HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Personal Folders Download to citation manager Author home page(s): Bartley P. Griffith Permission Requests Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mattsson, P. Articles by Griffith, B. P. Search for Related Content PubMed PubMed Citation Articles by Mattsson, P. Articles by Griffith, B. P. Related Collections Related Article

Ann Thorac Surg 1996;62:207-212
© 1996 The Society of Thoracic Surgeons

---

Original Articles: General Thoracic

Effect of Aminoguanidine and Cyclosporine on Lung Allograft Rejection

Divisions of Cardiothoracic Surgery, Immunopathology, Pathology, and Pharmacology, and Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania

	Abstract

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
Background. Aminoguanidine, a nitric oxide synthase inhibitor, has been shown to reduce the inflammatory allogeneic response. Here we used it in combination with cyclosporine to evaluate its effect on a clinically relevant immunosuppressive protocol.

Methods. Orthotopic left lung transplantation was performed in 120 rats, of which 24 were syngeneic Lewis to Lewis controls, and allogeneic transplantations were performed across major histoincompatibility barriers (ACI to Lewis). We studied synchronous histologic changes accompanying cytokines and nitric oxide synthase messenger RNA by reverse transcriptase polymerase chain reaction in the grafted lungs. Nitrate/nitrite, oxidized degradation products of nitric oxide, were measured in the whole blood, as were concentrations of cyclosporine. Lung tissue was immunohistochemically stained for nitric oxide synthase protein. Rats receiving allografts were either untreated (24) or received low-dose cyclosporine (232 ± 105 ng/mL blood by high-performance liquid chromatography), high-dose cyclosporine (2,046 ± 664 ng/mL), aminoguanidine alone (800 mg•kg^-1•day^-1 intraperitoneally), or aminoguanidine plus low-dose cyclosporine.

Results. The results suggest that aminoguanidine combined with low doses of cyclosporine can reduce the allogeneic response across major histoincompatibilities in rodent lung transplantation. Its biologic effect may not exclusively depend on the inhibition of nitric oxide synthase and may, by other means, reduce proinflammatory cytokines.

Conclusions. Aminoguanidine may be an effective adjuvant to conventional immunosuppression.

	Introduction

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
See also page 212.

Lung transplantation is complicated with a high incidence of rejection [1]. Chronic as well as repeated episodes of acute rejection are associated with inflammatory infiltrates that have been implicated in the development of obliterative bronchiolitis [2]. Nitric oxide (NO) has been shown to have a modulatory role in allograft rejection [3], and sole treatment with the nitric oxide synthase (iNOS) inhibitor AG in rodent heart transplant models with both minor and major histoincompatibility mismatches, and lung transplant models with major mismatches, have demonstrated reduced inflammation and longer survival of the graft [4–6]. In this study we were interested to know whether AG would have an adjuvant role to cyclosporine (CsA) in a rat lung transplant model across a major histoincompatibility barrier. We hoped to provide insights into the relative mechanism of AG action, relating grade of rejection to intragraft cytokines and iNOS, blood levels of NO metabolites NO₂^-/NO₃^-, and immunohistochemical stain of lung tissue for iNOS.

	Material and Methods

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
Animals
Viral and antibody free 250- to 325-g male Lewis and male ACI rats were purchased from Harlan-Sprague-Dawley, Indianapolis, IN. Animals were given standard rat food and water ad libitum and received care in compliance with the "Guide for the Care and Use of Laboratory Animals" published by the National Institutes of Health (NIH publication 85-23, revised 1985).

Lung Transplant Model
Transplantations were performed over a major histoincompatibility barrier using ACI as donor and Lewis as recipient. Lewis-to-Lewis combined grafting was used for syngeneic controls. Orthotopic left lung transplantation was performed as previously described [7, 8]. Briefly, while anesthetized with methoxyflurane, donor rats were anticoagulated with 1,000 units of intravenous heparin, after which their lungs were removed and flushed with 10 mL of cold heparinized lactated Ringer solution. Ischemic times of donor lungs ranged between 20 and 40 minutes. Recipient rats were intubated and, using rodent ventilators, anesthetized with a mixture of halothane (1% to 3%) and oxygen. The cuff technique was used for the pulmonary artery and vein anastomoses, whereas the bronchial anastomoses were completed with a 10-0 monofilament suture. Antibiotics in the form of clindamycin and ceftazidime were given to recipient rats preoperatively, and ceftazidime prophylaxis was maintained throughout the time period studied. Animals were euthanized by an overdose of thiopental on postoperative days (PODs) 1, 2, 4, and 6. By sacrifice, grafted and native lungs plus blood were removed for analysis.

