Cholesterol removal by peritoneal lavage with phospholipid-HDL apoprotein mixtures in hypercholesterolemic hamsters

Syrian hamsters were rendered hypercholesterolemic by supplementation of their diet with 1% cholesterol and 15% butter. The hamsters were injected intraperitoneally (i.p.) with about 20 mg of phospholipid liposomes containing trace amounts of [3H]cholesteryl linoleyl ether ([ 3H]CLE) alone or combined with 10 mg delipidated high-density lipoprotein (apoHDL). After 2 h the peritoneal cavity was washed repeatedly with up to 15 ml phosphate-buffered saline. 60%-70% of [3H]CLE were retained after i.p. injection without apoHDL, 30-50% in the presence of apoHDL. The amount of free cholesterol recovered in the peritoneal lavage was significantly higher when apoHDL was combined with 18:2 sphingomyelin or dilinoleyl phosphatidylcholine liposomes, when compared to either liposomes or apoHDL alone. It is suggested that supplementation of dialysate with HDL apolipoproteins and phospholipids in patients undergoing continuous peritoneal dialysis could be of use in a cholesterol depletion regimen.     

Echinococcus granulosus: development of an intermediate host mouse model for use in vaccination studies.

A mouse model has been developed to evaluate potential protective antigens which could render intermediate hosts resistant to a challenge infection with Echinococcus granulosus eggs. DBA/2J, CBA/J, Balb/cJ, C57/B16J and CF-1 mice were initially infected orally and parenterally with eggs, hatched eggs or activated oncospheres. Generally less than 1% of the oral dose established as cysts. Mean cysts counts were increased when Balb/cJ mice were injected intraperitoneally or intravenously with activated oncospheres. A challenge regime using 600 activated oncospheres injected intraperitoneally into adult Balb/cJ mice was subsequently adopted yielding means of 15-51 cysts per mouse. When activated oncospheres were injected intraperitoneally into Balb/cJ, DBA/2J and CF-1 mice, cysts were restricted to the peritoneal cavity. Activated oncospheres injected intravenously, however, lodged almost exclusively in the lung and thoracic cavity, except in DBA/2J mice where 55% lodged in the liver. This anatomical localization enabled the outcome of prior infection and challenge to be monitored separately. Prior infection rendered Balb/cJ mice fully resistant to subsequent challenge.     

Gas Exchange in Renal Failure: II—Pulmonary Gas Exchange during Peritoneal Dialysis

Blood gas analysis studies have been made in patients undergoing peritoneal dialysis. It has been shown that oxygen tensions are reduced when fluid has been run into the peritoneal cavity and that this fall in Pao₂ is reversed after running out the dialysate. The change in Pao₂ is greater with 2-litre than with 1-litre cycles.     

Delayed inhibition of DNA synthesis in mouse jejunum by low doses of actinomycin D

Actinomycin D (0.008 μg/gm of body weight) injected intraperitoneally every two hours, produced a prompt 50% inhibition of RNA synthesis in the jejunum of mice, and a delayed inhibition of DNA synthesis, that reached its maximum inhibition (68% of control values) 4.5 hours after the first injection of actinomycin D. Autoradiographic studies indicated that this low level of actinomycin D inhibited a step in the G₁ phase of the cell cycle, preventing the initiation, but not affecting the continuation, of DNA biosynthesis. The activity of DNA polymerase was not affected under these conditions. The results are substantially similar to those previously obtained with Ehrlich ascites cells growing in the peritoneal cavity of mice and can be interpreted as indicating that in the G₁ phase of dividing cells there is an actinomycin sensitive step whose inhibition prevents the entrance of cells into the DNA-synthesis phase.     

Peritonitis during continuous ambulatory peritoneal dialysis in children.

The use of continuous ambulatory peritoneal dialysis (CAPD) in children has proved beneficial. However, peritonitis remains the major complication. A review of the incidence of peritonitis in 55 children (mean age 9.6 years) who underwent CAPD between 1978 and 1984 showed that there were 67 episodes of peritonitis (1 per 9.4 patient-months) in 33 of the 55. Three patients accounted for 22 of the episodes. In all cases, treatment with antibiotics, given intraperitoneally, was successful. Cephalothin was routinely given for infections due to gram-positive organisms, tobramycin for infections due to gram-negative organisms. Peritonitis recurred in seven patients, of whom five had to have their catheters replaced because of associated chronic infections of the deep peritoneal cuff, the exit site or the catheter tunnel. Although peritonitis was a common complication of CAPD in this population, it did not affect the success of the technique.     

