Elevated Fab glycosylation of anti-hinge antibodies.

OBJECTIVE: Rheumatoid arthritis (RA) is characterized by systemic inflammation and the presence of anti-citrullinated protein antibodies (ACPAs), which contain remarkably high levels of Fab glycosylation. Anti-hinge antibodies (AHAs) recognize immunoglobulin G (IgG) hinge neoepitopes exposed following cleavage by inflammation-associated proteases, and are also frequently observed in RA, and at higher levels compared to healthy controls (HCs). Here, we investigated AHA specificity and levels of Fab glycosylation as potential immunological markers for RA. METHOD: AHA serum levels, specificity, and Fab glycosylation were determined for the IgG1/4-hinge cleaved by matrix metalloproteinase-3, cathepsin G, pepsin, or IdeS, using enzyme-linked immunosorbent assay and lectin affinity chromatography, in patients with early active RA (n = 69) and HCs (n = 97). RESULTS: AHA reactivity was detected for all hinge neoepitopes in both RA patients and HCs. Reactivity against CatG-IgG1-F(ab´)2s and pepsin-IgG4-F(ab´)2s was more prevalent in RA. Moreover, all AHA responses showed increased Fab glycosylation levels in both RA patients and HCs. CONCLUSIONS: AHA responses are characterized by elevated levels of Fab glycosylation and highly specific neoepitope recognition, not just in RA patients but also in HCs. These results suggest that extensive Fab glycosylation may develop in response to an inflammatory proteolytic microenvironment, but is not restricted to RA.

A multicenter, prospective, randomized trial evaluating a new hemostatic agent for spinal surgery.

STUDY DESIGN: A prospective, randomized trial comparing Proceed, a gelatin-based hemostatic sealant (treatment), with Gelfoam-thrombin (control) in stopping intraoperative bleeding during spinal surgery. OBJECTIVES: To determine the effectiveness and safety of Proceed. SUMMARY OF BACKGROUND DATA: Proceed has been tested in animal models to determine its safety and effectiveness as a hemostatic agent. The current study was conducted under a Food and Drug Administration-approved Investigational Device Exemption to evaluate the effectiveness and safety of Proceed in humans. METHODS: For this study, 127 patients undergoing spinal surgery were randomized into either the treatment or control group after standard surgical means to control bleeding had failed. The bleeding site was evaluated at 1, 2, 3, 6, and 10 minutes after the hemostatic agent was applied. The application was considered successful if the bleeding stopped within 10 minutes. Follow-up evaluation was performed at 12 to 36 hours, then at 6 to 8 weeks after surgery. RESULTS: Proceed stopped bleeding in 98% of the patients (first bleeding site only) within 10 minutes, as compared with 90% of the control patients (P = 0.001). At 3 minutes, successful hemostasis had been achieved in 97% of the Proceed group, as compared with 71% of the control group (P = 0.0001). There was no difference in the adverse event profile between the two groups. CONCLUSIONS: A significantly larger number of bleeding sites had achieved hemostasis with Proceed than with Gelfoam-thrombin at 1, 2, and 3 minutes after application. Proceed was as safe as Gelfoam-thrombin when used for hemostasis during spinal surgery procedures.

Comparative effects of isoproterenol and dopamine on myocardial oxygen consumption, blood flow distribution and total body oxygen consumption in conscious lambs with and without an aortopulmonary left to right shunt.

OBJECTIVES: We sought to study the effects of catecholamines on myocardial oxygen consumption (VO2), regional blood flows and total body VO2 in lambs with circulatory congestion. BACKGROUND: Catecholamines are often used to support cardiovascular function in children with circulatory congestion because they increase contractility as well as heart rate. However, these changes increase myocardial oxygen demand and thus can lead to a mismatch between myocardial oxygen supply and demand. Catecholamines can also change regional blood flows and VO2 unfavorably. METHODS: We infused isoproterenol (0.1 microg/kg body weight per min) and dopamine (10 microg/kg per min) and measured myocardial and total body VO2 and regional blood flows in chronically instrumented 7-week old lambs with and without a left to right shunt. RESULTS: Isoproterenol increased myocardial VO2, parallel to the increase in heart rate. However, myocardial blood flow and, consequently, oxygen supply also increased. This increase outweighed the increase in myocardial VO2, so that myocardial oxygen extraction decreased. Isoproterenol did not change blood flow distribution. Isoproterenol increased total body VO2; however, systemic oxygen supply increased even more, so that oxygen extraction decreased and mixed venous oxygen saturation increased. In contrast, dopamine had no or little effect on myocardial VO2 or blood flow distribution. CONCLUSIONS: We conclude that the catecholamines isoproterenol and dopamine do not lead to a mismatch between myocardial oxygen supply and demand, nor do they change blood flow distribution unfavorably in 7-week old lambs with a left to right shunt. We demonstrated that isoproterenol is superior to dopamine, because it shifts the balance between oxygen supply and consumption toward supply so that systemic oxygen extraction reserve increases.