A total of 120 animals were studied in six groups: group 1 (n = 24; syngeneic) no treatment; group 2 (n = 24; allogeneic) no treatment; group 3 (n = 24; allogeneic) low-dose CsA (2 mg•kg^-1•day^-1 injected subcutaneously); group 4 (n = 24; allogeneic) high-dose CsA (10 mg•kg^-1•day^-1 injected subcutaneously); group 5 (n = 12; allogeneic) AG (800 mg•kg^-1•day^-1 injected intraperitoneally); and group 6 (n = 12; allogeneic) AG (800 mg•kg^-1•day^-1 injected intraperitoneally) + low-dose CsA (2 mg•^-1•day^-1 injected subcutaneously).

Treatment
Previous work in our laboratory had shown that lungs transplanted from ACI to Lewis would be completely rejected within 6 days and that the grade of rejection should be dependent on the "clinical therapeutic" blood concentration of CsA relevant in humans at the same time as in our animal model and would achieve a moderate grade of rejection on the 4th and 6th PODs. Our selected high dose of CsA had been found to be more effective in limiting rejection in pilot experiments but was associated with very high CsA blood levels. In this study CsA/cremophore was administered in 0.9% sodium chloride (NaCl) as subcutaneous injections every 24 hours, with first dose given 10 to 20 minutes after transplantation. All rats receiving CsA were sacrificed for analysis 24 hours after the last CsA administration.

AG (Hemisulfate; Sigma Chemical Co, St. Louis, MO) was delivered through a catheter placed intraperitoneally. The AG powder was dissolved in 0.9% NaCl and injected at a dose of 200 mg/kg four times daily. The first AG injection was given 10 to 20 minutes postoperatively. All rats receiving AG were euthanized 6 hours after their last injection.

Analysis
HISTOLOGY.
Samples from grafted and native lungs were fixed in 10% buffered formalin and were paraffin embedded. Four-micrometer sections were stained with hematoxylin and eosin and were blindly graded on a 0 to 4 scale of rejection intensity:

• 0 No perivascular mononuclear infiltrates
  
• 1+ The perivascular mononuclear infiltrates localized to veins
  
• 2+ Distinct cuffs of 10 to 15 mononuclear cells surrounding arteries and veins
  
• 3+ Perivascular infiltrates as in 2+ with alveolar septal extension of the mononuclear cells
  
• 4+ Perivascular, septal, and peribronchiolar infiltrates with consolidation of the lung airspace filling by mononuclear cells and focal necrosis
  

CYTOKINE MESSENGER RNA ANALYSIS.
Tissue from 72 grafted lungs (three samples from each group at each of the studied PODs 1, 2, 4, and 6) were snap frozen and processed in parallel for RNA quantitation using radiolabeled primers [9]. Total RNA was extracted from tissue fragments with RNAzol (Cinna/Biotec, Houston, TX) and quantified spectrophotometrically. First strand DNA was synthesized by transcription from RNA in the presence of human placental RNAse inhibitor, 1 mmol/L deoxynucleoside triphosphates, oligo-dT primer, murine leukemia virus reverse transcriptase, and reverse transcriptase buffer. Oligonucleotides were designed from the published sequences of cytokine messenger RNAs and were custom-synthesized by the University of Pittsburgh DNA synthesis facility. The 5` primer is terminally labelled with g-32PdCTP and T4 kinase. The radiolabeled 5` primer is mixed with the unlabeled 5` and 3` primers to make the stock primer mix. The RT-polymerase chain reaction of the complementary DNA was performed according to the conditions described by Brenner and associates [10]. Amplification for various cytokines was carried out for 28 cycles on a model 480 thermal cycler (Perkin Elmer). The polymerase chain reaction product bands were quantified using a Betagen radioanalytic scanner. The results (cpm) obtained for each cytokine and iNOS were expressed as means ± standard deviations and normalized to ß-actin (ratio cytokine/actin). The cytokine messenger RNAs tested were interleukin-2 (IL-2), IL-4, IL-6, IL-10, and interferon-.

