ABSTRACT
BACKGROUND: Surgery is challenging during the coronavirus disease 2019 (COVID-19) pandemic. This study aimed to investigate the preoperative severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing in elective and emergency surgery and to calculate the patient contacts during hospital stay. MATERIALS AND METHODS: All surgeries defined by the German procedural classification (starting with a 5) from 1 June until 29 November 2020 were retrospectively evaluated regarding the preoperative SARS-CoV2 nasopharyngeal swab test. The results were then divided in emergency and elective surgeries. To show the personal contacts of the patients in a university hospital, we calculated the patient pathway within the department of urology and urosurgery for April 2020. Therefor we used the electronic patient records. RESULTS: Altogether 7745 surgical procedures in 5985 patients were performed, whereby 39 (0.5%) SARS-CoV2 tests were positive. 2833 (37%) surgical procedures were emergency cases and 4912 (63%) were elective procedures. 25 (0.9%) of the emergency group and 14 (0.3%) of the elective surgeries had a positive SARS-CoV2 test. The average number of contacts in the patient room was 12.83 (0-50) and 84.22 (0-249) at the ward level, not counting contacts with the clinic staff. CONCLUSIONS: Nearly 1% of the preoperative SARS-CoV2 tests of either emergency or elective surgeries tested positive in the 6 months prior to November 2020. Although the risk of undetected SARS-CoV2 infection appears to be low in terms of costs and personnel, preoperative screening is useful in high-risk areas to ensure further necessary surgeries, especially concerning cancer patients and to prevent virus spread in a hospital.
Subject(s)
COVID-19 , Coronavirus , COVID-19 Testing , Humans , Pandemics , Retrospective Studies , SARS-CoV-2ABSTRACT
Multidrug-resistant organisms (MDRO) have been developing as an emerging problem in allogeneic hematopoietic cell transplantation (HCT). Since no data are available on the course of MDRO colonization after HCT, we investigated in this retrospective, single-center study, persistence and clearance of MDRO after HCT. From June 2010 to December 2015, 121 consecutive HCT patients were included. Patients received a MDRO screening before conditioning as well as surveillance cultures after HCT. In MDRO-colonized patients, surveillance specimens were taken until MDRO were no longer detectable. Thirty-three patients (27%) were found to be colonized by at least one MDRO at any time point until day 100 post HCT. Day 100 (2-year) non-relapse mortality (NRM) and overall survival (OS) of MDRO-colonized (MDRO+) versus non-colonized (MDRO-) patients were essentially the same. NRM is 15% (21%) versus 15% (24%). Two-year OS is 60 versus 55% for MDRO+ versus MDRO- patients. Out of the 33 MDRO+ patients, 21 cleared the MDRO. Median time to non-detectability of MDRO was 6 months. In 12 patients, the MDRO persisted. There was a significant (p < 0.0001) survival difference between patients who cleared the MDRO versus those with MDRO persistence (2-year OS 80 vs 40%). Except for the length of antibiotic therapy as a potential risk factor for MDRO persistence after HCT, no other conventional factors could be identified. (a) colonization by MDRO per se had no negative impact on the outcome, (b) MDRO can be cleared by the majority of patients after allogeneic HCT, and (c) to increase the probability to clear MDRO, the use of antibiotics in MDRO+ patients should be reviewed critically.
