Reactive oxygen intermediates reduce inactivation of serotonin in isolated, perfused rat lungs

Reactive oxygen intermediates such as free radicals have been proposed to mediate lung injury. The present work examined whether or not enzymatically generated oxygen metabolites altered serotonin clearance. Isolated, plasma-perfused rat lungs were exposed to xanthine oxidase (XO) and hypoxanthine (HX). Pulmonary arterial pressure (Ppa) and lung weight were recorded. Fulminant edema was defined as a spontaneous weight increase exceeding 500 mg. Inactivation of serotonin was determined by superfusion bioassay. XO and HX reduced serotonin inactivation from 74 +/- 3% (mean +/- SEM) to 62 +/- 2%. This reduction was inhibited by the scavenger enzymes superoxide dismutase (SOD) and catalase and by allopurinol, an inhibitor of XO. Hydrostatic edema and perfusion per se did not decrease the pulmonary clearance of serotonin. XO and HX did not significantly alter Ppa. Fulminant edema developed in four of six lungs after exposure to XO and HX compared with none in the other groups. It was concluded that reactive oxygen intermediates inhibited serotonin inactivation in isolated rat lungs.     

Vascular smooth muscle structure and juvenile growth in rat intestinal venules

The morphological structure of individual vascular smooth muscle cells from intestinal venules was evaluated with a combination of quantitative scanning (SEM) and transmission (TEM) electron microscopy techniques. In addition, growth of individual venular smooth muscle cells and of the overall vessel wall was compared from measurements of these variables during the rapid juvenile growth spurt from ages 4 to 6 and 10 to 12 weeks in Wistar-Kyoto rats. SEM revealed that smooth muscle cells of intestinal venules in weanling rats are very long (379 +/- 91 [SD] microns) and wide (6.0 +/- 1.3 microns) and very little further cell enlargement occurs during rapid juvenile growth. TEM studies indicated that passive inner vessel diameter and total muscle layer cross-sectional area of both the largest and intermediate diameter venules of young rats, as well as the percentage of the total wall area as muscle tissue in each venule type, did not significantly increase during body growth. These observations indicate that both the intestinal venules and their smooth muscle cells reach mature dimensions at a very early stage of life. Comparison of intestinal vascular smooth muscle cell dimensions indicates that venular smooth muscle cells are much larger in both cell length and volume than comparable arteriolar smooth muscle cells.     

Blood flow velocity in pulmonary microvessels of bullfrog.

Flow velocity in the pulmonary microvessels of the exposed lung of bullfrogs was measured by means of a laser Doppler microscope of an oblique backward mode, together with a signal-analyzing system having a time sharing circuit triggered by the R-wave of the ECG. By these means, measurements of the changes of flow velocity contour in the cardiac cycle were made. Flow velocity was clearly pulsatile in response to cardiac cycles in all microvessels including capillaries. Flow velocities in the arteriole and venule consistently decreased for a short period after the R-wave (84 +/- 33 msec (mean +/- SD) in the arteriole and 130 +/- 31 msec in the venule, respectively) and rapidly increased up to a maximicronm value. The mean flow velocities in arterioles (diameter 50 +/- 17 micron) and venules (39 +/- 9 micronm) were 2.29 +/- 0.32 and 2.30 +/- 0.27 mm/sec. The amplitudes of pulsatile flow in these vessels were 0.83 +/- 0.31 and 0.63 +/- 0.16 mm/sec, respectively. In the capillary the times from the R-wave to the minimicronm and maximum values were variable. In some cases the velocity gradually increased without first decreasing and the increase sharply accelerated a certain time after the R-wave. The mean velocity in the pulmonary capillary and the amplitude of the pulsatile flow ere 1.78 +/- 0.31 and 0.37 +/- 0.12 mm/sec, resepctively. The ratios of the pulsatile amplitude to the mean velocity in the pulmonary capillary, venule and arteriole averaged 0.21, and 0.36, respectively.     

