Handling ability of gaseous microemboli of two pediatric arterial filters in a simulated CPB model.

OBJECTIVE: The purpose of this experiment was to compare the Sorin KIDS D131 and the Terumo Capiox AF02 pediatric arterial filters in a simulated CPB procedure to determine which filter is the better for clinical use. METHODS: The experimental circuit was primed with an 800 ml combination of lactated Ringer's solution and human blood (hematocrit (Hct) 30%). The two filters were tested under flow rates of 500, 1000, and 1500 ml/min at room temperature and their purge lines opened and closed as 5cc of air was injected into the circuit. RESULTS: As the flow rates increased, the number of gaseous microemboli (GME) being returned to the pseudo patient increased for both of the pediatric arterial filters. Having an open purge line increased the number of GME removed from the CPB circuit, caused less of a pressure drop than when closed and increased the total hemodynamic energy loss than when closed. Both of the filters performed and reacted similarly in decreasing GME, hemodynamic energy loss and pressure drop. The only minor difference was that the Capiox AF02 had slightly less stolen blood flow (109.5 ± 1.7 ml/min at 500 ml/min, 114.7 ± 1.1 ml/min at 1000 ml/min and 105.8 ± 4.2 ml/min at 1500ml/min) from the open purge line than the KIDS D131 (119.5 ± 2.5 ml/min at 500 ml/min, 128.3 ± 1.0 ml/min at 1000 ml/min and 126.3 ± 3.1 ml/min at 1500 ml/min). CONCLUSION: Our study confirmed that both the Sorin KIDS D131 and the Terumo Capiox AF02 were equivalent in their ability to remove significant numbers of GME, the amount of pressure drop and the total hemodynamic energy loss across the arterial filters at the various flow rates. An arterial filter is not an option, but a necessity for removing microemboli delivered to the patient.

Glucose consumption by rats decreases cytochrome P450 enzyme activity by altering hepatic lipids.

Although glucose is a ubiquitous nutrient, increased consumption of glucose decreases the metabolism of numerous drugs in humans and animals. To understand the mechanisms involved that cause decreased drug metabolism in rats that consume glucose in their water, enzyme activity and expression as well as determining the contribution of the lipids toward decreasing in vitro metabolic activity were investigated. Enzyme assays of hepatic CYP1A2, 2C6, 2C11 and 3A2 showed significant decreases in activity from glucose-treated rats compared to control. While immunodetection of CYP1A1, 2B1/2, 2C11, and 3A1/2 showed no significant difference in protein expression. Hepatic fatty acid synthase activity increased in the glucose-treated rats compared to controls. Studies with glucose-treated microsomal lipids reconstituted with microsomal proteins from control rats caused a significant decrease in benzyloxyresorufin O-dealkylase activity. The results presented here support the hypothesis that the activities of cytochrome P450 proteins are altered by modulating their catalytic activity as a result of the lipid environment rather than changing the level of expression of the individual enzymes.

Inhibition of liver microsomal cytochrome P450 activity and metabolism of the tobacco-specific nitrosamine NNK by capsaicin and ellagic acid.

The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3- pyridyl)-1-butanone (NNK), present in tobacco and tobacco smoke, is metabolically activated by microsomal enzymes. In this study, we examined the effect of capsaicin and ellagic acid on the in vitro metabolism of NNK by hamster and rat liver microsomes. Capsaicin is the principal component of Capsicum fruits used widely throughout the world as a food additive. Ellagic acid, with reported anticarcinogenic properties, is found in various soft fruits and nuts. Both capsaicin and ellagic acid inhibited the major pathways of NNK-reduction, N-pyridine oxidation and a-hydroxylation by hamster liver microsomes. Capsaicin inhibited NNK-reduction and a-hydroxylation and ellagic acid inhibited N-oxidation and a-hydroxylation by rat liver microsomes. The effects of capsaicin and ellagic acid on isozymes of cytochrome P450 were observed in the hydroxylation reactions of the metabolism of the steroid hormone testosterone. Results of these experiments indicated that both capsaicin and ellagic acid strongly inhibited the constitutive enzymes CYP 2A2, 3A1, 2C11, 2B1, 2B2 and 2C6. This study suggests that capsaicin and ellagic acid, as naturally occurring dietary constituents, possess antimutagenic and anticarcinogenic properties through the inhibition of xenobiotic metabolizing enzymes.