Subject(s)
Drug Resistance, Multiple, Bacterial , Drug Resistance, Multiple, Fungal , Hematopoietic Stem Cell Transplantation , Methicillin-Resistant Staphylococcus aureus , Pneumocystis Infections , Pneumocystis carinii , Staphylococcal Infections , Adult , Aged , Allografts , Female , Humans , Male , Middle Aged , Pneumocystis Infections/drug therapy , Pneumocystis Infections/epidemiology , Pneumocystis Infections/etiology , Retrospective Studies , Risk Factors , Staphylococcal Infections/drug therapy , Staphylococcal Infections/epidemiology , Staphylococcal Infections/etiologyABSTRACT
Diacylglycerol (DAG) was discovered as a potent lipid second messenger with protein kinase C (PKC) as its major cellular target more than 25 years ago. There is increasing evidence of significant complexity within lipid signaling, and the classical DAG-PKC model no longer stands alone but is part of a larger bioactive lipid universe involving glycerolipids and sphingolipids. Multiple layers of regulation exist among PKC- and DAG-metabolizing enzymes such as phosphatidylcholine (PC)-specific phospholipase D, and cross-talk exists between the glycerolipid and sphingolipid pathways, with PKC at the center. Currently, there is intense interest in the question of whether DAG derived from PC can function as a lipid second messenger and regulate PKC analogous to DAG derived from phosphatidylinositol-4,5-bisphosphate (PIP2). To address these issues and incorporate DAG-PKC and other signaling pathways into an expanded view of cell biology, it will be necessary to go beyond the classical approaches and concepts.
Subject(s)
Diglycerides/metabolism , Phospholipase D/metabolism , Protein Kinase C/metabolism , Signal Transduction , Animals , Ceramides/metabolism , Humans , Phosphatidic Acids/metabolismABSTRACT
We report a patient with encephalitis who had been diagnosed with an unspecific aetiology. During follow-up, pneumonia was identified due to Mycoplasma pneumoniae infection that could also be confirmed as causal for the brain inflammation. Despite the initially critical clinical situation, the patient's condition improved under specific antibiotic treatment. Pathophysiologic, differential diagnostic, and therapeutic implications are discussed, and guidelines for diagnosis are proposed.
Subject(s)
Meningoencephalitis/diagnosis , Military Personnel , Mycoplasma Infections/diagnosis , Mycoplasma pneumoniae , Adult , Anti-Bacterial Agents , Antibodies, Bacterial/cerebrospinal fluid , Antibodies, Viral/cerebrospinal fluid , Brain/pathology , Brain Stem/pathology , Cerebral Ventricles/pathology , Dexamethasone/therapeutic use , Diagnosis, Differential , Disease Progression , Dominance, Cerebral/physiology , Drug Therapy, Combination/therapeutic use , Encephalitis, Herpes Simplex/diagnosis , Encephalitis, Herpes Simplex/immunology , Humans , Magnetic Resonance Imaging , Male , Meningoencephalitis/drug therapy , Mycoplasma Infections/drug therapy , Mycoplasma pneumoniae/immunology , Neurologic Examination , Polymerase Chain Reaction , Quadriplegia/diagnosis , Quadriplegia/etiology , Thalamus/pathologyABSTRACT
BACKGROUND: Chronic alcohol ingestion leads to endotoxemia which is believed to play an important role in the pathogenesis of alcoholic liver disease (ALD). The purpose of this study was to determine if chronic ethanol consumption, in addition to affecting plasma endotoxin and cytokines, also affects the endotoxin-neutralizing capacity (ENC), sCD14, and sICAM-1, in patients with ALD. A second aim was to identify correlations between these latter parameters, endotoxin, and cytokines, especially IL-10. METHODS: Hospitalized patients with various degrees of ALD (n = 59), and 20 healthy volunteers were studied. Plasma endotoxin and ENC were determined using our kinetic Limulus amebocyte lysate test. Cytokines, sCD14, and sICAM-1 were measured by enzyme-linked immunosorbent assay. RESULTS: Patients with ALD exhibited a mild endotoxemia (p < 0.01) and a marked decrease in ENC (p < 0.0002). TNF-alpha (p < 0.05), IL-6 (p < 0.0001), sICAM (p < 0.005), and sCD14 (p < 0.0005) were significantly elevated in all patients with ALD, and IL-10 (p < 0.05) in patients with cirrhotic ALD. With the exception of IL-10, the cytokines correlated with each other and with sICAM-1. No correlations occurred between endotoxin, ENC, and sCD14, and between these and the cytokines and sICAM-1. Elevated levels of endotoxin correlate with acute excessive alcohol ingestion. No gender differences were observed. CONCLUSIONS: Acute alcohol intoxication rather than severe ALD results in significant endotoxemia. The limited capacity of plasma to neutralize endotoxin in liver injury seems to be an important factor in ALD which may be responsible for the release of endotoxin-induced mediators, such as cytokines, as well as s-ICAM-1, that are relevant in the pathogenesis of ALD.
