Bioartificial Liver Using Hepatocytes on Biosilon Microcamers: Treatment of Chemically Induced Acute Hepatic Failure in Rats

An artificial liver support procedure based on hemoperfusion via hepatocytes cultured on microcarriers is described. The efficiency of the system was assessed by the survival rate of rats treated with either lethal dosage of 7% CCl4 [30 ml/kg body weight (b.w.)] or D-galactosamine (2.5 g/kg b.w.). In CCl4-treated rats, hemoperfusion via empty microcarriers (n = 16) revealed no surviving animals, whereas the use of the bioartificial liver (n = 11) resulted in 80% (p less than 0.01) and 60% (p less than 0.05) survival 48 and 168 h after hepatotoxin, respectively. For the same time periods, the survival rate in D-galactosamine-intoxicated rats after hemoperfusion with hepatocytes (n = 20) was approximately 60% (p less than 0.05) and was only 5% in those of rats treated with empty microcarriers (n = 20). Sublethal dosage of 7% CCl4 (15 ml/kg b.w.) caused 25% mortality and prolonged (48 h) increase of activity of the liver enzymes and bilirubin levels in the serum of surviving animals. In these rats (n = 8) at the end of 3 h of hemoperfusion via hepatocytes, the bilirubin concentration decreased by 45% as compared with the control group (n = 6) treated with empty microcarriers. Moreover, by 48 h after intoxication, the use of the bioartificial liver resulted in more than a three-fold decrease in glutamate-oxaloacetate transaminase and a 10-fold decrease in glutamate-pyruvate transaminase serum activity as well as a fivefold decline in total and a ninefold decline in conjugated bilirubin levels as compared with the control animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Development of a Potassium-Specific Adsorbent for Direct Hemoperfusion

Abstract: Direct hemoperfusion (DHP) over an exchange resin for sodium is capable of rapidVVly lowering an vated serum potassium level but has not been used clin- ically because of subsequent electrolyte abnormalities. After the completion of detailed in vitro studies, we de- veloped a sodium/calcium/magnesium exchange resin mixture specifically to remove potassium from the blood. The mixture, which consisted of exchange resins for so- dium (48%), calcium (48%), and magnesium (4%), gave the most satisfactory results in vitro. DHP through these resin-mixture columns for 2 h reduced elevated plasma potassium levels (6.7 ± 1.1 to 3.5 ± 0.6 mEq/L, p < 0.001) in anephric dogs without including any significant change in the levels of sodium, calcium, magnesium, al- bumin, total protein, or cholesterol. The platelet losses and changes in the plasma free hemoglobin levels ob- served were similar to those found with activated char- coal hemoperfusion. These results suggest that this resin hemoperfusion may be useful in patients with acute hyperkalemia.     

丙酮酸乙酯对脓毒症休克犬肠黏膜屏障功能的保护作用

目的探讨丙酮酸乙酯（EP）对脓毒症休克犬肠黏膜屏障功能的影响。方法健康雄性杂种犬20只，内毒素（LPS）静脉注射复制犬脓毒症休克模型，随机分为对照组（8只）和EP组（12只）。对照组只接受林格液复苏。EP组另外给予丙酮酸乙酯首剂0．05g／kg．然后按0．05g·kg^-1·h^-1持续泵入。脓毒症休克模型建立前及建立后0、8、12和24h取血测定血浆二胺氧化酶（DAO）活性和血浆D-乳酸含量，试验24h处死动物后取小肠标本，进行肠黏膜炎性损伤病理学评分。结果EP组犬的肠黏膜炎性损伤程度病理学评分为2．33±0．25，明显轻于模型组的3．39±0．38，两组比较，差异有统计学意义（P〈0．05）。两组实验犬在休克后血浆D-乳酸含量和DAO活性逐渐升高，对照组较EP组升高更为明显（P〈0．05）。结论EP能显著改善肠组织灌流及功能指标，减轻肠黏膜的病理损害，对脓毒症休克时小肠有保护作用。     

Comparison of a pulsatile blood pump and a peristaltic roller pump during hemoperfusion treatment in a canine model of paraquat poisoning

