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[Current ascites therapy].

Ultrasonography detects ascites easily even in trace amounts. 80% of the cases are caused by hepatic disease, in the remaining 20% cancer, inflammation, pancreatic, renal, or cardiac disease can be found. The underlying disease should be investigated by few inexpensive laboratory test from serum, urine and ascites and by abdominal sonography. Hepatic ascites is caused by portal hypertension and disturbances of humoral factors. Sodium retention, peripheral, vasodilation, hyperdynamic circulation and progressive renal vasoconstriction lead to a stepwise deterioration of patients condition. Treatment with diuretics (furosemide, torsemide, or xipamide and spironolactone) and sodium-restriction (< 60 mval per day) control 85-90% of the cases with hepatic ascites. If this regimen fails, non-compliance, spontaneous bacterial peritonitis, hyponatremia or additional complications such as renal failure, Budd-Chiari syndrome or tumor should be considered. Ten to 15% of the patients develop refractory ascites and finally hepatorenal syndrome and have a poor prognosis. Early liver transplantation should be considered. Large volume paracentesis with albumin substitution is a therapeutic option in these patients. The transjugular intrahepatic portosystemic stent-shunt (TIPS) may be superior for patients with concurrent esophageal varices or hepatorenal syndrome. If TIPS is considered the patient should be referred to an experienced center. The peritoneo-venous shunt is restricted to rare indications. In the future, new drugs such as antagonists of endothelins or of the antidiuretic hormone may offer new therapeutic options.

Ascites↗

Peri-operative chylous ascites.

Chylous ascites occurs when lymphatics are disrupted due to primary lymphatic disease, infection, malignancy, or chronic liver disease. It may also occur following inadvertent interruption of abdominal lymphatics during surgery involving retroperitoneal dissection. It is suggested by some that during liver transplantation, severed hepatic lymphatics should be ligated or stented to avoid post-operative pleural and abdominal accumulation of chylous fluid. The occurrence of chylous ascites and the need to ligate lymphatics after orthotopic transplantation was assessed in 180 consecutive patients subjected to this procedure. Pre-operative chylous ascites present in one patient resolved following transplantation. Three patients who required retroperitoneal dissection to complete the biliary anastomosis via choledochojejunostomy or perform a hepatic artery graft developed post-operative chylous ascites which rapidly resolved without complications. These findings indicate special attention to transacted hepatic lymphatics is not required during orthotopic liver transplantation. Chylous ascites rarely occurs after liver transplantation and its transitory development is due to retroperitoneal dissection.

Aged↗

Alpha-1 acid glycoprotein is an immunosuppressive factor found in ascites from ovaria carcinoma.

BACKGROUND: Although the ascites of patients with ovarian carcinoma has been reported to contain immunosuppressive factors, the identity and source of this activity has not been fully elucidated. The objective of this study was to describe the purification of a single immunosuppressive protein, alpha-1 acid glycoprotein, from ovarian carcinoma ascites, identify its site of production, and describe a possible mechanism by which it inhibits lymphocytes. METHODS: Ascites from proteins from five patients with epithelial ovarian carcinoma first were differentially precipitated by size with different concentrations of polyethylene glycol and then separated on the basis of isoelectric focusing. The protein factions then were placed in a lymphocyte proliferation assay to determine immunosuppressive activity. Western blot analysis was used to identify alpha-1 acid glycoprotein as an immunosuppressive protein in ascites. Total RNA was extracted from ovarian and hepatic cell lines as well as primary and recurrent ovarian tumor samples. Reverse-transcriptase polymerase chain reaction then was utilized to identify the site of production of this protein. Purified alpha-1 acid glycoprotein was placed in lymphocyte culture and its effects on lymphocyte interleukin-2 (IL-2) production were measured by enzyme-linked immunoadsorbent assay. RESULTS: Addition of purified alpha-1 acid glycoprotein to the lymphocyte assay resulted in a 60% decrease in lymphocyte proliferation (P < 0.05). Alpha-1 acid glycoprotein transcript was not identified in ovarian tumor cells. The addition of purified alpha-1 acid glycoprotein to the lymphocyte culture resulted in a 65% decrease in IL-2 secretion into the media (P < 0.05). CONCLUSIONS: Alpha-1 acid glycoprotein is an immunosuppressive protein purified from ovarian carcinoma ascites. It is not expressed primarily by ovarian carcinoma cells. It appears to inhibit IL-2 secretion by lymphocytes.

Ascites↗

[Role of surgical therapy in the treatment of refractory ascites].