Subject(s)
Cytokines/blood , Endotoxemia/blood , Intercellular Adhesion Molecule-1/blood , Lipopolysaccharide Receptors/blood , Liver Diseases, Alcoholic/blood , Alcoholic Intoxication/blood , Endotoxemia/etiology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Interleukin-10/blood , Interleukin-6/blood , Liver Diseases, Alcoholic/complications , Solubility , Tumor Necrosis Factor-alpha/analysisABSTRACT
Signal transduction via protein kinase C (PKC) is closely regulated by its subcellular localization. In response to activation of cell-surface receptors, PKC is directed to the plasma membrane by two membrane-targeting domains, namely the C1 and C2 regions. This is followed by the return of the enzyme to the cytoplasm, a process shown recently to require PKC autophosphorylation (Feng, X., and Hannun, Y. A. (1998) J. Biol. Chem. 273, 26870-26874). In the present study, we examined mechanisms of translocation and reverse translocation and the role of autophosphorylation in these processes. By visualizing the trafficking of wild-type as well as mutant PKCbetaII in live cells, we demonstrated that in response to cell-surface receptor activation, the function of the C1 region is required but not sufficient for recruitment of the enzyme to the plasma membrane. The C2 region is also critical in anchoring the enzyme to the plasma membrane. Furthermore, the inability of a kinase-deficient PKC to undergo reverse translocation was restored by the addition of intracellular calcium chelators, suggesting a role for the C2 region in the persistent phase of translocation. On the other hand, the inability of a C2 deletion mutant (C1 region intact) to translocate in response to agonist was reversed in mutants lacking kinase activity or by mutation of the Ser(660) autophosphorylation site to alanine, suggesting that autophosphorylation of this site is required for opposing the action of the C2 region. Therefore, the membrane-targeting function of the C1 region is facilitated by the C2 region and appears to be opposed by autophosphorylation. Taken together, these findings provide novel evidence of the functional regulation of reversible PKC membrane localization by autophosphorylation, and they show that the dynamic translocation of PKC in response to agonists is tightly regulated in a collaborative fashion by the C1 and C2 regions in balance with the effects of autophosphorylation.
Subject(s)
Protein Kinase C/metabolism , Biological Transport , Cell Line , Cell Membrane/enzymology , Green Fluorescent Proteins , Humans , Luminescent Proteins/metabolism , Phosphorylation , Receptors, Cell Surface/metabolismABSTRACT
Previous subtyping of thromboxane A2 (TXA2) receptors in platelets and vascular smooth muscle cells was based on pharmacological criteria. Two distinct carboxy-terminal splice variants for TXA2 receptors exist and they couple to several different G protein alpha subunits including Galpha13, but it has not been established whether either or both isoforms interact with and signal through it. We sought to determine: (1) which TXA2 receptor isoforms exist in vascular smooth muscle, (2) if Galpha13 is present in vascular smooth muscle and (3) if Galpha13 interacts with either or both of the two TXA2 receptor isoforms as determined by changes in ligand binding properties and generation of intracellular signals. Both TXA2 receptor isoforms and Galpha13 were found in vascular smooth muscle cells. Both the alpha and beta isoforms of the TXA2 receptors were transiently transfected with or without Galpha13 into COS-7 (radioligand binding assays) or CHO cells (agonist induced Na+/H+ exchange). Co-expression of each receptor isoform with Galpha13 significantly (P<0.05) increased the affinity of each receptor for the two agonists, I-BOP and ONO11113, and decreased the affinity of the receptor for the antagonists, SQ29,548 and L657,925. I-BOP stimulated Na+/H+ exchange in vascular smooth muscle cells. Co-expression of Galpha13 with each TXA2 receptor isoform in CHO cells resulted in a significant (P<0.04) agonist induced increase in Na+/H+ exchange compared to cells not transfected with Galpha13. The results support the possibility that the previous classification of TXA2 receptor subtypes based on pharmacological criteria reflect unique interactions with specific G protein alpha subunits.