Glucose alters rat liver S9-mediated mutagenesis, metabolism and DNA-binding of aflatoxin B1.

The administration of 30% glucose in drinking water to rats for 48 h caused a significant increase in the hepatic S9-mediated mutagenicity of aflatoxin B1, in Salmonella typhimurium TA100 and in the binding of alfatoxin B1, to calf thymus DNA in vitro. These effects correlated with a reduction in the metabolism and detoxification of aflatoxin B1, by S9 from glucose-treated rats and suggest that the oral intake of sugar may affect the hepatocarcinogenicity of aflatoxin B1.

Effect of glucose administration on hamster liver S9-mediated mutagenesis, metabolism and DNA-binding of benzo[a]pyrene and aflatoxin B1.

Hamster liver S9 prepared from control animals and animals given 30% glucose in drinking water 48 h before killing was used in studies of benzo[a]pyrene (BaP) and aflatoxin (AFB1)-induced mutagenesis, metabolism of BaP and AFB1, and metabolite binding to calf thymus DNA. BAP-induced mutagenesis in Salmonella typhimurium TA100 was reduced 38.5% while AFB1-induced mutagenesis was increased 36% by S9 from glucose-treated hamsters. The reduction of [3H]BaP metabolite binding to calf thymus DNA in incubations with S9 from glucose-treated hamsters correlated with a decrease in unknown BP metabolite-deoxyribonucleoside adducts isolated by high performance liquid chromatography (HPLC). Differences in the 7R and 7S-diol epoxide-1 and 2 deoxyguanosine adducts of BaP between control and glucose-treated S9 were not observed. HPLC analysis of AFB1-DNA adducts showed a 25% increase in [3H]AFB1-N7-guanine in incubations of glucose-treated S9 with [3H]AFB1 and calf thymus DNA. HPLC analysis of the organosoluble fraction of incubations with [3H]BaP and [3H]AFB1 indicated a significant effect by glucose-treated S9 on metabolism. The effect of glucose on metabolism was further reflected in the reduction of both BaP and AFB1 metabolite conjugation with glucuronide and glutathione as determined by separation on an alumina column. These results indicate that the oral administration of 30% glucose in drinking water alters hamster liver S9-mediated mutagenesis and binding of BaP and AFB1 metabolites to DNA through an effect on the metabolism of these two carcinogens.

Inhibition of liver microsome-mediated mutagenesis, metabolism and DNA-binding of benzo[a]pyrene and benzo[a]pyrene 7,8-dihydrodiol in the rat following glucose administration.

Aroclor 1254-induced rat liver microsomes prepared from control and glucose-treated rats (30% glucose in drinking water 48 h prior to sacrifice) were used in studies of benzo[a]pyrene (BaP) and BaP 7,8-dihydrodiol (BaP 7,8-DHD)-induced mutagenesis in Salmonella typhimurium TA100. Microsome-dependent metabolism and metabolite binding of BaP and BaP 7,8-DHD to calf thymus DNA was also investigated. BaP-induced mutagenesis in TA100 was inhibited 27% and BaP 7,8-DHD-induced mutagenesis was inhibited 55% by microsomes from glucose-treated rats. [3H]BaP and [3H]BaP 7,8-DHD metabolite binding to DNA was inhibited 17% and 20%, respectively. High performance liquid chromatographic (hplc) analysis of enzyme-hydrolyzed DNA yielded 7R and 7S-diol epoxide-1 deoxyguanosine (BPDE-1:dG) adducts and BPDE-2:dG adducts of [3H]BaP and [3H]BaP 7,8-DHD. These adducts were inhibited 38% and 50%, respectively, by microsomes from glucose-treated rats. Hplc analysis of organosoluble metabolites of [3H]BaP and [3H]BaP 7,8-DHD showed an inhibition of metabolism of 28% and 50%, respectively, by microsomes from glucose-treated rats. The inhibition of metabolism correlated with the effect of glucose treatment on inhibition of BaP and BaP 7,8-DHD-induced mutagenesis and adduct formation. These results suggest that the mechanism by which glucose produces its effects on mutagenesis, DNA-binding and adduct formation is by an inhibition of microsome-mediated metabolism of BaP and BaP 7,8-DHD.