In 5-10% of cases ascites is not controlled by medical therapy and is defined refractory. These patients may be submitted to one of the four following surgical options: portal-systemic shunt, peritoneo-venous shunt, transjugular intrahepatic portal-systemic shunt, orthotopic liver transplantation. Although the portal-systemic shunt is efficient in clearing ascites, it does not improve the survival, which depends on liver function, and it is complicated by an important incidence of encephalopathy. Since the patients with refractory ascites and good hepatic risk are not usually many, it is possible to understand why derivative surgery has been disappointing with this indication. Although the peritoneo-venous shunt is associated with a significant rate of valve obstruction, it is an easy, effective and not expensive treatment. So, till now, it has been considered the first choice procedure of refractory ascites, if any situations, determinating the onset of postoperative complications, are not present. Recently a new method has been introduced in the therapy of portal hypertension, the transjugular intrahepatic portal-systemic shunt. This is a bloodless portal-systemic derivation and so it has caused great enthusiasm even if the available data are insufficient to give a definitive opinion on its role in management of ascites. Certainly the liver transplantation, which presents the great advantage to treat both the cirrhosis and its complications, seems to be the most rational therapy for these patients. However, at least for this moment, the well-known absence of organ donors makes still actual the palliative surgical measures.

Ascites↗

[Abdominal pain and ascites as manifestations of hereditary angioneurotic edema].

Hereditary angioneurotic edema (HAE) is an infrequent autosomal dominant disorder characterized by a decrease in the levels or a dysfunction of the complement C1 inhibitor factor (C1 inh). The clinical presentation varies widely and involves any area of the organism. Gastrointestinal involvement is usually as abdominal pain and may be accompanied by ascites. De novo diagnosis of HAE with abdominal pain and ascites as a form of presentation is difficult with differential diagnosis with abdominal pain of unknown origin. The appearance of ascites is rare with few cases reported in the literature. Both abdominal pain and ascites disappear a few days after initiation of medical treatment. Occasionally exploratory laparotomy has been required. A new case of abdominal pain and ascites as manifestations of HAE is herein reported.

Abdominal Pain↗

Activation of protein phosphatase 1alpha promoter in ascites hepatoma cells.

We have already reported the nucleotide sequence of 5'-flanking region of rat PP1alpha gene and identified the promoter region in NIH3T3 cells. Here we determined the promoter activity of PP1alpha gene in ascites hepatoma cells and compared with that of hepatocytes. Reporter analysis showed that the promoter activity was enhanced in an ascites hepatoma AH13 compared with normal hepatocytes at two regions, -37 to -198 bp and -492 to -848 bp upstream of the translation start site. At -37 to -198 bp region with high GC content, binding of Sp1 was markedly increased in all of five ascites hepatomas examined. At -492 to -848 bp region, two bands were clearly detected in two of the five ascites hepatomas, AH13 and AH7974F, by gel shift analysis. These results strongly suggest that the increase in PP1alpha mRNA expression in ascites hepatomas is at least in part due to the enhanced promoter activity of PP1alpha gene.

Animals↗

Phase I study of intraperitoneal metalloproteinase inhibitor BB94 in patients with malignant ascites.

This was an open Phase I study of i.p. matrix metalloproteinase inhibitor BB94 in patients with malignant ascites. The objective of the study was to determine the effect of increasing i.p. doses of BB94 with reference to the tolerance, safety, and pharmacokinetics of the compound. Twenty-three patients with malignant ascites had BB94 instilled into the peritoneal cavity after paracentesis. The compound was well tolerated; no serious adverse events were seen, and no specific toxicities were observed. High plasma concentrations were seen an hour after dosing, and BB94 was still present in the plasma at day 28 after treatment at levels in excess of the IC50 identified in preclinical studies. Five of the 23 patients neither reaccumulated ascites nor died up to 112 days after dosing. Seven patients died without reaccumulating ascites. Although the study was not designed to demonstrate clinical efficacy, the results were encouraging and support the further therapeutic evaluation of matrix metalloproteinase inhibitors in the management of malignant ascites.

Aged↗

Purification of a glycoprotein from mouse ascites fluid by immunoaffinity chromatography which is related to the major glycoprotein of murine leukemia viruses. Immunologic and structural comparison with purified viral glycoproteins.