Effects of exercise and isoproterenol on hemodynamics and myocardial VO2 in lambs with aortopulmonary shunts.

To compare hemodynamic changes induced by isoproterenol and exercise stress tests in individuals with and without left ventricular volume load, we studied 10 lambs with an aortopulmonary shunt [58 +/- 4% (SE) of left ventricular output] 2 wk after the shunt was created. Two studies, isoproterenol infusion at 0.1 microgram.kg-1.min-1 and treadmill exercise at 76 +/- 4% of predetermined maximal O2 consumption (VO2) were performed in each lamb in random order on different days. Identical experiments were performed in nine lambs without shunts. Isoproterenol and exercise induced similar changes in heart rate (43 +/- 5%); systemic (72 +/- 7%), pulmonary (35 +/- 3%), and shunt blood flows (8 +/- 6%); and stroke volume (NS) in shunt lambs. Aortic systolic pressure increased less during isoproterenol infusion than during exercise (7 +/- 3 vs. 27 +/- 5%), and left atrial pressure decreased during isoproterenol infusion (-23 +/- 4%) but changed in an opposite direction during exercise (7 +/- 6%). These changes were accompanied by a smaller increase in myocardial VO2 during isoproterenol infusion than during exercise (5.0 +/- 0.7 to 5.3 +/- 0.6 and 5.3 +/- 0.8 to 7.3 +/- 0.9 mumol.beat-1 x 100 g-1, respectively). In control lambs, stroke volume decreased during isoproterenol infusion despite an equal decrease in left atrial pressure as in shunt lambs. In conclusion, isoproterenol better stimulates the blood flow changes during exercise in shunt than in control lambs probably because of their higher ventricular filling pressures. In interpreting isoproterenol stress tests it should, however, be kept in mind that these changes are realized at a lower work load for the heart.

Left ventricular oxygen and substrate uptake in chronically hypoxemic lambs.

Myocardial oxygen demand may be increased in chronically hypoxemic lambs because of their increased heart rate. Therefore, we determined whether left ventricular (LV) oxygen supply, oxygen uptake and oxygen demand were proportionally increased in 6-wk-old lambs, after 4 wk of hypoxemia (n = 15), as compared with control lambs (n = 14). In addition, we determined whether LV glucose, pyruvate, lactate, FFA and ketoacids uptake were altered in hypoxemic lambs, because of alterations in arterial glucose, pyruvate and lactate concentrations, that may occur in hypoxemia. Hypoxemia was induced by the combination of an atrial septal defect and pulmonary stenosis. Arterial oxygen saturation was decreased in hypoxemic lambs (67 +/- 8 versus 91 +/- 3%, p < 0.001), Hb concentration was increased, so that arterial oxygen concentration was similar in both groups of lambs. Myocardial mass (61 +/- 13 versus 44 +/- 9 g, p < 0.001) and total myocardial blood flow (117 +/- 36 versus 62 +/- 27 mL.min-1, p < 0.001) were increased, mainly due to right ventricular hypertrophy. LV oxygen demand, estimated by the rate pressure product (2072 +/- 465 versus 1467 +/- 358 kPa.beat.min-1, p < 0.001), and oxygen uptake (723 +/- 223 versus 556 +/- 184 mumol.min-1 x 100 g-1, p < 0.05) were proportionally increased in hypoxemic lambs. LV oxygen supply increased linearly with oxygen uptake (r = 0.96) in all lambs, by adjustments in LV blood flow, which was increased in hypoxemic lambs (168 +/- 41 versus 134 +/- 45 mL.min-1 x 100g-1, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Myocardial substrate uptake in lambs with and without aortopulmonary shunts during strenuous exercise.