Effect of glucose administration on various rat hepatic constituents and on the spectral binding of hexobarbital and methadone to rat hepatic cytochrome P450.

The administration of glucose to rats for 48 h resulted in an increase in the duration of anesthesia produced by intraperitoneal injection of pentobarbital. The effect of this glucose treatment on hepatic glycogen and on microsomal protein, cytochrome P450 (P450), cholesterol, lipid and phospholipid content, as well as on the spectral binding of hexobarbital and methadone to microsomal P450, was examined. The glucose treatment appeared to have no effect on microsomal protein or P450 content. The binding of hexobarbital and methadone by P450 produced a type 1 spectrum in both control and glucose-treated animals. The glucose treatment resulted in a decrease in the spectral dissociation constant (Ks) and in the maximal spectral binding (delta Amax) of hexobarbital to P450 while with methadone there was an increase in Ks and a decrease in delta Amax. Total lipid, phosphatidylcholine and phosphatidylethanolamine were increased in the microsomal fractions from glucose-treated animals. The changes in the parameters for drug binding to microsomal P450 probably relate to the increased lipid content of the microsomes although changes in the proportion of P450 isoenzymes could be involved. The previously observed decrease in the microsomal metabolism of hexobarbital and methadone following glucose treatment may be due to decreased binding of these compounds to microsomal P450.

Change of deformability and Heinz body formation in G6PD-deficient erythrocytes treated with 5-hydroxy-6-desmethylprimaquine.

Glucose-6-phosphate deficient human erythrocytes were incubated with low concentrations of 5-hydroxy-6-desmethylprimaquine, a metabolite of primaquine in animals, for up to 18 hours under sterile conditions. These erythrocytes became less deformable than untreated erythrocytes. This decrease in deformability was closely associated with the extent and time course of formation of Heinz bodies in the G6PD-deficient erythrocytes. These results support the hypothesis that the in vivo formation of low concentrations of 5H6DPQ by metabolism of primaquine could be the cause of Heinz body formation and the hemolytic anemia seen when primaquine is administered to G6PD-deficient individuals.

Effects of an NADPH-generating system on primaquine degradation by hamster liver fractions.

1. Primaquine (PQ) often causes severe anaemia in individuals with glucose 6-phosphate dehydrogenase (G6PD) deficient erythrocytes, and metabolites have been implicated as the toxic substance. These studies present data identifying additional metabolites of PQ. 2. Two metabolites of primaquine (PQ) previously identified in human studies, namely, 6-methoxy-8-aminoquinoline (MAQ) and 8-(3-carboxy-1-methylpropylamino)-6-methoxyquinoline (PQC) were also formed on incubation of PQ with hamster liver fractions for up to 24 h without an NADPH-generating system. 3. The alcohol (PQAOH) and lactam (PQLT) derivatives of PQ were also formed on incubation with hamster liver fraction used in these studies. 4. The microsomal metabolism of PQ was decreased in presence of an NADPH-generating system, but not by SKF-525A or glutathione (GSH) indicating that the oxidative reactions were probably not due to the cytochrome P-450 system or free radical mechanisms.

