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Background Manufacturers of parenteral solutions adhere to European and US Pharmacopoeia standards to define safety and sterility. In response to excess cases of aseptic peritonitis in peritoneal dialysis patients using icodextrin-containing dialysate that met all pharmacopoeia standards, a global recall was issued in May, 2002. We aimed to establish the cause of aseptic peritonitis.Methods We analysed 186 reports of aseptic peritonitis between September, 2001, and January, 2003. Extensive physical, chemical, and microbiological investigations of recalled dialysate were done. We calculated dose-response curves for peptidoglycan-induced interleukin 6 elaboration in peripheral blood mononuclear cells (PBMCs) from healthy donors and for sterile peritonitis in rats.Findings Although its chemical constituents and concentrations of endotoxin were within pharmacopoeia specifications, the dialysis solution elicited an interlukin 6 response in vivo and in vitro. We identified peptidoglycan from thermophilic acidophilic bacteria (Alicyclobacillus acidocaldarius) as the contaminating proinflammatory substance. In the PBMC assay, strong dose-response relations were noted between peptidoglycan concentrations and interleukin 6. In rats injected with peptidoglycan, dose-dependent increases of intraperitoneal neutrophils and pyrogenic cytokines were recorded. We measured a positive relation between peptidoglycan concentrations in recalled dialysate and reports of aseptic peritonitis. After implementation of corrective actions, the rate of peritonitis returned to baseline.Interpretation Excess cases of aseptic peritonitis in peritoneal dialysis patients were due to peptidoglycan contamination of dialysate by Alicyclobacillus. This outbreak serves as an example of how contemporary parenteral products with microbial contaminants can be considered safe under current pharmacopoeia tests, but provoke adverse clinical effects.

Background:Addition of hyaluronan (HA) to the dialysis solution has been suggested as a means to protect the peritoneum from injury during peritoneal dialysis (PD). Methods:Concentrations of inflammatory mediators were determined in dialysate samples obtained from PD patients after 6-hour dwells with glucose-based (13.6 g/l) solution containing 0.1 and 0.5 g/l of exogenous high-molecular-weight HA. We additionally evaluated the effect of HA-supplemented dialysate, drained after dwell in PD patients, on function of human peritoneal mesothelial cells (MC) in in vitro culture. Results:Concentration of nitrites was significantly higher in HA 0.5 g/l supplemented dialysate (+43%, p < 0.05) as compared to control. Levels of monocyte chemoattractant protein (MCP-1), soluble intercellular adhesive molecule (s-ICAM), vascular endothelial growth factor (VEGF) and fibronectin were comparable in all the studied groups. However, when MC were exposed in in vitro conditions for 24 h to the studied dialysates, we observed that HA containing fluids inhibited the synthesis of MCP-1, s-ICAM, VEGF and fibronectin in these cells. HA-supplemented dialysate accelerated growth rate of in vitro proliferating MC. Conclusion:High-molecular-weight HA added to the dialysis fluid exerts anti-inflammatory and antifibrotic actions on the in vitro cultured MC and accelerates their growth rate what may be important for peritoneal healing during PD.

