Acute transverse myelitis caused by Coxsackie virus B5 infection.

A 6-year-old boy developed symptoms of rapidly progressive paraplegia, associated with bowel and urinary dysfunction, but without sensory loss. Magnetic resonance imaging (MRI) examination showed diffuse swelling of the lower spinal cord on T1-weighted images. Based on the clinical presentation and MRI findings, a diagnosis of acute transverse myelitis was made. The serum titer of neutralizing antibody against Coxsackie virus B5 rose from 1/4 on admission to 1/256 1 month later and Coxsackie virus B5 was isolated from stool samples. This case serves as a reminder that acute transverse myelitis can be a rare clinical manifestation of Coxsackie virus B5 infection.

Evaluation of several saccharides as osmotic agent for peritoneal dialysate.

OBJECTIVE: The carbonyl group of glucose reacts non-enzymatically with the amino group of protein to form advanced glycosylation end-products (AGEs). AGEs are found in the peritoneum of continuous ambulatory peritoneal dialysis patients, and this AGE formation is suspected to be one of the causes of impaired peritoneal function. In order to control AGE formation in the peritoneum, AGE formation and ultrafiltration in rats were examined with peritoneal dialysates using as osmotic agents saccharides that lack a carbonyl group, the saccharic acid lactobionate [molecular weight (MW) 358.30], the sugar alcohol maltitol (MW 344.32), and the nonreducing sugar nistose (MW 666.58). DESIGN: Bovine serum albumin (BSA) (25 mg/mL) was incubated with 18, 36, and 72 mg/mL maltitol, lactobionate, nistose, and glucose at 37 degrees C. After 3 or 6 weeks, amounts of furosine and N-(carboxymethyl) lysine were measured. A 20-mL intraperitoneal injection of a lactate-based dialysate (osmotic pressure 388 mOsm/kg) containing 4.34% maltitol, 4.52% lactobionate, or 8.4% nistose was given to Sprague-Dawley rats and, after 2, 4, or 6 hours, the quantity of effluent and levels of urea nitrogen and creatinine in the effluent and in serum were measured. RESULTS: No increases in furosine or N-(carboxymethyl) lysine were seen with maltitol, lactobionate, or nistose after 3 and 6 weeks of incubation with BSA; AGE formation was not observed. In the study in rats, the quantity of abdominal fluid after a 6-hour dwell was nistose > lactobionate > maltitol > glucose. No differences in dialysate-to-plasma ratios for urea nitrogen or creatinine were seen in any group. CONCLUSIONS: AGE formation in peritoneal tissue might be controlled by using saccharides with a modified carbonyl group as osmotic agents for peritoneal dialysates. Nistose is considered to yield the most efficient dialysis.

Suppression of N(epsilon)-(carboxymethyl)lysine generation by the antioxidant N-acetylcysteine.

OBJECTIVE: Accumulated evidence suggests that N(epsilon)-(carboxymethyl)lysine (CML), which is a dominant antigen of advanced glycation end-products (AGEs), is generated in the peritoneal cavity of patients undergoing continuous ambulatory peritoneal dialysis (CAPD), and that this process may be involved in the pathophysiology of the peritoneal injury found with CAPD treatment. Since CML is a sequential product of glycation and oxidation processes, CML generation could be suppressed by antioxidants. The aim of this in vitro study was to clarify the effect of N-acetylcysteine (NAC), an antioxidant, on CML generation from proteins under high glucose settings mimicking peritoneal dialysis solutions. DESIGN: Test proteins (bovine serum albumin/type I collagen) were incubated continuously for 16 weeks in glucose solutions (200 mmol/L) with or without NAC (2 mmol/L), and the generation time courses (8 and 16 weeks) of CML and furosine (the biomarker of the glycation products of the early Maillard reaction) were determined. RESULTS: In both proteins, furosine and CML were progressively generated in accordance with the duration of the incubation period. No apparent differences were found between solutions with and without NAC in furosine levels at the 8th and 16th weeks. However, the generation of CML was lower in the solution with NAC throughout the test periods. CONCLUSION: The results showed that NAC could suppress the generation of CML. This indicates the therapeutic potential of antioxidants for the glycoxidative stress-related peritoneal injury occurring during CAPD.