BLOOD LEVELS OF NO₂^-/NO₃^-.
Whole blood obtained from cardiac puncture by sacrifice was analyzed for NO₂^-/NO₃^- (oxidative degradation products of NO metabolism). Blood was deproteinized by treatment with NaOH and ZnSO. Whole blood NO₂^-/NO₃^- was determined using the high-performance liquid chromatography method described by Green and colleagues [11]. As with the cytokine analysis, a total of 72 samples were examined at similar time points.

IMMUNOHISTOCHEMISTRY.
Tissue specimens were snap frozen and embedded in Optimal Cutting Temperature Compound (OCT, Miles, Inc, Elkhart, IN). Sections were cut in a cryostat at 6 µm and dried overnight to ensure adherence to slides. A protein blocking reagent (Shandon Lipshaw, Pittsburgh, PA) was placed on the slides before the application of specific antibody. Mouse monoclonal fluorescein-conjugated anti-iNOS (Transduction Laboratories, Lexington, KY) was placed on the sections for 1 hour at a 1:50 and 1:100 concentration at room temperature. An irrelevant isotype-matched fluoresceinated control antibody (Dako Corp, Carpinteria, CA) was used as a negative control. After buffer wash, the slides were counterstained with a propidium iodide-containing mounting medium (Oncor, Inc, Gaithersburg, MD) and coverslipped. Slides were read and photographed using a Nikon Optiphot fluorescence microscope.

BLOOD LEVELS OF CSA.
Whole blood sampled through cardiac puncture by sacrifice was assayed by high-performance liquid chromatography using the method previously reported for our laboratory [12]. This high-performance liquid chromatography method measures the parent CsA in blood separate from any metabolites that might be in the sample.

STATISTICAL ANALYSIS.
Results are expressed as mean ± standard deviation, and the Student's t test was used to determine statistical significance.

	Results

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
Histology of Lungs
Isograft and native lungs revealed minimal inflammatory activity (Table 1). Treated and untreated lung allografts showed progressive rejection from PODs 1 through 6. Allograft recipients without treatment had totally rejected their allografts by POD 6 (grade 4 ± 0) (Fig 1A). Animals receiving low doses of CsA had a moderate degree of rejection on postoperative day 6 that was graded at 3.0 ± 0.2, whereas those receiving high doses of CsA had a mild score of 2.0 ± 0.1. Histologic rejection was slightly delayed in the AG-treated allografts, showing on POD 4 a similar result to that achieved with low-dose CsA (Fig 1B), but by the sixth POD all AG-treated animals fully rejected their grafts. The summed average scores for low-dose CsA plus AG were almost as good as for high-dose CsA (p = 0.3) and significantly better than low-dose CsA alone (p = 0.015) (Fig 1C).

View larger version (243K): [in this window] [in a new window]   Fig 1. . (A) Untreated lung allografts showed consolidated lung on postoperative day 4 with distorted airspaces and diffuse perivascular and interstitial mononuclear infiltrates with spillage of cells into airspaces. (B) On postoperative day 4, allograft lungs treated with aminoguanidine showed distinct and prominent perivascular and septal infiltrates that averaged a grade of 3.2. (C) On postoperative day 4, animals treated with low-dose cyclosporine demonstrated severe inflammation and were graded as severely rejecting (left). Those animals receiving aminoguanidine and cyclosporine were significantly improved (right).

View larger version (19K): [in this window] [in a new window]   Fig 2. . Lung nitric oxide synthase/actin was inhibited completely by cyclosporine (CsA) but only 30% by aminoguanidine (AG). Untreated and aminoguanidine-treated animals had modest elevations on postoperative day 4 and severely on postoperative day 6.

View larger version (23K): [in this window] [in a new window]   Fig 3. . Elevation of blood levels of nitrate/nitrite were only noted in untreated rats on postoperative day 4. (AG = aminoguanidine; CsA = cyclosporine.)

Immunohistochemical Stain for Nitric Oxide Synthase Enzyme
Nitric oxide synthase stained positive most apparent in inflammatory cells in untreated allografts on POD 4. Nitric oxide synthase was not detected in any CsA-treated animals but was rarely present in those receiving AG.