← Background Hyaluronan (HA) is a glycosaminoglycan found in connective tissues and tissue spaces, including the peritoneal cavity. In vivostudies in a rat model of peritoneal dialysis (PD) have shown that addition of HA to PD solution during an intraperitoneal dwell can alter peritoneal fluid transport and protect the peritoneal membrane from the effects of inflammation and repeated infusions of dialysis solution. The current study sought to evaluate the safety of intraperitoneal HA and its effect on peritoneal fluid and solute transport when administered during a dialysis dwell in humans.← Methods 13 PD patients were enrolled in a prospective, randomized crossover study involving three dialysis treatments using the following PD solutions: (1) a commercially available PD solution (Dianeal PD-4, 1.36% glucose; Baxter Healthcare Corporation, Alliston, Ontario, Canada); (2) Dianeal PD-4 containing 0.1 g/L HA, and (3) Dianeal PD-4 containing 0.5 g/LHA. Each 6-hour dialysis exchange was separated from the other exchanges by a 2-week washout period. Radioiodinated human serum albumin (RISA) was administered with the dialysis solution to evaluate intraperitoneal volume, net ultrafiltration (UF), and fluid reabsorption. Peritoneal clearances, dialysate/plasma ratios (D/P), and mass transfer area coefficients (MTACs) were determined for sodium, urea, creatinine, albumin, and glucose. Safety was evaluated by monitoring adverse events and changes in serum chemistries. Ten patients completed all three dialysis exchanges and two additional patients completed at least one treatment exchange.← Results There were no reported adverse events related to HA administration and no significant changes in serum chemistries. There were no significant differences in net UF or peritoneal volume profiles among the three treatments. Mean net UF calculated using residual volumes, estimated by RISA dilution, tended to be slightly higher during treatment with solution containing 0.1 g/L HA and 0.5 g/L HA [74 ± 86 (SE) and 41 ± 99 mL, respectively] compared to control treatment (–58 ± 129 mL). Although not statistically significant, there was a trend toward decreased fluid reabsorption during treatment with HA. Solute clearances, D/P ratios, and MTACs were similar for the three treatments. Serum levels of HA were also unaffected by the two treatment solutions.← Conclusions These data support the acute safety of HA when administered intraperitoneally with the dialysis solution to PD patients. Due to the small sample size and variability in net UF and fluid reabsorption, statistically significant differences were not demonstrated for these parameters. However, a trend toward decreased fluid reabsorption was observed, suggesting that HA may act by a mechanism similar to that observed in animal studies. Further studies are necessary to evaluate whether the beneficial effects of HA observed in animal studies can be shown in humans.

Objective L-2-Oxothiazolidine-4-carboxylate (OTZ), a cysteine precursor, is a substrate for intracellular glutathione synthesis. As shown previously, OTZ prevents free-radical induced cellular damage during in vitrosimulation of peritoneal dialysis. In the present study, we examined the effect of adding OTZ to peritoneal dialysis solution on peritoneal function and structure during lipopolysaccharide (LPS)-induced peritonitis in rats. In addition, we studied the effects of pretreatment with OTZ (given orally) on the effects of LPS-induced peritonitis in rats.Methods Peritonitis was induced in rats by adding LPS (5 μg/mL) to the dialysis fluid. For acute experiments, rats were exposed to a single infusion of dialysis solution containing LPS or to LPS plus 5 mmol/L OTZ; peritoneal cell counts and permeability were determined after 4 hours. Alternatively, rats were pretreated with OTZ added to the drinking water (0.1%) for 10 days prior to infusion of LPS. For chronic experiments, peritoneal dialysis was performed over a 3-week period in rats with implanted peritoneal catheters. On days 8, 9, and 10 of the experiment, the rats were infused intraperitoneally with solution containing LPS (5 μg/mL), or LPS plus 5 mmol/L OTZ, to induce acute peritonitis. At the end of dialysis (10 days after the episodes of peritonitis), peritoneal function was assessed using a peritoneal equilibration test (PET), and peritoneal biopsies were taken to assess effects on peritoneal morphology.Results In the acute experiments, exposure to LPS led to increased peritoneal cell counts (+61% vs control, p< 0.05) and enhanced permeability of the peritoneum, leading to a loss in ultrafiltration (–63%, p< 0.0005). The glutathione concentration in peritoneal leukocytes also decreased during acute peritonitis (–31%, p< 0.05). During LPS-induced peritonitis, OTZ prevented the increase in dialysate cell count and the decrease in cellular glutathione content. Simultaneous administration of OTZ did not prevent the increased peritoneal permeability induced by LPS. However, in rats pretreated with OTZ, LPS-induced permeability to protein was significantly lower than in the nontreated animals (0.049 ± 0.011 vs 0.087 ± 0.034, p< 0.05). In the chronic experiments, LPS-induced peritonitis did not lead to any functional differences in peritoneal transport at the end of 3 weeks of dialysis. However, LPS-induced peritonitis led to increased thickness of the peritoneum and neovascularization within peritoneal interstitium compared to peritonitis-free animals. In contrast to the LPS-treated animals, the peritoneum of the rats exposed to LPS in the presence of OTZ was of a thickness similar to that in the control rats.Conclusion Supplementation of dialysis fluid with OTZ prevented changes in cellular glutathione levels and dialysate cell counts during acute peritonitis in rats. During chronic dialysis in rats exposed to intermittent peritonitis episodes, OTZ prevented increased thickening and neovascularization of the peritoneum. Our results suggest this may help to protect the peritoneal membrane during episodes of peritonitis.

