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The phosphorylation of ribosomal proteins L14 and S3 in Krebs II ascites cells.

Krebs II ascites cells incubated in Earle's saline (lacking glucose and amino acids) contain ribosomes with proteins S6 and Lgamma phosphorylated, as do ascites cells grown in the peritonea of mice or hamster fibroblasts grown in Eagle's medium. When ascites cells were incubated in Eagle's medium (containing glucose and amino acids) there was extensive glycolysis, producing very acidic conditions, and ribosomal proteins S3 and L14 became phosphorylated whereas Lgamma became dephosphorylated. This altered pattern of phosphorylation could not be produced merely by incubating ascites cells in Earle's saline at a decreased pH, but a rather similar pattern was produced when Earle's saline was supplemented with amino acids (but with glucose still omitted). These results suggest that depriving ascites cells of glucose may induce the synthesis of a protein (or proteins), necessary for alteration of the pattern of phosphorylation of the ribosomal proteins.

Cells, Cultured↗

Acid stable protease inhibitor in ascites of ovarian carcinoma.

In patients with ovarian tumors, a novel protease inhibitor which is very stable in acid (AS-PI, acid stable protease inhibitor) was identified in the ascites and tumor fluid as well as in the urine and plasma. The highest AS-PI activity was observed in the tumor fluid of ovarian carcinomas (10.7 +/- 2.3 U/ml), followed by the ascites of ovarian carcinomas (8.2 +/- 4.2 U/ml). There was a significant difference in activity of the tumor fluid and ascites between malignant and benign tumors (p less than 0.005, p less than 0.05, respectively). The same antigenicity of AS-PI fractionated from ascites of ovarian carcinomas to urinary trypsin inhibitor was identified by double immunodiffusion and neutralization techniques. It migrated in the serum albumin fraction on immunoelectrophoresis. By gel filtration, the AS-PI in the ascites of ovarian carcinomas showed a molecular weight of 70000-80000. Two active components with molecular weights of 61300 +/- 2100 and 73300 +/- 2500 were detected by SDS polyacrylamide gel electrophoresis.

Chromatography, Gel↗

Role of altered systemic hemodynamics in the blunted renal response to atrial natriuretic peptide in rats with cirrhosis and ascites.

The natriuretic effect of pharmacological doses of atrial natriuretic peptide (ANP) is markedly reduced in cirrhosis with ascites. The current study, which includes two protocols, was carried out to investigate whether this phenomenon is related to the altered systemic hemodynamics present in cirrhosis. In protocol A, the administration of ANP (2.5 micrograms.kg-1 as a bolus followed by a constant infusion of 0.1 microgram.kg-1.min-1) to 10 rats with carbon tetrachloride-induced cirrhosis and ascites produced a significantly lower increase in diuresis (13.4 +/- 1.3 microliters/min) and natriuresis (2.3 +/- 0.3 mu Equiv/min) than in 10 control rats (56.3 +/- 1.4 microliters/min and 8.7 +/- 0.5 mu Equiv/min, respectively), indicating a renal resistance to the effect of ANP in this experimental model of cirrhosis. The reduction of arterial pressure induced by ANP was similar in both groups. However, since baseline mean arterial pressure was significantly lower in cirrhotic rats, the degree of hypotension during ANP infusion was also greater in this group of animals (82 +/- 3 vs. 109 +/- 2 mmHg). The aim of protocol B was to assess whether normalization of arterial pressure in cirrhotic rats increases the renal response to ANP. This protocol includes two groups of 10 rats with cirrhosis and ascites infused with a glucose solution containing norepinephrine (CT-NE rats) or angiotensin II (CT-AII rats) at doses to normalize arterial pressure and an additional control group of 10 cirrhotic rats with ascites receiving only glucose solution (CT rats). Angiotensin II, but not norepinephrine or glucose solution administration, was associated with a significant increase in urine volume and sodium excretion. During ANP infusion, CT rats showed a blunted diuretic and natriuretic response. In contrast, the ANP-induced increase in urine volume and sodium excretion observed in CT-NE (53.6 +/- 10.4 microliters/min and 9.3 +/- 2.2 mu Equiv/min) and CT-AII rats (98.3 +/- 11.6 microliters/min and 15.5 +/- 2.9 mu Equiv/m), was similar or even greater than that showed by the healthy rats of protocol A. The degree of hypotension during ANP administration was also similar (CT-NE, 104 +/- 2; CT-AII, 108 +/- 5 mmHg). These results suggest that the blunted response to pharmacological doses of ANP in cirrhosis with ascites is related to altered systemic hemodynamics of cirrhosis, which further deteriorates during the infusion of the peptide.