The rat urinary bladder: vasoactivity and macromolecular leakage in a new model

An in vivo model of the rat urinary bladder microcirculation has been developed and microcirculatory responses to agents which produce vasoconstriction, vasodilation, and macromolecular leakage have been characterized. The urinary bladder of anesthetized female Sprague-Dawley rats is exteriorized and positioned in a tissue bath with a single stay suture which does not penetrate the lumen of the bladder. All blood vessels and nerves from the animal remain intact. The tissue bath is filled with Krebs solution which is monitored and maintained at a temperature of 36 +/- 0.5 degrees and a pH of 7.4 +/- 0.5. In vivo television microscopy is used to monitor vascular diameter and flow changes and isothiocyanate-tagged bovine serum albumin fluorescence is used as an index of macromolecular leakage. Norepinephrine (10(-6) M) caused a statistically significant decrease in vascular diameters of both arterioles and venules while sodium nitroprusside (10(-7) M) significantly increased arteriolar and venular diameters, histamine (10(-4) M) caused no change in venular diameters but did induce a significant macromolecular leak from those vessels. Compound 48/80 (1 and 10 micrograms/ml) induced a significant dose-dependent macromolecular leakage from venules. However, only with the 10 micrograms/ml dose was there visually detectable mast cell degranulation. It is concluded that this rat urinary bladder model provides a stable, reproducible model of a smooth muscle microcirculatory bed in a controlled environment, which responds similarly to other microcirculations.     

Dose-dependent effects of fentanyl on indomethacin-induced gastric damage.

Whilst developing a rat model for studies of gastric protection, we noticed that the anaesthetic agent 'Hypnorm', containing the opiate fentanyl 0.315 mg/ml and the butyrophenone fluanisone 10 mg/ml, was itself protective against indomethacin-induced damage: unrestrained animals given indomethacin (20 mg/kg) subcutaneously had an ulcer score of 9 +/- 1 mm2, compared with 1 +/- 1 mm2 in animals pre-treated with Hypnorm (0.8 ml/kg) and then given indomethacin (p less than 0.01). Further investigation showed this effect to be due to fentanyl-inhibiting gastric acid secretion: doses of fentanyl (90 and 180 micrograms/kg) which decreased indomethacin-induced damage also caused a rise in intragastric pH from 2.7 +/- 0.6 in controls to 5.1 +/- 0.8 and 5.0 +/- 0.8, respectively. However, the response of fentanyl varied depending on the dose given: fentanyl, 3.6 micrograms/kg did not affect indomethacin-induced damage, 8 +/- 2 vs. 9 +/- 1 mm2; fentanyl, 18 micrograms/kg potentiated damage, 15 +/- 4 mm2 (p less than 0.05), whereas fentanyl, 90 micrograms/kg and 180 micrograms/kg decreased damage, 2 +/- 1 mm2 and 0.1 +/- 0.1 mm2, respectively (p less than 0.01). Neither the butyrophenone haloperidol (8.3 mg/kg) nor the alpha-adrenergic receptor antagonist phenoxybenzamine (3 mg/kg) protected against indomethacin-induced damage. We conclude that fentanyl affects intragastric pH and can both potentiate and protect against indomethacin-induced damage. Furthermore, the potentiation of gastric damage by fentanyl occurred at doses similar to those used for human anesthesia, so clinical studies are suggested.     

Heterogeneous microvascular coronary alpha-adrenergic vasoconstriction

We tested the hypothesis that humoral or neurogenic alpha-adrenergic activation in the coronary circulation would produce heterogeneous vascular reactions. To accomplish this, the epicardial coronary microcirculation was viewed through an intravital microscope using stroboscopic epi-illumination. Microvascular diameters were measured under control conditions during beta-adrenergic blockade (propranolol 1 mg/kg) and beta-adrenergic blockade with pacing; during coronary alpha-adrenergic activation in the presence of beta-adrenergic blockade with three doses of norepinephrine infusion (0.1, 0.5, and 1.0-2.0 micrograms/kg/min) or three frequencies of bilateral stellate nerve stimulation (2, 10, and 20 Hz); and during combined alpha- and beta-adrenergic blockade (phentolamine 2 mg/kg and propranolol 1 mg/kg). Diameters of both arterial and venous vessels were reduced during beta-adrenergic blockade but returned back to baseline with pacing. At the lowest level of norepinephrine infusion or frequency of bilateral stellate stimulation, microvessel constriction was not observed. At the higher doses of norepinephrine a -5.1 +/- 0.9% (1.0-2.0 micrograms/kg/min) and a -4.0 +/- 1.1% (0.5 micrograms/kg/min) decrease in diameter of arterial vessels greater than 100 microns in diameter were observed (p less than 0.05). At 10 Hz and 20 Hz of stellate stimulation, diameter decreased by -4.8 +/- 1.9% and -4.4 +/- 2.1%, respectively, in these relatively large vessels. Small coronary arterioles (less than 100 microns diameter) dilated significantly during the highest levels of nerve stimulation (9.2 +/- 2.5% increase in diameter) or infusion rate of norepinephrine (13.6 +/- 2.7% increase in diameter) (p less than 0.05). These constrictor and dilator responses were abolished following combined alpha- and beta-adrenergic blockade. Norepinephrine infusion resulted in a decrease in diameter of coronary veins and venules (7.2 +/- 1.3%) (p less than 0.05), whereas stellate stimulation did not significantly reduce venous and venular diameters. In summary, the coronary venous and venular vasculature responds to alpha-adrenergic activation from circulating norepinephrine but is not affected by stellate stimulation. In contrast, stellate stimulation and norepinephrine infusion elicit similar responses in the coronary arterial and arteriolar microvasculature. Constriction occurs in vessels greater than 100 microns in diameter, whereas dilation predominates in vessels less than 100 microns in diameter. Such heterogeneous arterial responses would undoubtedly result in a redistribution of coronary vascular resistance toward larger coronary arteries and arterioles.     