Background Hyaluronan (HA) is a major component of interstitial tissue that participates in fluid homeostasis, response to inflammation, and wound healing. Previous studies have shown that intraperitoneal administration of HA can affect peritoneal fluid transport during short peritoneal dialysis exchanges in anesthetized rats. We sought to investigate the effect of high molecular weight HA on peritoneal permeability in conscious rats during dialysis exchanges up to 8 hours in duration. In addition, we sought to investigate the absorption of HA from the peritoneal cavity, its accumulation in peritoneal tissues, and its metabolism in normal and uremic rats.Methods Experiments were performed on male Wistar rats infused with 30 mL peritoneal dialysis solution (Dianeal, Baxter Healthcare; Castelbar, Ireland) containing 10 mg/dL HA or with Dianeal alone (control). Peritoneal fluid removal (net ultrafiltration), permeability to glucose, creatinine, and total proteins, and tissue and blood levels of HA were determined in separate groups of rats at 1, 2, 4, 6, and 8 hours after intraperitoneal infusion. Hyaluronan appearance and disappearance from plasma were also studied for 24 hours in separate groups of normal and uremic rats.Results Net ultrafiltration was significantly greater (27%) in rats infused with HA at 4, 6, and 8 hours (p< 0.01) compared to controls. Transperitoneal equilibration of protein was reduced by 27% (p< 0.001) at 4 hours and by 30% (p< 0.01) at 8 hours. During the 8-hour exchange, peritoneal clearance of creatinine increased by 27% (p< 0.01), whereas the clearance of total protein decreased by 27% (p< 0.005). After 8 hours, 25.7% ± 3.1% of the administered HA was absorbed from the peritoneal cavity, peritoneal tissue HA concentration was increased by 117% (p< 0.001), and plasma HA levels increased by 435% (p< 0.001). Plasma HA levels returned to normal within 24 hours after intraperitoneal administration in both healthy and uremic rats.Conclusions Hyaluronan added to dialysis fluid is absorbed from the peritoneal cavity and accumulates in peritoneal tissues. Hyaluronan supplementation produces changes in peritoneal permeability, leading to higher net ultrafiltration and peritoneal creatinine clearance, whereas total protein clearance decreases. The HA that is absorbed from the peritoneal cavity appears to be rapidly metabolized in both healthy and uremic rats.

The effect of high-molecular-weight hyaluronan (HA) on peritoneal and systemic inflammation and peritoneal permeability to water and solutes was studied during endotoxin-induced peritonitis in rats. Acute peritonitis was induced by adding lipopolysaccharide (LPS) to the dialysis fluid (Dianeal 3.86; Baxter Healthcare, Ireland, Castlebar). HA was added to the dialysis solution in a concentration of 10 mg/dL. During 4- and 8-hour dwells of the dialysis fluid, we studied the intensity of peritoneal (dialysate) and systemic (blood) inflammation (dialysate cell count and differential, cytokine and HA levels), as well as the transperitoneal transport of solutes and water. In rats, the addition of LPS to the dialysis fluid induced changes in inflammatory reaction and transperitoneal transport similar to those seen in continuous ambulatory peritoneal dialysis patients with peritonitis. During peritonitis, the addition of HA to the dialysis fluid reduced the loss of ultrafiltration, which resulted in a greater peritoneal creatinine clearance during the 8 hours of dwell (29.9 ± 6.7 mL/8 h in the HA-LPS group versus 19.7 ± 7.8 mL/8 h in the LPS group; P< 0.05). Dialysate interferon-γ (INF-γ) levels during peritonitis were greater in HA-treated animals (536.8 ± 296.6 pg/mL in the HA-LPS group versus 169.8 ± 137.8 pg/mL in the LPS group; P< 0.05). Dialysate elastase activity increased during peritonitis (44.4 ± 9.3 versus 14.2 ± 4.1 U/mL in peritonitis-free rats); during peritonitis, the increase in dialysate elastase activity was less pronounced in the rats that had HA in the dialysate (27.3 ± 4.1 U/mL versus the LPS group; P< 0.01). We conclude that HA added to the dialysis fluid reduces loss of ultrafiltration during peritonitis in rats. In the presence of HA dialysate, INF-γ levels during peritonitis increased, whereas elastase activity decreased; these changes might improve the peritoneal immune reaction during peritonitis and at the same time prevent peritoneal membrane injury.