Increased myocardial fatty acid uptake during acute exercise could adversely affect myocardial O2 consumption in lambs with left-to-right shunts, which would be unfavorable in view of their decreased coronary blood flow reserve. Therefore, we studied myocardial substrate uptake (glucose, lactate, pyruvate, free fatty acids, triglycerides, beta-hydroxybutyrate, and acetoacetate) in 10 7-wk-old lambs with an aortopulmonary left-to-right shunt [61 +/- 3% (SE) of left ventricular output] and 9 control lambs during strenuous treadmill exercise. The hemodynamic reaction to exercise was similar in shunt and control lambs. The peripheral metabolic response to exercise was also similar in the two groups: glucose free fatty acids, and, most prominently, lactate concentrations increased. Myocardial O2 consumption increased but less in shunt than in control lambs because of a smaller increase in heart rate. In both groups myocardial lactate uptake increased substantially at the cost of other substrates, providing the heart with 40% of its oxidative metabolism. Fatty acid uptake was not different between the two groups. In conclusion, our data reveal no essential differences in myocardial substrate uptake between shunt and control lambs during a substantial circulatory load.

Myocardial blood flow and VO2 in lambs with an aortopulmonary shunt during strenuous exercise.

To determine how much myocardial O2 consumption (VO2) would increase during an additional load on the heart in shunt as compared with control lambs, we studied 12 7-wk-old lambs with an aortopulmonary left-to-right shunt (59 +/- 3% of left ventricular output, mean +/- SE) and 11 control lambs during exercise at 80% of their predetermined peak VO2 (VO2peak), at 12 +/- 1 days after surgery. During exercise, systolic aortic pressure increased by 25% in the two groups. Left atrial pressure and left ventricular stroke volume did not change significantly and remained considerably higher in shunt than in control lambs. Heart rate, however, increased less in shunt than in control lambs (163 +/- 8 to 235 +/- 8 vs. 107 +/- 7 to 230 +/- 8 beats/min). The same was true for left ventricular myocardial blood flow (245 +/- 19 to 391 +/- 27 vs. 128 +/- 10 to 320 +/- 45 ml.min-1 x 100 g-1) and myocardial VO2 (847 +/- 101 to 1,692 +/- 136 vs. 528 +/- 58 to 1,579 +/- 178 mumol O2 x min-1 x 100 g-1). We conclude that, despite the volume load, myocardial VO2 of shunt lambs does not increase to a greater extent than in control lambs during a considerable additional load on the heart.

Body fluid compartment volumes in chronically hypoxemic lambs.

Chronic hypoxemia is associated with a decreased growth rate and a decreased rate of cell division. In lambs with experimental cyanotic heart disease, the gain in body mass was decreased. In this study, we determined to what extent the lower body mass in these lambs was related to alterations in body fluid compartment volumes, specifically whether intracellular volume was decreased. Therefore, fluid compartment volumes were studied in nine lambs, after 3 to 4 wk of experimental cyanotic heart disease, and in 13 control lambs. Hypoxemic lambs had a lower arterial oxygen saturation (65 +/- 11 versus 91 +/- 2%, p < 0.001) and a higher Hb concentration (142 +/- 16 versus 101 +/- 8 g.L-1, p < 0.001). The lower body mass in hypoxemic lambs (10.5 +/- 2.3 versus 13.0 +/- 2.8 kg) could be mainly accounted for by a decrease in intracellular water volume (4.7 +/- 1.3 versus 6.6 +/- 1.5 L, p < 0.01). Total body water (753 +/- 27 versus 780 +/- 40 mL.kg-1) and extracellular water volume (307 +/- 25 versus 277 +/- 15 mL.kg-1) in hypoxemic lambs were not significantly different from those in control lambs, but intracellular water volume was decreased (445 +/- 27 versus 501 +/- 35 mL.kg-1, p < 0.01). The ratio of extracellular to intracellular water volume was higher in hypoxemic lambs (0.69 +/- 0.10 versus 0.55 +/- 0.04, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Blood volume and body fluid compartments in lambs with aortopulmonary left-to-right shunts.

