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We previously reported that granulocytes are able to produce superoxide (O(2) (-)), a highly reactive compound formed by the one-electron reduction of oxygen. The demonstration of O(2) (-) production was based on the observation that the reduction of extra-cellular cytochrome c by granulocytes was greatly diminished by superoxide dismutase, an enzyme catalyzing the conversion of O(2) (-) to hydrogen peroxide and oxygen. In the present report, studies concerning the effect of bacteria and serum on O(2) (-)-dependent cytochrome c reduction by granulocytes are described.In the absence of bacteria, the O(2) (-)-dependent reduction of extracellular cytochrome c by granulocytes under optimal assay conditions amounted to 9.2+/-2.8 SD nmol/3 x 10(6) cells/20 min. When bacteria (100 organisms/cell) were present, the O(2) (-)-dependent cytochrome c reduction under otherwise similar conditions increased by a factor of nearly four (34.5+/-9.4). There was no effect of albumin or catalase on cytochrome c reduction, and boiled dismutase had only a small effect. Omission of granulocytes or substitution of live cells by cells by cells killed by heat abolished O(2) (-)-dependent cytochrome c reduction. Bacteria killed by autoclaving were almost as effective as live bacteria in stimulating granulocyte O(2) (-) production. Measurements of particle uptake and O(2) uptake by granulocytes indicated that superoxide dismutase did not affect granulocyte metabolism nonspecifically, supporting the conclusion that the diminution of cytochrome c reduction in the presence of dismutase was due to the destruction of O(2) (-) by this enzyme. Stimulation of O(2) (-) production by bacteria was strongly dependent on the presence of serum in the incubation mixture. Serum heated to 56 degrees C for 45 min was as effective as unheated serum in stimulating O(2) (-) production in the presence of bacteria, but boiled serum had no effect. Other experiments suggested that incubation of bacteria with serum resulted in the release of a nonparticulate heat-labile substance capable of stimulating O(2) (-) production in the absence of bacteria. Certain characteristics of the O(2) (-)-dependent cytochrome c reduction by granulocytes were studied, including the dependence of this process on granulocyte, cytochrome c, and bacterial concentrations. In addition, O(2) (-)-dependent cytochrome c reduction was followed as a function of time. A constant rate was found with resting granulocytes. With bacteria the time course was more complex. A well-defined lag was followed by a fairly brief period of extremely vigorous cytochrome c reduction. During this period, the maximum rate of cytochrome c reduction exceeded the rate observed in the absence of bacteria by a factor of 12. The rate then decreased until by 40 min, it had slowed to the rate observed in the absence of bacteria. From the above results, it was concluded that the exposure of the granulocyte to bacteria plus serum initiates a process in which a defined quantity of O(2) (-) is formed in a rapid burst lasting 20-30 min. It is conceivable that the O(2) (-) generated by this process may be involved in the killing of bacteria by the granulocytes.

Superoxide-dependent cytochrome c reduction was used to measure Superoxide production by granulocytes from two patients with chronic granulomatous disease, from children of similar age wit...

Ethanolamine ammonia-lyase catalyzed the adenosylcobalamin-dependent deamination of 2-aminopropanol as well as that of ethanolamine. When the enzyme-coenzyme complex was frozen in liquid nitrogen while it was acting on 2-aminopropanol, a large, well defined EPR spectrum was demonstrated. The spectrum consisted of two components: a broad signal at g = 2.34, and an asymmetrical doublet with g values of 2.04 and 1.99. The over-all signal corresponded to about 1.3 spins per active site, the electron density being approximately equally divided between the species producing the g = 2.34 component and the one represented by the asymmetrical doublet. The configuration of the signal was the same regardless of whether the incubation was conducted aerobically or anaerobically. Both components of the signal were saturated by increasing power, the doublet being the more easily saturated component. EPR spectroscopy at two different microwave frequencies (X band and K band) showed that the doublet represented a single paramagnetic species whose signal was split by an interaction with another paramagnetic species in its vicinity. By its location, configuration, and the characteristics of the superimposed hyperfine structure, the g = 2.34 signal was assigned to the unpaired electron of cob(II)alamin. Experiments with isotopically labeled propanolamine permitted the doublet signal to be assigned to the 2-aminopropanol-1-yl radical. The major splitting of the doublet signal was attributed to a dipole-dipole interaction between the 2-amino-propanol-1-yl radical and the unpaired electron on cob(II)-alamin; assuming this explanation to be correct, the distance between the two spins was calculated to be ∼6 A. Rapid mixing, quick freeze experiments showed that the appearance of the paramagnetic species was a first order process with a rate constant of 7 s-1. Since the turnover number for propanolamine is 1 to 2 s-1, the process by which the paramagnetic species are generated is kinetically competent with respect to over-all catalysis. Once formed, the species disappeared with a half-time of about 1 to 2 min. Their disappearance is probably associated with the irreversible destruction of adenosylcobalamin by a side reaction not directly relevant to the catalytic mechanism. These results have been interpreted in terms of a mechanism whereby the early hydrogen transfer steps in the adenosylcobalamin-dependent rearrangement of propanolamine are as follows: [see PDF for equation]