The widespread use of calcium carbonate as a phosphate binder is limited by its tendency to develop hypercalcemia in some patients using effective dosages needed to control hyperphosphatemia. Most common continuous ambulatory peritoneal dialysis (CAPD) regimens using dialysis solutions containing 3.5 mEqlL of calcium result in net absorption of calcium from the dialysis solution and, hence limit the amount of oral calcium that can be administered. Peritoneal dialysis solutions with reduced calcium levels are needed for effective use of CaCO3 to control hyperphosphatemia in some dialysis patients.

Objective To assess the effect of insulin on the Na+/ K+-ATPase expression and activity in human peritoneal mesothelial cells (HPMC).Methods HPMC were isolated from the omental tissue of non-uremic patients, grown to confluence and rendered quiescent by serum deprivation for 24 hours. The activity of Na+/K+-ATPase was determined by measuring the ouabain-sensitive86Rb uptake. To assess whether the effect of insulin was related to changes in [Na+]i the sodium influx was measured with 22Na and the activity of Na+/K+ -A TPase was assessed in the presence of amiloride. Expression of Na+/K+ -A TPaseα1’ α2 and β1-subunit mRNAs was determined by RT/PCR.Results Exposure of HPMC to insulin resulted in a time and dose-dependent increase in the Na+/K+-ATPase activity. After 60 minutes the ouabain-sensitive 86Rb up take (cpm/104 cells) was increased from 6650±796 in control cells to 9763±1212 in HPMC exposed to 100 mU/ mL insulin (1.5-fold increase; n=4, P<0.05). In addition, incubation of HPMC with 100 mU/mL insulin resulted in a time-dependent increase in the 22Na influx. Pre-exposure of HPMC to 1 mM amiloride reduced the activity of Na+/K+-A TPase but did not block the stimulatory effect of insulin. RT/PCR analysis revealed that HPMC constitutively expressed α1 and β1-subunit mRNAs while the α2-subunit mRNA was barely detectable. Exposure of HPMC to insulin for up to 24 hours was not associated with any changes in the expression of either α1’ α2 or B1-subunit.Conclusion Insulin stimulates the Na+/K+-ATPase activity in HPMC in a time and dose-dependent manner. This effect appears to mediated by an increase in [Na+]i and is not related to alterations in Na+/K+-ATPase subunit mRNAs expression.

A one-step glc procedure was developed for the quantitative determination of valproic acid in plasma. After addition of internal standard (4-methyl valeric acid), plasma is buffered at pH 4.5 to avoid hydrolysis of valproic acid conjugate (s). Evaporation is avoided by extraction into a chloroform bead. The method is sufficiently sensitive to quantitate 0.8 g of the drug in 0.2 ml of plasma. The procedure has been thoroughly tested for precision and reproducibility through the determination of over two thousand samples.

The objective of this study was to determine whether a correlation exists between the rate of in vitro dissolution and bioavailability of levothyroxine sodium (T4) tablets. Dissolution versus time profiles for Synthroid, the Flint brand of levothyroxine sodium, and two competitors' tablets (brands A and B) were generated using an official dissolution apparatus (USP), and 0.05M phosphate buffer (pH 7.4) as the medium. These tablets were also utilized in single-dose crossover bioavailability studies in the hypothyroid dog model (n = 6). The average areas under the serum T4concentration versus time curve from 0 to 8h (AUC) for Synthroid, brand A, and brand B were 8.22, 6.32, and 8.70 ng-h/mL per dose (μg per kg body weight), respectively. Respective peak serum concentrations (Cmax) for each tablet formulation were 1.26, 1.07, and 1.36 ng/mL per dose. The corresponding dissolution rates, expressed as t50%, were 20.5, 3.06, and 14.1 min, respectively. Data analysis indicated no correlation between dissolution kinetic parameters and the bioavailability parameters AUC and Cmax. However, a linear relationship was observed between dissolution kinetics and both the time to reach maximal serum concentration (tmax) and the observed absorption rate constant (ka).