ZK 36-374, a stable analog of prostacyclin, prevents acute hypoxic pulmonary hypertension in the dog

Vasodilator therapy in pulmonary hypertension is limited by the lack of an agent selective for the pulmonary circulation. The effects of intravenous prostacyclin and two stable prostaglandin analogs, ZK 36-374 and CL 115,347, were assessed on the preconstricted pulmonary vasculature of the anesthetized dog. During hypoxic vasoconstriction ZK 36-374 (0.4 micrograms/kg per min) markedly reduced pulmonary artery pressure (26 +/- 3 to 13 +/- 1 mm Hg) (p less than 0.05) and pulmonary vascular resistance (6.2 +/- 1.1 to 2.8 +/- 0.2 mm Hg/liter per min) (p less than 0.01). There was no significant effect on cardiac output, aortic pressure or arterial blood gases. Pulmonary vasoconstriction induced by prostaglandin F2 alpha was similarly affected by ZK 36-374, and in this instance the aortic pressure was also reduced (158 +/- 11 to 129 +/- 11 mm Hg) (p less than 0.01). ZK 36-374 (0.2 micrograms/kg per min) was more effective in lowering hypoxic pulmonary vascular resistance (from 6.5 +/- 0.6 to 3.0 +/- 0.3 mm Hg/liter per min) than was prostacyclin (0.75 micrograms/kg per min) (from 6.3 +/- 0.6 to 4.2 +/- 0.4 mm Hg/liter per min) (p less than 0.05) and resulted in a smaller fall in aortic pressure (p less than 0.05). CL 115,347 (1.0 micrograms/kg per min) had no effect on the pulmonary vasculature during normoxia or when preconstricted by prostaglandin F2 alpha or hypoxia, but reduced aortic pressure and total systemic resistance (p less than 0.05). It appears to be a selective systemic vasodilator with no pulmonary vascular activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Role of bradykinin in ovine endotoxemia

We determined if the cardiopulmonary response to endotoxin (LPS) is mediated by bradykinin (BK). Sheep (n = 30) were prepared for chronic study with cardiopulmonary catheters, and chronic lung lymph fistulae. They were divided into the following study groups: A) BK infusion; B) LPS; C) LPS with angiotensin converting enzyme inhibitor (ACEI); D) ACEI alone; E) LPS with BK antagonist. Cardiac index and mean arterial pressure fell (6.9 +/- 0.4 to 5.3 +/- 0.3 L/min/m2 and 93 +/- 4 to 72 +/- 3 mmHg, respectively), and pulmonary lymph flow increased (10.5 +/- 3.0 to 17.8 +/- 4.3 ml/hr) during BK infusion. Addition of ACEI during BK infusion reduced the amount of BK necessary to induce a comparable response (125 to 2 micrograms/kg/hr). Administration of ACEI to LPS animals did not significantly alter their cardiopulmonary responses. BK antagonist did not change the response to LPS. Although the kallikrein-kinin pathway is believed to be activated during the septic response, our data do not lend support for the hypothesis that bradykinin is an important mediator of the cardiopulmonary changes.     