Abstract:  This study examined the treatment efficacy and the damage to the blood during hemoperfusion for treating paraquat poisoning using two blood pump mechanisms. Paraquat-poisoned animal models were prepared. A conventional hemodialysis machine, AK90, with a peristaltic roller pump and a cardiopulmonary support system, T-PLS, with a pulsatile blood pump were used during the animal experiments. A total of 12 dogs were treated with hemoperfusion using a charcoal column. Six dogs were treated with hemoperfusion and T-PLS, and the other six were treated with AK90. A paraquat dose of 30 mg/kg was administrated by an intravenous injection. Both pumps maintained blood flow rates of 125 mL/min measured by an ultrasonic flowmeter. For anticoagulation, heparin was administrated by an initial bolus (250 IU/kg) and a continuous injection (100 IU/kg/h). During the experiments, T-PLS and AK90 showed a similar toxin removal efficacy. Both devices decreased the plasma paraquat concentration to 10% of the initial dose within 4-h hemoperfusion. The two pumps showed similar hemolysis properties with acceptable levels. Although T-PLS was developed as a cardiopulmonary bypass system, it can also be used as a hemoperfusion treatment device.     

Implanted Right Atrial Catheters for Continuous Infusion of Solutions into Dogs

Silastic catheters were fabricated and aseptically implanted through the skin into the jugular vein of 64 dogs with the intravascular tip located in the right atrium. Solutions were infused through the catheter at 2 to 2.5 mL/h by a portable pump worn by the dog. Following 9.2 Gy total body irradiation (TBI) and allogeneic bone marrow transplantation (BMT), succinyl acetone, an experimental chemotherapeutic agent, was infused into 34 dogs. Hematopoietic growth factors were infused into an additional 30 dogs, two of which had 9.2 Gy TBI and an autologous BMT, and four of which had 4.0 Gy TBI and no BMT. All dogs received continuous oral and parenteral antibiotics while the catheters were in place. All catheters functioned well until electively removed (n= 28) or until the dogs died or were euthanized (n= 36) at 12 to 68 days after implantation. Mean length of catheter function was 30.3 ± 1.5 (SEM) days. No catheters were dislodged and there was no evidence of catheter-related blood loss or sepsis. Semiquantitative cultures of 5 catheters were negative, but Staphylococcus epidermidis was isolated from 3 of 7 catheters cultured in broth. Six dogs had thrombosis adjacent to the intravascular catheter tip. The catheters were well tolerated and facilitated successful long-term infusion of solutions into dogs.     

Moderate running exercise augments glycosaminoglycans and thickness of articular cartilage in the knee joint of young beagle dogs

The local influences of physical exercise on thickness and glycosaminoglycan (GAG) content of canine articular cartilage were measured by microspectrophotometry of Safranin O- and periodic acid-Schiff (PAS)-stained tissue sections. Female Beagle dogs were housed in individual cages (bottom 0.9 × 1.2 m) and divided into runner (n = 6) and control (n = 8) groups. The trainig program started at the age of 15 weeks. During the subsequent 10 weeks, the dogs were accustomed to running on a treadmill inclined 15° uphill. Thereafter, the dogs ran 1 h daily, 5 days a week, at a speed of 4 km/h for 15 weeks. At the age of 40 weeks, the dogs were killed, and the samples for histology were taken from 11 different anatomical locations of the right knee (stifle) joint. The thickness of the uncalcified cartilage increased 19–23% on the lateral condyle and patellar surface of the femur, whereas the enhancement was smaller in other parts of the trained cartilage. The calcified cartilage did not show thickness alterations. Total GAGs were augmented by 28% in the summits on the femoral condyles, more on the medial than lateral side. The increased GAGs appeared to be predominantly chondroitin sulphates and were localized in the intermediate, deep, and even in the calcified zones, whereas the superficial zone did not show changes. There was a concomitant increase of non-GAG oligosaccharides in the intermediate and deep zones, but not in the calcified cartilage. As judged from the enhanced GAG content and thickness, it is considered that moderate running exercise locally alters the biological properties of young articular cartilage at regions bearing the highest loading surplus.     

