Efficacy of a Platelet-Activating Factor Antagonist TCV-309 for Lung Preservation

To evaluate the efficacy of a platelet-activating factor (PAF) antagonist, TCV-309, for the prevention of lung reperfusion injury, an experimental study was performed by using a canine isolated lung reperfusion model. The canine lungs were preserved for 24 hours at 4°C. The animals were classified into 4 groups: Group A, domor lungs were flushed with a Euro-Collins (EC) solution only and preserved for 24 hours at 4°C as controls (n=7) ; Group B, donor dogs received intravenous TCV-309 at a dose of 30 μg/kg 15 minutes before heparin sodium administration, which was given prior to isolating the lungs and donor lungs were preserved for 24 hours at 4°C (n= 8); Group C, donor lungs were flushed with a EC solution containing TCV-309 (375 μg/500m1) and preserved for 24 hours at 4°C (n= 6), Group D, donor lungs were flushed and stored with a EC solution and then reperfused with allogeneous blood containing TCV-309, 0.75 μ g/ml of blood volume (n= 6). The pulmonary vascular resistance (mmHg/l/min) was significantly lower for Groups B, C and D than for Group A (A vs. B or C or D: 270. 8± 111.3 vs. 96.2± 78.3 (p <0.01) or 122.6± 75.4 (p <0.05) or 120. 3± 89.7 (p <0.05), respectively) (120 min). The airway pressure (AWP, cmH2O) was significantly lower for Groups B and C than for Group A (A vs. B or C or D: 21.0 ± 3.3 vs. 16.0 ± 2.2 (p <0.01) or 15.7 ± 2.2 (p <0.01) or 18.3 ± 1.4 (N.S.), respectively) (0 min). The AWP was significantly lower for Group D than for Group A (A vs. B or C or D: 30.4± 8.6 vs. 2 3.4 ± 7.0 (N.S.) or 21.5 ± 7.8 (N.S.) or 19.1 ± 2.0 (p < 0.01), respectively) (120 min). The static lung compliance (ml/cm H20) was significantly higher for Groups B, C and D than for Group A throughout the reperfusion. The blood oxygen tension(ΔPO2, mmHg)and lung water(%)were signifi一 cantly improved for Groups C and D, but not for Group A (ΔPO2, Avs.BorCorD:161.1ｱ66.2vs.246.7ｱ82.4(N.S.)or 309.9± 62.2 (p <0.05) or 307.2± 79.0 (p <0.05), lung water; 89.6 ± 0.8 vs. 87.3± 2.6 (N.S.) or 86.0 ± 1.7 (p < 0.0001) or 85.2 ± 2.3 (p <0.005). At the end of the harvest, the tissue level of lipid peroxidation (nmolMDA/mg protein) was significantly lower for Group B than for the others (B vs. others: 1.17 ± 0.2 3 vs. 1.51 ± 0.14 (p < 0.05) ). At the end of storage, the tissue level of myeloperoxidase (Units/g wet tissue) was significantly lower for Group C than for Groups A and D (C vs. A or B or D:13.5± 18.4 vs. 42.2± 14.2 (p <0.05) or 40.8± 52.2 (N.S.) or 42.6± 18.8 (p < 0.05)) . In conclusion, the PAF antagonist, TCV-309 , is effective for the prevention of ischemic reperfusion injury in 24-hour cold preserved canine lungs

Protective effect of FUT-175 on pulmonary function of xenografts in a guinea pig-to-rat lung perfusion model