Ethanolamine ammonia-lyase, an enzyme catalyzing the adenosylcobalamin-dependent deamination of ethanolamine, also catalyzes the conversion of l-2-aminopropanol to propionaldehyde and ammonia. In this reaction, tritium is transferred from enzyme·[5'-3H]adenosylcobalamin to the C-1 position of l-2-aminopropanol, as well as to the α carbon of the product aldehyde. The labeling pattern is consistent with the mechanism of hydrogen transfer deduced with other substrates. Tritium transfer also occurs between enzyme·[5'-3H]adenosylcobalamin and propionaldehyde in the presence of NH4+. Unlike the deamination of ethanolamine, the conversion of 2-aminopropanol to propionaldehyde and NH4+ is reversible, since tritiated 2-aminopropanol was isolated from reaction mixtures originally containing only propionaldehyde, ammonia, and enzyme·[5'-3H]adenosyl cobalamin. The partitioning of tritium derived from enzyme·[5'-3H]-adenosylcobalamin between l-2-aminopropanol and propionaldehyde was determined in reactions begun with l-2-aminopropanol as well as in reactions begun with propionaldehyde and ammonia. In the first case, the ratio [3H]propanolamine to [3H]propionaldehyde is 1.8:1; in the second case, the ratio is 0.3:1. This difference in the partitioning of tritium from the tritiated enzyme complex is consistent with the notion that there are at least two intermediates in the catalytic process, each exchanging tritium with coenzyme, which interconvert slowly with respect to the rate of tritium exchange.

MASSIVE liver necrosis is a rare entity, having an autopsy incidence of 0.15 per cent.1 It has been attributed to such conditions as viral hepatitis,2 drug hepatitis,1 carbon tetrachloride and chloroform poisoning3 and shock3 , 4 or associated administration of pressors.5 Interest in this disease has been rekindled by the recent spate of reports relating massive liver necrosis to halothane anesthesia.6 7 8 9 10 11 12 13 14 15 16 17 18 19 These reports have led to a comprehensive investigation, the National Halothane Study, which is currently being carried out under the auspices of the National Academy of Sciences–National Research Council to establish whether halothane is a cause of massive liver necrosis or . . .

Thyroid hormone administered in vivo increased carrier-mediated (atractyloside-sensitive) ADP uptake by rat liver mitochondria. 3 Days after a single large dose of triiodothyronine (20 μg/100 g of body weight), mitochondrial uptake of ADP measured at 6° was 2.35 ± 0.17 nmol/min per mg of protein, compared with an uptake of 1.81 ± 0.19 nmol/min per mg of protein in mitochondria from untreated rats ( P < 0.025). Cyanide (1.33 mM) had no effect on ADP uptake by mitochondria from either untreated or triiodothyronine-treated animals. Uptake of ADP by mitochondria from thyroidectomized rats treated with thyroxine for 7 days was 2.89 ± 0.40 nmol/min per mg in mitochondria from thyrotoxic rats (20 μg of thyroxine per 100 g per day) and 1.98 ± 0.22 nmol/min per mg in mitochondria from euthyroid rats (2 μg of thyroxine per 100 g per day) ( P < 0.025). Mitochondria from both untreated and thyroid hormone-treated rats displayed a highly significant linear correlation between ADP uptake and ADP-dependent (i.e., state 3 minus state 4) oxygen consumption. There was, however, no difference in respiratory control ratios between mitochondria from euthyroid and thyrotoxic animals. Administration of dinitrophenol (2 mg/100 g) also stimulated carrier-mediated ADP uptake, but respiratory control of mitochondria from dinitrophenol-treated animals was virtually abolished. Triiodothyronine in vitro , at concentrations of 100 and 0.1 nM, appeared to inhibit rather than stimulate the uptake of mitochondrial ADP. The relationship between these observations and the clinical manifestations of thyrotoxicosis is discussed from the point of view of the possible effects of increased mitochondrial ADP transport on oxidative phosphorylation and adenosyl nucleotide metabolism.