Ethanol enhances the functional recovery of stunned myocardium independent of K(ATP) channels in dogs

Chronic, intermittent exposure to small amounts of ethanol reduces myocardial infarct size in vivo. We tested the hypothesis that acute administration of ethanol enhances the functional recovery of stunned myocardium and that adenosine triphosphate-dependent potassium (K(ATP)) channels mediate this beneficial effect. Barbiturate-anesthetized dogs were instrumented for measurement of aortic and left ventricular pressure, +dP/dt(max), and subendocardial segment shortening (%SS) and were subjected to five 5-min periods of coronary artery occlusion, each separated by 5 min of reperfusion followed by a 3-h final reperfusion. In four groups (n = 7 each), dogs received 0.9% saline or ethanol (0.25, 0.5, or 1.0 g/kg over 30 min) in a random manner before occlusions and reperfusions. In other groups (n = 7 each), dogs received the K(ATP) channel antagonist glyburide (0.3 mg/kg, IV) 30 min before saline or ethanol (0.25 g/kg) was administered. Dogs receiving saline or glyburide alone demonstrated poor recovery of contractile function during reperfusion (%SS = 0.9% +/- 2.0% and 1.6% +/- 1.2% at 3 h, respectively). Recovery of %SS was enhanced in dogs receiving the 0.25- and 0.5-g/kg doses of ethanol (10.0% +/- 1.8% and 8.6% +/- 2.2% at 3 h, respectively) independent of alterations in hemodynamics or coronary collateral blood flow (radioactive microspheres). Glyburide did not affect improvement of recovery of stunned myocardium produced by ethanol (11.8% +/- 2.2% at 3 h). The results indicate that ethanol enhances the functional recovery of stunned myocardium independent of K(ATP) channels in vivo.     

FLT3 ligand promotes engraftment of allogeneic hematopoietic stem cells without significant graft-versus-host disease

BACKGROUND: Graft-versus-host (GVH) reactions contribute to stable engraftment of allogeneic hematopoietic stem cell transplants. It was hypothesized that the in vivo expansion of recipient dendritic cells (DC) with the administration of ligand for Flt3 (FL) could promote allogeneic engraftment after reduced-intensity conditioning by enhancing the GVH effect. METHODS: FL was first administered to three nonirradiated healthy dogs for 13 days at a dosage of 100 microg/kg/day. Next, nine dogs received 4.5 Gy total-body irradiation (TBI) and unmodified marrow grafts from dog leukocyte antigen (DLA)-identical littermates without posttransplant immunosuppression. FL was administered to the recipients at a dosage of 100 microg/kg/day from day -7 until day +5. RESULTS: In normal dogs, FL produced significant increases in monocytes (CD14+) and neutrophils in the peripheral blood, a marked increase in CD1c+ cells with DC-type morphology in lymph nodes, and increased alloreactivity of third-party responders to peripheral blood mononuclear cells in mixed lymphocyte reactions (P<0.001). Sustained engraftment was observed in eight of nine (89%) FL-treated dogs compared with 14 of 37 (38%) controls (P=0.02, logistic regression). All engrafted FL-treated dogs became stable complete (n=2) or mixed (n=6) hematopoietic chimeras without significant graft-versus-host disease (GVHD). Recipient chimeric dogs (n=4) were tolerant to skin transplants from their marrow donors but rejected skin grafts from unrelated dogs within 7 to 9 days (median, 8 days). CONCLUSIONS: In this study, the authors showed that FL administered to recipients promotes stable engraftment of allogeneic marrow from DLA-identical littermates after 4.5 Gy TBI without significant GVHD.     