Glucose effect on drug action, metabolism, and pharmacokinetic parameters in mice.

The glucose effect on hepatic drug metabolism (decreased) of barbiturates was maximum after 2 days of increased glucose intake as indicated by increased barbiturate sleep time in mice. However, this effect was not observed after 5 days of glucose treatment, and barbiturate sleep time was similar to the control after 6 days of treatment. Serum glucose and liver glycogen were, in general, not significantly different from control, even after chronic glucose intake, indicating that neither hypoglycemia nor alteration of liver glycogen levels were required for the glucose effect on drug action. However, in contrast to the decreased metabolism of barbiturate, there was increased metabolism of glucose in the glucose-treated animals. Brain levels of barbiturate in 48 hours glucose-treated mice were higher and declined at approximately half the rate of controls (Ke(G) 0.009 vs Ke(C) 0.015). A similar trend in barbiturate blood concentration indicated decreased metabolism of the barbiturate and/or decreased clearance of drug and metabolites. The glucose treatment altered the pentobarbital dose response curve, but there appeared to be no alteration of the sensitivity to insulin; exogenase insulin still produced significant hypoglycemia and prolonged barbiturate S.T. after 7 days of glucose treatment. Other factors may be involved in the glucose effect; increased permeability of the brain to barbiturate, decreased permeability to outflow so that brain concentrations remain higher for a long period of time.

Synthesis of certain hydroxy analogues of the antimalarial drug primaquine and their in vitro methemoglobin-producing and glutathione-depleting activity in human erythrocytes.

A number of hydroxy analogues of the antimalarial drug primaquine [8-[(4-amino-1-methylbutyl)amino]-6-methoxyquinoline] were synthesized and characterized by 1H NMR and mass spectra. Several of the compounds were found to be active in forming methemoglobin in human erythrocytes, particularly in those from glucose-6-phosphate dehydrogenase (G6PD) deficient subjects. Decreased levels of glutathione (GSH) in G6PD-deficient erythrocytes were also found with compounds that were active methemoglobin formers.

In vitro metabolism of the antimalarial agent primaquine by mouse liver enzymes and identification of a methemoglobin-forming metabolite.

From a mouse liver microsomal system, we have isolated and identified a methemoglobin-forming metabolite of primaquine (PQ). Evidence has been found for both O-dealkylation and hydroxylation of PQ to form a metabolite, 5,6-dihydroxy-8-(4-amino-1-methylbutylamino)quinoline, which is highly active in forming methemoglobin in both normal and glucose-6-phosphate dehydrogenase-deficient erythrocytes. It also actively decreases glutathione levels in glucose-6-phosphate dehydrogenase-deficient erythrocytes. The inhibitor SKF 525-A prevented metabolite formation while iproniazid and carbon monoxide did not inhibit metabolism completely but may have resulted in formation of a different unidentified metabolite. Mass spectrometry, HPLC, NMR, and other more indirect methods were used to help identify the metabolite. It was identified indirectly via a blue compound which results from extracting the actual metabolite from the incubation mixture with organic solvents under alkaline conditions in the presence of light. The blue compound was identified as a quinonimine in which the 8-amino side chain of PQ cyclizes to produce a third ring system.

Effects of orally administered activated charcoal on intestinal gas.

The effectiveness of activated charcoal in treating intestinal gas, following a gas producing meal, was compared with a placebo. Both the number of flatus events and breath hydrogen levels were measured. These experiments showed that orally administered activated charcoal was effective in preventing the large increase in the number of flatus events and increased breath hydrogen concentrations that normally occur following a gas-producing meal.

Metabolism of 8-aminoquinoline antimalarial agents.

