Significance of oxygen free radicals in the pathophysiology of hemorrhagic shock - A protocol.

Oxygen free radicals have been implicated as the deleterious agent in a variety of organ systems undergoing ischemia and subsequent reperfusion. Hemorrhagic shock represents a clinical situation that carries a high rate of morbidity and mortality despite adequate fluid resuscitation. Since this entity represents, in its most simplified sense, total body ischemia followed by reperfusion, it is likely that the generation of oxygen free radicals has some significance in the pathophysiology of this delayed morbidity. This is a research protocol, where rabbits will be subjected to severe hemorrhagic shock followed by adequate fluid resuscitation. In the first part of the experiment, free radical generation will be measured directly by Electron Paramagnetic Resonance (EPR) spectroscopy in various organ systems in rabbits before and during shock, and following resuscitation. In the second part, free radical scavengers will be introduced as an adjunct to fluid resuscitation in a group of rabbits subjected to hemorrhagic shock to see if mortality rates are affected. By acquiring a better understanding of the molecular mechanisms that may be responsible for the delayed morbidity in reperfusion injury in general, and hemorrhagic shock in particular, we will be able to better address the long-standing problem of multi system organ failure (MSOF) that often follows a successful resuscitation.

A rare case of hepatic duct injury from blunt abdominal trauma.

BACKGROUND: A 25 year-old male was brought to the emergency room following an apparent suicide attempt by jumping from the fourth floor. CASE REPORT: Patient had a large abdominal laceration in the right upper quadrant (RUQ). CT scan showed a sub-scapular hematoma of the liver. Due to the repeated episodes of hypotension, a laporotomy was performed and the left hepatic artery was ligated while the ductal injury was managed with a Roux-en-Y left hepatic jejunostomy and stent. Bile leakage was resolved post-operatively by day 5 and the patient was discharged home on day 13 after clearance from psychiatry. CONCLUSIONS: While non-iatrogenic extrahepatic biliary trauma is rare, a high degree of suspicion is essential, especially in cases like the one discussed in this report. Diagnosis can be difficult in patients undergoing observation.

Amelioration of ischemia-reperfusion injury in an isolated rabbit lung model using OXANOH.

OBJECTIVE: Acute respiratory distress syndrome (ARDS) remains a major cause of morbidity and mortality. Oxygen-free radicals (OFRs) produced during ischemia and reperfusion (IR) have been implicated as the final common pathway in the pathogenesis of this syndrome. Spin traps have been shown to decrease IR injury in several animal lung models. The hydroxylamine, OXANOH (2-ethyl-2,5,5-trimethyl-3-oxazolidine) has been proposed as an ideal spin trap that would trap extra- and intracellular OFRs producing the stable radical, OXANO• (2-ethyl-2,5,5-trimethyl-3-oxazolidinoxyl). Electron microscopy was used to investigate whether OXANOH would protect against IR injury in the rabbit lung. METHODS: OXANOH was obtained by hydrogenation of its stable radical, OXANO• using a safe laboratory technique. Several doses of OXANOH were tested to identify a nontoxic dose. Two quantitative methods were used based on the average surface area of the alveoli and average number of alveoli per unit surface area using scanning electron microscopy (SEM). A total of 20 animals were subjected to 2 hours of ischemia followed by 4 hours of reperfusion. On reperfusion, the 4 groups (N = 5) received no treatment, OXANOH, superoxide dismutase (SOD)/catalase, or oxypurinol. RESULTS: A therapeutic dose of 250 µmol/L of OXANO• was suggested in this in vitro model. All the 3 treatments showed significantly less injury compared to the control group and that SOD/catalase was significantly different from OXANOH and oxypurinol (P < .008). CONCLUSION: OXANOH ameliorated IR injury in the isolated rabbit lung, almost as effectively as SOD/catalase and oxypurinol.

Ischemia-reperfusion injury in the lung: quantitation using electron microscopy.

BACKGROUND: The primary objectives of this study were to determine the time course of ischemia-reperfusion injury in an isolated rabbit lung model and to quantify this damage using electron microscopic methodology coupled with statistical analyses. MATERIALS AND METHODS: Eight groups of isolated rabbit lungs (n = 5 per group) were subjected to predetermined periods of ischemia-reperfusion. Two hours of ischemia and 4 hours of reperfusion were concluded to be necessary to induce optimal ischemia-reperfusion injury in this model. Four other groups were subjected to 2 hours of ischemia followed by selected periods of reperfusion. These groups were compared to 4 control groups that were perfused for comparable time periods but without the initial ischemia. New quantitative methods were developed based on the average surface area of the alveoli and average number of alveoli per unit surface area, using scanning electron microscopic examination. RESULTS: Ischemia per se caused substantial damage. Restoration of volume and nutrients reversed this damage at 1 hour of reperfusion, but severe damage was evident at 4 hours of reperfusion, as reported by subjective and blinded examination. By using the new quantitative methods, there was a significant difference between the groups (P < .005) according to the time of post-ischemia-reperfusion, which correlated with the subjective evaluation of damage. CONCLUSIONS: These 2 new quantitative techniques provide an objective assessment of damage in the isolated rabbit lung model, suggesting that they warrant further consideration in similar studies of ischemia reperfusion injury.

Atrial natriuretic peptide protects against ischemia-reperfusion injury in rabbit hearts in vivo.

The aim of this study is to investigate whether atrial natriuretic peptide can mimic preconditioning to protect ischemia or reperfusion injury in rabbit hearts. New Zealand white rabbits were randomized into 3 groups: (1) Controls. Hearts received a 60 minute-occlusion of the left anterior descending artery, followed by a 180 minute-reperfusion. (2) Preconditioning. Two 5-minute periods of ischemia separated by a 10-minute reperfusion, followed by a 60-minute ischemia and a 180-minute reperfusion. (3) Atrial natriuretic peptide treatment. Bolus injection of exogenous atrial natriuretic peptide (2.5 microg/kg) given intravenously at 15 minutes prior to 60 minute-ischemia followed by a 180-minute reperfusion. Myocardial necrotic area and area at risk of necrosis were determined by triphenyltetrazolium chloride staining. Ratio of necrotic area to area at risk was 49.95% +/- 1.15%, 7.95% +/- 0.33%, and 8.36% +/- 0.61% in the controls, preconditioning group, and atrial natriuretic peptide group, respectively. Both preconditioning and atrial natriuretic peptide significantly reduced the size of infarct caused by ischemia (preconditioning vs controls, P < .05; atrial natriuretic peptide vs controls, P < .05). Atrial natriuretic peptide can mimic ischemic preconditioning to protect rabbit hearts from prolonged ischemia and reperfusion injury. It may be involved in the cardioprotective mechanisms of preconditioning.