Insulin reverses bupivacaine-induced cardiac depression in dogs

We tested the hypothesis that an insulin infusion would effectively treat bupivacaine-induced cardiac depression in dogs. In 24 mongrel dogs anesthetized with pentobarbital (5 mgkg(-1)h(-1), IV), 0.5% bupivacaine was administrated at a rate of 0.5 mgkg(-1)min(-1) until the mixed venous oxygen saturation decreased to 60% or less. The bupivacaine infusion induced a decrease in mean arterial pressure, cardiac output, and heart rate. The dogs were randomly assigned to one of four groups after the end of bupivacaine infusion. The Control (C, n = 6) and Glucose (G, n = 6) groups received an IV infusion of normal saline (2 mL/kg) and glucose (2 mL/kg of 50% dextrose in water) for 15 min, respectively. The Insulin-Glucose (IG, n = 6) group received an IV bolus of regular insulin (1 U/kg), plus a glucose infusion (2 mL/kg of 50% dextrose in water) for 15 min. The Insulin-Glucose-Potassium (IGK, n = 6) group received the same dose of insulin and glucose as the IG group, plus potassium (1-3 mEqkg(-1)h(-1)). Mean arterial pressure, cardiac output, heart rate, and mixed venous oxygen saturation recovered toward baseline level more rapidly in the IG and IGK groups than in the C group (within 5 min versus more than 20 min). These results suggest that the infusion of insulin and glucose might reverse bupivacaine-induced cardiac depression in dogs. Implications: We found that insulin and glucose rapidly reversed hemodynamic abnormality in dogs with bupivacaine-induced cardiac depression. This study implies a possible clinical application of insulin treatment for bupivacaine-induced cardiac depression.     

Rapid Hypothermic Aortic Flush Can Achieve Survival without Brain Damage after 30 Minutes Cardiac Arrest in Dogs

Background: Neither exsanguination to pulselessness nor cardiac arrest of 30 min duration can be reversed with complete neurologic recovery using conventional resuscitation methods. Techniques that might buy time for transport, surgical hemostasis, and initiation of cardiopulmonary bypass or other resuscitation methods would be valuable. We hypothesized that an aortic flush with high-volume cold normal saline solution at the start of exsanguination cardiac arrest could rapidly preserve cerebral viability during 30 min of complete global ischemia and achieve good outcome.

Methods: Sixteen dogs weighing 20-25 kg were exsanguinated to pulselessness over 5 min, and circulatory arrest was maintained for another 30 min. They were then resuscitated using closed-chest cardiopulmonary bypass and had assisted circulation for 2 h, mild hypothermia (34[degrees]C) for 12 h, controlled ventilation for 20 h, and intensive care to outcome evaluation at 72 h. Two minutes after the onset of circulatory arrest, the dogs received a flush of normal saline solution at 4[degrees]C into the aorta (cephalad) via a balloon catheter. Group I (n = 6) received a flush of 25 ml/kg saline with the balloon in the thoracic aorta; group II (n = 7) received a flush of 100 ml/kg saline with the balloon in the abdominal aorta.

Results: The aortic flush decreased mean tympanic membrane temperature (Tty) in group I from 37.6 +/- 0.1 to 33.3 +/- 1.6[degrees]C and in group II from 37.5 +/- 0.1 to 28.3 +/- 2.4[degrees]C (P = 0.001). In group I, four dogs achieved overall performance category (OPC) 4 (coma), and 2 dogs achieved OPC 5 (brain death). In group II, 4 dogs achieved OPC 1 (normal), and 3 dogs achieved OPC 2 (moderate disability). Median (interquartile range [IQR]) neurologic deficit scores (NDS 0-10% = normal; NDS 100% = brain death) were 69% (56-99%) in group I versus 4% (0-15%) in group II (P = 0.003). Median total brain histologic damage scores (HDS 0 = no damage; >100 = extensive damage; 1,064 = maximal damage) were 144 (74-168) in group I versus 18 (3-36) in group II (P = 0.004); in three dogs from group II, the brain was histologically normal (HDS 0-5).     

Effect of excitatory amino acid receptor blocker MK-801 on overall, neurologic, and morphologic outcome after prolonged cardiac arrest in dogs

