A novel configuration of bioartificial liver support system based on circulating microcarrier culture

The purpose of this investigation is to initiate a new bioartificial liver support system that utilizes circulating microcarrier cultures in the extracapillary space of a hollow fiber cartridge. The material exchange occurs on the membranes of the hollow fiber. Toxins are metabolized by the circulating cells on the microcarriers driven by a centrifugal pump. We inoculated 2-3 x 10E8 Hep G2 cells on 2.5 grams of Cytodex 3 microcarriers, and allowed them flowing in the extracapillary space of a modified plasma filter. 10% FCS Medium was pumped through the capillaries at different rates. Cells keep morphological integrity and functionality during the circulation. These preliminary results suggest that this configuration of a bioartificial liver support system offers a future investigation.     

Malignant B cells from patients with primary central nervous system lymphoma express stromal cell-derived factor-1

Although the pathogenesis of primary central nervous system lymphoma (PCNSL) remains unclear, it is hypothesized that specific chemokine-chemokine receptor interactions may attract malignant B lymphocytes into the CNS. Formalin-fixed, paraffin-embedded brain biopsy specimens from 40 patients with PCNSL were immunostained by an indirect immunohistochemical method incorporating antigen retrieval to detect the presence of B-cell chemokines, stromal cell-derived factor-1 (SDF-1; CXCL12) and macrophage inflammatory protein-3alpha (MIP-3alpha, CCL20), and the SDF-1 receptor, CXCR4. To assist in phenotyping of SDF-1 + cells, specimens were also stained for CD20 (B cells). Positive staining for SDF-1 was identified in all PCNSL cases and in tonsil. In biopsy specimens, SDF-1 expression was localized to resident brain cells and, in 80% of specimens, CD20+ malignant lymphocytes. Tumor cells also stained positively for CXCR4. In contrast, although expression ofMIP-3alpha was detected in tonsil, no expression of this chemokine could be demonstrated in PCNSL biopsy specimens. Our observations raise the possibility of targeting the SDF-1-CXCR4 signaling pathway as a potential treatment for PCNSL.     

Patient safety technology for microadsorbent systems in extracorporeal blood purification

Alternative technologies for extracorporeal blood purification systems based on microadsorbents in suspension are discussed. Principally, microadsorbents offer higher efficiency and flexibility when compared to conventional column-based adsorption systems. Systems already clinically employed (e.g., BioLogic DT) or close to clinical application (e.g., the microspheres-based detoxification system, MDS) are described. The MDS technology, in particular, is characterized by efficiency and a high degree of flexibility with respect to both the use of different adsorbents as well as the combination with hemodialysis/hemofiltration therapy. It was designed for continuous use in intensive-care units, but enables also the removal of low-density lipoprotein, fibrinogen, autoimmune antibodies, immune complexes, and other pathophysiologically relevant substances. Alternative anticoagulation regimes and safety systems on fluorescence sensor technology have recently been developed for the MDS and are presented in this paper.     

On-line hemodiafiltration does not induce inflammatory response in end-stage renal disease patients: results from a multicenter cross-over study

BACKGROUND: On-line hemodiafiltration (HDF) represents the supreme blood purification modality for end-stage renal disease (ESRD) patients. Large-volume infusion of on-line prepared substitution fluid may, however, expose patients to inflammatory contaminants. As a result, on-line HDF might aggravate chronic inflammation, which correlates with malnutrition, cardiovascular disease, and mortality among ESRD patients. METHODS: In a multicenter cross-over study, 27 ESRD patients were randomly assigned to treatment with on-line HDF and low-flux hemodialysis (HD). After 6 months, patients were crossed to the other treatment modality, and treatment continued for another 6 months. Both on-line HDF and low-flux HD were conducted with polysulfone membranes and ultrapure dialysis fluid. Samples were drawn at the end of each treatment period. RESULTS: Inflammatory parameters were elevated in the study population when compared to healthy controls. Induction of interleukin-1 receptor antagonist (IL-1Ra) and tumor necrosis factor alpha (TNF-alpha) was comparable for on-line HDF and low-flux HD, and there was no intradialytic increase in cytokine production. As a result, interleukin-6 (IL-6) plasma levels did not differ significantly between the two treatment modalities. Similarly, no difference between on-line HDF and low-flux HD was observed for C-reactive protein (CRP) and albumin. Markers of endothelial cell activation (soluble intercellular and vascular cell adhesion molecules sICAM-1 and sVCAM-1) as well as the cardiovascular risk marker cardiac troponin T (cTnT) remained elevated compared to healthy subjects, but showed no difference between the two treatment modalities. CONCLUSIONS: On-line HDF, as the most effective renal replacement therapy, does not provoke inflammatory response and is both safe and highly biocompatible.     