Angiotensin II↗

Management of cirrhotic ascites: physiological basis of diuretic action.

Ascites is a significant complication of cirrhosis that occurs in approximately 50% of patients. The mortality rate is high in patients with cirrhosis and ascites. Conventional interventions rest with dietary sodium restriction, diuretic use, large-volume paracentesis, peritoneovenous shunts and transjugular intrahepatic portosystemic shunts. The mainstay of therapy, however, is the judicious use of diuretics. This article reviews the physiological basis of diuretic use in patients with cirrhosis and ascites, as well as recent concepts on the pathogenesis of ascites formation. Through a better understanding of the pathophysiology of ascites formation and the mechanism of action of diuretics, improved extracellular fluid balance can be achieved in these patients.

Journal Article↗

Effect of Pfaffia paniculata (Brazilian ginseng) on the Ehrlich tumor in its ascitic form.

The roots of Pfaffia paniculata (Brazilian ginseng) have been indicated for the treatment of several diseases, among which the cancer. The purpose of this study was to investigate experimentally the possible antineoplastic effect of this root. Firstly, a toxicity study was performed in which the doses of 400 and 200 mg/Kg of the powdered root were administered by gavage for 10 days to BALB/cICB mice. The mice did not lose weight during the treatment. No increase in serum alanine-aminotransferase neither histopathological alteration (liver, kidney and spleen) was observed in mice treated with P. paniculata. The effect of this root on the ascitic Ehrlich tumor in BALB/cICB mice was then investigated. Male mice received, by gavage, once a day, 200 mg/Kg of the powdered root of P. paniculata or distilled water, as control, for 20 days. This protocol started 10 days before tumor inoculation with 5 x 10(6) cells i.p., and lasted until 10 days after. The ascitic tumor was evaluated by the quantification of the volume of the ascitic fluid, relative number of tumor cells and total number of tumor cells. A decrease in the total ascitic volume was observed in P. paniculata treated mice, that was followed by a numerical decrease in the total number of Ehrlich tumor cells. These results may indicate that P. paniculata anti-inflammatory effects were responsible by the decrease in the total ascitic fluid. In addition, the presence of tumor-cell inhibitory factors in P. paniculata roots is in agreement with other in vitro studies. The mechanisms of such tumor inhibition should be further investigated.

Alanine Transaminase↗

Spontaneous bacterial peritonitis in a patient with nephrogenic ascites during an episode of acute renal transplant rejection.

Spontaneous bacterial peritonitis (SBP) is a primary infection of asci tes without signs of perforation or penetration. It occurs most often in patients with liver cirrhosis but can also be diagnosed in patients with ascites from other causes. We report a kidney transplant recipient who developed nephrogenic ascites during an episode of acute rejection. The patient complained of fever, abdominal tenderness, and loose stools and showed all of the signs of peritonitis on physical examination. The patient's serum creatinine was elevated, and Duplex sonography of the graft was highly suggestive for acute rejection. Ascites puncture was performed. The ascitic fluid contained 4,000 leukocytes per microliter. No source of infection was detected, so the diagnosis of SBP was made. The patient was treated with ciprofloxacin intravenously and received low-dose steroid pulse therapy. The ascites culture grew Staphylococcus aureus that was highly sensitive to ciprofloxacin. The patient recovered rapidly. We could avoid laparotomy, which is associated with high mortality in patients suffering from SBP. No relapses of SBP occurred. Renal function has improved and remained stable.

Acute Disease↗

Relapsing ascites and uremia due to urinary bladder leakage.

A 62-year-old female was admitted for a new episode of ascites, complicated by uremia. She had been treated 6 years earlier for bladder cancer with partial cystectomy and postoperative radiotherapy. At the work-up for both previous episodes of ascites, 6 and 24 months earlier, no definite diagnosis was made, and each time the ascites disappeared spontaneously. Also, at the third episode, the uremia resolved. Finally, explorative laparoscopy demonstrated urinary leakage from the superior portion of the bladder. This feature of vanishing and relapsing ascites is explained by intermittent covering by overlying abdominal structures and subsequent spontaneous healing. The pseudo-renal failure is caused by reabsorption of urine through the peritoneum. Only nine cases of recurring ascites and eight cases of bladder perforation after radiotherapy have been described in the literature.