Saccharides as osmotic agents in peritoneal dialysate: determination of molecular weight essential for more efficient fluid removal.

Glucose has widely been used as an osmotic agent in continuous ambulatory peritoneal dialysis (CAPD) but owing to its low molecular weight (MW), ultrafiltration (UF) in a 6-hour dialysate exchange does not appear sufficient. Thus, in the present study, a search was made to find saccharides that would ensure optimal UF, exceeding that obtained with glucose in a 6-hour dwell period. Rats were intraperitoneally infused with peritoneal dialysate (20 mL/body) containing the same molar concentrations (75.5 mM, 126 mM, or 204 mM) of various osmotic agents including glucose (monosaccharide, MW = 180.16) at concentrations of 1.36%, 2.27%, and 3.86%; maltose (disaccharide, MW = 342.30) at concentrations of 2.58% and 4.32%; maltotriose (trisaccharide, MW = 504.44), at concentrations of 3.81% and 6.36%; and maltopentaose (pentasaccharide, MW = 828.72) at concentrations of 6.26% and 10.4%. Intraperitoneal-fluid volume was measured at 1, 2, 3, 4, and 6 hours. With glucose as the osmotic agent, maximal UF was noted at 2 to 4 hours. UF was more efficient at 6 hours with maltotriose or maltopentaose as the osmotic agent. Even with a trisaccharide or pentasaccharide (molecular weights of 500 to 830), UF at 6 hours was found more efficient, as evidenced by the delay in peritoneal absorption. Fluid removal in 6 hours should be greater when using a dialysate containing oligosaccharides such as maltotriose or maltopentaose in place of glucose.

In vitro and in vivo generation and kinetics of glycosylation products in peritoneal dialysis effluents.

There are indications that advanced glycosylation end-products (AGEs) may affect in some manner the functions of the peritoneum. Only a few reports discuss the actual generation of glycosylated proteins and the intraperitoneal kinetics involved during continuous ambulatory peritoneal dialysis (CAPD). To demonstrate the formation of AGEs and their time-courses in peritoneal dialysis (PD) effluents, measurements were made of furosine, carboxymethyl lysine (CML), and pentosidine as glycosylation markers in vitro and in vivo using PD effluents obtained every 2 hours for up to 8 hours from 3 nondiabetic CAPD patients. Furosine and CML were found to be generated relatively early (within 6 hours), and their de novo formation was markedly enhanced subsequent to glucose addition. Furosine and CML production increased in proportion to glucose concentration. Pentosidine production did not change with time for up to 24 hours, and was not induced by glucose during this period. Carboxymethyl lysine production appeared to be suppressed in vitro with the addition of serum protein. Furosine in the intraperitoneal dialysate was initially the same as that in the plasma, but increased to twice as much in just 2 hours in vivo. Production of CML increased, but apparently was suppressed with leakage of plasma protein into the dialysate; this possibly may have been due to dilutional or antiglycation effects in the plasma. No stimulation of pentosidine production could be detected in the intraperitoneal dialysate even at 8 hours. The initiation of glycosylation of peritoneal proteins, with generation of furosine and CML but not pentosidine, is clearly shown by the present results to occur relatively early in the intraperitoneal dialysate, i.e., within a matter of hours.

Kinetic analysis of furosine and pentosidine in CAPD patients.

Advanced glycation end products (AGEs) have been noted in the peritoneal tissue of continuous ambulatory peritoneal dialysis (CAPD) patients, and this may cause an increase in membrane permeability. In vivo and in vitro kinetic analysis was carried out on furosine and pentosidine, early and late glycation products. Plasma furosine and pentosidine were measured by HPLC in patients with renal dysfunction with or without diabetes mellitus (DM) and dialysate pentosidine and furosine in CAPD patients. Only those dialysis patients without residual renal function were used in this study. Plasma furosine was remarkably high in DM, hemodialysis (HD), and CAPD patients. Plasma pentosidine appeared to depend on renal function and was not influenced by diabetic condition. Plasma pentosidine was significantly higher in CAPD than HD patients. A weak positive correlation was noted between dialysate and plasma furosine and pentosidine, indicating the main source of furosine and pentosidine in PD effluent to be plasma. Serial dialysate sampling showed furosine and pentosidine to increase linearly. Mean dialysate/plasma (D/P) of furosine and pentosidine were 0.043 and 0.012, respectively. Protein-bound product size and in situ formation of furosine in the peritoneal cavity would be the reason for these differences in D/P. In situ formation of early glycation products in the peritoneal cavity may be concluded to take place in CAPD patients, and high plasma pentosidine may lead to its accumulation in tissue, resulting, possibly, in pathological change.