	Comment

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
Nitric oxide is a molecule that has recently been demonstrated to have protean physiologic effects [13]. Our group in Pittsburgh has had a long interest in NO and early on showed that large amounts of NO are produced during alloimmunity in response to inflammatory cytokines of the T-helper lymphocyte family [14–16]. In a rat cardiac transplant model, iNOS was induced into endothelial cells, the myocytes, and more significantly in macrophages associated with the inflammatory immunoresponse [17]. The macrophage is at the center of the alloimmunity network. When stimulated, it is a major source of iNOS [18]. It is the professional antigen-presenting cell and major cell along with the lymphocyte present during lung rejection. Because nitric oxide had been shown to have conflicting immune effects, which included lymphoproliferation of cytotoxicity [19], we were uncertain whether administration of a selective inhibitor AG would affect the response [20]. Work in myocyte cultures with stimulated macrophages has shown that NO was cytotoxic and caused cardiac myocytes to die of apoptosis [17]. Worrall and colleagues [4] have recently demonstrated that AG could prolong cardiac allograft survival in minor histoincompatible rodents. In that study AG alone appeared useful and was clearly associated with a reduction in early inflammation. We were aware of an as yet unpublished work from the St. Louis group that suggested similar benefits in minor and major mismatched rodent lung transplants [5]. We sought to confirm their finding in our model of rodent lung transplantation across the major histoincompatible barrier represented by the ACI-to-Lewis rodents and wondered whether or not, when combined with CsA, AG could show an adjunctive role. We were skeptical that AG would in fact be useful when associated with CsA for, in addition to CsA's established inhibitory function on T-lymphocytes, it recently had been shown to dose-dependently inhibit NO production in lipopolysaccharide-stimulated macrophages [21, 22].

In our study syngeneic Lewis isografts demonstrated minimal elevation of cytokines and no increase in iNOS message or protein. The histologic changes were minor, and there were no elevations of NO degradation products in the blood. As expected, untreated ACI-to-Lewis allografts showed progressive mild, moderate, and severe rejection through the sixth posttransplantation day. We had chosen this model for a serious allogeneic response and for the high and low doses of CsA to provide a range of control with immunosuppression. Following the paradigm suggested from other solid organs, inflammatory cytokines were detected early in the lung allograft and expression was dense between days 4 and 6. This was associated with significant increases in iNOS message as detected by RT-polymerase chain reaction from lung tissue. We were able to detect significant iNOS protein by immunohistochemical staining that was especially dense on the fourth POD. As in the previous rodent heart study [4], blood levels of NO₂^-/NO₃^- rose above normal on the fourth POD and then fell by day 6, when the graft had become completely rejected. The results showed that AG alone could, as it had in a less incompatible heart allograft model [4], reduce the rejection grade early after transplantation and delay complete rejection. Another study confirmed the relative inability of iNOS inhibition to prevent fully rejected allografts [23].

This suggests that inhibition of the arginine NO pathway is insignificant alone to prevent rejection; however, the improvement in histology on POD 4 with AG equaled that achieved by low-dose CsA, which underscores the important contributory role of NO in transplant immunity. Because AG is a preferential inhibitor of the inducible iNOS isoform, it did not surprise us that there was staining, albeit minimal, present for iNOS. We were, however, not anticipating the extent to which AG alone would be associated with suppression of iNOS gene. The overall antiinflammatory effect of AG clearly is dominated by its iNOS inhibition, which reduced NO available to promote recent additional proinflammatory TH1 cytokines and lessened its local cytotoxic effects. We reasoned that less secondary cytokines ultimately resulted in less iNOS gene detected. We confirmed the striking inhibitory effects of CsA on NO as there was complete inhibition of the iNOS message, no staining for the enzyme, and no evidence of NO degradation as the blood levels of its metabolites were normal. We were interested to find that cytokine inhibition was significant and mostly CsA dose-dependent. The control of rejection with the higher CsA dose followed its general inhibitory iNOS and cytokine pattern, whereas the lower dose of CsA, in spite of being associated with the complete inhibition of NO, fared less well with the cytokine inhibition and control of immunosuppression.