Journal Article↗

The usefulness of PCR amplification of the IS6110 insertion element of M. tuberculosis complex in ascitic fluid of patients with peritoneal tuberculosis.

Background: The diagnosis of tuberculous peritonitis (TP) may be difficult and elusive. The present study was designed to demonstrate the diagnostic usefulness of a nested polymerase chain reaction (PCR) assay, specific for the IS6110 insertion element of M. tuberculosis complex, in patients with ascites who were suspected of having TP in order to achieve a more timely diagnosis and treatment. Methods: Three HIV-negative patients suffering from fever and ascites were evaluated for suspected TP. Specimens were obtained from ascitic fluid, bone marrow, and peripheral blood and analyzed by both conventional methods and nested PCR for the presence of bacilli. Response to antituberculous treatment was considered as the final criterion for diagnosis of peritoneal tuberculosis. Results: All three patients had an excellent response to antituberculous therapy. Our PCR-based protocol detected M. tuberculosis complex DNA in the ascitic fluid of all patients, whereas conventional methods failed to establish the disease. Furthermore, in one patient, M. tuberculosis was also detected in both bone marrow and peripheral blood. Conclusions: PCR amplification of the IS6110 sequence of M. tuberculosis complex in ascitic fluid is a useful tool when peritoneal tuberculosis is suspected. However, its validity still needs to be established.

Journal Article↗

Purification of rat liver and mouse ascites DNA-dependent RNA polymerase I.

Three forms of RNA polymerase were assayed in nuclei and nucleoli isolated from rat liver and from Krebs II ascites cells. Assays of rat liver nuclei in the absence of exogenous DNA showed polymerase I accounted for 72% of the total activity, polymerase II for 17%, and polymerase III for 11%. The total activity in ascites nuclei was similar but the ratios of polymerase activities were different: polymerase I, 53%; polymerase II, 41%; and polymerase III, 6%. These values may reflect differences in the transcriptional activity of the nuclei. After isolation of nucleoli, both rat liver and ascites polymerase I accounted for 85% of enzyme activity. When exogenous calf-thymus DNA was added to nucleoli, there was a greater than 50% increase in activity suggesting that less than one-half of the polymerase I present was bound to endogenous template. Polymerase I was solubilized from either rat liver or ascites nucleoli by sonication at high ionic strength and subsequently purified by ion filtration, phosphocellulose, sucrose gradient centrifugation, and DNA-cellulose chromatography. The essentially homogenous ascites enzyme had a specific activity of 86 units/mg when assayed with native calf-thymus DNA and of 876 units/mg when assayed with poly(deoxycytidylic acid). Electrophoresis of the enzyme in sodium dodecyl sulfate indicated the presence of six subunits with molecular weights of 205 000, 125 000, 51 000, 44 000, 26 000 and 16 000. After the same purification procedure, the rat liver enzyme had a similar specific activity (98 units/mg) on native calf thymus and 362 units/mg on poly(deoxycytidylic acid).

Animals↗

Some inhibitory effects of (--)-emetine on growth of Ehrlich ascites carcinoma.

(-)-Emetine has little or no effect on O(2) consumption of Ehrlich ascites-cell suspensions or on the viability of transplanted Ehrlich ascites-tumour cells exposed to, or incubated with, the drug in vitro before inoculation into new-host mice. (-)-Emetine administered as a single injection to mice bearing Ehrlich ascites-tumour cells slows the growth rate of the tumour. The subcutaneous or intraperitoneal injection of the drug depresses protein and DNA synthesis of the tumour cells in vivo in a reversible manner. Mice bearing ascitic sarcoma 180 or Ehrlich ascites-tumour cells when given a course of treatment with (-)-emetine or (-)-O-methyltubulosine have a substantially lower tumour load than untreated controls and correspondingly longer survival times.

Animals↗

Polyamines and their biosynthetic enzymes in Ehrlich ascites-carcinoma cells. Modification of tumour polyamine pattern by diamines.