A left-to-right shunt is accompanied by an increased plasma and blood volume. Since this is likely realized through renin/aldosterone-mediated salt and water retention, other body fluid compartments may be changed too. Therefore, we studied blood volume and body fluid compartments by a single-injection, triple-indicator dilution technique in nine 8-wk-old lambs with an aortopulmonary left-to-right shunt (55 +/- 3% of left ventricular output; mean +/- SEM) and in 11 control lambs, 2.5 wk after surgery. Systemic blood flow was maintained at the same level as in control lambs, but the aortic pressure of the shunt lambs was lower. Blood volume in shunt lambs was larger than in control lambs (110 +/- 6 vs. 84 +/- 7 ml/kg, P < 0.001) through an increase in plasma volume, which correlated significantly with the magnitude of the left-to-right shunt (r = 0.81, P < 0.01). Red blood cell volume was equal to that of control lambs. Evidence was obtained that the increase in plasma volume was induced by a transient increase in renin (8.0 +/- 2.2 vs. 1.6 +/- 0.2 nmol.l-1.h-1; P < 0.02) and aldosterone (0.51 +/- 0.14 vs. 0.24 +/- 0.09 nmol/liter) concentrations. Interstitial water volume, however, was not significantly different from that in control lambs. The amount of intravascular protein was significantly higher than in control lambs (5.0 +/- 0.3 vs. 3.5 +/- 0.2 g/kg body mass, P < 0.001). There were no significant differences in intracellular and total body water volumes between the two groups. We conclude that the increased amount of intravascular protein confines the fluid retained by the kidneys to the vascular compartment.

Distribution of systemic blood flow in lambs with an aortopulmonary shunt during strenuous exercise.

We studied regional blood flows with radioactive-labeled microspheres in 12 7-wk-old lambs with an aortopulmonary left-to-right shunt [59 +/- 3% (SE) of left ventricular (LV) output] and in 11 control lambs, at rest and during exercise at 80% of predetermined peak O2 consumption. At rest, systemic blood flow was similar in the two groups. Blood flow to the heart and diaphragm was substantially higher in the shunt than in the control lambs. Blood flow to the other organs was not significantly different between the two groups. During exercise, systemic blood flow increased substantially but less in shunt (81%) than in control lambs (134%). Blood flow to the heart and diaphragm increased, that to the heart still being higher in shunt than in control lambs. Blood flow to the brain did not change, whereas that to the kidneys and splanchnic organs decreased to the same extent (25%) in shunt and control lambs. Intrahepatic and intrarenal blood flow redistribution in the shunt lambs persisted during exercise. In conclusion, myocardial blood flow is not increased at the expense of one particular organ, nor is it associated with an essential change in exercise-induced redistribution in shunt lambs.

Myocardial carbohydrate, ketone, and fatty acid uptake in conscious lambs with aortopulmonary shunts.

A left to right shunt increases myocardial work and is often accompanied by increased catecholamine levels. Because both increased myocardial work and increased catecholamine levels may induce increased fatty acid utilization, which could increase resting myocardial oxygen consumption and therefore unfavorably affect coronary reserve, we studied myocardial uptake of glucose, pyruvate, lactate, beta-OH-butyrate, acetoacetate, FFA, and triglycerides in 12 7-wk-old lambs with aortopulmonary left to right shunts (58 +/- 2% of left ventricular output, mean +/- SEM) and in 10 control lambs 2 wk after surgery. Despite the shunt, systemic blood flow in the shunt lambs was maintained at the same level as in the control lambs. This was accomplished by an increased heart rate and stroke volume. Furthermore, the shunt was accompanied by an increased myocardial oxygen consumption in the shunt lambs (834 +/- 70 versus 528 +/- 43 mumol O2.min-1 x 100 g-1; p less than 0.05). There were no significant differences in arterial substrate concentrations between the two groups. The same was true for arteriovenous differences across the myocardium, with the exception of lactate, which was substantially higher in shunt than in control lambs (72 +/- 25 versus 18 +/- 23 mumol/L; p less than 0.05). As a consequence, myocardial lactate uptake in the shunt lambs was increased 15-fold (18 +/- 6 versus 1 +/- 2 mumol.min-1 x 100 g-1; p less than 0.02), whereas uptake of the other substrates merely paralleled the increased myocardial blood flow.(ABSTRACT TRUNCATED AT 250 WORDS)

Maximal exercise capacity and oxygen consumption of lambs with an aortopulmonary left-to-right shunt.