The toxicity of cyclohexanone, used as a solvent cement in polyvinyl chloride medical devices, was assessed in Wistar and Gunn rats. The Gunn rat was used because it has a negligible activity of UDP glucuronosyltransferase toward bilirubin and certain other aglycones. Cyclohexanone was administered iv for 28 consecutive days to Wistar and Gunn rats in two doses (50 and 100 mg/kg), using solutions containing 0.25 and 0.50 g per 100 ml, respectively, at a constant volume of 20 mg/kg. Saline (0.9% NaCI) was used as the control. Daily observations for signs of toxicity showed no adverse effects in Wistar or Gunn rats injected with either dose. Daily weight changes of control and test animals were similar. Ophthalmologic examinations revealed no treatment-related structural lesions. No adverse effects were noted when the data from the hemogram or clinical chemistry parameters were evaluated. Gross pathological and histopathologic assessment showed no alterations due to cyclohexanone treatment. Urinary excretions of total and glucuronide conjugates of cyclohexanol were similar for Wistar and Gunn rats; less than 1% was excreted as free cyclohexanone and cyclohexanol. It is concluded that the Gunn rat is capable of forming glucuronides of cyclohexanol and that cyclohexanone at these doses has a negligible toxic potential.

Objective Some patients develop a mild acidemia during treatment with amino acid-based peritoneal dialysis solutions due to hydrogen ion produced by metabolism of lysine, arginine, and methionine. In this study we modified the formulation of such a solution by reducing these amino acids and adding anionic amino acids so as to provide minimal net acid production.Design A modified formula (MF) was compared to a conventional formula (CF) of the solution in a randomized crossover study in 12 stable continuous ambulatory peritoneal dialysis patients. Patients were given each solution for 14 days without a wash-out period. Each patient replaced one or two dextrose dialysis exchanges with amino acid solution, depending upon oral protein intake and body weight. Total intake (oral protein plus amino acids absorbed) was equivalent to 1.1 -1.3 g protein/kg body weight/day.Plasma bicarbonate and urea were assessed at the beginning and end of each 14-day period.Results In the group as a whole, without regard to the order in which the solutions were given, patients had a decrease in serum bicarbonate with CF and an increase in bicarbonate when they received MF. Similar trends were observed regardless of the order in which the solutions were administered. Serum urea did not differ between the two solutions.Conclusion The results suggest that patients are less prone to develop acidemia when receiving MF as opposed to CF. Further studies will be necessary to determine the long-term effects and the relative nutritional benefits of the two solutions.

Patients on CAPD using calcium carbonate (CaC03) as phosphate binder might benefit from low -calcium (Ca) concentration dialysis solutions; however, no data are available for the effects of this regimen on Ca metabolism. We studied 10 patients on stable CAPD regimens with standard dialysis solutions (Ca 7 mg/dL) who were taking CaC03 to control hyperphosphatemia (mean daily doses 4.5±2.4 g). Hypercalcemic episodes had been recorded in 6 patients. Standard dialysis solutions were replaced with solutions containing 5 mg/dL of Ca. Calcium and phosphate peritoneal mass transfer (MT), serum concentrations of total Ca, ionized Ca (Ca++), phosphate, intact PTH, and mid-molecular PTH, were evaluated before and 48 hours after change of dialysate. The switch to low-Ca solutions was accompanied by significant changes in calcium mass transfer (Ca MT) (+9.84±48.22 versus −96.74±48.32 mg/day, p<.001). Ca MT was significantly (p<.05) correlated with the serum/ dialysate Cagradient. There was no difference in phosphate MT. Serum Ca++ significantly (p<.05) decreased from 5.20±0.32 to 4.88±0.36 mg/dL, and intact PTH significantly increased (81.5± 139 versus 112.4± 168 pg/mL, p<.05). It is concluded that dialysis solutions with Ca 5 mg/dL result in a negative peritoneal Ca MT and can be useful to prevent and treat hypercalcemia in CAPD patients taking CaC03 as phosphate binder. A careful monitoring of ionized calcium, PTH, and phosphate is suggested when an extensive and long-term use of this solution is considered.

