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Ann Thorac Surg 2006;81:2202-2206
© 2006 The Society of Thoracic Surgeons

Original article: Cardiovascular

Cerebral Endothelial Nitric Oxide Synthase Expression is Reduced After Very Low Flow Bypass

Vesa Anttila, MD a , d , Helen Christou, MD b , Ikuo Hagino, MD a , Yusuke Iwata, MD e , Bret A. Mettler, MD a , Angeles Fernandez-Gonzalez, PhD c , David Zurakowski, PhD c , Richard A. Jonas, MD e , *

a Department of Cardiovascular Surgery, Children's Hospital, Boston, Massachusetts
b Department of Medicine, Children's Hospital, Boston, Massachusetts
c Department of Biostatistics, Children's Hospital, Boston, Massachusetts
d Department of Surgery, University Hospital of Oulu, Oulu, Finland
e Department of Cardiovascular Surgery, Children's National Medical Center, Washington, District of Columbia

Accepted for publication January 4, 2006.

* Address correspondence to Dr Jonas, Department of Cardiovascular Surgery, Children's National Medical Center, 111 Michigan Avenue, Washington, DC 20010 (Email: rjonas{at}cnmc.org).


   Abstract
Top
 Abstract
Introduction
 Material and Methods
 Results
 Comment
 Acknowledgments
 References
 
BACKGROUND: In previous studies we have shown that delayed capillary reperfusion after low flow bypass predicts neurologic injury. In this acute study, we hypothesized that low flow reduces endothelial nitric oxide synthase (eNOS) expression, which may lead to more profound inflammatory response and delayed capillary perfusion.

METHODS: Twelve piglets (13.2 ± 0.7 kg) had a cranial window placed over the parietal cerebral cortex for direct examination of the microcirculation using intravital fluorescence microscopy. Animals were cooled to 15°C or 34°C on cardiopulmonary bypass (pH stat, hematocrit 30%, pump flow 100 mL/kg/minute) followed by 2 hours of low flow (50 mL/kg/minute) or very low flow (10 mL/kg/minute). Rhodamine staining was used to observe adherent and rolling leukocytes in postcapillary venules. The eNOS protein expression was determined by Western immunoblotting.

RESULTS: High temperature and low flow rate correlated with significantly reduced eNOS expression (p < 0.01). Univariate comparisons based on Student t tests indicated that eNOS protein levels were lower at 34°C than at 15°C (0.7 ± 0.6 vs 1.7 ± 0.5, p < 0.01) and at 10 mL/kg per minute compared with 50 mL/kg per minute (0.8 ± 0.7 vs 1.6 ± 0.5, p = 0.03). Moreover, two-way analysis of variance revealed that temperature (F = 21.6, p < 0.001) and flow rate (F = 13.8, p = 0.005) were independent multivariate predictors of eNOS expression. During low flow bypass, eNOS was inversely correlated with numbers of adherent (p = 0.002) and rolling (p = 0.006) leukocytes, following an exponential decay curve closely.

CONCLUSIONS: eNOS expression is reduced after very low flow bypass, particularly at a higher bypass temperature. This is associated with delayed capillary reperfusion. Reduced eNOS is also associated with increased white cell activation which may lead to greater neurologic injury.


   Introduction
Top
 Abstract
 Introduction
Material and Methods
 Results
 Comment
 Acknowledgments
 References
 
Endothelium-derived nitric oxide (NO) is produced by the endothelial NO synthase (eNOS) through oxidative conversion of L-arginine to L-citrulline. When formed by the vascular endothelium NO diffuses to the adjacent cells and activates soluble guanylate cyclase, which in turn mediates many of the beneficial effects of NO. In vascular smooth muscle, NO is a potent vasodilator and regulates regional blood flow [1–3]. In addition, NO also inhibits platelet adherence and aggregation, reduces adherence of leukocytes to the endothelium, and suppresses proliferation of vascular smooth muscle cells (4–7). Studies using animal models of cerebral ischemia have demonstrated that eNOS and vascular NO play a prominent role in maintaining cerebral blood flow and preventing neuronal injury [3, 8–10].

