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PubMed1998

Comparison of In Vitro Monoclonal Antibody Production Methods with an In Vivo Ascites Production Technique.

Economic, technical, legislative, and ethical issues influence decisions concerning alternatives to the use of animals in biomedical research. Since the development of cell hybridoma technology, production of ascites in mice has been a popular technique for generating high concentrations of monoclonal antibodies. However, the availability of several in vitro methods that can be performed in most laboratories make these alternative methods of monoclonal antibody production an attractive option. To evaluate the practicality of the use of in vitro techniques, three tissue culture methods and a technique for production of ascites were compared on the basis of yield, material costs, and time requirements. Analysis of results revealed that the time and material costs to produce 100 mg of monoclonal antibody 7.16.4 by inducing ascites in irradiated mice was similar or slightly more than that for tissue culture in standard plastic flasks and hollow fiber cartridge bioreactors; however, tissue culture in gas permeable bags was slightly less expensive than these methods in terms of cost and time. When ascites production in irradiated mice was compared with tissue culture in plastic flasks or gas permeable bags for monoclonal antibody 225, the in vitro methods were approximately five times more expensive than the ascites production technique. Information reported here outlines factors that should be considered and evaluated when choosing a monoclonal antibody production method. Furthermore, these results documented that in vitro technology for the production of monoclonal antibodies can be adapted by most conventional laboratories to provide sufficient resources for the most commonly performed experimental protocols.

Journal Article↗

PubMed1976

Evidence that approximately eighty per cent of the soluble proteins from Ehrlich ascites cells are Nalpha-acetylated.

Analysis of soluble Ehrlich ascites proteins by the Sanger procedure revealed methionine, alanine, valine, and glycine as the major NH2-terminal amino acids. The average monomer weights of these proteins calculated from the yields of NH2-terminal amino acids was 144,000. In contrast, the average monomer weight of Ehrlich ascites soluble proteins calculated from the data obtained after electrophoresis in polyacrylamide gels containing sodium dodecyl sulfate was 32,500. The explanation for the disparity in the estimates of average monomer weight obtained by the procedures appears to be that extensive blocking of alpha-NH2 groups by acetate occurs in these proteins, i.e. of the acetate present in the acidic peptides isolated from proteolytic digests of ascites proteins, 23.2 nmol/mg of protein appears to originate from N-acetyl amino acids. These results suggest that approximately 80% of the soluble proteins from Ehrlich ascites cells contain acetate at their NH2-terminal residues. The extensive N-acetylation of proteins does not appear to be limited to Ehrlich ascites cells and may be characteristic of eukaryotic proteins.

Acetates↗

PubMed1998

Ascites free-water dynamics in decompensated cirrhosis: effects of an acute Hemaccel infusion.

BACKGROUND: Plasma-expanders (PE) are commonly employed to reduce the hemodynamic effects of large-volume paracentesis in patients with decompensated cirrhosis. The aim of this paper is to investigate the effects of PE acute administration on ascites dynamics in cirrhotic patients. METHODS: Wash-out intraabdominal pressure (IAP), ascites volume and free-water peritoneal clearance (FWPC) were evaluated in six decompensated cirrhosis by means of a methylene-blue (MB) dilution method. The method is based on the compartmental analysis of MB peritoneal clearance and has been validated by previous deuterium oxide studies. Immediately after the MB dilution test, a rapid Hemaccel i.v. infusion (500 ml 3.5% in 15 min) was perform-ed in all subjects, thereafter carrying out a second MB test. The obtained results were analyzed by two-way variance analysis. RESULTS: IAP and ascites volume values were not appreciably modified by the PE treatment. Following the Hemaccel infusion, only a fair reduction of FWPC values (from 88.33+/-7.78 to 76.98+/-8.09 ml/min) was observed. CONCLUSIONS: The results obtained indicate that, at least at the employed doses, Hemaccel acute administration has a low impact on the ascites dynamics in decompensated cirrhotics. These data cannot therefore support the hypothesis of a therapeutic use of PE in ascitic patients, but to avoid excessive plasma volume reductions in association with a large volume paracentesis.

Journal Article↗

PubMedDate not supplied

Circadian influence on the immunization of mice with live Bacillus Calmette-Guérin (BCG) and subsequent challenge with Ehrlich ascites carcinoma.