1. Ehrlich ascites-carcinoma cells contained relatively high concentrations of spermidine and spermine, but the putrescine content of the washed cells was less than 10% of that of higher polyamines. 2. Ascites-tumour cells likewise exhibited high activities of L-ornithine decarboxylase (EC 4.1.1.17), S-adenosyl-L-methionine decarboxylase (EC 4.1.1.50), spermidine synthase (EC 2.5.1.16) and spermine synthase. 3. During the first days after the inoculation, the polyamine pattern of the ascites cells was characterized by a high molar ratio of spermidine to spermine, which markedly decreased on aging of the cells. 4. Various diamines injected into mice bearing ascites cells rapidly and powerfully decreased ornithine decarboxylase activity in the carcinoma cells, apparently through a mechanism that was not a direct inhibition of the enzyme in vitro. Cadaverine (1,5-diaminopentane) and 1,6-diaminohexane were the most potent inhibitors of ornithine decarboxylase among the amines tested. 5. Chronic treatment of the mice with diamines resulted in a virtually complete disappearance of ornithine decarboxylase activity, and after 24h a significant decline in spermidine accumulation. 6. Cadaverine appeared to be an especially suitable compound for use as an inhibitor of the synthesis of higher polyamines, at least in Ehrlich ascites cells, since this diamine also acted as a competitive inhibitor for putrescine in the spermidine synthase reaction without being incorporated into the higher polyamines.

Adenosylmethionine Decarboxylase↗

Phospholipid metabolism in Ehrlich ascites tumor cells. I. Turnover rate of phosphatidylinositol.

1. Radioactive precursors, 32 PI, [1-14C]glycerol, and [1-14C]acetate, were individually injected into the peritoneal cavity of mice bearing Ehrlich ascites tumor, and the rates of incorporation into phospholipid fraction of Ehrlich ascites tumor cells were estimated. Although no distinct difference in specific activities was observed between phosphatidylinositol and other phospholipid classes as regards the incorporation of [1-14C]acetate of [1-14C]glycerol, a higher rate of incorporation of 32Pi into phosphatidylinositol was observed. The specific activity of phosphatidylinositol reached more than ten times that of phosphatidylcholine in the first hour. 2. The radioactivities incorporated into the phospholipids of Ehrlich ascites tumor cells and liver were estimated after simultaneous injection 32Pi and [2-3H]inositol. The incorporation of 32Pi into phosphatidylinositol of liver was similar in specific activity to those of other phospholipids. The ratio (3H/32Pi) of phosphatidylinositol only slightly in the ascites tumor cells, while an appreciable decrease of the ratio was observed in the liver during the first 3 hr. 3. These results suggest that phosphatidylinositol synthesis through pathways other than de novo synthesis is rapid in ascites tumor cells.

Acetates↗

Binding and cytotoxicity of Ricinus communis lectins to HeLa cells, Sarcoma 180 ascites tumor cells and erythrocytes.

The binding of Ricinus communis lectins to HeLa cells, Sarcoma 180 ascites tumor cells and human erythrocytes was studied in detail. Scatchard plots of binding of 125I-lectins to these cells gave biphasic lines except for HeLa cells at 0 degree C. The association constants of lectins for the three cell types at 37 degrees C were lower than those at 0 degree C. The numbers of total binding sites were estimated to be 7 to 16 X 10(7) per HeLa cell, 3 to 4 X 10(7) per Sarcoma 180 ascites tumor cell and 0.4 to 1 X 10(6) per erythrocyte. A fraction, 16 to 27% of the total amount of cell-bound lectin at 37 degrees C, appeared to be bound irreversibly as judged by non-removal on washing with 0.1 M lactose, whereas no lectin was irreversibly bound at 0 degree C. In the case of erythrocytes, no lectin became irreversibly bound even at 37 degrees C. The toxicity of lectins on HeLa cells and Sarcoma 180 ascites tumor cells was investigated. The toxicity of ricin D was 50 times for Sarcoma 180 ascites tumor cells and 140 times for HeLa cells as much as that for castor bean hemagglutinin. As to the sensitivities of both cell types to these lectins, it became apparent that Sarcoma 180 ascites tumor cells were more susceptible than HeLa cells.

Animals↗

Variation in protein concentrations in acute and chronic ascites in Dirofilaria immitis-infected dogs.

Microfilaremic dogs, developing ascites acutely following a reaction to diethylcarbamazine therapy, had similar protein concentrations in their ascitic fluid and plasma. In contrast, in dogs chronically infected with Dirofilaria immitis, the protein concentrations of ascitic fluid were found to be significantly lower than plasma protein concentrations. The acute development of ascites in such dogs is associated with high ascitic protein levels.