Proteins immunologically related to the major glycoprotein from murine leukemia viruses, gp70, are expressed in the tissue and body fluids of unmanipulated mice in the absence of virus particle production. Cross-reactive gp70 was isolated from ascites fluid of NZB and (NZB X NZW)F1 mice utilizing immunoaffinity chromatography. An enriched antibody preparation bound to Sepharose was used as an immunosorbent to purify gp70 over 600-fold in one step with a 30% recovery. The gp70s from ascites fluid were compared with gp70s purified from laboratory virus strains Friend, Rauscher, Moloney, and Scripps leukemia viruses. Direct antibody binding studies showed that the viral gp70s were more closely related to each other than to the gp70s from ascites fluid. Furthermore, the gp70s from ascites fluid were more reactive with antibody to xenotropic virus than were the gp70s from the laboratory viruses tested. Tryptic peptide map analysis of radioiodinated proteins showed that gp70 from Scripps virus is closely related to gp70 from Moloney virus in primary structure and less closely related structurally to gp70 from Rauscher virus and gp70 from NZB ascites fluid.

Animals↗

Characterization of an ovarian cancer activating factor in ascites from ovarian cancer patients.

Ascites from ovarian cancer patients contain potent growth-promoting activity toward human ovarian cancer cells both in vitro and in vivo. This activity is associated with rapid increases in cytosolic free calcium ([Ca2+]i) as a consequence of phosphoinositide hydrolysis. In this study, we describe the purification, characterization, and identification of an ovarian cancer activating factor (OCAF) from ascites of ovarian cancer patients. We have isolated OCAF by a combination of solvent extraction, silica gel chromatography, and TLC. Mass spectral analysis, phospholipase sensitivity, and gas chromatographic behavior of purified OCAF indicate that OCAF is composed of various species of lysophosphatidic acid (LPA), including LPAs with polyunsaturated fatty acyl chains (linoleic, arachidonic, and docosahexaenoic acids). However, OCAF is more potent than sn-1 palmitoyl, oleoyl, or stearoyl LPA in increasing [Ca2+]i in ovarian cancer cells. The ability of OCAF to alter [Ca2+]i is sensitive to the effects of lipoxidase, whereas the activity of sn-1 oleoyl, stearoyl, or palmitoyl LPA is not, suggesting that polyunsaturated bonds in the fatty acyl chain of OCAF may account for its increased ability to activate ovarian cancer cells. Furthermore, a sn-2 linoleoyl LPA generated by phospholipase A1 treatment of synthetic phosphatidic acid is much more active than are sn-1 palmitoyl, stearoyl, or oleoyl LPA in increasing [Ca2+]i in ovarian cancer cells. Taken together, these data suggest that the ability of OCAF to increase cellular calcium may reside in the structure and/or location of the fatty acyl chain of LPA. Purified OCAF, at concentrations similar to those present in ascites from ovarian cancer patients, was sufficient to induce proliferation of ovarian cancer cells, as indicated by thymidine incorporation, reduction of 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, or colony formation. However, even at optimal concentrations of OCAF, proliferation was lower than that induced by FCS or ascites from ovarian cancer patients, indicating that, although OCAF may be a major regulator of ovarian cancer cells in vivo, it is not the sole mediator present in ascites, and it likely functions in concert with other growth factor activities.

Ascites↗

Interpretation of ascitic fluid data.

Except in the case of chylous or hemorrhagic effusion, the gross appearance of ascitic fluid is not especially helpful in the investigation of ascites of unknown origin. The differential diagnosis of ascites rests on more objective data obtained from chemical and cytologic analyses. Both routine and special ascitic fluid tests are listed here, and test results are interpreted as they apply to four major disease states.

Abdominal Neoplasms↗

Passage of radioactive erythrocytes from the peritoneal cavity into the blood stream during experimental ascites.

Intraperitoneal injection of red cells tagged with radioiron into dogs with experimental ascites demonstrated that such cells were rapidly transferred into the circulating blood. When the experimental animals were not actively producing ascitic fluid, 43.4, 67.0, and 56.4 per cent respectively, of the administered radioactive red cells passed to the blood in 72 hours. In the same three dogs during active ascitic fluid formation, 25.9, 51.2, and 38.8 per cent of the administered radioactivity was removed in a similar period. The amount of radioactivity in the blood stream, consequent on the passage of red cells from the peritoneal cavity into the circulation, becomes nearly constant in 48 hours, whereas for radioactive plasma proteins the plateau is attained in 24 hours (Fig. 1). In normal dogs (16), the passage of red cells from the peritoneal cavity was complete in 72 hours, while in ascitic dogs, 5 to 47 per cent of the injected tagged red cells remained behind in the peritoneum after the same period.

Animals↗

Ehrlich ascites tumor cells expressing anti-sense glutaminase mRNA lose their capacity to evade the mouse immune system.