Microspheres based detoxification system: in vitro study and mathematical estimation of filter performance.

Because of the closed plasma (secondary) circuit in the Microspheres based Detoxification System (MDS), a convective blood purification system, the same amount of filtrated plasma is backfiltrated into the blood circuit. Therefore, there is no direct way to determine the ultrafiltrate production rate, which is an important factor of efficiency. The only possible way to estimate the filtration properties of the filter is to consider pressure values. In this study the pressure distribution in the filter was investigated in vitro. To explain the results and to calculate inaccessible parameters, a mathematical model was established which also considered the asymmetric behaviour of the filter membrane. The result was a linear pressure gradient, agreement with the measurements was reasonably good (calculated primary pressure loss differs <13% from measured value when using mean measured filter resistance as model parameter). Linear pressure distribution offers the possibility of easily calculating the filtration length, a parameter which can be used to estimate the filter condition. The comparison between calculated filtration and backfiltration rates offers an instrument of control for these values.     

K-antigen identification, hemolysin production, and hemagglutination types of Escherichia coli O6 strains isolated from patients with urinary tract infections

Serotyping of 1918 Escherichia coli strains isolated in significant cell numbers from the urine of patients with urinary tract infections (UTI) revealed the presence of 117 O6 strains. The K antigens were identified by means of K-specific phages and serological methods. The phages used included a K1 phage pool (phi 1, A-E) and the separate phages phi 2, phi 5, phi 7, phi 12 and phi 13. The presence of H antigens, type 1 fimbriae formation, hemolysin production and mannose-resistant hemagglutination (MRHA) ability with human A, sheep, calf and pig erythrocytes were also analyzed. Six different MRHA types were defined and discussed in relation to the O6:K:H serotype. Remarkably, E. coli O6 strains were found to possess a whole arsenal of virulence factors (K antigens, MRHA, hemolysin). The most common serotypes - O6:K2:H1/H- (26), O6:K5:H1/H- (35) and O6:K13:H1/H- (20) - differed from each other in some cases in both MRHA type and hemolysin production.     

Biocompatibility of a new high-permeability modified cellulose membrane for haemodialysis

The biocompatibility and solute permeability characteristicsof a high-permeability modified cellulose membrane (Hemophan-HP^®)(He-HP) were compared with those of two synthetic membranes(poly(ethylene-co-vinyl alcohol) (EVAL) and poly(acrylonitrile-co-sodiummethallyl sulphonate) (AN69)) and Cuprophan^® in a multicentre,four-way cross-over clinical trial. Cuprophan^® membranescaused significant complement activation, leukopenia, and granulocyteelas-tase release. He-HP membranes demonstrated a lesser effect,which was similar to that observed for the EVAL membrane, althoughless than that seen with the AN69 membrane. A similar orderfor the four membranes was seen for their effect on platelets.Cuprophan^® membranes provided superior small-molecule removalto the other three membranes. In contrast, Cuprophan^® wasessentially impermeable to ß₂-microglobulin, whereasHe-HP, EVAL, and AN69 allowed the removal of 60–90 mgof ß₂-microglobulin per treatment. However, a decreasein the plasma concentration of ß₂-microglobulin wasobserved only with the AN69 membrane, most probably as a resultof the ability of that membrane to adsorb proteins. Our resultsdemonstrate that high-permeability membranes of comparable biocompatibilityto some synthetic membranes can be fabricated from cellulosederivatives.     

Rapid typing of Escherichia coli K antigens using bacteriophages

E. coli capsular (K) antigens are important virulence factors contributing to the development of urinary tract infections (UTI). Serotyping of these antigens is laborious and depends on the availability of respective antisera which are difficult to prepare because of the low immunogenicity of these polysaccharide antigens. The application of specific K phages are a big advantage. Two collections of E. coli strains (500 from Rostock, 214 from Erfurt) isolated from patients with UTI have been investigated with a set of ten K specific bacteriophages including phi 1, phi 2, phi 5, phi 7, phi 9, phi 12, phi 13, phi 20, phi 30 and phi 36. The K1 antigen has been found to be the most frequent capsular antigen (16.4% in Rostock; 12.2% in Erfurt) followed by K13/K20 in Erfurt (8.9%) and K5 in Rostock (8%) and Erfurt (7.5%). The collection of phages proved to be suitable for the detection of UTI associated K antigens covering the most common O serogroups found in UTI. The method appears to be simple, non-laborious and applicable for routine use.