Mouse cachexia induced by trehalose dimycolate from Nocardia asteroides

Trehalose dimycolate (TDM) isolated from Nocardia asteroides induced in mice a severely wasted condition known as cachexia. Intraperitoneal injection of mice with five 10 micrograms doses of TDM in mineral oil at intervals of 2 d killed 90% of the animals within 26 d. Death followed a precipitous weight loss and an inflammatory process in the peritoneal cavity. When mice were injected intraperitoneally with a single 10 micrograms dose of TDM, 48 h later, they had begun to lose weight and exhibited extreme hypertriglyceridaemia and hypoglycaemia. Tumour necrosis factor (or cachectin) was detected in the plasma from animals injected with TDM. This cytokine released by mononuclear phagocytes may be involved in the induction of cachexia by TDM.     

Potential use of tuftsin in treatment of candida peritonitis in a murine model

A major complication of continuous ambulatory peritoneal dialysis (CAPD) is peritonitis caused by Candida albicans. Increasing the activity of the peritoneal macrophages, the predominant cell type found in the peritoneal cavity, may be a promising treatment for this infection. Tuftsin was found to increase thioglycollate-elicited mouse peritoneal macrophage activity. 2x10(-7) M tuftsin enhanced two-fold cell association with radiolabelled candida, superoxide aniom production, and killing activity. Thus, a model consisting of mice undergoing peritoneal dialysis was developed in order to study the use of tuftsin as a therapeutic drug against peritoneal candidiasis. Administration of tuftsin (50 micrograms/mouse) before candidiasis induction with a lethal dose of candida (7x10(8) candida per mouse) improved mouse survival up to 70%, compared with 10% in the control group. The potential of tuftsin as a treatment for candidiasis was shown when the infection was induced with a sublethal dose of candida. Daily intraperitoneal injections of tuftsin (50 micrograms) to the sublethally infected mice caused a significant decrease in the number of candida recovered from the peritoneal cavity and from the blood (from 700 +/- 190 to 110 +/- 26 CFU/ml and from 100 +/- 26 CFU/ml to 17 +/- 8 CFU/ml, respectively). In addition, a larger number of peritoneal macrophages with greater phagocytic and killing activity were found in the tuftsin-treated mice. The effect of tuftsin may promote its potential use in the therapy of peritonitis in patients undergoing chronic peritoneal dialysis.     

Uptake of radioactive thymidine and cytidine by Ehrlich ascites tumor cells in different stages of growth

Summary In Strong A female mice, the Ehrlich ascites tumor inoculated into the peritoneal cavity grows exponentially for the first 7 days with a doubling time of about 36 hours. The tumor enters then into a late stage during which the number of tumor cells in the peritoneal cavity does not increase. The uptake of intraperitoneally injected thymidine decreases from the exponential to the late stage, mostly because of a decrease in the fraction of cells in DNA synthesis. During the exponential phase, the uptake of thymidine is a function of the amount of radioactive thymidine injected per tumor cell, the utilization decreasing with increasing cell dose. The uptake of intraperitoneally injected cytidine decreases slightly with time after inoculation although the fraction of tumor cells in RNA synthesis remains constant.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his 80th birthday.This investigation was supported by U.S.P.H.S. Grant CA-05667. The author is a U.S.P.H.S. Research Career Development Awardee.     

