Charles F. Babbs

Free radicals and the etiology of colon cancer

This hypothesis paper reviews diverse evidence suggesting that intracolonic production of oxygen radicals may play a role in carcinogenesis. The hypothesis began to evolve when the author made the chance discovery that 1/10,000 dilutions of feces generated detectable quantities of highly reactive hydroxyl radicals (HO.). The rate of HO. formation, detected using DMSO as a molecular probe, was quite remarkable, corresponding to that which would be produced by over 10,000 rads of gamma irradiation per day, absorbed in the periphery of the fecal mass adjacent to the mucosa. The relatively high concentrations of iron in feces, together with the ability of bile pigments to act as iron chelators that support Fenton chemistry, may very well permit efficient HO. generation from superoxide and hydrogen peroxide produced by bacterial metabolism. Such free radical generation in feces could provide a missing link in our understanding of the etiology of colon cancer: the oxidation of procarcinogens either by fecal HO., or by secondary peroxyl radicals (ROO.) to form active carcinogens or mitogenic tumor promotors. Intracolonic free radical formation may explain the high incidence of cancer in the colon and rectum, compared to other regions of the GI tract, as well as the observed correlations of a higher incidence of colon cancer with red meat in the diet, which increases stool iron, and with excessive fat in the diet, which may increase the fecal content of procarcinogens and bile pigments.

Oxygen radicals in ulcerative colitis

This article reviews the pathophysiologic concept that superoxide and hydrogen peroxide, generated by activated leukocytes, together with low-molecular-weight chelate iron derived from fecal sources and from denatured hemoglobin, amplify the inflammatory response and subsequent mucosal damage in patients with active episodes of ulcerative colitis. The putative pathogenic mechanisms reviewed are as follows: (1) Dietary iron is concentrated in fecal material owing to normally limited iron absorption. (2) Mucosal bleeding, characteristic of ulcerative colitis, as well as supplemental oral iron therapy for chronic anemia, further conspire to maintain or elevate mucosal iron concentration in colitis. (3) Fenton chemistry, driven especially by leukocyte-generated superoxide and hydrogen peroxide, leads to formation of hydroxyl radicals. (4) The resultant oxidative stress leads to the extension and propagation of crypt abscesses, either through direct membrane disruption by lipid peroxidation or through generation of secondary toxic oxidants such as chloramines. (5) Chemotactic products of lipid peroxidation, including 4-hydroxynonenal, provide positive feedback to accelerate this inflammatory/oxidative process, leading to acute exacerbations of the disease. (6) Other oxidized products, such as oxidized tryptophan metabolites, created by free radical mechanisms in or near the mucosa, may act as carcinogens or tumor promotors that contribute to the exceedingly high incidence of colon carcinoma in patients suffering from chronic ulcerative colitis. In this way, self-sustaining cycles of oxidant formation may amplify flare-ups of inflammation and mucosal injury in ulcerative colitis. This concept, if proved correct by subsequent research, would provide a rationale for several novel clinical approaches to the management of ulcerative colitis, including use of SOD mimetics, iron chelators, and chain-breaking antioxidants.

Intrapulmonary epinephrine during prolonged cardiopulmonary resuscitation: Improved regional blood flow and resuscitation in dogs

Blood flow to vital organs was measured at five-minute intervals during 20 minutes of cardiopulmonary resuscitation (CPR) and ventricular fibrillation in two groups of anesthetized dogs (n = 15 per group). The relationship between organ blood flow and restoration of circulation after 20 minutes was assessed with no additional treatment in Group I and with intrapulmonary epinephrine in Group II. Cardiac output and organ blood flow did not vary significantly in Group I. In Group II, intrapulmonary epinephrine significantly improved blood flow to the myocardium, the brain, and the adrenals. A mean myocardial blood flow of less than 0.13 mL/min/g resulted in no survival, while a flow of greater than 0.16 mL/min/g resulted in survival. These studies show that a critical level of myocardial blood flow is required to restore ability of the heart to function as a pump after prolonged CPR, and that a drug that increases flow improves resuscitation efforts.

