Pyrogen transfer across high- and low-flux hemodialysis membranes

The extent to which bacterial products from contaminated dialysate enter a patient's blood depends upon the type and permeability of the hemodialysis membrane in use. This study was performed to assess the transfer of pyrogenic substances across both high- and low-flux membranes (DIAPES, Fresenius Polysulfone, Helixone, Polyamide S). All experiments were carried out in the saline-saline model. The dialysate pool was contaminated either with purified lipopolysaccharide (LPS) (250 and 500 EU/mL) or with sterile bacterial culture filtrates (20 EU/mL), and in vitro dialysis was performed under diffusive and convective conditions. A significant transfer of endotoxin was observed for both low- and high-flux DIAPES challenged with either LPS or with bacterial culture filtrates. Under identical conditions, no transfer of endotoxins was detectable across Fresenius Polysulfone and Helixone upon challenge with purified LPS. With bacterial culture filtrates, endotoxin concentrations for Polyamide S and Fresenius Polysulfone were about 10% and 1%, respectively, of those measured for DIAPES, whereas no transfer of endotoxin was detectable for Helixone. Using an alternative assay (induction of interleukin-1 receptor antagonist, IL-1Ra, in whole blood), only the DIAPES membrane showed the passage of cytokine-inducing substances. Thus, when saline is present in both the blood and dialysate compartments (i.e., the situation during predialysis priming procedures), dialysis membranes differ profoundly with respect to their permeability to endotoxins.     

The Translated Amino Acid Sequence of an Insertion in the Hepatitis E Virus Strain 47832c Genome,But Not the RNA Sequence,Is Essential for Efficient Cell Culture Replication

The hepatitis E virus (HEV) can cause hepatitis E in humans. Recently, the occurrence of HEV strains carrying insertions in their hypervariable genome region has been described in chronically infected patients. The insertions originate from human genes or from the HEV genome itself. Although their distinct functions are largely unknown, an involvement in efficient cell culture replication was shown for some strains. The HEV strain 47832c, originally isolated from a chronically infected transplant patient, carries a bipartite insertion composed of HEV genome duplications. Here, several mutants with deletions and substitutions of the insertion were generated and tested in cell culture. Complete deletion of the insertion abolished virus replication and even a single glycine to arginine substitution led to reduced cell culture growth. A mutant encoding a frameshift of the inserted sequence was not infectious, whereas a mutant carrying synonymous codons in this region replicated similar like the wild type. Substitution of the insertion with the S17 insertion from HEV strain Kernow C1-p6 did not result in viable virus, which might indicate strain- or cell type-specificity of the insertions. Generally, the translated amino acid sequence of the insertion, but not the RNA sequence, seems to be responsible for the observed effect.     

Cytokine induction in patients undergoing regular online hemodiafiltration treatment

End-stage renal disease (ESRD) patients are known to suffer from chronic inflammation as the result of an ongoing subacute cytokine induction, which may contribute considerably to dialysis-related, long-term morbidity and mortality. Preparation of infusate from cytokine-inducing dialysis fluid and its administration in large quantities as well as the use of high-flux membranes bear the risk of aggravating the chronic inflammatory response among online hemodiafiltration (online HDF) patients. In order to assess the inflammatory risk associated with online HDF, we compared the cytokine induction profile of ESRD patients receiving either online HDF or low-flux hemodialysis (low-flux HD). Specifically, we measured spontaneous and lipopolysaccharide (LPS)-stimulated tumor necrosis factor alpha (TNFalpha) and interleukin-1 receptor antagonist (IL-1Ra) release during ex vivo incubation of whole blood. Ultrapure dialysis fluid and polysulfone membranes were used for both treatment modalities. LPS-stimulated release of TNFalpha and IL-1Ra was elevated for both online HDF and low-flux HD patients compared to healthy individuals (TNFalpha: 2,336 +/- 346 and 2,192 +/- 398 versus 1,218 +/- 224 pg/106 white blood cells [WBC]; IL-1Ra: 2,410 +/- 284 and 2,326 +/- 186 versus 1,678 +/- 219 pg/106 WBC). Likewise, spontaneous production of TNFalpha, but not IL-1Ra, was higher in online HDF and low-flux HD patients than in normal controls (37 +/- 32 and 22 +/- 19 versus 0.8 +/- 0.3 pg TNFalpha/106 WBC). There was no difference in spontaneous and LPS-stimulated cytokine release between both dialysis groups. In addition, intradialytic cytokine induction was not significant for either treatment modality as spontaneous and LPS-stimulated cytokine release were not increased postdialysis. These findings indicate that online HDF does not contribute to chronic leukocyte activation and, consequently, does not place ESRD patients at greater risk with respect to inflammatory morbidity and mortality.     

Hemagglutinating ability and fimbrial antigens of Escherichia coli strains isolated from urine

The close connection between mannose-resistant hemagglutination (MRHA) and adhesion to uroepithelial cells of urinary E. coli with regard to the pathogenesis of urinary tract infection (UTI) prompted us to examine the hemagglutinating ability of 1499 E. coli strains from urine using human blood group OP1 erythrocytes. In 317 strains (21.2%), an MRHA was found. There were no significant differences in the prevalence of MRHA related to the isolation time and admitting hospital. A correlation was found between MRHA and the presence of P fimbriae in the strains investigated. Another association appears to exist between certain O:K:H serovars and distinct fimbrial antigens which had been serologically identified. The F11 antigen was detected most frequently and proved to be present in strains of serovars O1:K1:H-, O1::K1:H7, O2:K1:H-, O2:K1:H4, O2:K1:H7, and O15:K1:H7. The F8 antigen was strongly associated with serovar O18:K5:H-. O18:K5:H1 and O6:K5:H1 were apparently related to cross-reacting F14 antigens.     

Particle leakage in extracorporeal blood purification systems based on microparticle suspensions

AIM: The newly developed 'Microspheres based Detoxification System' (MDS) designed for any extracorporeal adsorption therapy uses microparticles as adsorbents characterized by a size of 1-20 microm in diameter which are recirculated in the secondary (filtrate) circuit connected to a hollow fiber filter located in the primary (blood) circuit. In the case of a leakage or rupture in the hollow fiber filter, microspheres can enter patients' blood circuits and cause embolic episodes in different organs with varying degrees of clinical relevance. Aim of this study was to determine the amount of particles infused to a patient during a long-term treatment under different failure conditions of the filter. METHODS: The filters were prepared by cutting single hollow fibers. Fresh-frozen plasma (FFP) and a mixture of glycerol and water were used as a medium together with microparticles potentially used in the MDS. The amounts of particles transferred from the filtrate into the primary circuit were measured. RESULTS: The analysis of particle transfer in the case of a single cut hollow fiber inside the membrane results in particle volumes of up to 26 ml calculated for 10 h. CONCLUSION: Particle leakage in microparticle suspension based detoxification systems can lead to considerable particle transfer to the patient. Therefore, a particle detection unit which is able to detect critical amounts of particles (<1 ml particle volume/treatment) in the extracorporeal blood line is necessary for patient safety.