A survey of the literature suggests continued interest in modifying the composition of peritoneal dialysis solutions. Osmotic agents such as glucose polymers and short-chain polypeptides may find a role in peritoneal dialysis fluids as partial substitutes for dextrose. New solutions containing amino acids should serve as more than sinks for uremic toxins by providing nutritional support and by ameliorating uremic lipid abnormalities. Increasing emphasis is being paid to biocompatibility during the development of new peritoneal dialysis fluids.

Objectives To assess whether dialysate containing short-chain polypeptides is well tolerated in continuous ambulatory peritoneal dialysis (CAPD) patients and to determine its effect on fluid and solute transport, plasma amino acid levels, and biochemical parameters.Design Two -treatment, two-period cross -over design.Setting Renal Unit, Academic Medical Center, Amsterdam and Renal Unit, University Hospital, Gent.Patients Two groups of 10 stable CAPD patients.Intervention All patients received a trial solution (1.36% glucose and 1% peptides, 381 mOsm/kg) and a control solution (2.27% glucose, 404 mOsm/kg) in randomized order. The patients were examined on four consecutive days in which two dwell periods on days 1 and 3 of either 4 (Group I) or 8 hours (Group II) were performed.Results The peptide solution was well tolerated in all patients. In addition, no differences were found in the parameters for the effective peritoneal surface area and the intrinsic permeability, implying that no irritating effect of the peptide solution was present. Net ultrafiltration was not different in Group I: -43±125 versus 86±125 mL (mean±SEM) and marginally lower in Group II: -94±64 versus 51±64 mL, despite the lower osmolality of the trial solution compared to the control solution. Glucose absorption was higher than the peptide absorption in all patients: Group I: 66±10% versus 57±13% (p = 0.0003); Group II: 80±5% versus 72±11% (p = 0.006). No differences in plasma amino acid profiles could be detected.Conclusion Short-chain polypeptides are absorbed less than glucose and can be used as an osmotic agent in CAPD patients. However, longer-term studies are needed to evaluate possible additional effects of peptides on the nutritional status of CAPD patients.

Objective To evaluate the changes in biocompatibility of peritoneal dialysis solutions during intraperitoneal dwell.Design We studied the effect of the drained dialysates at time 0 and after 30, 60, 120, 240, and 360 minutes of intraperitoneal dwell on the growth of peritoneal mesothelial cells and fibroblasts and the synthesis of proteins by these cells. On one day the patients were dialyzed with glucose-based Dianeal and on alternate days with an amino acid-containing solution based on Travasol.Patients Dialysates were collected from 4 patients during continuous ambulatory peritoneal dialysis (CAPD) training.Results Unused dialysis solutions containing glucose or amino acids inhibit growth of mesothelial cells and fibroblasts. Dialysates obtained after 30 or 60 minutes of intraperitoneal dwell support the growth of these cells in a way similar to 10% fetal calf serum, but dialysates drained after a longer dwell of 120 360 minutes had a stronger effect on growth of these cells than did serum. All glucose-based dialysates stimulate the synthesis of collagen in mesothelial cells, whereas they reduce the synthesis of non-collagen proteins. All glucose-based dialysates reduce the synthesis of collagen and noncollagen proteins in fibroblasts compared with the production of these proteins in the presence of serum.Conclusion Changes in the properties of the dialysis solutions during their intraperitoneal dwells do not seem to increase their biocompatibility. Indeed, excessive mitogenic effect and the stimulation of collagen synthesis of the dialysates may induce pathological changes in the peritoneum.