Hypoxia has been associated with both the up regulation and down regulation of steady-state eNOS messenger ribonucleic acid (mRNA) expression [11, 12]. Exposure of human or bovine endothelial cells to low oxygen tension results in a profound decrease in the transcript for eNOS and a corresponding fall in eNOS protein levels [13]. In contrast, the upregulation of eNOS mRNA has been detected in hypoxic bovine aortic endothelial cells [14].

In addition to eNOS, other isoforms of NOS play a role during cerebral ischemia. Excitotoxic and ischemic insults excessively activate neuronal NOS (nNOS), resulting in concentrations of NO that are toxic to surrounding neurons [3, 8]. Inducible NOS (iNOS), which is not normally present in tissue, is induced after ischemia and contributes to secondary late-phase damage [15]. In contrast to nNOS and eNOS, the iNOS expression is delayed starting 6 to 12 hours after ischemic insult [16].

Little is known about the expression of NOS isoforms in the cerebral microcirculation during cardiopulmonary bypass. We hypothesized that ischemic injury to cerebral endothelial cells during cardiopulmonary bypass causes a decreased production of NO by decreasing expression of eNOS within endothelial cells. This hypothesis is further supported by the delayed or no-reflow phenomenon observed in our previous intravital microscopy (IVM) study [17].

Using IVM in our previous work, we demonstrated the mechanism of regional microvascular ischemia, increased leukocyte activation, and endothelial cell injury could result from critically reduced flow causing microvascular ischemic vasoconstriction. We hypothesized that endothelial cell hypoxic injury results in reduced eNOS expression.


   Material and Methods
Top
 Abstract
 Introduction
 Material and Methods
Results
 Comment
 Acknowledgments
 References
 
Surgical Preparation
Twelve juvenile (6–7 weeks) Yorkshire piglets with a mean body weight 13.2 ± 0.7 kg were included in the study. All animals received humane care in accordance with the "Principles of Laboratory Animal Care" formulated by the National Society for Medical Research and the "Guide for the Care and use of Laboratory Animals" prepared by the Institute of Laboratory Animal Resources, National Research Council, and published by the National Academy Press, revised in 1996.

After sedation with an intramuscular injection of ketamine (20 mg/kg) and xylazine (4 mg/kg), the piglets were intubated and ventilated with 21% oxygen to achieve an arterial partial pressure of carbon dioxide (PCO 2) of 35-40 mm Hg. After fentanyl (25 µg/kg intravenous [IV]) induction, anesthesia was maintained through continuous infusion of fentanyl (25 µg/kg/hour), midazolam (0.2 mg/kg/hour), and pancuronium (0.2 mg/kg/hour) through a right femoral vein cannula (19G Intracath; Becton Dickinson, Sandy, UT) advanced into the inferior vena cava. Temperature probes were placed into the esophagus and rectum. Continuous blood pressure monitoring was performed through a left superficial femoral artery cannula (19G Intracath; Becton Dickinson) advanced into the abdominal aorta. A right anterolateral thoracotomy was performed in the third intercostal space. After systemic heparinization (300 IU/kg IV), an 8F arterial cannula (Bio-Medicus; Medtronic Inc, Eden Prairie, MN) was placed into the abdominal aorta through the right femoral vein and a 28F venous cannula (Harvey; Bard, Tewksbury, MA) was inserted into the right atrium for cardiopulmonary bypass (CPB). The piglets were positioned prone in a stereotactic frame, and a cranial window (15 x 15 mm) was created over the parietal cerebral cortex with an electric drill. After incision of the dura, the surface (pial) vessels were visualized with the cranial window closed with a glass cover slip.

Experimental Protocol
Twelve piglets entered into the experimental protocol. Temperature of either 15°C or 34°C was maintained during a reduced flow state of 10 or 50 mg/kg per minute. Every combination of temperature and flow rate (2 x 2 = 4 settings) was performed in three separate experiments.

Arterial pressure was monitored continuously throughout each experiment and recorded every 15 minutes. Hemoglobin, hematocrit, glucose, lactate, partial pressure of oxygen (PO 2), pCO2, and pH were measured every 15 minutes on CPB (Stat Profile 9, Nova,Waltham, MA).