Non-specific immunostimulation with Bacillus Calmette-Guérin (BCG) is of current interest in the treatment of cancer. The main objective of the series of experiments described in this paper was to evaluate the influence the host's circadian system has on a. the stimulation of the immune system with BCG and b. the subsequent efficiency of that stimulated immune system against the Ehrlich Ascites Carcinoma (EAC). There was a circadian rhythm in the length of survival time in non-immunized mice challenged with the EAC. Mice receiving an EAC challenge during the middle of the light period survived significantly longer than those challenged with the EAC around the time of transition from dark to light. Mice immunized with BCG and challenged with EAC also demonstrated a circadian rhythm in the length of survival 30 days after EAC challenge with 86% survivors in the mice treated at 10(00) and 60% survivors in the mice treated at 07(00). The same relationship was also observed 70 and 80 days after EAC challenge. Eighty days after EAC challenge, a circadian rhythm was apparent in the frequency of solid tumors at the site of the initial EAC injection. The highest incidence of solid tumors occurred at 13(00). A circadian rhythm was found in the increase in body weight between the first and second BCG or saline injections. Rectal temperatures recorded on the 8th, 12th and 16th day after EAC challenge were characterized by circadian rhythmicity. In the mice without development of ascites, the peak temperature consistently occurred at 01(00). In the mice with ascites there was a phase advance in the rectal temperature rhythm of 3 h so that the peak in the rhythm consistently occurred at 22(00). In the mice with ascites a further finding was an increasing hypothermia as the ascites continued to develop; however, this hypothermia was not detectable during the time of the peak (10(00)) in the temperature rhythm. The mice which did not die by the 80th day after EAC challenge were challenged again with 5.0 x 10(6) EAC cells, and during the next 46 days circadian variations were observed in the numbers of mice which survived. Similar changes were observed during an additional 46 days after a third EAC challenge of 41.5 x 10(6) cells.

Animals↗

PubMed1984

Nitrogen movement between host and tumor in mice inoculated with Ehrlich ascitic tumor cells.

Tumors function as a nitrogen trap, and they compete with the host for nitrogen compounds. In experiments with whole animals infected with Ehrlich ascitic tumor cells, the glutamine, glutamate, asparagine, and aspartate concentrations were determined for host plasma, ascitic liquid, and tumor cells, throughout the period of tumor growth. Concentration gradients of glutamine or asparagine were created from the host tissues towards the ascitic liquid. The countergradient step from ascitic liquid to tumor cells may be overcome by an active transport process with an apparent Km for glutamine of 3.1 X 10(-4) M. On the other hand, a reverse flux of glutamate and aspartate was seen to take place from cells to plasma. In vitro incubations of tumor cells with near physiological concentrations of glutamine, or asparagine plus glucose, confirmed the host-to-tumor nitrogen movement previously deduced from the relative amino acid concentrations in plasma, ascitic liquid, and tumor cells. The ammonemia detected in tumor-bearing mice at the terminal stage could result from the hydrolysis of glutamine, which was rapidly metabolized by the tumor cells.

Ammonia↗

PubMed1983

Relationship between antitumor effect and metabolites of 5-fluorouracil in combination treatment with 5-fluorouracil and guanosine in ascites sarcoma 180 tumor system.