Subject(s)
Muscle, Smooth, Vascular/metabolism , Receptors, Thromboxane/metabolism , Animals , Blotting, Western , Bridged Bicyclo Compounds, Heterocyclic/pharmacology , CHO Cells , COS Cells , Cell Line , Cloning, Molecular , Cricetinae , DNA, Complementary/biosynthesis , Fatty Acids, Unsaturated/pharmacology , GTP-Binding Proteins/chemistry , GTP-Binding Proteins/genetics , Hydrazines/pharmacology , Protein Isoforms/genetics , Protein Isoforms/metabolism , Receptors, Thromboxane/antagonists & inhibitors , Receptors, Thromboxane/genetics , TransfectionABSTRACT
A cDNA for a thromboxane A2 (TXA2) receptor was cloned from an SV40 transformed African Green Monkey kidney cell line (COS-7). The sequence is 98% homologous with the isoform of the human TXA2 receptor and has agonist and antagonist ligand binding characteristics that are not significantly different from the human receptor. Stimulation of the COS-7 cells with the TXA2 receptor agonist, ONO 11113 resulted in a significant increase in cAMP formation that was blocked by a receptor antagonist. The results raise the question of the utility of the COS-7 cell line for studies of cloned and expressed TXA2 receptor signalling mechanisms.
Subject(s)
Receptors, Thromboxane/genetics , Amino Acid Sequence , Animals , COS Cells , Cloning, Molecular , Conserved Sequence , Cyclic AMP/metabolism , DNA, Complementary/chemistry , Molecular Sequence Data , Polymerase Chain Reaction , RNA/analysis , RNA/isolation & purification , Receptors, Thromboxane/antagonists & inhibitors , Receptors, Thromboxane/chemistry , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Thromboxane A2/analogs & derivatives , Thromboxane A2/pharmacologyABSTRACT
We have studied the binding of two G-protein-regulated phospholipase C (PLC) enzymes, PLCs-beta1 and -beta2, to membrane surfaces using sucrose-loaded bilayer phospholipid vesicles of varying compositions. Neither enzyme binds appreciably to pure phosphatidylcholine vesicles at lipid concentrations up to 10(-3) M. PLC-beta1 and PLC-beta2 bind vesicles composed of phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine (molar ratio 1:1:1) with an approximate Kd of 10(-5) M. Inclusion of 2% PtdIns(4,5)P2 in these vesicles had no effect on the affinity of this interaction. As reported by others, removal of the C-terminus of PLC-beta1 and PLC-beta2 produces catalytically active fragments. The affinity of these truncated proteins for phospholipid vesicles is dramatically reduced suggesting that this region of the proteins contains residues important for membrane binding. Inclusion of G-protein alpha- and betagamma-subunit activators in the phospholipid vesicles does not increase the binding of PLC-beta1 or PLC-beta2, and the magnitude of G-protein-mediated PLC activation observed at low phospholipid concentrations (10(-6) M) is comparable to that observed at concentrations at which the enzymes are predominantly membrane-bound (10(-3) M). PLC-beta1 and -beta2 contain C2 domains but Ca2+ does not enhance binding to the vesicles. Our results indicate that binding of these enzymes to membranes involves the C-temini of the proteins and suggest that activation of these enzymes by G-proteins results from a regulated interaction between the membrane-bound proteins rather than G-protein-dependent recruitment of soluble enzymes to a substrate-containing phospholipid surface.