Cardiopulmonary Bypass Technique
The CPB circuit consisted of a roller-pump, membrane oxygenator (Minimax; Medtronic Inc, Anaheim, CA) and sterile tubing with a 40 µm arterial filter. Phlebotomy performed on the operative day of fresh whole blood from donor pigs was transfused into the prime as required to increase hematocrit 30%. Methylprednisolone (30 mg/kg), furosemide (0.25 mg/kg), and sodium carbonate (10 mL) were added to the prime. Full bypass flow was set at 100 mL/kg per minute. The pH-stat strategy management was determined by experimental protocol. After CPB commencement, the animals were perfused for 10 minutes at normothermia (esophageal temperature 37°C) followed by 40 minutes of cooling on CPB to an esophageal temperature of 15°C or 34°C as per protocol. Ventilation was stopped after the establishment of CPB. After cooling for 40 minutes, low flow perfusion at 10 mL/kg per minute or 50 mL/kg per minute flow rate was initiated for 120 minutes. After completion of the low flow period, the animals were sacrificed by an intravenous injection of Fatal-Plus (0.11 mL/kg; Vortech Pharmaceuticals, Dearborn, MI). A brain biopsy was performed through the IVM burr hole. This tissue was analyzed for quantification of NOS using Western immunoblotting.

Intravital Fluorescence Microscopy
An epifluorescence microscope (Model MZ FLIII, Leica, Heerbrugg, Switzerland) containing a 100 W mercury gas discharge lamp with a rapid filter exchanger was placed over the cranial window. A green filter (536–556 nm excitation/ >590 nm emission wavelength) was used for visualization of the rhodamine-labeled leukocytes.

The microscope images from the charge-coupled device (CCD) video camera (Dage-300-RC; Dage-MTI) were time-stamped using a time-code generator (VTG-33, For-A) and transferred to a high-resolution 12-inch monitor (Dage HR-1000; Dage-MTI, Michigan City, IN) with videotaping. A Scion LG-3 frame grabber card (Scion Corp, Frederick, MD), along with a computer-assisted image analysis system (NIH Image, National Institutes of Health, Bethesda, MD), was used for subsequent offline analysis with final image magnification being 400x.

For the assessment of leukocyte-endothelial cell interactions, the circulating leukocytes were labeled with 2 mL of 0.2% rhodamine-6G (Sigma Chemical, St. Louis, MO). The number of adherent (adherent leukocytes/100-µm vessel length) and rolling leukocytes (rollers/100-µm vessel length per minute) in a 20 to 30 µm postcapillary venule were observed using a green filter set to excite rhodamine fluorescence.

Intravital fluorescence microscopy was performed at baseline, at 10 minutes of normothermic CPB, at 20 minutes of cooling, at the end of cooling, and at every 15 minutes during low flow period. The duration of brain tissue epi-illumination was limited to less than one minute and was shut off between video recordings to avoid thermal injury.

Quantitation of eNOS Protein: Western Analysis
Sampled brain tissue was rapidly frozen with subsequent homogenization in lysis buffer. Twenty micrograms protein extracts were separated on an SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. Western immunoblotting was performed using an antihuman eNOS (NOS type III) antibody (BD Biosciences) in a 1:1000 dilution. The intensity of the normalized specific bands was quantified using NIH Image software.

Statistical Analysis
The Kolmogorov-Smirnov goodness-of-fit test indicated that eNOS and all other continuous variables followed a normal distribution. Therefore, repeated-measures analysis of variance (ANOVA) was used to evaluate changes over time and to compare rates of change between the groups [18]. Temperature and flow rate were evaluated by F-tests using ANOVA to determine if they were independently associated with eNOS. One-way ANOVA with a Bonferroni adjustment was used to detect differences between the experimental groups at different time points. The Pearson product-moment correlation coefficient (r) was used to measure the linear association between intravital microscopy data and eNOS quantification, temperature, and flow rate. Various linear and nonlinear models were compared for describing the relationship between eNOS and leukocyte counts during low flow bypass with R2 used as a criterion of fit [19]. Statistical analysis was performed using the SPSS package (version 12.0; SPSS Inc, Chicago, IL). For all comparisons, a two-sided value of p less than 0.05 was regarded as statistically significant. Data are presented in terms of the mean and standard deviation (SD).


   Results
Top
 Abstract
 Introduction
 Material and Methods
 Results
Comment
 Acknowledgments
 References
 
There were no statistically significant differences at baseline (before CPB) between the experimental groups regarding hematocrit, blood gas, mean arterial pressure, or intravital microscopy parameters.