Excitatory amino acids accumulating in the brain during ischemia may cause selective neuronal damage postischemia. This hypothesis was tested in a series of studies using MK-801, an N-methyl-D-aspartate (NMDA) receptor blocker, in a reproducible outcome model of prolonged cardiac arrest in dogs. After normothermic ventricular fibrillation cardiac arrest, the dogs were resuscitated with closed-chest femoral veno-arterial cardiopulmonary bypass. At 4 h they were separated from bypass, ventilation was controlled for 20 h, and intensive care was continued to 96 h. In Study I, ventricular fibrilation cardiac arrest (no-flow) was 17 min; starting immediately with reperfusion, MK-801 1200 mg/kg (n = 5) or an equal volume of placebo (n = 5) was infused over 12 h in blinded, randomized fashion. In Study II, the duration of the no-flow period was reduced to 15 min, and MK-801 2400 mg.kg-1 (n = 4) or placebo (n = 4) was infused. In Study III, no-flow lasted for 15 min, and MK-801 2400 mg/kg was started 30 min before ventricular fibrillation (n = 4); comparison was with Study II controls. In all three studies, MK-801 plasma concentrations peaked at greater than 50 ng/ml and were 15-30 ng/ml over 12 h. All 22 dogs of experiments within protocol survived with severe brain damage. MK-801 delayed return of pupillary reactivity, EEG activity, consciousness, and respiration, necessitating longer periods of controlled ventilation. Neurologic deficit scores, overall performance categories, and brain and heart morphologic damage scores at 96 h did not differ between placebo and MK-801 pretreatment or post-treatment groups. These negative outcome results after prolonged cardiac arrest do not negate the hyperexcitability hypothesis of selective vulnerability, but suggest the existance of additional mechanisms of secondary brain damage.     

Failure of motor evoked potentials to predict neurologic outcome in experimental thoracic aortic occlusion

Motor evoked potential monitoring was tested as an alternative to somatosensory evoked potential monitoring in evaluating spinal cord function during thoracic aortic occlusion in dogs. Twenty-seven animals underwent 60 minutes of cross-clamping of the proximal descending thoracic aorta with (n = 18) or without (n = 9) cerebrospinal fluid drainage. Spinal cord blood flow was measured with microspheres, and neurologic outcome was evaluated at 24 hours with Tarlov's scoring system. Cerebrospinal fluid drainage improved neurologic outcome (p less than 0.05). Motor evoked potentials recorded over the lumbar spinal cord were lost in 9 of 20 dogs with ischemic cord injury and were not lost in any of the 7 dogs that were neurologically normal. Somatosensory evoked potential were lost in 19 of 20 paraplegic/paraparetic dogs and lost in 3 of 7 normal dogs (p less than 0.01). After reperfusion, motor evoked potentials returned in all nine neurologically injured dogs that lost the potentials and were still present at 24 hours. Changes in amplitude, latency, or time until loss or return of motor evoked potentials or somatosensory evoked potentials did not predict neurologic injury. Loss of somatosensory evoked potentials had a high sensitivity (95%) but had low specificity (67%) because of peripheral nerve ischemia. Loss of motor evoked potentials recorded from the spinal cord had high specificity (100%) but a low sensitivity (46%) and was therefore not a reliable predictor of neurologic injury. Return of motor evoked potentials during reperfusion did not correlate with functional recovery. Motor evoked potentials stimulated in the cortex and recorded from the spinal cord had low overall accuracy (59%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of prolonged surgical trauma on the extravascular lung water and central blood volume in the dog

Extravascular lung water (EVLW) and central blood volume (CBV) were measured in 13 dogs with a double-indicator dilution technique (dye-cold), the indicators being detected intravascularly. Animals in a control group (n = 5) were mechanically ventilated for 8-15 h after baseline measurements. Another group of animals (n = 8) were subjected to extensive lymph duct cannulations, including a thoracoabdominal incision, and the dogs were followed for 8 to 18 h postoperatively. All dogs (n = 13) then received a dose of oleic acid intravenously in order to create lung damage. A positive fluid balance was maintained throughout the experiment. Basal EVLW was 8.8 ml/kg (n = 13) (similar in both groups), and did not change significantly in either group before oleic acid. Basal CBV was 18.5 ml/kg (n = 13); it increased (P less than 0.05) in the control group and decreased (P less than 0.05) in the surgery group during a 8-15-h period. EVLW was doubled (P less than 0.001) and CBV decreased slightly 2 h after oleic acid administration. The lung damage was similar in both groups, and was accompanied by increased pulmonary vascular pressures and marked reductions in arterial oxygen tension and thoracic compliance. The findings suggest that an earlier proposed dog model for the simultaneous measurement of EVLW and lymph flow can be used in long-term studies on lung fluid balance.     