Subject(s)
Calcium/pharmacology , GTP-Binding Proteins/metabolism , Isoenzymes/metabolism , Liposomes , Phosphatidylinositol Phosphates/pharmacology , Phospholipids/metabolism , Type C Phospholipases/metabolism , Animals , Cattle , Humans , Kinetics , Macromolecular Substances , Phospholipase C beta , Recombinant Proteins/metabolism , Structure-Activity Relationship , SucroseABSTRACT
It has been postulated that synthetic membranes, such as polysulfone membranes, are rather impermeable for endotoxin or endotoxin fragments and can be used for sterile filtration of dialysate. It has never been investigated, however, whether endotoxin permeability may be different in commercially available polysulfone membranes. In vitro, we found a significantly different permeability for endotoxin in two standard dialyzers and one test dialyzer with high-flux polysulfone membranes. In contrast to the F-60 dialyzer with a very low permeability for endotoxin, a stepwise increasing load of endotoxin concentration in the dialysate compartment of the PN 1913 test dialyzer and Primus 1350 polysulfone dialyzer was followed by a stepwise increase of endotoxin in the blood compartment. A significant transfer across the membranes was found when the endotoxin concentration in the dialysate compartment was > 10 ng/mL in the PN 1913 and > 0.5 ng/mL in the Primus 1350. In the latter, about 0.5% of the endotoxin concentration of the dialysate compartment was found in the blood compartment. The data suggest that manufacturers have to evaluate the performance and other properties of their synthetic membranes in detail.
Subject(s)
Biocompatible Materials , Endotoxins/pharmacokinetics , Membranes, Artificial , Polymers , Renal Dialysis/instrumentation , Sulfones , Dialysis Solutions/chemistry , Endotoxins/administration & dosage , Escherichia coli , Osmolar ConcentrationABSTRACT
AIM: Of this study was to define the significance of approximate ultrasound explorations of the thyroid gland for the clinical screening for thyroid diseases. METHOD: 918 patients in the medical service of a general hospital were screened for thyroid diseases by means of clinical, functional and sonomorphological methods. A sonographic screening scan with a 5 MHz probe provided data concerning structure and dimension of thyroid gland. Maximal sagittal diameter (SDm, normal range < 18 mm) was taken as a measure for thyroid size. RESULTS: Echogenicity has no essential meaning for thyroid dysfunction screening, but is indispensable for the detection of circumscript or general echo deficiency. SDm has proved to be a simple and well reproducible measure for thyroid size. Among 41 patients with thyrotoxicosis 40 had enlarged SDm, which means a sensitivity of 97.6%. However, a high rate of false positives makes TSH-screening of all enlarged thyroid glands obligatory. Moreover, SDm allows the estimation of a lack of thyroid mass as the most common cause of hypothyroidism. CONCLUSION: We recommend that an approximate ultrasound exploration of the thyroid gland including measuring of SDm should be part of every diagnostic procedure for thyroid disease, as it is precise and not time consuming.
Subject(s)
Mass Screening , Thyroid Diseases/prevention & control , Adult , Aged , Female , Goiter, Nodular/diagnostic imaging , Goiter, Nodular/prevention & control , Humans , Hyperthyroidism/diagnostic imaging , Hyperthyroidism/prevention & control , Hypothyroidism/diagnostic imaging , Hypothyroidism/prevention & control , Male , Middle Aged , Predictive Value of Tests , Reference Values , Thyroid Diseases/diagnostic imaging , Thyroid Function Tests , Thyroid Gland/diagnostic imaging , UltrasonographyABSTRACT
At birth maternal infection with the Varizella-Zoster-Virus (VZV) is very rare but poses a truly life-threatening risk to the newborn. The neonatal mortality rate is up to 20-30%, if the maternal VZV-infection occurs between day 4 ante partum and day 2 post partum. The mortality can be decreased, if labour is successfully delayed by tocolytic agents until VZV-antibodies produced by maternal immune response have passed the placental barrier. There are indications that the mortality may also be lowered by passive immunisation of the newborn, but further research is needed. We recommend strongly to check VZV-IgG-antibodies-titres promptly, if VZV-infection is suspected. Labour then should be delayed by tocolytic agents as described in 2 case reports to allow maternal IgG-antibodies to cross the placental barrier to the foetus.