Forskolin derivatives. I. Synthesis, and cardiovascular and adenylate cyclase-stimulating activities of water-soluble forskolins.

Water-soluble forskolin and 7-deacetylforskolin derivatives with an aminoacetyl, a 3-aminopropionyl, or a 4-aminobutyryl group at the 6- or 7-position were prepared, and their positive inotropic as well as vasodilative activities were evaluated in anesthetized dogs. 7-Deacetylforskolin (2) and 7-deacetyl-1-silylforskolin (6) were converted to the corresponding 7-chloroacylderivatives (3, 7, 10), which were reacted with amines to obtain 7-aminoacyl-7-deacetylforskolins (4a-f, 9a, b, 11). The 7-acyl substituents migrated to the 6-position with sodium hydroxide in acetonitrile-water to afford 6-aminoacyl-7-deacetylforskolins (12a-f). The 7-position of 12a, d-f was selectively acetylated with acetyl chloride to obtain the corresponding 6-aminoacylforskolins (13a-d). Among the 6-aminoacylforskolins, 6-(3-dimethylaminopropionyl)forskolin (13b) and 6-(4-dimethylaminobutyryl)forskolin (13d) exhibited potent positive inotropic and vasodilative activities comparable to those of forskolin (1). The activities of 13b and 13d were approximately ten times more potent than those of 7-aminoacyl- and 6-aminoacyl-7-deacetylforskolins (4a-f, 9a, 12a-c, f). 6-Dimethylaminoacetylforskolin (13a) and 6-(3-diethylaminopropionyl)forskolin (13c) were less potent than 1. The effects of the soluble forskolins on adenylate cyclase activity were also examined in vitro. 6-Aminoacylforskolins (13a-d) exhibited potent adenylate cyclase-stimulating activity, comparable to that of 1.

Incorporation of bile acid of low concentration into model and biological membranes studied by 2H and 31P NMR.

We have analyzed the manner of incorporation of bile acid into lipid bilayers and resultant perturbation of the bilayer structure with lower bile acid/lipid ratios relevant to the physiological conditions (approximately 1 mM) by 2H and 31P NMR methods, as an aid to understanding the possible role as an endogenous tumor promoter in colon cancer besides the primary physiological function of solubilizing lipids. On the basis of the 2H quadrupole splittings of [6,6,7,7,8-2H5]deoxycholate and [11,11,12,12-2H4]chenodeoxycholate in the presence of lamellar multibilayers of egg yolk lecithin, these bile acids were found to be incorporated in such a manner that the B-D rings lie parallel with the normal of the bilayers when the ratio of the bile acid to lipid is low (less than 0.11). When the ratio is increased, these bile acid molecules are not dispersed entirely in the bilayer but aggregate to form micelles with lipids. Further, we studied the resultant perturbation of the multibilayers of egg yolk lecithin analyzed by using the 2H quadrupole splitting of [18,18,18-2H3]stearic acid as a probe and by 31P chemical shift anisotropy. We found that the bilayer structure is retained even at the bile acid-to-lipid ratio of 0.25, although a small amount of an isotropic phase appeared such as small vesicles and micelles. The molecular ordering of fatty acyl chains was rather enhanced by the presence of 1 mM deoxycholate in erythrocyte ghosts as seen from the 2H quadrupole splitting of [16,16,16-2H3]palmitic acid, although deoxycholate caused hemolysis in this condition. The former observation can be explained by the way the lipid-protein interaction is modified by deoxycholate located in the interface between the lipids and proteins.