As a highly reactive substance produced in biological systems by the one-electron reduction of oxygen, superoxide (O(2) (-)) seemed a likely candidate as a bactericidal agent in leukocytes. The reduction of cytochrome c, a process in which O(2) (-) may serve as an electron donor, was found to occur when the cytochrome was incubated with leukocytes. O(2) (-) was identified as the agent responsible for the leukocyte-mediated reduction of cytochrome c by the demonstration that the reaction was abolished by superoxide dismutase, an enzyme that destroys O(2) (-), but not by boiled dismutase, albumin, or catalase. Leukocyte O(2) (-) production doubled in the presence of latex particles. The average rate of formation of O(2) (-) in the presence of these particles was 1.03 nmol/10(7) cells per 15 min. This rate, however, is only a lower limit of the true rate of O(2) (-) production, since any O(2) (-) which reacted with constituents other than cytochrome c would have gone undetected. Thus. O(2) (-) is made by leukocytes under circumstances which suggest that it may be involved in bacterial killing.

Methylmalonyl-CoA mutase-dependent cleavage of coenzyme B12 took place in the presence of succinyl-CoA, one of the substrates of the enzyme, and in the presence of malonyl-CoA, a competitive inhibitor (Ki 0.12 mm). Other competitive inhibitors of the reaction, including acetyl-CoA, propionyl-CoA, glutaryl-CoA, and CoA itself, did not replace malonyl-CoA in this cleavage reaction. With both succinyl-CoA and malonyl-CoA, the cleavage of coenzyme was progressive with time, without evidence for an initial burst. The cleavage reaction did not depend on oxygen, since it occurred under anaerobic as well as aerobic conditions. The adenosyl derivative formed by the cleavage of 5'-deoxyadenosylcobalamin in the presence of each of the CoA esters was identified chromatographically as 5'-deoxyadenosine. Malonyl-CoA did not appear to be chemically altered during the course of the reaction; experimental difficulties precluded a similar investigation of the fate of succinyl-CoA. These experiments have shown that under certain circumstances, methylmalonyl-CoA mutase is able to cleave the carbon-cobalt bond of coenzyme B12 with the production of 5'-deoxyadenosine. The findings are consistent with the view that the currently proposed mechanism of coenzyme B12-dependent rearrangements, which involves cleavage of the carbon-cobalt bond followed by abstraction of a substrate hydrogen atom to form 5'-deoxyadenosine, may be generally valid for all reactions of this class.

Vitamin K-dependent Factor VII synthesis in rat-liver slices was not affected by puromycin although protein synthesis was inhibited by 98 %. The results indicate that vitamin K is required for the formation of Factor VII from a polypeptide precursor.

The previous observation (Babior, B. M., J. Biol. Chem., 244, 449 (1969)) that the tritium isotope effect for the coenzyme B12-dependent deamination of ethanolamine was smaller than the deuterium isotope effect prompted a reexamination of the various isotope effects observed in this reaction. Measurements of the rates of deamination of [1-D]- and [1,1-D2]ethanolamine showed that no secondary isotope effect was detectable within the limits of experimental error. Activation parameters for the deamination of unlabeled ethanolamine and [1,1-D2]ethanolamine at 23° were as follows: for the unlabeled substrate, Ea = 3.9 kcal per mole and S = -9.7 e.u.; for the deuterated substrate, Ea = 5.5 kcal per mole and S = -9.4 e.u. The similarity of these values implies that the rate-limiting step does not change on passing from the unlabeled to the deuterium-labeled substrate. Measurement of the tritium isotope effect for each of the two individual hydrogen transfer steps of the reaction gave the following results: for the first step (transfer of hydrogen from substrate to coenzyme), kt/kh = 4.7; for the second step (transfer of hydrogen from coenzyme to product), kt/kh = 160 and kt/kd = 22. The over-all deuterium isotope effect was found to be 7.4, confirming previous results. From these and other observations the following conclusions were drawn: (a) the rate-limiting step is the transfer of hydrogen from coenzyme to product; (b) an irreversible step occurs between the first hydrogen transfer step and the second; and (c) exchange between free and enzyme-bound coenzyme B12 during the course of the reaction is slow.