Why then was the association of AG and low-dose CsA as effective in inhibiting iNOS gene, iNOS enzyme, and cytokines as high-dose CsA? When AG was added to low-dose CsA, the control of rejection was similar to that achieved by high-dose CsA, and it was significantly better than that obtained with low-dose CsA alone. It is likely that some amount of NO was produced in the inflammatory reaction in spite of our ability to demonstrate it in the CsA-treated animals. The fact that the proinflammatory cytokines were substantially lowered when AG was added to the low-dose CsA suggests that the AG was effective in blocking the lymphoproliferative and cytotoxic effects of small amounts of NO. It is possible, of course, that AG had heretofore unrecognized effects in the inflammatory process that resulted in a non-NO inhibitory suppression of cytokine activity. If the lung transplant model can be adapted to mice, it would seem most reasonable then to follow these studies with similar ones in the recently available iNOS-deficient animals. Additionally, it would be of interest whether the competitive inhibitor of this enzyme, methylated L-arginine analogue N^G monomethyl-L-arginine, will provide a similar beneficial effect on lung transplantation alone and when combined with CsA.

It is interesting that although our mainstay immunosuppressants, such as FK506 and CsA, have provided tremendous ability to control proliferation of lymphocytes, there are multiple pathways of escape. It might well be that blockade of NO to some extent by AG might prove useful, as suggested in this study, in swinging the modulatory or buffering action of NO in allogeneic immunity toward reduced inflammation and nonspecific control of the destructive phase of the allogeneic antigenic response.

	Acknowledgments

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
Supported by National Heart, Lung and Blood Institute grant 5 RO1 HL48091-03 and grant R37-AI-16869.

We express gratitude to Joanne Gizzi for her considerable assistance with the preparation of the manuscript.

	Footnotes

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 
Presented at the Thirty-second Annual Meeting of The Society of Thoracic Surgeons, Orlando, FL, Jan 29-31, 1996.

Address correspondence to Dr Griffith, Division of Cardiothoracic Surgery, University of Pittsburgh Medical Center, C700 PUH, 200 Lothrop St, Pittsburgh, PA 15213-2582.

	References

Top Footnotes Abstract Introduction Material and Methods Results Comment Acknowledgments References

 

1. Keenan RJ, Konishi H, Kanshi K, et al. Clinical trial of tracrolimus versus cyclosporine in lung transplantation. Ann Thorac Surg 1995;60:580–5.[Abstract/Free Full Text]
2. Bando K, Paradis IL, Similo S, et al. Obliterative bronchiolitis after lung and heart-lung transplantation. J Thorac Cardiovasc Surg 1995;110:4–14.[Abstract/Free Full Text]
3. Langrehr JM, Hoffman RA, Billiar TR, Lee KKW, Schraut WH, Simmons RL. Nitric oxide synthesis in the in-vivo allograft response: a possible regulatory mechanism. Surgery 1991;100:335–42.
4. Worrall NK, Lazenby WD, Misko TP, et al. Modulation of in-vivo alloreactivity by inhibition of inducible nitric oxide synthase. J Exp Med 1995;181:63–70.[Abstract/Free Full Text]
5. Shiraishi T, DeMeester SR, Worrall NK, et al. Inhibition of inducible nitric oxide synthase ameliorates rat lung allograft rejection. J Thorac Cardiovasc Surg (in press).
6. Winlaw DS, Schyvens CG, Smythe GA, et al. Selective inhibition of nitric oxide production during cardiac allograft rejection causes a small increase in graft survival. Transplant 1995;60:77–82.[Medline]
7. Keenan RJ, Duncan AJ, Yousem SA, et al. Improved immunosuppression with aerosolized cyclosporine in experimental pulmonary transplantation. Transplant 1992;53:20–5.[Medline]
8. Mizuta T, Kawaguchi A, Nakahara K, Kawashima Y. Simplified rat lung transplantation using a cuff technique. J Thorac Cardiovasc Surg 1989;97:578–81.[Abstract]
9. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched with ribonuclease. Biochemistry 1979;18:5294–8.[Medline]
10. Brenner CA, Tam AW, Nelson PA. Message amplification phenotyping (MAPPing): a technique to simultaneously measure multiple mRNAs from small numbers of cells. BioTechniques 1989;7:1096–101.[Medline]
11. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem 1982;126:131–8.[Medline]
12. Ptachcinski RJ, Venkataramanan R, Rosenthal JT, Burckart GJ, Taylor RJ, Hakala TR. Cyclosporine kinetics in renal transplantation. Clin Pharmacol Ther 1985;38:296–300.[Medline]
13. Moncada S, Higgs A. The L-arginine nitric oxide pathway. N Engl J Med 1993;329:2002–12.[Free Full Text]
14. Hoffman RA, Langrehr JM, Billiar TR, et al. Alloantigen-induced activation of rat spleenocytes is regulated by the oxidative metabolism of L-arginine. J Immunol 1990;145:2220–6.[Abstract]
15. Hoffman RA, Langrehr JM, Simmons RL. Nitric oxide synthesis: a consequence of alloimmune interaction. Xeno 1994;2:5–7.
16. Hoffman RA. Reactive nitrogen intermediates in immunology. Methods: A Companion to Methods in Enzymology (in press).
17. Corbett JA, Tilton RG, Chang K, et al. Induction of myocardial nitric oxide synthase by cardiac allograft rejection. J Clin Invest 1994;94:714–21.
18. Stuehr OJ, Marlet FA. Mammalian nitrate biosynthesis: mouse macrophages produce nitrite and nitrate in response to Escherichia coli lipopolysaccharide. Proc Natl Acad Sci USA 1985;82:7738–42.[Abstract/Free Full Text]
19. Billiar TR. Nitric oxide. Novel biology with clinical relevance. Ann Surg 1995;221:339–49.[Medline]
20. Griffith MTD, Messent M, MacAllister RJ, Evans TW. Aminoguanidine selectively inhibits inducible nitric oxide synthase. Br J Pharmacol 1993;118:2963–8.
21. Buckart V, Imai Y, Kalhmann B, Kolb N. Cyclosporine A protects pancreatic islet cells from nitric oxide-dependent macrophages cytotoxicity. FEBS Lett 1992;313:56.[Medline]
22. Conde M, Andrade J, Bedoya FJ, et al. Inhibitory effect of cyclosporin A and FK 506 on nitric oxide production by cultured macrophages. Evidence of a direct effect on nitric oxide synthase activity. J Immunol 1995;84:476–81.
23. Yang X, Chowdhury N, Cai B, et al. Induction of myocardial nitric oxide synthase by cardiac allograft rejection. J Clin Investig 1994;94:714–21.