Heredity and hypertension: impact on metabolic characteristics.

This study was performed to evaluate the possible role of heredity in the clinical characteristics of hypertension. Metabolic, endocrine, and renal measurements were compared in subjects with normal blood pressure who had a family history of hypertension (n = 60) with those of subjects with normal blood pressure who did not have a family history of hypertension (n = 48). The groups were matched for age (mean, 44 +/- 2 years and 45 +/- 2 years) and blood pressure (127 +/- 1/77 +/- 1 mm Hg and 127 +/- 2/77 +/- 1 mm Hg). The following parameters were higher in the patients with a family history of hypertension than in those without. Plasma insulin concentrations (14.1 +/- 1.1 vs 10.8 +/- 1.0 microU/ml; p less than 0.05), insulin-glucose ratio (0.15 +/- 0.01 vs 0.11 +/- 0.010; p less than 0.05), norepinephrine concentrations (315 +/- 24 pg/ml vs 208 +/- 20 pg/ml; p less than 0.01), plasma renin activity (2.1 +/- 0.2 ng Angl/ml/hr vs 1.6 +/- 0.2 ng Angl/ml/hr; p less than 0.02), total cholesterol levels (217 +/- 8 mg/dl vs 197 +/- 0.3 mg/dl; p less than 0.05), creatinine clearance (125 +/- 9 ml/min vs 96 +/- 8 ml/min; p less than 0.01), and albumin excretion rate (3.2 +/- 0.3 micrograms/min vs 2.6 +/- 0.3 micrograms/min; p = 0.1). Moreover, patients with a family history of hypertension had smaller increases in systolic blood pressure during treadmill exercise (55 +/- 3 mm Hg vs 64 +/- 3 mm Hg; p less than 0.03). There were no differences in echocardiographic left ventricular mass index between the groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stimulation of collagenase production in human synovial fibroblast cultures by poly (I). poly (C)

Poly (I). poly (C) was found to induce collagenase secretion in both "normal" and "rheumatoid" synovial fibroblast cultures. Induction was time dependent, was maximal at 20-50 micrograms/ml poly (I). poly (C) and was dependent on de novo synthesis. Induction was prevented by hydrocortisone (0.1 microgram/ml) but inhibition of prostaglandin E production by diclofenac (0.2 microgram/ml) or ibuprofen (0.1 microgram/ml) did not affect collagenase production. Collagenase induction was accompanied by stimulation of hyaluronic acid and prostaglandin E production. This in vitro system may represent a model for the study of pathological secretory activities in the inflamed joint.     

Role of molecular charge in disruption of the blood-brain barrier during acute hypertension

Acute hypertension disrupts the blood-brain barrier and may neutralize the negative charge on cerebral endothelium. The goal of this study was to determine the effects of molecular charge on permeability of the blood-brain barrier during acute hypertension. Intravital fluorescent microscopy and fluorescein-labeled dextrans were used to evaluate disruption of the blood-brain barrier during acute hypertension in rats. Disruption of the blood-brain barrier was quantitated by calculating clearance of neutral dextran and of anionic dextran sulfate in two groups of rats. Pressure in pial venules, which are the primary site of disruption of the blood-brain barrier during acute hypertension, was measured using a servo-null device. When systemic arterial pressure was increased from 87 +/- 5 (mean +/- SEM) to 188 +/- 5 mm Hg, clearance of neutral dextran increased from 0.04 +/- 0.01 to 4.38 +/- 0.72 ml/sec x 10(-6). When systemic arterial pressure was increased from 91 +/- 4 to 181 +/- 3 mm Hg, clearance of anionic dextran sulfate increased from 0.02 +/- 0.01 to only 0.70 +/- 0.23 ml/sec x 10(-6). Increases in pial venular pressure were similar in the two groups. Thus, similar increases in systemic arterial pressure and pial venular pressure during acute hypertension produce less disruption of the blood-brain barrier to anionic dextran sulfate than neutral dextran. The findings suggest that 1) the net negative charge of cerebral vessels may be preserved during acute hypertension, and 2) molecular charge is an important determinant of the severity of disruption of the blood-brain barrier during acute hypertension.     