The effect of halothane anesthesia on myocardial necrosis, hemodynamic performance, and regional myocardial blood flow in dogs following coronary artery occlusion

The effect of halothane anesthesia on myocardial necrosis resulting from coronary artery ligation was examined in 28 anesthetized mongrel dogs. In 18 dogs, the left anterior descending coronary artery (LAD) was ligated immediately proximal to the first apical diagonal branch, and 1 h later the dogs were assigned randomly either to receive halothane, 0.5-1.0% inspired in room air for 12 h (n = 10) or to awaken without further intervention (control, n = 8). Infarct size was measured by staining the myocardium with triphenyl tetrazolium chloride 24 h after LAD ligation. Infarct size in halothane-treated dogs was 17.8 +/- 2.0% of the left ventricle, compared with 27.3 +/- 3.3% in control dogs (P less than 0.05). Myocardial salvage was present transmurally but was greatest in epicardial regions. In 10 additional dogs, hemodynamic variables (heart rate, arterial pressure, left ventricular end-diastolic pressure, peak left ventricular dP/dt, tension-time index, and rate-pressure product) were measured or calculated, and radionuclide-labeled microspheres were injected for measurement of cardiac output and regional myocardial blood flow (RMBF). Thirty minutes after LAD ligation and after initial hemodynamic measurements and microsphere injection, these dogs were assigned randomly to receive either halothane, 1.0%, inspired in room air (n = 5) or no intervention (control, n = 5). After 15 min of halothane inhalation (45 min after LAD ligation in control dogs), measurements were repeated. Halothane inhalation reduced heart rate, arterial pressure, and indexes of left ventricular contractile and pump performance. During halothane treatment, RMBF declined in normal myocardium but not in ischemic regions, while neither normal nor ischemic zone RMBF changed in control dogs. Systemic vascular resistance was unchanged in either group. Thus, halothane was associated with a 35% smaller myocardial infarct, transmural myocardial salvage, reduced heart rate, reduced left ventricular contractile and pump performance, reduced RMBF to nonischemic regions, and unchanged RMBF in the ischemic myocardium.     

Rapid hypothermic aortic flush can achieve survival without brain damage after 30 minutes cardiac arrest in dogs

BACKGROUND: Neither exsanguination to pulselessness nor cardiac arrest of 30 min duration can be reversed with complete neurologic recovery using conventional resuscitation methods. Techniques that might buy time for transport, surgical hemostasis, and initiation of cardiopulmonary bypass or other resuscitation methods would be valuable. We hypothesized that an aortic flush with high-volume cold normal saline solution at the start of exsanguination cardiac arrest could rapidly preserve cerebral viability during 30 min of complete global ischemia and achieve good outcome. METHODS: Sixteen dogs weighing 20-25 kg were exsanguinated to pulselessness over 5 min, and circulatory arrest was maintained for another 30 min. They were then resuscitated using closed-chest cardiopulmonary bypass and had assisted circulation for 2 h, mild hypothermia (34 degrees C) for 12 h, controlled ventilation for 20 h, and intensive care to outcome evaluation at 72 h. Two minutes after the onset of circulatory arrest, the dogs received a flush of normal saline solution at 4 degrees C into the aorta (cephalad) via a balloon catheter. Group I (n = 6) received a flush of 25 ml/kg saline with the balloon in the thoracic aorta; group II (n = 7) received a flush of 100 ml/kg saline with the balloon in the abdominal aorta. RESULTS: The aortic flush decreased mean tympanic membrane temperature (Tty) in group I from 37.6 +/- 0.1 to 33.3 +/- 1.6 degrees C and in group II from 37.5 +/- 0.1 to 28.3 +/- 2.4 degrees C (P = 0.001). In group 1, four dogs achieved overall performance category (OPC) 4 (coma), and 2 dogs achieved OPC 5 (brain death). In group II, 4 dogs achieved OPC 1 (normal), and 3 dogs achieved OPC 2 (moderate disability). Median (interquartile range [IQR]) neurologic deficit scores (NDS 0-10% = normal; NDS 100% = brain death) were 69% (56-99%) in group I versus 4% (0-15%) in group II (P = 0.003). Median total brain histologic damage scores (HDS 0 = no damage; > 100 = extensive damage; 1,064 = maximal damage) were 144 (74-168) in group I versus 18 (3-36) in group II (P = 0.004); in three dogs from group II, the brain was histologically normal (HDS 0-5). CONCLUSIONS: A single high-volume flush of cold saline (4 degrees C) into the abdominal aorta given 2 min after the onset of cardiac arrest rapidly induces moderate-to-deep cerebral hypothermia and can result in survival without functional or histologic brain damage, even after 30 min of no blood flow.     

