5.7 pmol/mg protein. Dopamine, although less potent, produced a similar response. Preincubation with a DA1 receptor antagonist, SCH 23390, inhibited the increase in cyclic AMP levels by 90%. Such inhibition did not occur with the alpha- and beta-receptor antagonists phenoxybenzamine and propranolol, respectively. This investigation demonstrates that adenylate cyclase-coupled DA1 receptors are present in porcine and canine trabecular meshwork tissue.

Effects of selective dopamine-1 receptor activation on intraocular pressure in man.

The lack of specific agonists and antagonists has, until recently, precluded investigation of a role for dopamine receptors in the control of intraocular pressure. In the present study, we have examined the effects of fenoldopam, a novel selective dopamine1 (DA1) receptor agonist, on intraocular pressure, in eight healthy human volunteers. Fenoldopam, infused intravenously at 0.5 micrograms kg-1 min-1, increased intraocular pressure from 14.6 +/- 0.9 to 17.6 +/- 1.4 mmHg (P less than 0.05) while a control saline infusion had no effect. Pupil diameter and blood pressure did not change. In the same subjects, i.v. norepinephrine or angiotensin II both increased intraocular pressure--from 13.8 +/- 1.4- to 17.6 +/- 1.4 mmHg and from 13.4 +/- 1.3- to 17.5 +/- 1.7 mmHg respectively (P less than 0.05), and mean arterial pressure by about 20 mmHg. These data suggest that: (1) DA1 receptor activation can modulate intraocular pressure; (2) the intraocular pressure effects of the DA1 receptor agonist, fenoldopam, are independent of changes in systemic blood pressure, in contrast to those of norepinephrine or angiotensin II where intraocular and systemic blood pressures increase in parallel; (3) the ability of a DA1 receptor antagonist to lower intraocular pressure merits investigation.

Immunochemical characterization of human lung epoxide hydrolases.

Immunochemical techniques were used to investigate the biochemical properties of human lung epoxide hydrolases. Two epoxide hydrolases with different immunoreactive properties were identified. These two epoxide hydrolases were found in both cytosolic and microsomal cell fractions. Immunotitration of enzyme activity showed that enzymes that catalyze the hydration of benzo(a)pyrene 4,5-oxide react with antiserum to rat microsomal epoxide hydrolase; those that hydrate trans-stilbene oxide do not. Immunotitration and Western blot experiments showed that microsomal and cytosolic benzo(a)pyrene 4,5-oxide hydrolases have significant structural homology. Immunohistochemical staining of human lung benzo(a)pyrene 4,5-oxide hydrolase showed that the enzyme is localized primarily in the bronchial epithelium. No cell type-specific localization was observed. An enzyme-linked immunosorbent assay was developed which allows direct quantitation of benzo(a)pyrene 4,5-oxide hydrolase protein. Levels of enzyme protein detected by this assay correlated well with enzyme levels determined by substrate conversion assays.

Multiple epoxide hydrolases in human lung.

Benzo(a)pyrene 4,5-oxide (BPO) hydrolase activity and trans-stilbene oxide hydrolase activity were measured in human lung samples obtained from 12 different patients. These activities, attributable to different epoxide hydrolases, were both found in significant amounts in the cytosolic as well as the microsomal cell fractions. The catalytic properties of human lung BPO hydrolases were further investigated, and similarities between the cytosolic and microsomal forms of this enzyme were noted. Similar Km values were found for both forms (22 microM for the microsomal enzyme, 37 microM for the cytosolic enzyme). Both forms responded in similar fashion to a number of enzyme inhibitors in vitro, and the IC50 values for each inhibitor were the same for both forms. Both forms of the enzyme were inhibited uncompetitively by trichloropropene oxide, and the Kii value for the cytosolic hydrolase (11 microM) was similar to that obtained for the microsomal hydrolase (15 microM). These two forms of BPO hydrolase in the human lung are very similar, possibly identical, enzymes.