Role of veins and cerebral venous pressure in disruption of the blood-brain barrier

The goal of this study was to determine whether increases in cerebral venous pressure contribute to, and may account for, disruption of the blood-brain barrier during acute hypertension and hyperosmolar stimuli. We studied the relation between pial venous pressure and disruption of the blood-brain barrier during acute arterial hypertension, superior venae cavae occlusion, and superfusion with hyperosmolar arabinose. Sprague-Dawley rats were studied using intravital fluorescent microscopy and fluorescein-labelled dextran (mol. wt. = 70,000). Disruption of the blood-brain barrier was characterized by the appearance of microvascular leaky sites and quantitated by the clearance of fluorescein dextran. We measured pressure (servo null) in pial arterioles and venules 40-60 micron in diameter. Acute hypertension, occlusion of the superior venae cavae, and hyperosmolar arabinose produced leaky sites primarily in venules. Acute hypertension increased arteriolar pressure and also venular pressure, from 7 +/- 1 (mean +/- SE) to 28 +/- 2 mm Hg. Clearance of fluorescein dextran increased from 0.03 +/- 0.01 to 2.90 +/- 0.40 ml/sec X 10(-6). Occlusion of the superior venae cavae increased pial venous pressure from 7 +/- 1 to 30 +/- 3 mm Hg, and clearance of fluorescein dextran, from 0.02 +/- 0.01 to 3.10 +/- 0.59 ml/sec X 10(-6). In contrast to acute hypertension, there was a decrease in arterial and pial arteriolar pressure during occlusion of the superior venae cavae. Thus, similar increases in venous pressure during acute hypertension and superior venae cavae occlusion, despite directionally opposite changes in arterial and arteriolar pressure, produced similar disruption of the blood-brain barrier.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of stromelysin and collagenase in synovial fluid from patients with rheumatoid arthritis and posttraumatic knee injury.

OBJECTIVE. To quantify stromelysin and collagenase in synovial fluid (SF) from patients with rheumatoid arthritis (RA) or traumatic knee injury. METHODS. Stromelysin and collagenase were measured in the SF of 33 patients with RA or posttraumatic knee injury, using specific double-antibody sandwich enzyme-linked immunosorbent assays. Stromelysin was fractionated from representative SF, and the molecular form was identified by immunoblot analysis. RESULTS. The stromelysin concentration was approximately 20-fold higher than the collagenase concentration in the fluids from patients with RA and approximately 8-fold higher in the fluids from patients with traumatic injury. For both metalloproteinases, there was a higher enzyme concentration in RA SF than in the SF from patients with trauma (stromelysin 40.1 +/- 26 micrograms/ml [mean +/- SD] in RA SF, 8.5 +/- 15 micrograms/ml in trauma SF; collagenase 2.2 +/- 3.3 micrograms/ml in RA SF, 1.1 +/- 2.3 micrograms/ml in trauma SF). The majority of the stromelysin within the SF bound to reactive red-agarose and was identified as prostromelysin based on electrophoretic mobility and immunoblotting with monospecific antibodies. CONCLUSION. The finding of high levels of stromelysin in SF from patients with RA supports the proposal that this enzyme may play a role in the connective tissue degradation observed in this disease.     

Combined dose ratios of dopamine and dobutamine and right ventricular performance after cardiac surgery.

The effect of combined administration of different dose ratios of dobutamine (DB) and dopamine (DA) (DB/DA ratio of 1:1; 1.5:0.5; 2:0; 0.5:1.5; and 0:2), with the added dose kept constant (10 micrograms/kg/min-20 micrograms/kg/min), on right ventricular function (measured by the thermal washout method with the aid of a rapid-response thermistor) was determined in ten patients after cardiac surgery (between 12 and 24 h after surgery). The following values represent the mean +/- SD of DB only and of the DB/DA-equal combination vs DA only. The DB/DA-equal or DB-dominant combination increased the right ventricular ejection fraction vs DA only (0.39 +/- 0.12 [p less than 0.01] and 0.37 +/- 0.11 [p less than 0.05], respectively, vs 0.32 +/- 0.12) and the stroke volume index (43 +/- 12 ml/m2 [p less than 0.01] and 41 +/- 15 ml/m2, respectively, vs 38 +/- 14 ml/m2) and decreased right ventricular end-diastolic pressure (RVEDP) (10 +/- 4 mm Hg [p less than 0.01] and 11 +/- 4 mm Hg [p less than 0.05], respectively, vs 13 +/- 5 mm Hg) and pulmonary capillary wedge pressure (10 +/- 4 mm Hg [p less than 0.01] and 12 +/- 5 mm Hg [p less than 0.05], respectively, vs 14 +/- 6 mm Hg) to the same degree as DB alone. The DB/DA-equal or DB-dominant combination did not induce tachycardia (heart rate, 105 +/- 11 [p less than 0.05] and 95 +/- 14 beats per minute, respectively, vs 90 +/- 17 beats per minute) or have any effect on the right ventricular end-diastolic volume index (RVEDVI) (115 +/- 30 ml/m2 and 117 +/- 33 ml/m2, respectively, vs 127 +/- 42 ml/m2). Moreover, the diastolic parameters of the right ventricle (the ratio of RVEDVI/RVEDP: 15 +/- 8 [p less than 0.05] and 13 +/- 7, ml/mm Hg/m2, respectively, 11 +/- 5 ml/mm Hg/m2) decreased as the ratio of DA increased. This change in the diastolic properties of the right ventricle might have been caused by release of norepinephrine in the myocardium by DA or by improved coronary perfusion with DB. The DB/DA-equal and DB-dominant combinations were superior to DB or DA alone and to the DA-dominant combination in obtaining enhanced right ventricular performance.     