Leukocyte-Endothelial Cell Interactions
At the end of the cooling period, more adherent leukocytes were seen at 34°C than at 15°C in the postcapillary venules (p = 0.02). At this time point, the temperature positively correlated with the number of adherent leukocytes (Pearson r = 0.72, p < 0.01).

Using multivariable analysis temperature was an independent predictor of the number of rolling and adherent leukocytes (p < 0.01). During the entire 120 minutes of low flow bypass, a statistically greater number of rolling and adherent leukocytes were seen at 34°C as compared with 15°C (p < 0.05).

Temperature was positively correlated with the number of adherent (r = 0.75, p = 0.008) and rolling leukocytes (r = 0.85, p < 0.001) during the full 120 minutes of low flow bypass.

eNOS Western Analysis
Univariate comparisons based on Student t tests indicated that eNOS protein levels were lower at 34°C than 15°C (0.7 ± 0.6 vs 1.7 ± 0.5, p < 0.01) and at 10 mL/kg per minute compared with 50 mL/kg per minute (0.8 ± 0.7 vs 1.6 ± 0.5, p = 0.03) (Fig 1). Moreover, two-way ANOVA revealed that temperature (F-test = 21.6, p < 0.001) and flow rate (F-test = 13.8, p = 0.005) were both independent multivariate predictors of eNOS expression. The ANOVA indicated no significant interaction (F-test = 1.20, p = 0.31), indicating that eNOS levels were lower at the higher bypass temperature of 34°C independent of flow rate, and that eNOS levels are higher at the higher flow rate of 50 mL/kg per minute independent of temperature. Conversely, an inverse correlation was seen by an increasing mean number of rolling and adherent leukocytes during low flow CPB and a reduced eNOS (Pearson r = –0.723 and r = –0.71, respectively, both p < 0.01).


Figure 1
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  		Fig 1. Piglet brain eNOS protein levels after critically low flow bypass. (A) Representative Western blot of three independent experiments (NS = non-specific band). (B) Quantitative analysis of eNOS protein levels (arbitrary units) in piglet brains in the four experimental conditions. Condition 1: temperature 34°C, flow 10 mL/kg per minute; condition 2: temperature 34°C, flow 50 mL/kg per minute; condition 3: temperature 15°C, flow 10 mL/kg per minute; condition 4: temperature 15°C, flow 50 mL/kg per minute. Data are expressed as the mean and standard deviation compared with condition 1 from triplicate experiments. (* p < 0.05, statistically significant difference compared with three other conditions; eNOS = endothelial nitric oxide synthase.)

 
The relationship between eNOS and leukocyte count during low flow bypass was best described by a nonlinear exponential decay function of the general form, y = Cekx, where e is the base of the natural logarithm, approximately 2.718. For adherent leukocytes, the exponential model had good fit (R2 = 0.64) and was described as a curve by the equation, y = 7.23e(–0.44x) (Fig 2A). A linear fit to the data produced an R2 of only 0.51. Similarly, nonlinear regression yielded an exponential decay function with the equation y = 3.6e(–1.1x) as the best model for describing the inverse relationship between eNOS and rolling leukocytes during low flow (R-squared = 0.57) (Fig 2B). Again, linear regression yielded a lower R2 value (R2 = 0.49) suggesting that the nonlinear curve fit the data better. Modeling these data indicate that leukocyte counts are eNOS dependent and follow a nonlinear relationship.


Figure 2
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  		Fig 2. (A) Inverse relationship between eNOS protein levels and adherent leukocyte counts during low flow as modeled using nonlinear regression based on the following exponential equation: y = 7.23exp(–0.44x), where x denotes eNOS protein level, y represents the predicted number of adherent leukocytes, and e is the base of the natural logarithm. (B) Inverse relationship between eNOS protein levels and rolling leukocyte counts during low flow as modeled using nonlinear regression based on the following exponential equation: y = 3.6e(–1.1x), where x denotes the eNOS protein level, y represents the predicted number of adherent leukocytes, and e is the base of the natural logarithm. (eNOS = endothelial nitric oxide synthase.)