Glutaminase (EC 3.5.1.2) is a key enzyme in rapidly proliferating cells. Using anti-sense technology, an Ehrlich ascites tumor cell line (0.28AS-2) with reduced glutaminase activity has been obtained. We investigated the in vivo growth characteristics of the 0.28AS-2 cells. When injected i.p. into normal Swiss albino mice, the 0.28AS-2 cells were unable to grow. On the contrary, when injected into nude mice, they developed into solid tumors. Mice inoculated with 0.28AS-2 cells kept immunologic memory and rejected a second inoculation with parental Ehrlich ascites tumor cells. Expression of both polymorphic epithelial mucin-1 (MUC-1) and the enzyme N-acetyl-alpha-D-galactosaminidase, proteins implicated in host immune system escape, were markedly diminished in 0.28AS-2 cells. Study of the immune system response in mice inoculated with 0.28AS-2 cells revealed an increase in splenic CD18 cells and the presence of a large number of activated F4/80+ macrophages in the ascites cavity. These features, not observed in mice inoculated with parental Ehrlich ascites tumor cells, indicate that a distinctive, strong immune response occurred in animals inoculated with 0.28AS-2 cells. Our results suggest that inhibition of glutaminase expression using anti-sense technology induces phenotypic changes in Ehrlich ascites tumor cells that allow the development of an effective anti-tumor immune response, which makes the cells unable to develop in vivo tumors.

Animals↗

Mouse Lewis lung carcinoma and hepatoma ascites treatment by combination of liposome chemotherapy and non-specific immunotherapy.

Liposomes have been used as biological carriers of anti-tumor drugs, and their potential use has been tested in mouse Lewis lung carcinoma and hepatoma ascites tumor models. Ara-C3 given by the intraperitoneal i.p. route either at 35 mg/kg in a single dose or at 2.5 mg/kg/dose with 5 doses/day for 3 days had no effect on the average survival time of i.v. implanted LLC. However, the same single dose of Ara-C encapsulated in positively charged MLV significantly improved the average survival time of LLC-bearing mice. MTX was chosen as a test drug for the treatment of hepatoma ascites. Non-encapsulated MTX given at either 3 or 30 mg/kg by the i.p. route had little effect on the average survival of i.p.-implanted hepatoma ascites. However, MTX encapsulated in SUV at a 3 mg/kg dose by the i.p. route significantly improved the average survival time of tumor-bearing mice. A combination of chemotherapy and non-specific immunotherapy has also been tested with these 2 tumor models. Two non-specific microbial immune stimulators, Bacillus Calmette Guérin (BCG) and Corynebacterium parvum (CP) were tested by both the i.v. and i.p. routes. A combination of BCG therapy with non-encapsulated anti-tumor drugs was not effective for either of the tumor models. A combination of BCG therapy with liposome therapy appeared to improve the average survival time of LLC-bearing mice. In particular, BCG treatment by the i.p. route in combination with liposome therapy resulted in a 20% long-term survival rate among treated mice. However, BCG therapy by either route in combination with SUV encapsulated MTX therapy had no effect on the average survival time of hepatoma ascites-bearing mice. Immunostimulation with CP at a given dose appears to be superior to BCG therapy for both tumor models. In the treatment of LLC, injection of CP by either the i.v. or i.p. route appears to be equally effective in combination with liposome therapy. However, for the treatment of hepatoma ascites, CP was only effective by the i.p. route, in combination with liposome therapy.

Animals↗

Chylous ascites following treatment for gynecologic malignancies.

BACKGROUND: Chylous ascites is a rare complication following abdominal radiation or para-aortic lymph node dissection in the management of gynecologic malignancies. Treatment options include dietary restriction with addition of medium-chain triglycerides, serial paracenteses, total parenteral nutrition, and somatostatin. Current opinion advocates that surgical exploration and peritoneo-venous shunts be reserved for refractory cases. CASES: Two patients developed chylous ascites, one after completion of surgical staging and chemoradiation for stage IIB squamous carcinoma of the cervix and one following para-aortic lymph node dissection for recurrent malignant mixed mullerian tumor of the endometrium. In both cases resolution of the chylous ascites followed placement of a peritoneo-venous shunt. CONCLUSIONS: Chylous ascites should be considered in the differential diagnosis of ascites in patients with gynecologic malignancy treated with radiation or para-aortic lymph node dissection.

Aged↗

Lack of protein-mediated alpha-tocopherol transfer between membranes in the cytoplasm of ascites hepatomas.