The preovulatory increase in ovarian collagenase activity in the rat is independent of prostaglandin production

In the present study, we have examined the role of gonadotropins and prostaglandins in the preovulatory increase of ovarian collagenase activity in the rat. Whole ovaries of immature PMSG-primed rats (20 IU) were removed before and 8 h after the rats were treated with human (h) CG, Nembutal, and/or indomethacin. The ovaries were homogenized in a solution containing Triton X-100 (0.25%) and centrifuged. Collagenase was extracted by resuspending the pellets in buffer containing 100 mM CaCl2, heating to 60 C for 6 min, and centrifuging. The supernatants were treated with dithiothreitol (2 mM) and iodoacetamide (5 mM) to inactivate collagenase inhibitors. Collagenase activity was measured as the percent digestion of 3H-type I collagen/100 microliters aliquot of ovarian sample. At zero time (52 h after PMSG), ovarian collagenase activity was 4.2 +/- 1.2% digestion (mean +/- SEM, n = 3). In ovaries collected 8 h after the endogenous LH surge or 8 h after the administration of 10 IU hCG at time zero, collagenase activity rose to 19.6 +/- 2.1 (n = 6) and 22.5 +/- 1.7% digestion (n = 11), respectively. Indomethacin (1.5 mg/100 g BW) administered 30 min after hCG, produced no change in collagenase activity (24.8 +/- 2.5% digestion, n = 7) although the expected increase in ovarian prostaglandin E after hCG treatment was blocked. When the endogenous LH surge was blocked with Nembutal (3 mg/100 g BW), collagenase activity in 8-h ovaries was 6.8 +/- 1.1% digestion (n = 10). The Nembutal block of the preovulatory collagenase increase was overcome by administration of hCG (8-h ovarian enzyme activity = 22.7 +/- 3.2% digestion, n = 8). These observations demonstrate that hCG stimulates ovarian collagenase activity and that this stimulation is not dependent on prostaglandin synthesis.     

Effect of heparin and warfarin on chronic hypoxic pulmonary hypertension and vascular remodeling in the guinea pig

Chronic hypoxia produces pulmonary hypertension and pulmonary vascular remodeling. Heparin partially prevents the rise in right ventricular pressure and vascular remodeling in chronically hypoxic mice. To determine if this is due to the anticoagulant property of heparin or another property, we compared the effect of oral warfarin given at an anticoagulating dose (0.5 mg/kg/day) to heparin given by continuous infusion at a dose that does not prolong the partial thromboplastin time (PTT) (20 units/kg/h) on hypoxic pulmonary hypertension and vascular remodeling in the guinea pig. Normoxic control animals either untreated or treated with heparin or Coumadin were all alike in blood gases, pulmonary vascular resistance, right heart weights, and pulmonary histology. Hypoxia (10% 0(2) for 10 days) induced similar and significant increases in mean pulmonary artery (PA) pressure in both the hypoxic control and warfarin groups (19 +/- 1 mm Hg (mean +/- SEM) in both groups versus 11 +/- 0.1 mm Hg in the normoxic control group; p less than 0.05). Total pulmonary vascular resistance (TPR) was also increased from 0.041 +/- 0.002 in the normoxic control group to 0.087 +/- 0.007 and 0.071 +/- 0.003 mm Hg/ml/min/kg in the hypoxic control and warfarin groups, respectively (p less than 0.05). Whereas anticoagulation with warfarin did not protect the guinea pig from developing pulmonary hypertension, heparin markedly reduced PA and TPR (15 +/- 1 mm Hg and 0.052 +/- 0.002 mm Hg/ml/min/kg, respectively; p less than 0.05 versus hypoxic control or warfarin).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of dobutamine on lung microvascular fluid flux in sheep with "sepsis syndrome"