This article has been cited by other articles:

				D. Hongo, J. S. Bryson, A. M. Kaplan, and D. A. Cohen Endogenous Nitric Oxide Protects against T Cell-Dependent Lethality during Graft-versus-Host Disease and Idiopathic Pneumonia Syndrome J. Immunol., August 1, 2004; 173(3): 1744 - 1756. [Abstract] [Full Text] [PDF]

				S. M. Wildhirt, S. Weismueller, C. Schulze, N. Conrad, A. Kornberg, and B. Reichart Inducible nitric oxide synthase activation after ischemia/reperfusion contributes to myocardial dysfunction and extent of infarct size in rabbits: evidence for a late phase of nitric oxide-mediated reperfusion injury Cardiovasc Res, August 15, 1999; 43(3): 698 - 711. [Abstract] [Full Text] [PDF]

				P. E. SILKOFF, M. CARAMORI, L. TREMBLAY, P. MCCLEAN, C. CHAPARRO, S. KESTEN, M. HUTCHEON, A. S. SLUTSKY, N. ZAMEL, and S. KESHAVJEE Exhaled Nitric Oxide in Human Lung Transplantation . A Noninvasive Marker of Acute Rejection Am. J. Respir. Crit. Care Med., June 1, 1998; 157(6): 1822 - 1828. [Abstract] [Full Text] [PDF]

				S. N. Mitruka, S. M. Pham, A. Zeevi, S. Li, J. Cai, G. J. Burckart, S. A. Yousem, R. J. Keenan, and B. P. Griffith Aerosol cyclosporine prevents acute allograft rejection in experimental lung transplantation J. Thorac. Cardiovasc. Surg., January 1, 1998; 115(1): 28 - 37. [Abstract] [Full Text] [PDF]

This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Personal Folders Download to citation manager Author home page(s): Bartley P. Griffith Permission Requests Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mattsson, P. Articles by Griffith, B. P. Search for Related Content PubMed PubMed Citation Articles by Mattsson, P. Articles by Griffith, B. P. Related Collections Related Article