Transfer-stimulating activity for alpha-tocopherol and the concentration of alpha-tocopherol and peroxidized lipids in rat ascites hepatoma cells were compared with those from normal and regenerating liver. The ability of supernatants from ascites hepatomas (AH-13, AH-60C, AH-109A) to enhance the transfer of alpha-tocopherol was much lower than that from normal livers. The alpha-tocopherol per mg protein of supernatant from ascites hepatomas was lower than that from normal liver. Regenerating liver showed almost the same values as normal liver in activity to stimulate the transfer of alpha-tocopherol and alpha-tocopherol content of the supernatant. By gel filtration, about 60% of alpha-tocopherol in the supernatant of normal liver was detected in the fractions containing the 30 K protein, which stimulates transfer of alpha-tocopherol between membranes, whereas no significant amount of alpha-tocopherol was detected in 30 K protein fractions of AH-60C supernatant. Little stimulating activity for alpha-tocopherol transfer was detected in AH-60C, AH-109A and AH-13. All ascites hepatomas tested contained less arachidonic acid and docosahexaenoic acid than normal and regenerating liver. An absorption peak with maximum intensity at 233 nm, which is due to conjugated dienes, was observed in UV-absorption spectra of ascites hepatoma total lipids, indicating that peroxidized lipids accumulate in these cells. With normal and regenerating liver, no significant peak due to conjugated dienes was detected.

Animals↗

Bioflavonoid regulation of ATPase and hexokinase activity in Ehrlich ascites cell mitochondria.

(1) The mitochondrial ATPase (EC 3.6.1.3) Ehrlich ascites cell mitochondria, was inhibited by D-glucose under physiological concentrations of ATP. The generation of ADP by the mitochondrial bound hexokinase, seems to be the reason for the D-glucose inhibitory effect. Reversal of the inhibitory effect of ADP on Ehrlich ascites cell mitochondria ATPase by an ATP-regenerating system was achieved. (2) Dissociation of mitochondrial bound hexokinase from the mitochondria eliminated the inhibitory effect of D-glucose. Rebinding of the hexokinase to the mitochondria regenerated the D-glucose inhibitory effect on Ehrlich ascites cell mitochondria ATPase. (3) Bioflavonoids such as quercetin inhibit the mitochondrial hexokinase activity, but do not change the mitochondrial ATPase activity of isolated Ehrlich ascites tumor cell mitochondria. (4) The inhibitory effect of bioflavonoids on mitochondrial bound hexokinase activity is shown to be dissociable from the ascites tumor cell mitochondria and seems to be associated with regulatory rather than catalitic sites of the enzyme.

Adenosine Diphosphate↗

Calcium permeability of Ehrlich ascites tumour cell plasma membrane in vivo.

Passive Ca2+ entry into Ehrlich ascites tumour cells has been investigated. Passive equilibrium of Ca2+ takes place in ascites tumour cells only under conditions of exhaustive energy depletion. The specific Ca2+ ionophore A23187 does not affect Ca2+ entry into ascites tumour cells under active metabolic conditions, but it increases the rate of Ca2+ equilibration in ascites tumour cells in the early stages of energy depletion. The results of the present experiments lead to the conclusion that in ascites tumour cell plasma membrane Ca2+ permeability is not a limiting step in the regulation of intracellular calcium content, while the energy-dependent Ca2+ extrusion is the main mechanism that prevents uncontrolled intracellular Ca2+ increase. The results taken together support the hypothesis that increased Ca2+ influx into the cell, caused by plasma membrane alteration, is responsible for permanently elevated mitotic activity and for deranged metabolic behavior of these neoplastic cells.

Anaerobiosis↗

RNA synthesis in Yoshida ascites tumor cells after ultraviolet and gamma irradiation.

The residual RNA synthesis in Yoshida ascites tumor cells following exposure to ultraviolet radiation was insensitive to rifampicin AF/013, an inhibitor of initiation of RNA synthesis in eukaryotic cells. On the other hand, residual RNA synthesis in the ascites tumor cells after gamma-irradiation was inhibited by rifampicin AF/013. In vitro synthesis of RNA on ultraviolet-irradiated ascites chromatin likewise was resistant to rifampicin AF/013. Ultraviolet irradiation was found to decrease the number of RNA chain initiation sites on ascites chromatin considerably whereas the number of initiation sites on ascites chromatin increased considerably after gamma irradiation. These results indicate that ultraviolet irradiation may preferentially affect the DNA segments participating in initiation of RNA synthesis.

Animals↗