Simple high-performance liquid chromatographic method for determination of losartan and E-3174 metabolite in human plasma, urine and dialysate

A simple high-performance liquid chromatographic (HPLC) method was developed for the determination of losartan and its E-3174 metabolite in human plasma, urine and dialysate. For plasma, a gradient mobile phase consisting of 25 mM potassium phosphate and acetonitrile pH 2.2 was used with a phenyl analytical column and fluorescence detection. For urine and dialysate, an isocratic mobile phase consisting of 25 mM potassium phosphate and acetonitrile (60:40, v/v) pH 2.2 was used. The method demonstrated linearity from 10 to 1000 ng/ml with a detection limit of 1 ng/ml for losartan and E-3174 using 10 μl of prepared plasma, urine or dialysate. The method was utilized in a study evaluating the pharmacokinetic and pharmacodynamic effects of losartan in patients with kidney failure undergoing continuous ambulatory peritoneal dialysis (CAPD).     

Study on the ultrastructure of the peritoneal stomata in humans.

In 16 human specimens the topography and organization of stomata and mesothelial cells of the diaphragmatic, pelvic wall and anterior abdominal wall peritoneum were studied by transmission electron microscopy, scanning electron microscopy and the image processing technique. The mesothelial cells were organized into two discrete populations, cuboidal cells and flattened cells. The stomata were found only among cuboidal cells, either on the muscular portion or on the tendinous portion of the diaphragm. The size and shape of stomata, which were arranged in a cluster or a strip, were often irregular. The average area of a stoma on the muscular portion was 10.43 +/- 1.61 microns2, on the tendinous portion 7.93 +/- 1.67 microns2. Stomata opened to submesothelial connective tissue, under which numerous lymphatics were observed. Stomata were not discovered in the pelvic and anterior abdominal wall peritoneum. In animal experiments intraperitoneally injected trypan blue particles were rapidly removed from the peritoneal cavity through stomata of the diaphragmatic peritoneum in rabbits. It is suggested that stomata may be the main pathway for draining matter from the peritoneal cavity and that the diaphragmatic peritoneum shows the strongest absorption in all parts of the peritoneum.     

Mouse Virulence of K(L) Antigen-Containing Strains of Escherichia coli

The mouse virulence of two K antigen-containing (L variety) strains of Escherichia coli (serotype O2:K1) isolated from human septicemia, and of their variants which lacked K antigen, was studied. The strains containing envelope antigen (K+) were highly virulent when injected intracerebrally or when suspended in mucin and injected intraperitoneally. After intraperitoneal injection of E-107 K+ (but not K−), there was a marked initial growth in the peritoneal cavity followed by bacteremia and infection of all the organs examined. In the mucin-enhanced lethal infection, this growth continued until death of the animal; in the nonlethal infection, growth ceased and the count dropped quickly after approximately 5 hr. Host defenses were depressed greatly by intraperitoneally, but not intravenously, administered mucin. Bacteria were most virulent when injected intraperitoneally. In vitro phagocytosis of the K+ bacteria required opsonins not needed for phagocytosis of the smooth K− variants. Opsonins were found in immunized rabbit and normal mouse sera. Immune rabbit sera contained antibodies with anti-K specificity which were opsonic in vitro and highly protective in vivo when administered passively. There appears to be a lesser anti-O opsonic and protective activity involving one of the strains (E-107 K+), and colonial morphology, agglutination, and absorption tests indicated a low amount of K antigen on this organism. No anti-O opsonic or protective activity could be shown involving the other strain (E-102 K+). When standard serological typing procedures were used, these two strains appeared to be identical serologically, but they differed greatly in sensitivity to immune rabbit serum in phagocytosis experiments in vitro.     

A Multifunctional Mesothelin Antibody-tagged Microparticle Targets Human Mesotheliomas

Pleural and peritoneal mesotheliomas (MMs) are chemoresistant tumors with no effective therapeutic strategies. The authors first injected multifunctional, acid-prepared mesoporous spheres (APMS), microparticles functionalized with tetraethylene glycol oligomers, intraperitoneally into rodents. Biodistribution of APMS was observed in major organs, peritoneal lavage fluid (PLF), and urine of normal mice and rats. After verification of increased mesothelin in human mesotheliomas injected into severe combined immunodeficient (SCID) mice, APMS were then functionalized with an antibody to mesothelin (APMS-MB) or bovine serum albumin (BSA), a nonspecific protein control, and tumor targeting was evaluated by inductively coupled plasma mass spectrometry and multifluorescence confocal microscopy. Some APMS were initially cleared via the urine over a 24 hr period, and small amounts were observed in liver, spleen, and kidneys at 24 hr and 6 days. Targeting with APMS-MB increased APMS uptake in mesenteric tumors at 6 days. Approximately 10% to 12% of the initially injected amount was observed in both spheroid and mesenteric MM at this time point. The data suggest that localized delivery of APMS-MB into the peritoneal cavity after encapsulation of drugs, DNA, or macromolecules is a novel therapeutic approach for MM and other tumors (ovarian and pancreatic) that overexpress mesothelin.     