Patients with villous adenoma of the colon may sustain substantial fluid and electrolyte losses resulting from a voluminous mucous diarrhea composed of the secretion of the tumor. Such a patient is described, and fluid and electrolyte disturbances in this case and in others reported in the literature are reviewed. The symptoms are rapidly progressing weakness and gastrointestinal complaints. The patient may present in collapse, with confusion or coma. On rectal examination, the tumor may be felt or the mucous diarrhea may be observed. These patients display a combination of dehydration, hyponatremia and hypokalemia. Azotemia may be severe even in the absence of renal disease. Early in the illness they are either alkalotic or in normal acid-base balance; as fluid losses become more severe, acidosis supervenes. The present patient was treated with large amounts of fluids and potassium followed by resection of the tumor.

When a mixture of coenzyme B12 (5'-deoxyadenosylcobalamin) and ethanolamine ammonia-lyase is denatured with hot propanol after a 30-sec incubation, 1.5% of the coenzyme is released in a form in which the carbon-cobalt bond is dissociated. The fraction of dissociated coenzyme rises to 3.6% when substrate or product is present in the incubation. In the absence of enzyme, the coenzyme remains intact. Reversibility of dissociation is suggested by the observation that dissociation of the coenzyme is decreased by two-thirds when the enzyme is denatured with tetrahydrofuran. If the process were irreversible, the same amount of dissociated cofactor would be released regardless of the method of denaturation, while with a reversible reaction the amount of dissociated cofactor released on denaturation could vary depending on the relative susceptibility to denaturation of the (enzyme-intact coenzyme) and (enzyme-dissociated coenzyme) complexes. The adenosyl fragment derived from coenzyme dissociated in the presence of substrate is chromatographically identical with 5'-deoxyadenosine. In the absence of substrate, a number of products are observed, including adenosine 5'-aldehyde and a compound which behaves like 5',8-cycloadenosine. These results suggest that (a) rupture of the carbon-cobalt bond is an integral part of the reaction mechanism; (b) carbon-cobalt bond rupture and hydrogen transfer are homolytic processes; and (c) enzyme-bound 5'-deoxyadenosine is an intermediate in the reaction.

An enzyme which catalyzes the aromatization of cyclohexanecarboxyl coenzyme A has been isolated from whole liver as well as from liver mitochondria. Incubation of the soluble, partially purified enzyme with cyclohexanecarboxyl-CoA produces cyclohexene-1-carboxyl-CoA, benzoyl-CoA, and a polar compound which has not been further characterized. All three isomers of cyclohexenecarboxyl-CoA are converted to benzoyl-CoA by the liver enzyme, but evidence is presented indicating that only cyclohexene-1-carboxyl-CoA lies on the path from cyclohexanecarboxyl-CoA to benzoyl-CoA. Aromatization takes place either in air or anaerobically in the presence of artificial electron acceptors. The enzyme system is inactivated by low pH and activity is restored by the addition of flavin adenine dinucleotide.

Summary Carrier-mediated ADP uptake by liver mitochondria from myxedematous thyroidectomized rats was compared with that of mitochondria from thyroidectomized animals rendered euthyroid or hyperthyroid by exogenous thyroxine (T 4 ). Uptake by mitochondria from thyroidectomized rats treated with 0, 2, 5, and 20 μg T 4 /100 g body weight/day for 6 days was 1.24 ± 0.16, 2.26 ± 0.14, 3.82 ± 0.09, and 8.20 ± 0.58 nmoles ADP/min/mg protein respectively (mean ± SE; p 4 /100 g/d, a physiological replacement dose. These findings provide further evidence that the calorigenic action of thyroid hormone may be mediated by an action on the mitochondrial ADP-ATP carrier.