Myocardial perfusion pressure: A predictor of 24-hour survival during prolonged cardiac arrest in dogs

Myocardial perfusion pressure, defined as the aortic diastolic pressure minus the right atrial diastolic pressure, correlates with coronary blood flow during cardiopulmonary resuscitation (CPR) and predicts initial resuscitation success. Whether this hemodynamic parameter can predict 24-h survival is not known. We examined the relationship between myocardial perfusion pressure and 24-h survival in 60 dogs that underwent prolonged (20 min) ventricular fibrillation and CPR. Forty-two (70%) animals were initially resuscitated and 20 (33%) survived for 24 h. Myocardial perfusion pressure was significantly greater when measured at 5, 10, 15 and 20 min of ventricular fibrillation in the resuscitated animals than in the non-resuscitated animals (P less than 0.01). Likewise, the myocardial perfusion pressure was also greater in the animals that survived 24 h than in animals that were resuscitated, but died before 24 h (P less than 0.02). Myocardial perfusion pressure measured after 10 min of CPR was 11 +/- 2 mmHg in animals never resuscitated, 20 +/- 3 mmHg in those resuscitated that died before 24 h and 29 +/- 2 mmHg in those that survived 24 h (P less than 0.05). A myocardial perfusion pressure at 10 min of CPR of 20 mmHg or less is an excellent predictor of poor survival (negative predictive value = 96%). Myocardial perfusion pressure is a useful index of CPR effectiveness and therefore may be a useful guide in helping to optimize resuscitation efforts.

Optimum compression to ventilation ratios in CPR under realistic, practical conditions: a physiological and mathematical analysis

OBJECTIVE To develop and evaluate a practical formula for the optimum ratio of compressions to ventilations in cardiopulmonary resuscitation (CPR). The optimum value of a variable is that for which a desired result is maximized. Here the desired result is assumed to be either oxygen delivery to peripheral tissues or a combination of oxygen delivery and waste product removal. METHOD Equations describing oxygen delivery and blood flow during CPR as functions of the number of compressions and the number of ventilations delivered over time were developed from principles of classical physiology. These equations were solved explicitly in terms of the compression/ventilation ratio and evaluated for a wide range of conditions using Monte Carlo simulations. RESULTS As the compression to ventilation ratio was increased from 0 to 50 or more, both oxygen delivery and the combination of oxygen delivery with blood flow increased to maximum values and then gradually declined. For variables typical of standard CPR as taught and specified in international guidelines, maximum values occurred at compression/ventilation ratios near 30:2. For variables typical of actual lay rescuer performance in the field, maximal values occurred at compression/ventilation ratios near 60:2. CONCLUSION Current guidelines overestimate the need for ventilation during standard CPR by two to four-fold. Blood flow and oxygen delivery to the periphery can be improved by eliminating interruptions of chest compression for these unnecessary ventilations.

[11] Detection and quantitation of hydroxyl radical using dimethyl sulfoxide as molecular probe

Generation of clearly harmful amounts of hydroxyl radicals in biological systems can be studied using DMSO as a molecular probe. DMSO is oxidized by HO. to form the stable, nonradical compound methanesulfinic acid, which is not normally found in living systems and which can be easily extracted from tissue and measured spectrophotometrically. The present method provides a simple, inexpensive assay for methanesulfinic acid in biological materials. As little as 10 nmol of sulfinate can be detected, and interference from diverse biological compounds is minimal. Additionally, there is no interference from a large excess of dimethyl sulfoxide, which is necessary if the assay is to be applied directly to tissues pretreated with DMSO. When straightforward cleanup procedures are utilized, there is minimal interference from glutathione or sulfate, and potentially troublesome interference from detergentlike substances can usually be minimized. Owing to its relative specificity for sulfinic acids at acid pH, the diazonium coupling reaction can thus be exploited to provide an efficient and inexpensive means of detecting methanesulfinic acid in DMSO-pretreated biological materials. The results provide a direct chemical means for measuring cumulative HO. generation.