 

   Comment
Top
 Abstract
 Introduction
 Material and Methods
 Results
 Comment
Acknowledgments
 References
 
Few previous studies have used intravital fluorescence microscopy for direct visualization of the cerebral microcirculation during CPB in porcine model. Using intravital microscopy, microvascular (including capillary) blood flow can be directly visualized and analyzed after labeling plasma with fluorescent dyes. Combining IVM with quantitative analysis of eNOS provides the opportunity to study microvascular disturbances associated with eNOS expression.

This study has demonstrated that mildly hypothermic (34°C) cardiopulmonary bypass with critically reduced flow rate is associated with significantly decreased expression of eNOS compared with deep hypothermic (15°C) bypass. This result suggests that more profound hypothermia preserves expression of eNOS by endothelial cells. This finding is consistent with previous reports from other groups, which have documented that hypoxia results in down regulation of eNOS expression [13]. Hypothermia appears to help to maintain the capability of the vascular endothelium to produce nitric oxide.

There is now considerable evidence indicating that an acute inflammatory response occurs after CPB, and that it is more profound when normothermic or mild hypothermic temperatures are used [20, 21]. The current study confirms our previous finding that higher bypass temperature results in a greater inflammatory response as detected by an increased number of rolling and adherent leukocytes in the postcapillary venules (22). The number of leukocytes was found to correlate inversely with eNOS expression and followed an exponential relationship with moderately good fit according to R-squared criteria. A mechanism whereby NO is likely to mitigate leukocyte adherence is based on the avidity of NO for interacting with a superoxide free radical. Endogenous NO competes with superoxide dismutase to inactivate basally produced superoxide radical [23, 24]. Loss of NO after NOS inhibition may lead to an increased level of superoxide radical which has been shown to be a proadherent molecule in a variety of microcirculatory beds [25, 26]. Our finding that reduced eNOS protein levels resulted in an increase in activated leukocytes in cerebral pial venules is consistent with similar previous findings in noncerebral tissues [27] as well as in the cerebral microcirculation [5]. However, this finding reflects the anti-inflammatory effect of NO.

In conclusion, this study shows that an ischemic insult to cerebral endothelial cells results in reduced eNOS expression, and that this is associated with increased white cell activation which may lead to greater neurologic injury. Deep hypothermic bypass maintains eNOS expression resulting in less activated neutrophlis reflecting a less severe inflammatory response.[4][6][7]


   Acknowledgments
Top
 Abstract
 Introduction
 Material and Methods
 Results
 Comment
 Acknowledgments
References
 
Doctor Anttila was supported by The Academy of Finland and The Finnish Medical Foundation.


   References
Top
 Abstract
 Introduction
 Material and Methods
 Results
 Comment
 Acknowledgments
 References
 

