PubMed · 15949141
The source did not provide an abstract. Follow the original record for more information.
Explore connections, maps & timelines
Ulrich Seydel. 2005. Editorial.. https://doi.org/10.1179/096805105x46600
Cite the original work for its findings. Save a collection to share your selection of sources.
KEEP EXPLORING
BACKGROUND: Applications of fluorescence-activated cell sorting (FACS) are ideally performed under aseptic conditions so that isolated cells can be successfully cultured, transplanted, or processed for the isolation of protein and nucleic acids. However, modern "off-the shelf" flow cytometers are suboptimally designed for these purposes because nonsterile instrument hardware components directly contact sample-harboring fluids, compromising their sterility. METHODS: We have described the design and modular modification of a cytometer with a sterile and disposable FACS fluid handling system that meets requirements of high-speed FACS and good manufacturing practice. This system was tested for functionality and its ability to maintain a clean and sterile fluid environment. RESULTS: Our data have shown that this new fluidic subsystem completely replicated the intended function of the manufacturer's standard fluid handling system, and isolates the fluid from contaminants such as bacteria and fungus, endotoxins, mycoplasma, and helicobacter. CONCLUSIONS: FACS has emerged as a powerful tool used to study and manipulate stem cells. However, if stem cell discoveries are to be fully utilized in clinical transplant medicine, aseptic instrument configurations must be developed. For this purpose, we have designed a disposable sterile fluid handling system.
Studies aimed at dissecting the complex pathophysiology of sepsis with multiple organ failure have traditionally focused on lipopolysaccharide of gram-negative bacteria, which is widely regarded as the classical endotoxin. However, gram-positive sepsis now accounts for up to 50% of all cases, calling for a shift of focus. Peptidoglycan (PepG) is the major cell wall component of gram-positive bacteria and has been increasingly recognized as an important proinflammatory molecule. During gram-positive infections, PepG reaches the circulation by bacterial breakdown or translocation from the intestine. Administration of PepG induces all the classical features of infectious illness and endotoxemia and may cause systemic inflammation with organ failure in animal models. Its potency, however, is crucially dependent on various features of its complex structure. PepG interacts with the innate immune system through receptors mainly expressed on monocytes/macrophages but may induce inflammatory changes in other cell types as well. Among the most extensively studied receptor systems are the nucleotide-binding oligomerization domains, the toll-like receptors, and the PepG recognition proteins. Based on the current available literature, we would like to propose that PepG must be regarded as an endotoxin in its own right and to encourage further work in the field of PepG signaling.
BACKGROUND: On-line hemodiafiltration is gaining popularity due to increasing evidence of clinical benefits however it also requires strict attention to hygiene and safety as notable quantities of liquid are reinfused into the patient. Although most centers are improving their attention to water quality, a frequent concern is the inadvertent or accidental contamination of water and whether the redundant safety controls are sufficient to protect the patient. In the present study, in order to simulate a worst-case safety condition, we tested in vitro the reliability of paired hemodiafiltration - (PHF), under low, moderate and high bacterial contamination of the water supply. Tests were performed using various bacterial concentrations (range 85-2000 cfu/mL) of Pseudomonas Aeruginosa. Samples were analyzed from different sites throughout the entire on-line hemodiafiltration circuit for bacteria endotoxin, fungus and ability to stimulate whole blood production of TNFalfa. RESULTS: In the in vitro contamination study, with the three bacterial concentrations tested at various points of the circuit, bacteria were below the level of detection and endotoxins were < 0.01 UE/mL. Addition of dialysate samples taken after the first stage of microfiltration, as well as after the first and second stage of ultrafiltration and incubated with whole blood were not associated with stimulated production of TNFalfa . CONCLUSIONS: PHF appeared to be a safe and feasible method for on-line hemodiafiltration even in the unforeseen presence of bacterial contamination of the feed water or water distribution system.