Journal Article↗

Ehrlich ascites tumour cells show tissue-specific adherence and modify their shape upon contact with embryonic fibroblasts and myotubes.

Adhesiveness of Ehrlich ascites tumour (EAT) cells to glass, to mouse peritoneal membrane, living and aldehyde-fixed mouse embryo fibroblasts and chick embryo fibroblasts, myoblasts and myotubes was investigated. The ascitic EAT cells (and leukaemia L1210 cells) did not adhere to glass and peritoneum but readily adhered to embryo fibroblasts, myoblasts and myotubes. The attachment was followed by cell spreading and migration. Fixation of fibroblasts or myogenic cells with aldehydes did not prevent ascitic cells from attaching but reduced the rate of spreading. Only direct interaction of ascitic cells with embryo myoblasts or fibroblasts induced changes in tumour cell adhesiveness followed by cell spreading and locomotion. These results are discussed in relation to an observation that ascitic cells growing as a cell suspension intraperitoneally grow as a solid tumour when injected subcutaneously.

Animals↗

Detection of feline coronavirus antibody, feline immunodeficiency virus antibody, and feline leukemia virus antigen in ascites from cats with effusive feline infectious peritonitis.

To investigate the usefulness of ascites as a material for viral tests in cats with effusive feline infectious peritonitis (FIP), we attempted to detect anti-feline coronavirus antibody, anti-feline immunodeficiency virus antibody, and feline leukemia virus antigen in ascites from 88 cats clinically suspected with effusive FIP. In each of these three viral tests, all cats positive for serum antibody/antigen were also positive for ascitic antibody/antigen, while cats negative for serum antibody/antigen were also negative for ascitic antibody/antigen. This finding indicates that ascites is useful for these viral tests.

Animals↗

[Alterations in the isozyme spectrum and the activity of lactate dehydrogenase in development of Ehrlich ascites tumor].

By means of polyacrylamide gel disc electrophoresis lactate dehydrogenase belonging only to the M-type was observed in mice liver tissue, muscles, ascite cells and ascites fluid in the course of development of Ehrlich ascites tumor. In the tissues studied and in the tumor the LDH type was not altered in the course of development of the tumor. At the same time the LDH activity was slightly decreased in muscles; in the terminal period it was increased in liver tissue; in ascitic cells or fluid it was increased in the course of development of the tumor. In blood serum the M-fraction was the main type of LDH. The M4 isozyme accounted for about 80% of the total LDH activity. The remainding 20% were distributed approximately equally between the other four isozymes. As the disease proceeded redistribution of isozymes occurred in the direction of the M-type. About 70% of the H-type LDH was found in heart; isozymes H4 and H3M accounted for about 35% each of the total LDH-activity. Direct relation was not observed between the alteration in the enzymatic activity and the LDH isozyme spectrum in tumor and tissues of animals with developing Ehrlich ascites tumor.

Animals↗

[ASCITES IN HEPATIC CIRRHOSIS: RECOGNITION INVESTIGATIONAND TREATMENT]

Hepatic cirrhosis is the most common cause of ascites. It is caused by liver failure leading to complex interrelated circulatory and renal changes resulting in retention of sodium and water and portal hypertension localising that sodium and water in the peritoneum. Ascites is an important development in cirrhosis as it implies a generally poor long term prognosis. Investigation is important as ascites is not always dueto cirrhosis, may bethe consequence of complications of cirrhosis such as hepatocellular carcinoma, and may be associated with infection which is fatal if untreated. Most patients respond to treatment with sodium restriction and diuretic drugs. This treatment takes time, and increasingly doctors use therapeutic paracentesis with sodium restriction and diuretics to prevent recurrence of ascites. Paracentesis, however, is not without complications, and it is particularly important to give colloid replacement to prevent hypovolaemia which can lead to renal failure. Patients who do not respond to this treatment may be helped by a TIPSS procedure or a peritoneovenous shunt. However, these patients usually have very poor liverfunction and the possibility of fiver transplantation should be considered. Infection is a very serious complication of ascites (spontaneous bacterial peritonitis) and carries a generally poor prognosis.Antibiotic prophylaxis is important to prevent recurrence and liver transpiantation shoulcl be considered.

Journal Article↗