Combination therapy with microwave coagulation and intracavitary irradiation for bladder cancer. Technique and preliminary clinical results

Twenty-five patients with transitional cell carcinoma of the bladder have been treated with combined therapy consisting of microwave regional coagulation and intracavitary irradiation. A remote-controlled after-loading system was utilized for the radiation therapy. The follow-up period ranged from 6 to 19 months with an average of 11.4 months. Tumor stages were Tis (n = 2), Ta or T1 (n = 17), T2 (n = 2), T3 (n = 3) and T4 (n = 1), and grades were G1 (n = 10), G2 (n = 11) and G3 (n = 4). In 23 patients (92%), there was no endoscopic or histologic evidence of tumor after the initial treatment. Heterotopic recurrences were found after 2 or 3 months in 3 patients who received microwave regional coagulation or intracavitary regional irradiation. Additional intracavitary whole bladder mucosal irradiation was performed for 10 patients with multiple tumors and frequent recurrent tumors. Nine patients had no recurrence (average follow-up 11 months). Our preliminary findings indicate that combination therapy of microwave coagulation and intracavitary irradiation is a useful treatment for bladder cancer.     

Morphological and hemodynamic changes of the lung after injection of 5% ethanolamine oleate into dogs

We examined the pulmonary hemodynamics and morphology after injection of a sclerosing solution of 5% ethanolamine oleate (EO) into 24 normal dogs. EO of 0.5 ml/kg (n = 5), 1.0 ml/kg (n = 6), and 3.0 ml/kg (n = 7) was injected through the jugular vein into the right atrium for pathological examination and gravimetric study of the lung, while monitoring the pulmonary hemodynamics for 12 h. Normal saline of 3.0 ml/kg was injected into the remaining 6 control dogs, using the same method. Cardiac output significantly decreased immediately after injection of the sclerosant in all dogs given 0.5 ml/kg, 1.0 ml/kg and 3.0 ml/kg injections of EO; however, there was a tendency toward recovery from 6 h after injection in dogs given 0.5 ml/kg and from 9 h in dogs given 1.0 ml/kg. Pulmonary hypertension just after injection and hypoxia at 9-12 h occurred only when 3.0 ml/kg was injected. Irreversible pulmonary hemorrhage was present in the excised lungs in 4 of 7 dogs given 3.0 ml/kg, while there were no significant lesions in the other dogs. The lung water content in cases of 1.0 and 3.0 ml/kg injections was significantly higher than that in the controls, while there was no significant difference between those given 0.5 ml/kg and of the controls. The findings obtained in this study suggest that EO less than 0.5 ml/kg used for sclerosing esophageal varices seems to have little untoward influence on pulmonary hemodynamics and morphology.     

Endotoxin filtration and immune stimulation improve survival from gram-negative sepsis

Polymyxin B, when bound to a polystyrene fiber (PMX-F), has been used experimentally as an extracorporeal blood filter to reduce serum lipopolysaccharide levels, which are believed to be responsible for physiologic alterations in the septic state. To validate our theory that a combination of PMX-F, systemic antibiotics, and immune stimulation would improve survival, 78 rats were given intravenous doses of Escherichia coli (range, 4.6 to 6.2 X 10(8) colony-forming units/ml). They were then randomized into groups receiving either systemic gentamicin (n = 10); pretreatment with muramyl dipeptide (n = 11); or extracorporeal hemoperfusion through either a sham column (n = 8), PMX-F-packed column with systemic gentamicin (n = 8); or PMX-F-packed column with systemic gentamicin and muramyl dipeptide pretreatment (n = 8). Thirty-three control rats received no treatment. Sham hemoperfusion (13%) and control (21%) rats had the lowest survival rate, although increased improvement was noted in rats treated with gentamicin (30%) or the combination of PMX-F filtration and gentamicin (50%). The most significant improvements occurred in rats pretreated with muramyl dipeptide (53%) and in rats treated with a combination of PMX-F, gentamicin, and muramyl dipeptide (88%). These data show that lipopolysaccharide filtration and nonspecific immune stimulation are additive to antibiotic therapy and are useful as adjunctive measures in the multimodal treatment of experimental gram-negative bacterial infection.     