The effect on pulmonary fluid balance of adrenergic receptor agonist agents commonly employed in clinical sepsis has not been well characterized. Therefore, we tested the hypothesis that dobutamine would increase pulmonary microvasular fluid flux in experimental sepsis-induced lung injury. To define the effects of this synthetic catecholamine on pulmonary lymph flow (QL), we infused dobutamine in sheep at two doses in sequence (5 micrograms/kg/min and 10 micrograms/kg/min) before and after the induction of intraperitoneal sepsis which resulted in the development of lung microvascular injury. In the nonseptic state, cardiac output increased at both 5 micrograms/kg/min and 10 micrograms/kg/min (22 and 36 percent, respectively), while QL was unchanged from baseline (for 5 micrograms, delta QL = +0.44 +/- 1.35 ml/15 min; not significant) (for 10 micrograms, delta QL = -0.20 +/- 1.0 ml/15 min; not significant). Values for the ratio of lymph/plasma total protein levels [( L/P]TP) fell modestly in the nonseptic study at both doses (p less than 0.05). With established sepsis syndrome, QL increased from the nonseptic baseline study (2.99 +/- 1.8 to 7.01 +/- 3.95 ml/15 min; p less than 0.05), without change in [L/P]TP ratios or the calculated microvascular hydrostatic pressure. (Pmv) During sepsis, dobutamine infusion was again associated with an increase in cardiac output at both the 5 micrograms/kg/min (+29 percent) and 10 micrograms/kg/min (+33 percent) doses, while QL increased modestly only with the lower dose of dobutamine infused (5 micrograms/kg/min, delta QL = 1.80 +/- 2.2 ml/15 min; p less than 0.05). In this model of sepsis-induced lung injury, dobutamine increased systemic flow without substantially augmenting QL.     

Once-daily dosing of a new ultrasustained-release theophylline preparation

Theophylline plasma levels and profiles were evaluated in patients with chronic obstructive pulmonary disease during once-daily dosing of an ultrasustained-release theophylline preparation (Theo-1; capsules filled with microgranules containing 400 mg anhydrous theophylline). In a first study, 6 patients received a single morning dose of 800 mg (a) in the fasting state, and (b) with a protein-fat-rich breakfast in a random order, and the systemic theophylline availability was evaluated for 48 h. No significant differences were found either in Cmax (a: 7.0 +/- 3.2 micrograms/ml; b: 7.6 +/- 2.6 micrograms/ml), or in Tmax (a: 11.7 +/- 6.1 h; b: 10.2 +/- 3.6 h). Elimination half-life was in a 11.4 +/- 4.4 h and in b 12.9 +/- 4.8 h (p less than 0.05). In a second study, the steady-state theophylline levels were measured during a 24-hour dosage interval on day 8 after intake of 800 mg at 8 a.m. in 16 patients and at 8 p.m. in 11 patients. Plateau-shaped plasma concentration-time curves were obtained, with small fluctuations between the peak (Cmax) and trough (Cmin) levels: [100(Cmax-Cmin)/Cmin] was 83 +/- 40% after morning dose, and 54 +/- 26% after evening dose (p less than 0.05). Cmax was 12 +/- 5 and 11 +/- 4 micrograms/ml, respectively (NS). Tmax was 9 +/- 3 and 11 +/- 3 h, respectively (NS). The FDA fluctuation for the 37 patients was 48 +/- 20%. In a third study, the dose-plasma concentration relationship was evaluated in steady state in 6 patients receiving 400, 800 and 1,200 mg for 3 days each. The trough plasma concentrations were 2.6 +/- 0.9, 6.2 +/- 2.1 and 10.2 +/- 3.1 micrograms/ml, respectively. Six hours after drug intake the plasma levels were 5.0 +/- 1.6, 10.6 +/- 2.5 and 15.4 +/- 4.2 micrograms/ml, respectively; and 12 h after drug intake, 4.9 +/- 1.4, 11.6 +/- 2.4 and 14.5 +/- 3.7 micrograms/ml, respectively. In conclusion, we found in these studies that with once-daily dosing of the ultrasustained-release preparation Theo-1, plateau-shaped 24-hour theophylline plasma levels could be achieved. The relationship between daily dosage and theophylline plasma levels was linear intraindividually but showed an important interindividual variation. No consistent interference by food intake was found and no serious side effects occurred within therapeutic plasma levels.     