The antitumor activity of 5-fluorouracil (FUra) against ascites Sarcoma 180 was significantly enhanced by coadministration of guanosine, and slightly by adenosine, but not by cytidine or uridine. In advanced ascites Sarcoma 180, guanosine also enhanced the action of FUra, but adenosine, uridine, and cytidine did not. The potentiation of antitumor activity by guanosine was reversed by addition of cytidine. The antitumor activity of FUra was significantly potentiated when guanosine was administered either 0 to 15 min before or 5 min after FUra. Changes in metabolites of FUra after potentiation by guanosine were investigated. Total radioactivity in the plasma was significantly decreased 10 min after the combined administration of [6-14C]FUra (3 mg/kg i.p.) and guanosine (100 mg/kg i.p.) in comparison with that of [6-14C]FUra alone and was slightly decreased by coadministration of [6-14C]FUra and adenosine. Conversely, it was significantly increased by uridine or cytidine. The decrease in total radioactivity in the plasma caused by guanosine was completely reversed by addition of cytidine. FUra, 5-fluorouridine, alpha-fluoro-beta-ureidopropionic acid, and alpha-fluoro-beta-alanine were found in the plasma. Intact FUra accounted for about 55% of the total radioactivity. The proportion of metabolites of [6-14C]FUra was not changed by coadministration of [6-14C]FUra and guanosine, adenosine, or cytidine, but the proportion of FUrd was increased by uridine. In the ascitic fluid, the total radioactivity derived from [6-14C]FUra was decreased by its combined administration with guanosine, and it was reversed by addition of cytidine. This pattern was similar to that in the plasma. The main FUra compound was intact FUra itself (90%), and 5-fluorouridine accounted for 1% of the total radioactivity in the ascitic fluid. On the other hand, total radioactivity of [6-14C]FUra in the tumor cells was significantly and slightly increased by guanosine and adenosine, respectively. Total radioactivity after [6-14C]FUra in combination with uridine or cytidine was less than that after [6-14C]FUra alone. Incorporation of [6-14C]FUra into RNA was increased about 3.7 times by its combination with guanosine in comparison with FUra alone, and it was increased 2.0, 0.6, and 0.7 times by adenosine, uridine, and cytidine, respectively. Moreover, FUra-nucleotides were significantly increased by guanosine. The increased radioactivity in RNA and FUra-nucleotides of tumor cells caused by guanosine was completely reversed by cytidine. These changes in incorporation into tumor cells were comparable to those in antitumor activity against ascites Sarcoma 180. The potentiation of antitumor activity of FUra by guanosine was considered to be due to an increase in incorporation of FUra into FUra-nucleotides and RNA in the tumor cells.

Adenosine↗

PubMed1993

Malignant ascites. A comparison of peritoneovenous shunting and nonoperative management.

BACKGROUND: Effective palliation of malignant ascites remains a difficult management problem. METHODS: Eighty-five patients with malignant ascites were studied. Forty-two patients had peritoneovenous shunts (PVS) inserted (16 LeVeen, 17 single-valve Denver, 9 double-valve Denver). RESULTS: Shunt patency was not related to the type of shunt, type of cancer, or any characteristic of the ascitic fluid. Ascites was controlled in 64% of patients with shunts and serum albumin levels were preserved. Survival and quality of life were not significantly different in comparison with those of patients treated by abdominal paracentesis. CONCLUSIONS: PVS allowed many patients to be treated successfully outside the hospital and are indicated in carefully selected cases.

Abdomen↗

PubMedDate not supplied

Opsonic activity of human ascitic fluid: a potentially important protective mechanism against spontaneous bacterial peritonitis.

The opsonic activity of 60 ascitic fluids from 47 patients was measured using a standard opsonophagocytic assay. Curve analysis of the opsonic activity compared to the ascitic fluid concentration of total protein, total hemolytic complement, C3 and C4 yielded correlation coefficients of 0.84 (p less than 0.001), 0.84 (p less than 0.001), 0.94 (p less than 0.001) and 0.92 (p less than 0.001), respectively. There appeared to be a threshold of concentration for each protein below which there was no killing of bacteria. Cirrhotic ascites had significantly (all p less than 0.001) lower concentrations of total protein and complement and less opsonic activity than noncirrhotic ascites (including malignant, cardiac and miscellaneous types). Perhaps it is the dilution of crucial antimicrobial proteins below a threshold which predisposes to spontaneous bacterial peritonitis.

Ascites↗

PubMed1989

Penetration of cefoperazone into ascites.

The pharmacokinetics of cefoperazone was studied in eleven cirrhotic patients with ascites after i.v. administration of a single dose of 15 mg.kg-1 (n = 7) or after three doses of 15 mg.kg-1 given at 12 h intervals (n = 4). The concentrations of cefoperazone in serum and ascitic fluid were determined by HPLC. The peak serum cefoperazone concentration after a single i.v. injection of 15 mg.kg-1 was 96.0 mg.l-1. The serum elimination half-life was longer (5.0 h) than in normal subjects. The penetration of cefoperazone into ascites was satisfactory (32.3% and 58.3% after single and repeated injections, respectively). Ascitic fluid concentrations of cefoperazone exceeded 5.4 mg.ml-1 from 0.5 to 6 h after the single i.v. injection, levels which are well above the MIC of most pathogens found in spontaneous bacterial peritonitis. Adjustment of the dose of cefoperazone in cases of severe hepatic insufficiency does not appear to be necessary provided that renal function is normal.