Preclinical Assessment of Adjunctive tPA and DNase for Peritoneal Dialysis Associated Peritonitis

A major complication of peritoneal dialysis is the development of peritonitis, which is associated with reduced technique and patient survival. The inflammatory response elicited by infection results in a fibrin and debris-rich environment within the peritoneal cavity, which may reduce the effectiveness of antimicrobial agents and predispose to recurrence or relapse of infection. Strategies to enhance responses to antimicrobial agents therefore have the potential to improve patient outcomes. This study presents pre-clinical data describing the compatibility of tPA and DNase in combination with antimicrobial agents used for the treatment of PD peritonitis. tPA and DNase were stable in standard dialysate solution and in the presence of antimicrobial agents, and were safe when given intraperitoneally in a mouse model with no evidence of local or systemic toxicity. Adjunctive tPA and DNase may have a role in the management of patients presenting with PD peritonitis.     

Continuous ambulatory peritoneal dialysis: no longer experimental.

Many patients with end-stage renal disease have now been maintained for 5 years or more with continuous ambulatory peritoneal dialysis (CAPD). Viewed initially as an experimental alternative to be used only when hemodialysis was not feasible, CAPD is now seen as the treatment of choice in an increasing number of situations. CAPD is suitable for self-care. The main concern in the early years--peritonitis--is now less frightening and less frequent (one episode occurring every 18 patient-months as compared with every 8 initially), and this has allowed chronic complications of CAPD, such as malnutrition and loss of the peritoneum''s capacity for ultrafiltration, to come to light. As would be expected, among patients of advanced age and those who have heart disease or diabetes, survival rates tend to be lower than among other CAPD patients. However, hypertension seems to be more easily controlled, pre-existing anemia can be significantly ameliorated, and young children grow more normally than they do with hemodialysis. Diabetes-related changes in vision stabilize in most CAPD patients, and control of the blood glucose level is good; insulin is administered intraperitoneally. CAPD is thus showing itself to be a feasible form of long-term treatment for end-stage renal disease.     

Quantitation of dextran 70 in peritoneal dialysate from patients administered 7.5% polyglucose

A method using gel permeation chromatography was evaluated for the quantitation of dextran 70 in dialysate samples containing polyglucose. Dialysate samples containing dextran 70 and polyglucose were pretreated using the enzyme α-amylase to selectively hydrolyze the α(1–4)-linked polyglucose, while leaving the α(1–6)-linked dextran 70 intact. Following sample deproteinization with trichloroacetic acid, dextran 70 was quantitated using gel permeation chromatography with refractive index detection. This method was evaluated for accuracy, precision, specificity, linearity, range, and analyte stability. Adequate method linearity with a correlation of >0.999 was established over the range of dextran 70 concentration from 1 to 0.025 mg/ml. Method precision was approximately 2% R.S.D. and accuracy (% recovery) was approximately 98–100% in the typical sample concentration range (1–0.5 mg/ml). This method was applied to the determination of intraperitoneal fluid kinetics in continuous ambulatory peritoneal dialysis (CAPD) patients administered daily night-time intraperitoneal exchanges with either 7.5% polyglucose or 4.25% dextrose. Dextran 70 was added to the dialysis solutions to yield an initial concentration of 1 mg/ml. Dialysate samples were collected at various times over a 10-h dwell-time and assayed for dextran 70. Intraperitoneal volume profiles based on dextran 70 concentrations and drain volumes were then calculated for each dialysis solution.     