We determined maximal exercise capacity and measured hemodynamics in 10 6-wk-old lambs with an aortopulmonary left-to-right shunt [S, 57 +/- 11%, (SD)] and in 9 control lambs (C) during a graded treadmill test 8 days after surgery. Maximal exercise capacity (3.7 +/- 0.2 km/h and 10 +/- 5% inclination vs. 4.0 +/- 0.9 km/h and 15 +/- 0% inclination, P less than 0.02) and peak oxygen consumption (25 +/- 7 vs. 34 +/- 8 ml O2.min-1.kg-1, P less than 0.02) were both lower in the shunt than in the control lambs. This was due to a lower maximal systemic blood flow in the shunt lambs (271 +/- 38 vs. 359 +/- 71 ml.min-1.kg-1, P less than 0.01). Despite their high maximal left ventricular output, which was higher than in the control lambs (448 +/- 87 vs. 359 +/- 71 ml.min-1.kg-1, P less than 0.05), the left-to-right shunt could not be compensated for during maximal exercise because of a decreased reserve in heart rate (S: 183 +/- 22 to 277 +/- 38 beats/min; C: 136 +/- 25 to 287 +/- 29 beats/min) and in left ventricular stroke volume (S: 1.8 +/- 0.3 to 1.6 +/- 0.4 ml/kg; C: 1.0 +/- 0.3 to 1.3 +/- 0.2 ml/kg). We conclude that exercise capacity of shunt lambs is lower than that of control lambs, despite a good left ventricular performance, because a part of the reserves for increasing the left ventricular output is already utilized at rest.

Effects of nitroprusside on myocardial blood flow and oxygen consumption in conscious lambs with an aortopulmonary left-to-right shunt.

We studied the effect of vasodilation on regional myocardial blood flow and oxygen consumption of the left ventricular free wall by infusing 10 micrograms/kg/min sodium nitroprusside into 7-week-old conscious lambs with and without aortopulmonary left-to-right shunts. Resting myocardial oxygen consumption in the 13 shunt lambs was significantly higher than in the nine control lambs (989 +/- 104 [+/- SEM] vs. 432 +/- 41 mumol/min/100 g left ventricle). This was achieved by a significantly higher left ventricular myocardial blood flow (294 +/- 33 vs. 143 +/- 16 ml/min/100 g left ventricle) because the arteriovenous oxygen concentration difference across the left ventricular free wall was similar in shunt and control lambs. Infusion of nitroprusside did not significantly change myocardial oxygen consumption and regional myocardial blood flows at 10 and 50 minutes after the onset of the infusion. This occurred despite a substantial drop in aortic and left atrial pressures and stroke volume, which decreases wall stress as well as external work of the left ventricle. Heart rate, however, increased significantly. We postulate that, during infusion of nitroprusside, the potential decrease in myocardial oxygen consumption due to a decrease in wall stress and external work of the left ventricle is neutralized by the consequences of the increased heart rate. In view of this and because of its hemodynamic effects, we do not consider sodium nitroprusside useful in the treatment of circulatory congestion in patients with left-to-right shunts, normal arterial pressures, and normal systemic blood flows.

Circulatory effects of sodium nitroprusside in the conscious lamb with an aortopulmonary left to right shunt.

We studied the effect on the circulation of reducing peripheral vascular resistance by infusing sodium nitroprusside into lambs of three different age groups (subgroup A, 11-26 days, subgroup B, 32-52 days, and subgroup C, 61-88 days of age) with and without an aortopulmonary left to right shunt. Infusion of 10 micrograms/kg/min nitroprusside over 2 h decreased aortic and left atrial pressures markedly and increased heart rate, whereas systemic, pulmonary, and left to right shunt blood flows hardly changed. Within 30 min after the onset of infusion, the hemodynamic variables stabilized. Aortic and left atrial pressures were still below control levels at that time. The different flows remained the same and heart rate, after an initial fall, increased again. The pattern of hemodynamic changes was not influenced by age or the presence of an aortopulmonary left to right shunt. Based on this study, we do not advocate sodium nitroprusside administration in case of a left to right shunt with normal arterial pressure and systemic blood flow.