Objective To study how the presence of osmotic solutes in medium affects growth of the peritoneal mesothelial cells and fibroblasts and how osmotic solutes influence the production of factors regulating growth of these cells.Design The proliferation of mesothelial cells and fibroblasts was evaluated by measuring the incorporation of 3H-thymidine into the cells. Cells were exposed to osmotic solutes; the concentration of the latter in the medium was continuously lowered over the time of the experiment to simulate changes of their concentration in the dialysate. The synthesis of factors influencing the proliferation of the mesothelial cells or fibroblasts, by mesothelial cells or fibroblasts themselves, or by peritoneal leukocytes, was tested by the characteristics of the “conditioned” medium. The conditioned medium was produced by exposing standard medium to mesothelial or fibroblasts monolayer or to peritoneal leukocytes over 24 hours; following filtration it was applied to growing test cells for the study of growth factors.Results The effect of osmotic solutes on the growth of mesothelial cells is less inhibitory when their concentration is gradually lowered over the time of the study, compared to previous findings with a constant concentration. Peritoneal leukocytes produce growth factors for mesothelial cells and fibroblasts. Glucose and amino acids inhibit production of peritonealleukocyte-derived growth factors for mesothelial cells, while glycerol increases synthesis of such growth factors for fibroblasts. Mesothelial cells produce factors stimulating the proliferation of mesothelial cells and fibroblasts. In the presence of glycerol or amino acids synthesis of mesothelium derived growth factors for fibroblasts is augmented. Finally, fibroblasts produce factors that inhibit the proliferation of the mesothelial cells, and this effect is potentiated in the presence of amino acids.Conclusions Cytotoxicity of the osmotic solutes measured by the inhibition of growth of the mesothelial cells or their increased damage is significantly reduced during in vitro kinetic study when the concentration of these solutes is gradually lowered. Presence of osmotic solutes in the medium affects synthesis of growth factors derived from mesothelium, fibroblasts, or peritoneal leukocytes, which affect the proliferation of mesothelial cells or fibroblasts.

Objective To assess effects of the inflammatory cytokines (IL-1-beta, TNF-alpha, TGF-beta 1) and dialysate effluent on synthesis of hyaluronic acid by human peritoneal mesothelial cells (HMC) in in vitro culture.Methods Dialysate effluent was collected after the overnight dwell of DianeaI 1.5% from patients during CAPD training. HMC were obtained from omentum from nonuremic donors or were harvested from the dialysate effluent from CAPD patients. Synthesis of hyaluronic acid was studied on monolayers of HMC, which were deprived of serum 48 hours priortoexperiment. Effects of cytokines were tested in a medium with low serum concentration (0.1%) or in medium mixed (1:1 v/v) with the autologous dialysate. Hyaluronic acid level in medium was measured with radioimmunoassay.Results Cytokines enhanced synthesis of hyaluronic acid by HMC, and the strongest effect was induced by IL-1. Effluent dialysate stimulates synthesis of hyaluronic acid stronger than 10% FCS. Effluent dialysate and IL-1 synergistically enhance synthesis of hyaluronic acid by HMC.Conclusion Effluent dialysate from CAPD patients stimulates production of hyaluronic acid by HMC and acts synergistically with cytokines.

Objective To study the mechanism(s) of potassium transport into human mesothelial cells (HMC) exposed to osmotic solutes.Design Using potassium analog 86Rb, we evaluated its intracellular transport through three pathways: 1. blocked by ouabain; 2. blocked by furosemide but not by ouabain; 3. blocked by neither furosemide nor ouabain. Experiments were performed in a normotonic medium (control) or in a medium supplemented with osmotic solutes (glucose, glycerol, mannitol). Both the acute and chronic effects of osmotic solutes on potassium transport were studied.Results The acute exposure of mesothelial cells to osmotic solutes modifies the intracellular transport of potassium through all studied channels, and the effect is specific for every solute. In mesothelial cells exposed over 7 days to glucose (90 mM), the intracellular transport via ouabain and furosemide-blocked channels is decreased, whereas it is increased through the third pathway. Total intracellular accumulation of 86Rb (potassium) ions in mesothelial cells cultured in a medium supplemented with various concentrations of glucose is decreased, and this effect is proportional to the concentration of glucose in the medium.Conclusions The intracellular transport of potassium in mesothelial cells is regulated through at least three independent mechanisms. Acute or chronic exposure of mesothelial cells to a hypertonic medium affects the intracellular accumulation of potassium, an d this effect is specific for the various osmotic solutes.

A GLC method for the determination of meprobamate in water, plasma, and urine is described. The procedure is based on alkaline hydrolysis of meprobamate followed by silylation of the hydrolysis product to give a trimethylsilyl derivative. Quantitation of the drug was effected using a homolog of the hydrolysis product as an internal standard, which is also silylated during the procedure. The method is specific, sensitive, and reproducible and has been used for the routine analysis of biological samples obtained from meprobamate-treated dogs and humans.