  1. Loscalzo J. Nitric oxide and vascular disease N Engl J Med 1995;333:251-253.[Free Full Text]
  2. Rudic RD, Sessa WC. Nitric oxide in endothelial dysfunction and vascular remodelingclinical correlates and experimental links. Am J Hum Genet 1999;64:673-677.[Medline]
  3. Samdani AF, Dawson TM, Dawson VL. Nitric oxide synthase in models of focal ischemia Stroke 1997;28:1283-1288.[Abstract/Free Full Text]
  4. Huang PL, Huang Z, Mashimo H, et al. Hypertension in mice lacking the gene for endothelial nitric oxide synthase Nature 1995;377:239-242.[Medline]
  5. Gidday JM, Park TS, Shah AR, Gonzales ER. Modulation of basal and postischemic leukocyte-endothelial adherence by nitric oxide Stroke 1998;29:1423-1429.[Abstract/Free Full Text]
  6. Hudetz AG, Wood JD, Kampine JP. Nitric oxide synthase inhibitor augments post-ischemic leukocyte adhesion in the cerebral microcirculation in vivo Neurol Res 1999;21:378-384.[Medline]
  7. Lindauer U, Dreier J, Angstwurm K, et al. Role of nitric oxide synthase inhibition in leukocyte-endothelium interaction in the rat pial microvasculature J Cereb Blood Flow Metab 1996;16:1143-1152.[Medline]
  8. Iadecola C. Bright and dark sides of nitric oxide in ischemic brain injury Trends Neurosci 1997;20:132-139.[Medline]
  9. Huang Z, Huang PL, Ma J, et al. Enlarged infarcts in endothelial nitric oxide synthase knockout mice are attenuated by nitro-L-arginine J Cereb Blood Flow Metab 1996;16:981-987.[Medline]
  10. Dalkara T, Morikawa E, Panahian N, Moskowitz MA. Blood flow-dependent functional recovery in a rat model of focal cerebral ischemia Am J Physiol 1994;267:H678-H683.[Medline]
  11. McQuillan LP, Leung GK, Marsden PA, Kostyk SK, Kourembanas S. Hypoxia inhibits expression of eNOS via transcriptional and posttranscriptional mechanisms Am J Physiol 1994;267:H1921-H1927.[Medline]
  12. Liao JK, Zulueta JJ, Yu FS, Peng HB, Cote CG, Hassoun PM. Regulation of bovine endothelial constitutive nitric oxide synthase by oxygen J Clin Invest 1995;96:2661-2666.[Medline]
  13. Phelan MW, Faller DV. Hypoxia decreases constitutive nitric oxide synthase transcript and protein in cultured endothelial cells J Cell Physiol 1996;167:469-476.[Medline]
  14. Arnet UA, McMillan A, Dinerman JL, Ballermann B, Lowenstein CJ. Regulation of endothelial nitric-oxide synthase during hypoxia J Biol Chem 1996;271:15069-15073.[Abstract/Free Full Text]
  15. Endres M, Laufs U, Liao JK, Moskowitz MA. Targeting eNOS for stroke protection Trends Neurosci 2004;27:283-289.[Medline]
  16. Iadecola C, Zhang F, Casey R, Clark HB, Ross ME. Inducible nitric oxide synthase gene expression in vascular cells after transient focal cerebral ischemia Stroke 1996;27:1373-1380.[Abstract/Free Full Text]
  17. Duebener LF, Sakamoto T, Hatsuoka S, et al. Effects of hematocrit on cerebral microcirculation and tissue oxygenation during deep hypothermic bypass Circulation 2001;104:I260-I264.[Medline]
  18. Wallenstein S, Zucker CL, Fleiss JL. Some statistical methods useful in circulation research Circ Res 1980;47:1-9.[Abstract/Free Full Text]
  19. McCullagh P, Nelder JA. Generalized linear models. 2nd ed. New York: Chapman & Hall; 1989.
  20. Chello M, Mastroroberto P, Romano R, Ascione R, Pantaleo D, De Amicis V. Complement and neutrophil activation during cardiopulmonary bypassa randomized comparison of hypothermic and normothermic circulation. Eur J Cardiothorac Surg 1997;11:162-168.[Abstract]
  21. Wagner FM, Schiller W, Dilg G, Depner C, Welz A, Lacour-Gayet F. Direct visualization of the influence of normothermic as opposed to hypothermic cardiopulmonary bypass on the systemic microcirculation in neonatal piglets J Thorac Cardiovasc Surg 2004;127:1781-1788Cardiol Young 2001;11:532-8.22. Anttila V, Hagino I, Zurakowski D, Lidov HG, Jonas RA. Higher bypass temperature correlates with increased white cell activation in the cerebral microcirculation.[Abstract/Free Full Text]
  22. Rubanyi GM, Ho EH, Cantor EH, Lumma WC, Botelho LH. Cytoprotective function of nitric oxideinactivation of superoxide radicals produced by human leukocytes. Biochem Biophys Res Commun 1991;181:1392-1397.[Medline]
  23. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA. Apparent hydroxyl radical production by peroxynitriteimplications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A 1990;87:1620-1624.[Abstract/Free Full Text]
  24. Gaboury J, Woodman RC, Granger DN, Reinhardt P, Kubes P. Nitric oxide prevents leukocyte adherencerole of superoxide. Am J Physiol 1993;265:H862-H867.[Medline]
  25. Serrano Jr CV, Mikhail EA, Wang P, Noble B, Kuppusamy P, Zweier JL. Superoxide and hydrogen peroxide induce CD18-mediated adhesion in the postischemic heart Biochim Biophys Acta 1996;3:191-202.
  26. Kubes P, Suzuki M, Granger DN. Nitric oxidean endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci U S A 1991;88:4651-4655.[Abstract/Free Full Text]



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