N-acetylcysteine restores isoflurane-induced preconditioning against myocardial infarction during hyperglycemia

BACKGROUND: Hyperglycemia generates reactive oxygen species and prevents isoflurane-induced preconditioning. The authors tested the hypothesis that scavenging reactive oxygen species with N-acetylcysteine will restore protection against myocardial infarction produced by isoflurane in vivo. METHODS: Barbiturate-anesthetized dogs (n = 45) were instrumented for measurement of systemic hemodynamics. Myocardial infarct size and coronary collateral blood flow were measured with triphenyltetrazolium staining and radioactive microspheres, respectively. All dogs were subjected to a 60-min left anterior descending coronary artery occlusion followed by 3 h of reperfusion. Dogs were randomly assigned to receive an infusion of 0.9% saline or 15% dextrose in water to increase blood glucose concentrations to 600 mg/dl (hyperglycemia) in the absence or presence of isoflurane (1.0 minimum alveolar concentration) with or without pretreatment with N-acetylcysteine (150 mg/kg i.v.) in six experimental groups. Isoflurane was discontinued, and blood glucose concentrations were allowed to return to baseline values before left anterior descending coronary artery occlusion. RESULTS: Myocardial infarct size was 27 +/- 2% (n = 8) of the left ventricular area at risk in control experiments. Isoflurane significantly (P < 0.05) decreased infarct size (13 +/- 2%; n = 7). Hyperglycemia alone did not alter infarct size (29 +/- 3%; n = 7) but abolished the protective effect of isoflurane (25 +/- 2%; n = 8). N-Acetylcysteine alone did not affect infarct size (28 +/- 2%; n = 8) but restored isoflurane-induced cardioprotection during hyperglycemia (10 +/- 1%; n = 7). CONCLUSIONS: Acute hyperglycemia abolishes reductions in myocardial infarct size produced by isoflurane, but N-acetylcysteine restores these beneficial effects. The results suggest that excessive quantities of reactive oxygen species generated during hyperglycemia impair isoflurane-induced preconditioning in dogs.     

Donor fascia in urogynaecological procedures: a canine model

OBJECTIVE: To explore the in vivo characteristics of donor fascia used in urogynaecological procedures, in a canine model. MATERIALS AND METHODS: Two experiments were conducted. In the first, donor fascia grafts were obtained from 12 dogs, the grafts freeze-dried and half were irradiated. The grafts were used for sacrocolpopexy and suburethral slings in each of five dogs. The dogs were killed at 2, 6 and 12 weeks after graft implantation, the grafts retrieved and assessed using tensilometry. In the second experiment, unirradiated sacrocolpopexy grafts were implanted in eight dogs; four grafts were placed under no tension and four under moderate tension. At 8 weeks, the grafts were retrieved and assessed by tensilometry. Measures of strength in both experiments included the ultimate tensile strength, ultimate strain and stiffness. All measures were compared using Kruskal-Wallis nonparametric tests in both studies. RESULTS: In the first experiment, a significant minority (23%) of grafts had complete loss of strength. Measures of graft strength did not vary when analysed according to donor dog, host dog, history of graft irradiation, duration of implantation or location of graft. In the second experiment, grafts placed under no tension tended to have lower tensile strength (chi2(1) = 3.125, P = 0.077), lower stiffness (chi2(1) = 3.125, P = 0.077) and lower ultimate strain (chi 2(1) = 3.182, P = 0.074). CONCLUSION: Graft irradiation as an isolated variable did not predispose grafts to failure in vivo. Biomechanical factors at the implantation site are likely to play a critical role in determining ultimate graft strength.