A study was carried out on the effect of a large number of B12 analogues on the coenzyme B12-dependent enzyme ethanolamine deaminase. The analogues tested included MeB12, EtB12, AB12, RB12, BB12, IB12, DB12, UB12, CNB12, and OHB12. DB12 was found to be capable of replacing the coenzyme. In contrast, all the rest of the analogues were inhibitors. With all the inhibitors except IB12, the inhibition was progressive, with the reaction velocity decaying exponentially with time to a final, constant value. The extent of inhibition by IB12 was independent of time; kinetics showed that this inhibition was competitive with respect to the coenzyme. Inhibition by MeB12 was also found to be competitive with DMBC, when its effect of the initial reaction rate was considered. Studies of the time-dependent increase in the extent of inhibition observed with MeB12 showed that the rate constant for this process (the deactivation constant) followed saturation kinetics with respect to the concentration of inhibitor. A similar dependence of the deactivation constant on the inhibitor concentration was observed with RB12. The time-dependent decrease in the reaction rate caused by this derivative was found to be irreversible. Kinetic studies performed at the end of the decay period showed that RB12 behaved as a competitive inhibitor with respect to the coenzyme, but that its Ki at the end of this period was much lower than the Ki estimated for the beginning of the reaction. Furthermore, the Vm for ethanolamine deaminase at the end of the decay period was only one-sixth of the Vm of the uninhibited enzyme. Light scattering studies and studies of the effect of enzyme concentration on the characteristics of the decay indicated that the inhibition is not associated with polymerization or depolymerization of the enzyme. Experiments with tritiated MeB12 showed that the progressive inhibition induced by this derivative is not accompanied by cleavage of the carbon-cobalt bond. Titration experiments with RB12 have shown that the progressive inhibition is fully expressed at an inhibitor to enzyme ratio of 2:1. These results can be explained by a model in which the presence of inhibitor induces a change in the enzyme from the form isolated in the purification to a form with a lower Vm and a much greater affinity for the inhibitor. The abbreviations used are: MeB12, methylcobalamin; EtB12, β-hydroxyethylcobalamin; AB12, β-(2-tetrahydropyryloxy)ethylcobalamin; RB12, 1-O-methyl-5-deoxyribosylcobalamin; BB12, δ-(9-adenyl)butylcobalamin; IB12, 5'-deoxyinosylcobalamin; DB12, 2',5'-dideoxyadenosylcobalamin; UB12, 5'-deoxyuridylcobalamin; CNB12, cyanocobalamin; OHB12, hydroxocobalamin; DMBC, 5'-deoxyadenosylcobalamin.

Ethanolamine ammonia lyase catalyzes the coenzyme B12-dependent conversion of ethanolamine to acetaldehyde and NH3. When the enzyme-coenzyme complex is frozen in liquid nitrogen during the act of catalysis, an electron spin resonance signal equivalent to about 0.04 spin per active site can be detected. By saturation experiments, the signal consists of two components, a broad derivative signal composed of two peaks (g = 2.34 and g = 2.08) assigned to the cobalt of B12r, and a narrow signal (g = 2.007) assigned to a free radical. With [1,1-D2]ethanolamine as substrate, the shape and position of the signal was unchanged, but its size was increased. In the absence of substrate, a signal was still observed, but the concentration of unpaired electrons was reduced. The broad metal signal was unchanged except for a decrease in size. The radical signal was narrower (10 gauss, compared with 18 gauss in the presence of substrate) and was shifted (g = 2.003) with respect to the substrate-dependent signal, suggesting that the radical in the absence of substrate may have been different from the substrate-dependent radical. These observations support the proposal that the mechanism of action of this coenzyme B12-dependent enzyme involves homolysis of the carbon-cobalt bond of the coenzyme.

The uptake of intrinsic factor-cobalamin complex by guinea pig ileal brush borders is inhibited by particle-free sonicates of distal but not proximal brush border preparations. This finding suggests that sonication releases intrinsic factor receptor from ileal brush borders in a form which is not sedimented by centrifugation for 1 h at 105 000× g .