DETERMINATION OF ENOLASE ISOZYMES IN VARIOUS ADRENAL GLAND TUMOURS

Enolase isozymes (alpha enolase and gamma enolase) in the extracts of adrenal tumours (phaeochromocytoma, adenoma of primary aldosteronism and Cushing's syndrome, and neurinoma) were determined by means of enzyme immunoassay systems. The mean +/- SEM, respectively, of alpha and gamma enolase levels were 2.5 +/- 0.37 microgram/mg protein and 3.2 +/- 0.69 micrograms/mg protein for 9 phaeochromocytomas, 15.2 +/- 3.1 microgram/mg protein and 0.65 +/- 0.18 microgram/mg protein for three adenomas with primary aldosteronism, 10.8 +/- 3.0 micrograms/mg protein and 0.23 +/- 0.02 micrograms/mg protein for five adenomas causing Cushing's syndrome, and 3.8 +/- 0.88 micrograms/mg protein and 0.30 +/- 0.15 micrograms/mg protein for three neurinomas. Thus, the gamma enolase concentration in the extract of phaeochromocytoma was higher than that of other adrenal tumours. The serum level of gamma enolase was determined in 36 patients with adrenal tumours and 26 normal controls by radioimmunoassay. The mean +/- SEM for gamma enolase level was 5.4 +/- 0.3 ng/ml in normal controls, 9.1 +/- 0.9 ng/ml for 10 patients with phaeochromocytoma, 6.3 +/- 0.3 ng/ml for 11 with primary aldosteronism, 5.5 +/- 0.4 ng/ml for 11 with Cushing's syndrome, and 5.1 +/- 0.7 ng/ml for four with neurinoma. Thus, patients with phaeochromocytoma had a significantly higher serum gamma enolase levels than did those with tumours derived from adrenal cortex and normal controls. In patients with phaeochromocytoma, serum gamma enolase levels showed a significant positive correlation with urinary adrenaline levels (P less than 0.05), and after resection the elevated level of gamma enolase fell significantly (P less than 0.05) and returned to normal.(ABSTRACT TRUNCATED AT 250 WORDS)     

Human chorionic gonadotrophin relaxation of human pregnant myometrium and activation of the BKCa channel

The uterorelaxant effect of human chorionic gonadotropin (hCG) is regarded as an important mediator in maintenance of uterine quiescence during pregnancy with clinical potential for tocolysis, the mechanisms of which are unknown. The large conductance calcium-activated K(+) channel (BK(Ca)) is ubiquitously encountered in human uterine tissue and plays a significant role in modulating myometrial cell membrane potential and excitability. The objective of this study was to investigate the involvement of BK(Ca) channel function in the response of human myometrial cells to hCG. Single electrophysiological BK(Ca) channel recordings from freshly dispersed myocytes were obtained in the presence and absence of increasing hCG concentrations. Isometric tension studies, investigating the effects of hCG on isolated myometrial contractions, in the presence and absence of the BK(Ca) channel blocker, iberiotoxin, were performed. The hCG significantly increased the open-state probability of these channels in a concentration-dependent manner [control 0.036 +/- 0.01; 1 IU/ml hCG 0.065 +/- 0.014 (P = 0.262); 10 IU/ml hCG 0.111 +/- 0.009 (P = 0.001); and 100 IU/ml hCG 0.098 +/- 0.004 (P = 0.007)]. In vitro functional studies demonstrated that hCG exerted a significant concentration-dependent relaxant effect on human myometrial tissue. This effect was significantly attenuated by preincubation with iberiotoxin (P < 0.05). These findings outline that activation of BK(Ca) channel activity may explain the potent uterorelaxant effect of hCG.