Background: FUT-175 (nafamostat mesilate) has a variety of pharmacological effects; in addition to its stable potent serine protease inhibitory activity, it exerts far stronger anti-complement activity than other protease inhibitors. Here, we evaluated the protective effect of FUT-175 on pulmonary function of xenografts in an ex vivo guinea pig-to-rat lung perfusion model, using a device for analyzing pulmonary function in small animals. Methods: Animals were divided into three groups (n = 6 each), Isograft (Group I), Xenograft (Group X), and Xenograft with FUT-175 (Group XF). In the latter, 10 mg of FUT-175 was added to the extracorporeal circuit before perfusion with xenogeneic blood was started. The following parameters were serially measured in these three groups: complement activity causing 50% hemolysis (CH50 units) in the perfusion blood either before or during perfusion, pulmonary arterial pressure, dynamic pulmonary compliance, and airway resistance. In addition, Hematoxylin and Eosin staining of the lungs and assays of rat IgM, IgG, and anti-C3 deposition were carried out after perfusion. Results: The duration of satisfactory pulmonary function after the start of perfusion was significantly increased in Group XF. CH50 in Group XF decreased significantly than in Group X. In addition, FUT-175 suppressed both the increase in pulmonary arterial pressure and airway resistance, and the decrease in dynamic pulmonary compliance. In Group XF, intraalveolar hemorrhage and the thickening of the arterial wall were not observed. Groups X and XF showed deposition of IgM, IgG, and C3 at the endothelium of the pulmonary arteries but less in Group I. Conclusions: This study suggests that FUT-175 inhibited complement activation including the alternative pathway and improved lung xenograft pulmonary function. FUT-175 ameliorates hyperacute rejection in a guinea pig-to-rat ex vivo xenogeneic lung perfusion model

Expression of Keratinocyte Growth Factor and Its Receptor in Rat Tracheal Cartilage: Possible Involvement in Wound Healing of the Damaged Cartilage

Keratinocyte growth factor (KGF) is involved in the development and regeneration of a variety of tissues. To clarify the role of KGF in cartilage wound healing, we examined the expression of KGF and its receptor (KGFR) immunohistochemically in the wound healing area of rat tracheal cartilage, and the direct effect of recombinant KGF on the proliferation and differentiation of primary cultures of rat chondrocytes. KGF was found in the cytoplasm of both chondrocytes and perichondrial cells. On the other hand, KGFR was detected only in the plasma membrane of chondrocytes. Although the expression of KGF was similar in the cartilage and perichondrial area before and after injury, KGFR expression was induced after injury and limited to proliferating chondrocytes. The staining pattern of KGF and KGFR was same in the mature and the immature rat tracheal cartilage. Moreover, in vitro experiments using primary cultured chondrocytes revealed that KGF at 200 ng/ml significantly increased the number of chondrocytes (~1.5-fold), and significantly reduced acid mucopolysaccharide production. These results indicate that KGF stimulates chondrocyte proliferation, suggesting that KGF could therapeutically modulate the wound healing process in the tracheal cartilage

Gene transfer of tumor necrosis factor inhibitor improves the function of lung allografts

AbstractBackgroundTumor necrosis factor is an important mediator of lung transplant acute rejection. Soluble type I tumor necrosis factor receptor binds to tumor necrosis factor-α and -β and inhibits their function. The objectives of this study were to demonstrate efficient in vivo gene transfer of a soluble type I tumor necrosis factor receptor fusion protein (sTNF-RI-Ig) and determine its effects on lung allograft acute rejection.MethodsThree groups of Fischer rats (n = 6 per group) underwent recipient intramuscular transfection 24 hours before transplantation with saline, 1 × 1010 plaque-forming units of control adenovirus encoding β-galactosidase, or 1 × 1010 plaque-forming units of adenovirus encoding human sTNF-RI-Ig (Ad.sTNF-RI-Ig). One group (n = 6) received recipient intramuscular transfection with 1 × 1010 Ad.sTNF-RI-Ig at the time of transplantation. Brown Norway donor lung grafts were stored for 5 hours before orthotopic lung transplantation. Graft function and rejection scores were assessed 5 days after transplantation. Time-dependent transgene expression in muscle, serum, and lung grafts were evaluated by using enzyme-linked immunosorbent assay of human soluble type I tumor necrosis factor receptor.ResultsRecipient intramuscular transfection with 1 × 1010 plaque-forming units of Ad.sTNF-RI-Ig significantly improved arterial oxygenation when delivered 24 hours before transplantation compared with saline, β-galactosidase, and Ad.sTNF-RI-Ig transfection at the time of transplantation (435.8 ± 106.6 mm Hg vs 142.3 ± 146.3 mm Hg, 177.4 ± 153.7 mm Hg, and 237.3 ± 185.2 mm Hg; P = .002, .005, and .046, respectively). Transgene expression was time dependent, and there was a trend toward lower vascular rejection scores (P = .066) in the Ad.sTNF-RI-Ig group transfected 24 hours before transplantation.ConclusionsRecipient intramuscular Ad.sTNF-RI-Ig gene transfer improves allograft function in a well-established model of acute rejection. Maximum benefit was observed when transfection occurred 24 hours before transplantation