To determine whether the molecular configuration of vitamin B(12) influences the attachment of intrinsic factor-vitamin B(12) complex to ileal microvillous membrane receptor sites, we have examined the kinetics of uptake of intrinsic factor-bound cyanocobalamin by brush borders and microvillous membranes isolated from guinea pig ileum, and have compared this uptake with that of intrinsic factor alone and with that of intrinsic factor complexed with various analogs of cyanocobalamin. We first studied the kinetics of binding of cyanocobalamin and other cobamides to human gastric intrinsic factor. The binding of cyanocobalamin showed saturation kinetics and, at relatively high concentrations of cyanocobalamin, a Scatchard plot of binding was linear. The dissociation constant for the intrinsic factor-cyanocobalamin complex was 0.066 nM. When the binding of various vitamin B(12) analogs to intrinsic factor was determined by competition experiments, the analogs could be separated into two categories: those with affinities similar to that of cyanocobalamin and those with affinities much lower than that of cyanocobalamin. The affinity of cyanocobalamin for intrinsic factor was not altered by various substitutions at the -CN position, while removal of a single amido group on the corrin ring of substitution of the dimethylbenzimidazole base greatly reduced affinity. Removal of the base totally abolished binding. These findings, confirming those reported by others, are consistent with the concept that the cyanocobalamin molecule fits into a "pocket" in the intrinsic factor molecule, with the nucleotide base facing inward and the -CN side of the planar corrin ring facing outward. We then investigated the attachment of intrinsic factor-bound cyanocobalamin to ileal receptor. Attachment to microvillous membranes showed saturation kinetics with a dissociation constant of 0.25 nM. Attachment was rapid and was 70% complete within 5 min; the second-order rate constant for attachment was 1.3 x 10(6) M(-1) s(-1). The half-time for dissociation of intrinsic factor-bound cyanocobalamin from the ileal receptor was approximately 35 min. Free intrinsic factor inhibited the attachment of intrinsic factor-bound cyanocobalamin, but the rate of attachment of free intrinsic factor was slower than that of intrinsic factor bound to cyanocobalamin. When intrinsic factor was complexed with various analogs of cyanocobalamin, the affinities of these complexes for ileal microvillous membranes were similar to that of intrinsic factor-bound cyanocobalamin. These findings suggest that the molecular configuration of vitamin B(12) is not a major determinant in the interaction between intrinsic factor-bound vitamin B(12) and its ileal receptor site.

A study was conducted on the effect of warfarin on factor VII formation by a cell-free system from rat liver. Administration of warfarin to the rat from which the liver is obtained results in a marked decrease in factor VII formation by the liver homogenate. The rate of fall of factor VII formation was considerably more rapid than the rate of disappearance of factor VII from warfarin-treated rats. Restoration of factor VII formation upon the administration of vitamin K to warfarinized rats was also investigated. Factor VII was not made by homogenates prepared from livers removed from the rat immediately after administration of vitamin K, although slices prepared from such livers were able to make the clotting factor. However, homogenates prepared from livers taken a half hour or longer after giving vitamin K were able to make factor VII. These results suggest that vitamin K must be modified before it can participate in clotting factor formation, and that under the condition of these experiments this modification can take place in slices but not in homogenates.

EPR spectroscopy of a mixture of ethanolamine deaminase, 5′-deoxy-adenosylcobalamin and deuterated ethanolamine frozen during the act of catalysis revealed the appearance of a signal at g = 2.09. This signal disappeared when the reaction was permitted to continue until the substrate was exhausted, but reappeared upon subsequent addition of substrate. No signal was observed when enzyme or coenzyme was omitted from the reaction mixture, but small signals did appear when substrate was omitted. There were significant differences between these latter signals and the signal obtained from a sample containing all three components. This evidence is consistent with the hypothesis that species with unpaired electrons are involved in the mechanism of action of ethanolamine deaminase.

The kallikrein-bradykinin system was studied in patients with chronic alcoholic liver disease. Kallikreinogen levels were depressed in patients with cirrhosis when compared with levels in ...

No one knows at present whether halothane is toxic to the liver. For several years a study¶ to determine the answer to this question has been going on. It is now completed and a summary report has ...

Upon deamination of chirally labeled [2-2H, 3H]ethanolamine by the adenosylcobalamin-dependent ethanolamine ammonia-lyase (EC 4.3.1.7) from Clostridium sp., chirality of the labeled carbon atom is lost. This result is consistent with a mechanism involving the 1-amino-1-hydroxyethane2-yl radical as a catalytic intermediate.

The kallikrein-kinin system was studied in 10 postgastrectomy patients, 5 with the dumping syndrome and 5 patients without symptoms. After drinking 220 ml of 45% glucose, none of the asymp...

A patient had massive ascites caused by chronic pancreatitis. Formation of the ascites was accompanied by little pain. The fluid was hemorrhagic and had a high protein content, but could be distinguished from effusions of other etiologies by a very high amylase content. The ascites subsided spontaneously after three weeks.