The disposition kinetics of tri-(2-ethylhexyl)trimellitate (TEHTM), a new plasticizer for polyvinyl chloride (PVC) plastic, was studied in rats following intravenous administration of (14C-carbonyl]tri-(2-ethylhexyl)trimellitate using an oil in water emulsion as the vehicle. The distribution half-life, elimination half-life, and clearance values estimated from the plasma concentration of radioactivity data obtained following iv administration of 10.5 mg/kg of TEHTM (59.9 μCilkg), were 46.2 min, 5.34 d, and 40.5 ml/kg • h, respectively. Following iv dosage of 15.6 mg/kg of TEHTM (28.0 μCilkg), significant accumulation of radioactivity was found in the liver, lungs, and spleen, with liver accounting for 72% of the administered dosage at 24 h. Excretion of TEHTM and its biotransformation products was slow, with 21.3% of the administered radioactivity found in the feces and 2.8% in the urine during the 14-d collection period. Biliary excretion seems to be the major route of elimination of TEHTM. The pharma-cokinetic data gathered in the present investigation are compared to di-(2-ethylhexyl)phthalate (DEHP), a widely used plasticizer for PVC.

Objective To assess the in vitrosynthesis rate of hyaluronan (HA) by human peritoneal mesothelial cells and peritoneal fibroblasts in the presence of effluent dialysate from continuous ambulatory peritoneal dialysis (CAPD) patients.Methods We used primary cultures of human peritoneal mesothelial cells and peritoneal fibroblasts from nonuremic patients to study the effect of interleukin-1 β (11–1 β) and pooled effluent dialysate, from noninfected and infected CAPD patients, on the synthesis of HA by the studied cells. We also tested the effect of the exogenous HA on the synthesis rate of that glycosaminoglycan. We studied the correlation between HA concentration in effluent dialysate and the stimulatory effect of that solution on in vitrosynthesis of HA by mesothelium.Results Peritoneal fibroblasts produce more HA than mesothelial cells. Noninfected effluent dialysates or dialysates from CAPD patients with peritonitis stimulate synthesis of HA by mesothelial cells and fibroblasts. Interleukin-1 β has a stimulating effect, which was synergistic with effluent dialysates, on the synthesis of HA by mesothelium and peritoneal fibroblasts. A weak correlation was demonstrated between the level of HA in effluent dialysate and the stimulatory effect of that dialysate on in vitrosynthesis of HA by mesothelial cells.Conclusions Peritoneal fibroblasts are a more potent source of HA than are mesothelial cells, but probably the latter are the main source of HA in drained dialysate. Although effluent dialysates contain factors that stimulate the production of HA by mesothelium, there is weak correlation between that stimulatory effect and the actual HA concentration in the dialysate, which, in some patients, might suggest low “responsiveness” of the membrane.

Surface tension and light-scattering studies on aqueous solutions of sodium glycocholate gave CMC values of 0.0129 and 0.0138 mole/1., respectively. Light-scattering studies in 0.1 MNaCl gave a weight-average micellar weight of about 1950, indicating that four monomers of glycocholate aggregate to form the primary micelle. Preferential broadening of the angular methyl signals in the NMR spectra suggests that glycocholate micelles are formed by back-to-back hydrophobic association of the steroid nuclei. The signal due to the glycine methylene protons in the hydrophilic region of the molecule showed little broadening. Solubility studies showed that testosterone is solubilized by glycocholate below the apparent binary CMC. The solubilizing capacity is quite low. The complex association between testosterone and glycocholate suggests mixed micelle formation.

A method is proposed for determining the bioavailability of drugs whose elimination from a one-compartment body model occurs by one or more apparent first-order processes in parallel with one capacity-limited elimination process. Plasma concentration-time data obtained after intravenous administration of the drugs are necessary to calculate the equation parameters(K1, Km,andVm)needed to measure the extent of absorption after oral administration and to calculate the absorption rate constant using a modified Wagner-Nelson equation. Using simulatedC,tdata,it is shown that the error involved in calculating the fraction of a dose absorbed after oral administration by assuming linear kinetics is a function of the absorption rate constant and the extent of absorption.