Adult↗

PubMed1975

Pancreatic ascites. A case report and review of the literature.

A case of pancreatic ascites is reported and compared with 55 previously reported cases. A 42-year-old black male chronic alcoholic presenting with abdominal pain was found at operation to have chronic pancreatitis with no pseudocyst formation or overt duct disruption, in contrast to the majority of cases reported. The diagnosis and differentiation from cirrhosis of the liver were based on the operative findings, elevated serum amylase level, ascitic fluid amylase value, and protein content. Surgical exploration alone has proven beneficial--the patient has done well in the past 2 years with no recurrence of the ascites and continued weight gain. The clinical course was compatible with pancreatitis although the radiographic and angiographic studies were not diagnostic. It is suggested that the clinical entity of pancreatic ascites occurs more often than reported and a workup for it should be done even in the face of unconvincing radiographic and angiographic evidence.

Adult↗

PubMed1981

Intraperitoneal coagulation in chronic liver disease ascites.

Twenty patients with the ascites of chronic liver disease were investigated for in vivo evidence of active coagulation within ascites by detection of fibrin monomer. Increased levels of fibrin monomer, increased fibrin/fibrinogen degradation products, and absent or low levels of fibrinogen in the ascites of all patients tested confirmed the presence of intraperitoneal coagulation when ascites is present.

Ascites↗

PubMed1990

Serum-ascites albumin gradients in nonalcoholic liver disease.

Several studies performed in alcoholics with advanced liver disease have demonstrated a positive correlation between the serum-ascites albumin gradient (SAAG) and measured portal venous pressure. A single study performed in 15 patients with exudative malignant ascites and 29 patients with alcoholic liver disease demonstrated that a SAAG of less than 1.1 was essentially diagnostic of a malignant origin of the ascites. In an effort to confirm and extend these observations to individuals with nonalcoholic liver disease, 24 patients with nonalcoholic liver disease and 11 with alcoholic liver disease undergoing orthotopic liver transplantation (OTLx) were studied. At the time of liver transplantation, each had their serum and ascitic fluid albumin levels determined, the gradient calculated, and their portal venous pressure (PVP) as well as the corrected portal venous pressure (PPc) measured directly. A significant correlation (r = 0.624) between the PPc and the SAAG was found in the 11 alcoholics (P less than 0.05). No such correlation existed for those with nonalcoholic liver disease (r = 0.398). Moreover, a SAAG less than 1.1 was found in three of nonalcoholics with cirrhosis in the absence of an abdominal malignancy. We conclude that (1) the SAAG and PPc are statistically related to each other in individuals with alcoholic liver disease but not in those with a nonalcoholic cause for cirrhosis, and (2) SAAG less than 1.1 is not diagnostic of abdominal malignancy but can occur in those with advanced nonmalignant hepatic disease.

Adult↗

PubMed1983

Immunoregulatory properties and LP-pattern of human ovarian carcinoma ascites and pleural effusions of nonmalignant origin.

The immunoregulatory activity of ascitic fluids obtained from patients with ovarian carcinoma, and pleural effusions collected from patients with various nonmalignant diseases, was tested. The following assays were performed: in vivo, PFC-response of mouse splenocytes to SRBC; in vitro, antibody-dependent cellular cytotoxicity (ADCC), and phagocytic activity of human peripheral blood mononuclear cells (PBMoC). The migration ability of mouse splenocytes in the presence of ascitic fluids was also evaluated. Parallelly, lipid (triglyceride and cholesterol) and lipoprotein (high-density/HDL/, low-density/LDL/, and very-low-density/VLDL/) content in ascites and pleural effusions, were determined. Both types of fluids suppressed the PFC-response and inhibited phagocytosis of 51Cr-labeled sheep erythrocytes by PBMoC, but augmented ADCC of PBMoC against sheep erythrocytes. An elevation of VLDL and a decrease of HDL and LDL in ascitic fluids and pleural effusions, as compared to normal human sera, were found. There were similar changes in the cholesterol content of these lipoprotein fractions, i.e., an increase of VLDL- and a decrease of HDL- and LDL-cholesterol.