Two Case Reports of Successful Withdrawal of Mycofenolate Mofetil After Living Donor Lobar Lung Transplantation

Background: Lung transplantation cases have immunosuppression maintained using a calcineurin inhibitor, anti-metabolites, and steroid. Case Report: We report 2 clinical cases in which anti-metabolites (mycophenolate mofetil) were successfully withdrawn after living donor lobar lung transplantation by monitoring immune function using the ImmuKnow® assay. In the first case, a 43-year-old woman underwent living donor lobar lung transplantation for pulmonary alveolar proteinosis. Two healthy relatives donated a lower lobe each. Immunosuppression was maintained using tacrolimus, mycophenolate mofetil, and steroid. Six months posttransplantation, she developed invasive pulmonary aspergillosis. During anti-fungal treatment, we withdrew mycophenolate mofetil and tacrolimus trough levels were kept around 8 ng/mL. Despite the resulting low-level immunosuppression, the ImmuKnow assay showed immune function to be in the moderate range with tacrolimus and steroid alone, encouraging us to maintain this strategy to avoid recurrence of invasive pulmonary aspergillosis. In the second case, a 24-year-old man underwent living donor lobar lung transplantation for cystic fibrosis. Two healthy relatives donated a lower lobe each. Immunosuppression was maintained using tacrolimus, mycophenolate mofetil, and steroid. Five months posttransplantation, he developed persistent Pseudomonas aeruginosa pneumonia derived from the paranasal sinuses. Under ImmuKnow assay monitoring, mycophenolate mofetil was withdrawn, but immune function was maintained within the moderate range using tacrolimus and steroid alone. Discussion: Respiratory function in both cases was maintained; no findings of bronchiolitis obliterans syndrome were noted during this period. To the best of our knowledge, no reports have described successful anti-metabolite withdrawal in lung transplantation with ImmuKnow monitoring. Immune evaluation by ImmuKnow could offer a useful method to monitor and control immune status, particularly among recipients susceptible to infection, revealing that moderate immune function could be maintained using tacrolimus and steroid in living donor lobar lung transplantation

Surgical Management of Spontaneous Haemopneumothorax

We report 2 cases of spontaneous haemopneumothorax treated surgically. Case 1, a 19-year-old man was referred to our hospital because of trapped lung after continuous tube drainage for 3 weeks. The chest roentgenogram showed a left haemopneumothorax with collapsed lung. At thoracotomy, 200g of clot blood was evacuated and decortication was performed. Case 2, a 20-year-old man was admitted complaining of chest pain and dyspnea. X-ray examination showed a moderate lung collapse and homogenous density. Chest tube drainage yealded 480ml of blood. However, bleeding was continuous and expansion of the right lung was poor, emergency thoracotomy was carried out and 950g of clot blood was obtained. Recovery was uneventful and complete expansion of the right lung was observed. From our experience, early thoracotomy for spontaneous haemopneumothorax is recommended

The Efficacy of Postoperative Chemotherapy with Cisplatinum and Pepleomycine for Esophageal Cancer