Some of the most effective antimalarial agents are derivatives of 8-aminoquinoline. The metabolic products of many of these compounds appear to be toxic to the erythrocytes of certain human subjects, especially those deficient in glucose-6-phosphate dehydrogenase. Although a number of studies have been conducted over many years, the metabolism of most of these compounds has not been determined. These studies are reviewed.Adult dogs dosed with tritium-labelled primaquine were observed to excrete approximately 16% of the injected radioactivity in the urine within 8 hours. Organic extracts of the urine were fractionated by thin-layer chromatography and the metabolic pattern obtained. Some primaquine was excreted along with at least five metabolites including 5-hydroxy-6-methoxy-8-(4-amino-1-methylbutylamino)quinoline (5HPQ) and a small amount of 6-hydroxy-8-(4-amino-1-methylbutylamino)quinoline (6HPQ). The 5HPQ could form a quinoneimine-type compound which may be a methaemoglobin-forming compound. This and other metabolites isolated from urine were found to be active methaemoglobin formers in in vitro studies. In vitro metabolism of primaquine by mouse liver enzymes also produced compounds capable of methaemoglobin formation. One of these had a blue colour when exposed to alkaline conditions, air, and light, and mass spectral data and nuclear magnetic resonance analysis indicated a structure similar to a 5,6-dihydroxy derivative of primaquine. However, the chemical structure of the metabolite was not identified in these studies.

Excretion and disposition of [14C]carbaryl in pregnant, non-pregnant and foetal tissues of the rat after acute administration.

1. Non-pregnant or pregnant Sprague-Dawley rats on the 18th and 19th days of gestation were injected i.p. with a tracter dose (2.8 microCi/kg) of either [ring-14C]carbaryl or [carbonyl-14C]carbaryl. Distribution of total 14C was examined in foetal, maternal and non-pregnant rat tissues. Pregnancy alters the disposition and excretion of carbaryl. 2. Carbaryl crossed the placenta and was rapidly distributed in all foetal tissues. Highest concentrations were seen in foetal kidney. At 8 h after injection, foetal brain, heart and lung all contained more 14C, on a weight basis, than their maternal organ counterparts. Elimination from the whole foetus was biphasic, and after 8 h approx. 3% of the dose was still present in the whole foetus. 3. Significantly more 14CO2 was exhaled by the pregnant rat during 8 h than by non-pregnant. Urinary excretion of 14C after dosage with [ring-14C]carbaryl was significantly less in pregnant than in non-pregnant rats. 4. Kinetically, the tissue distribution of 14C from carbaryl or metabolites was biphasic in pregnant and non-pregnant animals. [14C]carbaryl concn. declined rapidly for 1 to 2 h. After 2 h the 14C levels from animals dosed with [ring-14C]carbaryl declined more slowly. 5. The pattern of 14C distribution was more complicated after injection of [carbonyl-14C]carbaryl. The 14C activity increased in the animal tissues after 2 h, in contrast to animals dosed with [ring-14C]carbaryl. Non-pregnant animals treated with [carbonyl-14C]carbaryl did not show a similar pattern of distribution. Carbamylated tissue proteins may, after time, release some bound carbonyl-14C label, causing the increase in 14C activity seen between 2 and 8 h. However, this does not necessarily imply uptake of the intact carbamate.

Influence of glucose (in vivo or in vitro) on duration of narcotic analgesics and on the kinetics of drug metabolism.

The duration of analgesia of the narcotics, methadone, morphine and codeine was prolonged by glucose treatment. This prolongation was associated with a decrease in in vitro metabolism of the narcotics. Chlorpromazine metabolism was not significantly inhibited by glucose treatment, indicating that glucose exerts some selectivity in the extent to which it inhibits various oxidative metabolic pathways. Michaelis-Menten type kinetics showed a mixed type of competitive and noncompetitive inhibition of methadone metabolism upon oral administration of glucose or in vitro addition of glucose to an enzyme system prepared from mouse liver. Other factors may be involved, such as the possibility that glucose might increase the permeability of the brain to barbiturate. However, in glucose-treated mice decreased metabolism of the barbiturate and narcotics seemed to be the major factor in the prolongation of their duration of action.