Diminished Selenium Levels in Hemodialysis and Continuous Ambulatory Peritoneal Dialysis Patients

In this cross-sectional study, selenium (Se) levels in the sera of 35 hemodialysis (HD) patients and 34 patients undergoing continuous ambulatory peritoneal dialysis (CAPD) for more than 3 months were compared with the serum Se levels of 34 healthy volunteers. The observed Se levels of 100.8 ± 51.9 μg/L in the sera of the HD patients and of 65.5 ± 32.1 μg/L in the sera of the CAPD patients were significantly lower than the 134.9 ± 81.2 μg/L of the controls, with p = 0.002 and 0.02, respectively. Furthermore, the Se levels were significantly higher in the HD rather than the CAPD patients (p = 0.01). In the spent dialysate effluent fluid of 32 of the CAPD patients Se was undetectable, in the remaining two CAPD patients the Se levels were 1.9 and 4.6μg/l, respectively. The low Se levels of HD and CAPD patients as compared to healthy persons are attributed to diminished Se retention due to chronic oxidative stress.     

Severe hypoglycaemia due to combined use of parenteral nutrition and renal dialysis.

A 24-year-old woman who had sustained serious injuries in a road traffic accident required renal dialysis daily and was fed intravenously with a solution containing 25% dextrose. Subsequently insulin had to be added to the parenteral fluid to maintain blood glucose concentrations at physiological values. On one occasion parenteral feeding was continued until dialysis was started; she became comatose and the plasma glucose concentration was found to be less than 1 mmol/l (18 mg/100 ml). She responded rapidly to a 50 ml intravenous bolus of 50% dextrose. When parenteral feeding and dialysis are used simultaneously glucose passes across the semi-permeable membrane from the blood to the dialysate so that hypoglycaemia may occur. Insulin added to the parenteral fluid further decreases blood glucose concentrations. Stopping parenteral feeding 30-45 minutes before dialysis is started eliminates this danger of hypoglycaemia.     

Preliminary description of lesions in juvenile largemouth bass injected with largemouth bass virus

Juvenile largemouth bass Micropterus salmoides were intraperitoneally injected with largemouth bass virus (LMBV), a member of the genus Ranavirus, family Iridoviridae. Moribund fish which had been injected with 10(6.2) tissue culture infectious doses, 50% endpoint (TCID50), were sampled 4 d after injection; other largemouth bass injected with this dose died between 3 and 5 d after injection. Fish injected with 10(2.8) TCID50 of LMBV were also examined after 4 d and had lesions similar to those of fish injected with the high dose. Clinical signs included darker pigmentation, inflammation and necrosis at the site of injection, distended abdomen, corkscrew swimming, and lateral recumbency. Internally, fish had focally pale livers, bright red spleens and reddened intestinal ceca. Histologically acute fibrinous peritonitis affected the surface of all organs in the peritoneal cavity, but deeper portions of organs appeared normal. There was also necrosis of the gastrointestinal mucosa. Except for the injection site, lesions were confined to the peritoneal cavity.     

Inflammatory process caused by intraperitoneal injection of polyester in the rat: chemotactic activity in lavage fluid from the cavity

Minced polyester threads introduced into peritoneal cavity of guinea pigs or rats cause a granulomatous inflammation with evidence of macrophage stimulation. Chemotactic agents play an important role in the inflammatory reaction; they may be exogenous and/or endogenous. These are released locally by the cells involved in inflammation. In this paper the chemotactic effects of the peritoneal fluids from rats bearing the polyester inflammatory process, have been studied on PMN cells "in vitro". The peritoneal cavity fluids were obtained by washing the cavity of untreated rats or rats intraperitoneally injected with polyester, 1, 3, 7, 14 days after the intraperitoneal injection. The chemotactic response was assayed by employing modified chemotaxis Boyden chambers (Blind Well Neuro Probe) and polymorphonuclear leukocytes from normal or treated rats. Quantification of the migration was calculated by chemotactic index (A/B) (B = random migration, A = chemotaxis). The results demonstrated that the peritoneal fluids taken 3 and 7 days after the intraperitoneal polyester injection, elicit an evident chemotaxis response greater than that showed by peritoneal fluids from control rats. It is suggested that chemotactic factors can be produced and released by mononuclear cells involved in the inflammatory process.