From May 1984 to August 1991, 58 patients without preoperative adjuvant therapy underwent resection of the esophagus for esophageal cancer. Three weeks after esophagectomy, one cycle of postoperative chemotherapy, consisting of intravenously infused cisplatinum at a dose of 70mg/m2 on day 1 and intramuscular pepleomycine at a dose of 5mg/body from day 1 to day 5, was administered in the 24 patients. In all patients receiving chemotherapy, mild fatigue or poor appetite occurred after drug administration, but severe drug toxicity, such as bone marrow depression, gastrointestinal bleeding or pulmonary fibrosis, did not occur. Eighteen patients (75%) died from cancer. The 3-year survival rate was 22.9% and the 5-year surival rate was 17.1%. In the patients who underwent curative operations, the 3-year and 5-year survival rates were 46.2% and 27.7%. However, there was no significant difference in the survival rates between the patients with postoperative chemotherapy and patients with esophagectomy alone. We conclude that one cycle of postoperative chemotherapy with cisplatinum and pepleomycin does not affect the survival of patients undergoing esophagectomy for esophageal cancer

Surgery for Early Esophageal Carcinoma

Early esophageal carcinomas are clinicopathologically evaluated in 11 patients whose carcinomas are limited to the epithelium (ep) and the mucosa (mm). Most of ep and mm carcinomas were detected by mass-screening without any symptom. Even when mass examination for gastric lesion is attempted, precise examination of the esophagus should be made at the same time. It is assured that a ep and mm esophageal carcinoma ensured satisfactory outcome. It is emphasized that early detection is only a way to improve the surgical outcome of the treatment for esophageal carcinomas

Class II MHC Antigen Expression in Bronchial Lavage Cells in a Canine Lung Allograft Model using FK506 Immunosuppression

Footnotes: Katsunobu Kawahara, MD., The First Department of Surgery, Nagasaki University School Medicine, Sakamoto-machi 7-1 #852, Nagasaki, Japan, TEL 0958-47-2111, FAX 0958-47-5034 Abbreviations: MHC: Major histocompatibility antigens, CsA: Cyclosporine A, BAL: Bronchoalveolar lavage, APC: Antigen presenting cell To assess the effect of FK506 on class II MHC antigen expression on lymphocytes recovered from bronchoalveolar lavage (BAL) in dogs following left lung allotransplantation, flow cytometric anlysis with OKIa-1 monoclonal antibody was performed. Dogs were divided into two groups: Group 1 (n = 6), control group (dogs without surgery); Group 2 (n = 23), dogs recieving left lung allotransplantation and immunosuppression using FK506 (0.1mg/kg/day intramuscularly). No significant difference existed in the percentage of OKIa-1 positive lymphocytes recovered from control animals (31.2 ± 11.2%) vs. normal allografts in Group 2 (34.8 ± 8.5%, n = 16). With rejection, however, the percentage of OKIa-1 positive lymphocytes increases significantly in Group 2: 56.2 ± 10.3% in mild rejection (n = 6) and 91.4 ± 4.3% in moderate or severe rejection (n = 4) (p < 0.01). Chest radiographs appeared normal in allografted lungs with histologically mild rejection. The percentage of the OKIa-1 positive lymphocytes in BAL did not significantly change during the first, second, or third week following transplantation, ((32.2 ± 6.4 (n = 5), 36.4 ± 4.2 (n = 4), and 35.8 ± 12.3 (n = 7), respectively)) in the allografted lungs without rejection. FK506 does not affect the class II MHC antigen expression of lymphocytes recovered from BAL in canine allografted lungs without rejection. Furthermore, this compound does not change in class II antigen expression seen with allograft rejection

Measurements of the Wind generated by Desert Environment Wind Tunnel (Constant Speed Wind and Periodically Varying Wind)

Wind characteristics generated by the wind tunnel named Desert Environment Wind Tunnel, which can generate arbitrary pattern of wind speed variation by changing the angle of pitch of the blower blades, were investigated with both of a hot-wire anemometer and a ultrasonic anemometer. Distributions of the wind velocity components (v, w) in a plane perpendicular to the mainstream (u-direction), whose wind speed varies like a sine wave, were measured downstream at x = 1.5m from the exit of wind tunnel nozzle. The experimental results revealed that the flow expands when it accelerates and, in reverse, the flow contracts then slowing down