Comparative circulatory effects of isoproterenol, dopamine, and dobutamine in conscious lambs with and without aortopulmonary left-to-right shunts.

We studied the effect on the circulation of the catecholamines isoproterenol, dopamine, and dobutamine in chronically instrumented lambs with aortopulmonary left-to-right shunts (ages 11 to 87 days) and without shunts (ages 8 to 97 days). Infusion of 0.1 microgram/kg/min isoproterenol or 10 micrograms/kg/min dobutamine markedly increased heart rate and systemic and pulmonary blood flows, while stroke volume and the left-to-right shunt flow did not change. Since pulmonary blood flow increased and the left-to-right shunt flow did not change, the left-to-right shunt fraction decreased with the infusions of isoproterenol and dobutamine. The hemodynamic changes during the infusion of isoproterenol and dobutamine occurred immediately after the start of infusion and stabilized within a few minutes. The pattern of hemodynamic changes was not influenced by the presence of an aortopulmonary left-to-right shunt or by age. Infusion of 10 micrograms/kg/min dopamine caused only small hemodynamic changes. This study shows that heart rate and systemic blood flow in the lamb are closely related. Furthermore, it demonstrates that despite an increased systemic blood flow, left-to-right shunt flow does not change after infusion of isoproterenol and dobutamine. Any decision as to which positive inotropic agent might be preferred at an early age should await experimental work concerning the effect of these agents on the myocardial oxygen demand and on the distribution of the systemic blood flow.

Myocardial blood flow and VO2 in conscious lambs with an aortopulmonary shunt.

We measured myocardial blood flow (QLVFW) and O2 consumption of the left ventricular free wall (VO2, LVFW) in 13 chronically instrumented, 7-wk-old lambs with an aortopulmonary left-to-right shunt (S) and in 10 control lambs without a shunt (C). The measured VO2, LVFW was compared with the calculated values obtained by two predictive indexes, the rate-pressure product (RPP) and the pressure-work index (PWI). Measured VO2, LVFW in S lambs was significantly higher than in C lambs [983 +/- 104 (SE) vs. 475 +/- 57 mumol X min-1 X 100 g LV-1, P less than 0.001]. This was achieved by the significantly higher QLVFW (294 +/- 33 vs. 145 +/- 15 ml X min-1 X 100 g LV-1, P less than 0.002), since the arteriovenous O2 difference across the left ventricular free wall was similar in both groups of lambs. Total coronary blood flow per unit body mass in S lambs was higher than in C lambs (14.1 +/- 1.5 vs. 5.6 +/- 0.6 ml X min-1 X kg-1, P less than 0.001), not only because of the increased VO2, LVFW per unit muscle mass, but also because of the increased total heart weight (102.4 +/- 6.4 vs. 81.2 +/- 4.9 g, P less than 0.02). Correct estimation of VO2, LVFW by means of the RPP and the PWI was only possible in C lambs. In S lambs the estimated values were significantly lower than the measured ones.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparative study of four methods for quantifying Doppler ultrasound waveforms from the femoral artery.

Currently, there is no agreement as to the best method for quantifying Doppler ultrasound recordings from peripheral arteries in order to detect occlusive disease. The four methods assessed in this study are: the pulsatility index, height-width index, path length index, and a Laplace transform function index. Recordings of the Doppler ultrasound spectral waveforms from the common femoral artery of 232 limbs were digitized to obtain the maximum velocity waveforms. This data was analyzed and the various indices were computed and then compared with the arteriographic grades. The effect of distal disease was also examined. The diagnostic accuracy of each index was determined from receiver operating characteristics curves. We concluded that all four indices were capable of detecting significant aortoiliac disease with approximately equal diagnostic accuracy of 90-95% but that pulsatility index had the advantages of simplicity and ease of calculation.