Multiresolution detection of spiculated lesions in digital mammograms

We present a novel multiresolution scheme for the detection of spiculated lesions in digital mammograms. First, a multiresolution representation of the original mammogram is obtained using a linear phase nonseparable two-dimensional (2-D) wavelet transform. A set of features is then extracted at each resolution in the wavelet pyramid for every pixel. This approach addresses the difficulty of predetermining the neighborhood size for feature extraction to characterize objects that may appear in different sizes. Detection is performed from the coarsest resolution to the finest resolution using a binary tree classifier. This top-down approach requires less computation by starting with the least amount of data and propagating detection results to finer resolutions. Experimental results using the MIAS image database have shown that this algorithm is capable of detecting spiculated lesions of very different sizes at low false positive rates.

Role of iron ions in the genesis of reperfusion injury following successful cardiopulmonary resuscitation: Preliminary data and a biochemical hypothesis

Presented is a rationale for use of a new class of drugs, the iron chelating agents, in advanced cardiac life support (ACLS) to prevent late deaths and brain damage following successful cardiopulmonary resuscitation. The relevant biochemical hypothesis states that free iron ions, liberated from bound intracellular stores during ischemia, catalyze initiation of free radical mediated reactions that propagate through membrane lipids and proteins. Progressive ultrastructural damage may result, ultimately causing deterioration of function and death. Chelation of intracellular iron by deferoxamine, a commercially available drug that distributes to the intracellular space and has a great affinity for iron ions, may prevent such reactions. A hypothesis concerning relevant pathological chemistry is developed in detail.

Changes in expired end-tidal carbon dioxide during cardiopulmonary resuscitation in dogs: A prognostic guide for resuscitation efforts

Expired end-tidal carbon dioxide (PCO2) measurements made during cardiopulmonary resuscitation have correlated with cardiac output and coronary perfusion pressure when wide ranges of blood flow are included. The utility of such measurements for predicting resuscitation outcome during the low flow state associated with closed chest cardiopulmonary resuscitation remains uncertain. Expired end-tidal PCO2 and coronary perfusion pressures were measured in 15 mongrel dogs undergoing 15 min of closed chest cardiopulmonary resuscitation after a 3 min period of untreated ventricular fibrillation. In six successfully resuscitated dogs, the mean expired end-tidal PCO2 was significantly higher than that in nine nonresuscitated dogs only after 14 min of cardiopulmonary resuscitation (6.2 +/- 1.2 versus 3.4 +/- 0.8 mm Hg; p less than 0.05). No differences in expired end-tidal PCO2 values were found at 2, 7 or 12 min of cardiopulmonary resuscitation. A significant decline in end-tidal PCO2 levels during the resuscitation effort was seen in the nonresuscitated group (from 6.3 +/- 0.8 to 3.4 +/- 0.8 mm Hg; p less than 0.05); the successfully resuscitated group had constant PCO2 levels throughout the 15 min of cardiac arrest (from 6.8 +/- 1.1 to 6.2 +/- 1.2 mm Hg). Changes in expired PCO2 levels during cardiopulmonary resuscitation may be a useful noninvasive predictor of successful resuscitation and survival from cardiac arrest.

Quantitation of the hydroxyl radical by reaction with dimethyl sulfoxide

This investigation was conducted to validate the use of dimethyl sulfoxide (DMSO) as a quantitative molecular probe for the generation of hydroxyl radicals (HO.) in aqueous systems. Reaction of HO. with DMSO produces methane sulfinic acid as a primary product, which can be detected by a simple colorimetric assay. To evaluate this method for estimating total HO. production, we studied three model systems, including the Fenton reaction, gamma irradiation of water, and ultraviolet photolysis of hydrogen peroxide, for which the theoretical maximum yield of HO. could be calculated and compared to measured DMSO oxidation. The results confirm that 0.05 to 1 M DMSO may be used to capture nearly all of the expected HO. radicals formed. Thus, methane sulfinic acid production from DMSO holds promise as an easily measured marker for HO. formation in aqueous systems pretreated with DMSO.