Animals↗

PubMed1976

Clinical significance of pancreatic ascites.

Recurrence of pancreatic ascites in a patient after two courses of peritoneal lavage suggested pancreatic ductal disruption as the etiolgy, and this was confirmed at operation. Peritoneal lavage in two other patients with pancreatic ascites without ductal disruption or cysts was associated with resolution ascites. The use of peritoneal lavage in patients with acute or chronic pancreatic ascites may guide the selection of those requiring early surgical treatment.

Acute Disease↗

PubMed1985

Consequences of intraperitoneal bile: bile ascites versus bile peritonitis.

Recent experience with patients with bile ascites and bile peritonitis prompted a review of other case histories in the medical literature of these conditions. The clinical courses of 24 patients with bile ascites and 34 with bile peritonitis were reviewed. Bile ascites occurred most often as a postoperative complication of biliary tract operations and also occasionally after trauma. Clinical signs were minimal except for abdominal distention, and operations were delayed for an average of 30 days. Peritoneal fluid was sterile in the 11 patients studied. In contrast, bile peritonitis occurred most commonly after spontaneous perforation of the gallbladder or hepatic ducts but also after trauma. All patients had severe signs of peritoneal irritation, and operation was performed earlier, at a mean of 4 days after onset of symptoms. Of 11 patients with specimens of their peritoneal fluid cultured, 6 had sterile fluid and 5 had bacteria. Although both bile salt concentration and bacteria have been implicated in the development of bile peritonitis rather than bile ascites, our understanding of the mechanisms involved is still incomplete.

Adolescent↗

PubMed1988

Protein leak from normal vasculature due to human malignant ascites.

Malignant ascites is an accumulation of protein rich fluid (a filtrate of whole blood) in the peritoneal cavity of patients with abdominal malignancies. The normal peritoneal microvasculature of the cremaster muscle of rats, with the nerve and blood supply intact, was visualized before and after exposure of the tissue to human malignant ascites fluid and to human plasma. In vivo fluorescent microscopy was used to quantitate leakage of fluorescent-tagged albumin. Exposure of the abluminal side of the vasculature to malignant ascitic fluid and plasma causes significant protein leakage from the small veins to the interstitial space. This suggests that the continued production of malignant ascites may be caused by a positive feedback system, which is related to factors present in a normal plasma filtrate. These factors can induce leakage of protein by an effect on the abluminal side of the normal peritoneal microvasculature.

Adenocarcinoma↗

PubMed1991

Malignant small bowel obstruction and ascites: not a contraindication to percutaneous gastrostomy.

Percutaneous gastrostomy (PG) with gastropexy was performed for relief of malignant small bowel obstruction in 12 patients with extensive ascites. Abdominal paracentesis was performed before PG in nine patients and after PG in one patient. Gastrostomy catheters were inserted without complication in all patients. Clinical follow up revealed that pericatheter leakage of ascitic fluid and skin excoriation occurred only in the three patients who did not have paracentesis performed before PG. No dislodgement of gastrostomy catheters occurred but mild peritonitis was noted in one patient. Our experience suggests that although in the past extensive ascites was a relative contraindication for PG, these patients can now be successfully treated with a combination of ultrasound-guided paracentesis to reduce pericatheter leakage of ascitic fluid, and gastropexy to prevent catheter dislodgement.

Abdomen↗

PubMed1983

Immunological studies of ascitic fluid in patients with cirrhosis of the liver and hepatitis B surface antigen.

Sixteen cirrhotic patients with ascites and HBs antigen (HBsAg), were chosen for this study. Serum and ascitic fluid samples were tested for Ag, total proteins, electrophoretic pattern and immunoglobulin IgG, IgA, IgM and IgE. Transmission of HBsAg through the peritoneal membrane was confirmed. All protein fractions and immunoglobulins were presented in the ascitic fluid. Seven patients with ascites due to intra-abdominal neoplasms were also studied as controls. Significant differences were observed and are discussed.

Abdominal Neoplasms↗