Subject(s)
Chickenpox/congenital , Pregnancy Complications, Infectious/diagnosis , Adolescent , Adult , Antibodies, Viral/blood , Chickenpox/immunology , Chickenpox/therapy , Female , Herpesvirus 3, Human/immunology , Humans , Immunity, Maternally-Acquired/immunology , Immunization, Passive , Immunoglobulin G/blood , Immunoglobulin M/blood , Infant, Newborn , Pregnancy , Pregnancy Complications, Infectious/immunology , Pregnancy Complications, Infectious/therapy , Risk Factors , TocolysisABSTRACT
A number of problems may be involved in the detection of endotoxin in plasma of patients using LAL (Limulus amebocyte lysate). When collecting blood or processing samples, contamination with endotoxin or its adsorption to material must be avoided. In our laboratory a kinetic LAL microtiter assay was developed that takes into account plasma-related interferences with the LAL endotoxin reaction by performing an internal standardization in each sample. Negative results do not absolutely exclude the involvement of endotoxins in the underlying disease. High levels of endotoxins do not necessarily reflect the severeness of the clinical status of the patient. Due to nonendotoxin-specific reactions with some complete lysates, false-positive levels may result, e.g., following immunoglobulin therapy. In spite of these limitations, the LAL test remains a valuable tool in the evaluation of gram-negative infections.
Subject(s)
Endotoxins/blood , Limulus Test , Sepsis/diagnosis , Humans , Predictive Value of Tests , Prognosis , Sepsis/bloodSubject(s)
Disabled Persons , Rehabilitation , Germany, East , Holistic Health , Humans , National Health Programs , Public Policy , Research , Research Support as Topic , UniversitiesABSTRACT
Four phytopharmaceutics (Carnivora, Pascotox forte-Injectopas, Esberitox N, Iascador M), which sometimes cause side effects after parenteral administration (fever, rigor, nausea), were examined for their endotoxin content by the kinetic turbidometric Limulus-amebocyte-lysate (LAL) microtitre test. Contaminations of over 10(5) EU/ml (endotoxin units; 1 EU = 0.1 ng of the FDA standard EC-5) were found in correlation with the clinical picture. In one preparation (Carnivora) very high endotoxin levels were always found; contaminations were only occasionally found in the others. These endotoxin measurements are supported by tests of endotoxin-dependent parameters in in-vivo experiments (reduction in leukocytes, acute death in hyperreactive mice). These findings underline the urgent need for a widening of the regulations on testing for pyrogens to include those parenteral preparations which now do not have to be tested because of their small volume (less than 15 ml).
Subject(s)
Endotoxins/analysis , Plant Extracts/adverse effects , Plant Extracts/analysis , Plant Proteins , Animals , Droseraceae , Drug Contamination , Humans , Leukocyte Count/drug effects , Limulus Test , MiceABSTRACT
Recently, there has been some concern that high-flux membranes may expose dialysis patients to the risk of endotoxin transfer secondary to backfiltration within the dialyzer. To evaluate the safety of high-flux polysulfone dialyzers, we examined in an in vitro recirculation system whether lipopolysaccharides (LPS) and lipid A respectively penetrate from the dialysate to the blood compartment and vice versa using a F-60 hemofilter (Fresenius AG). For the detection of endotoxin, a sensitive, kinetic limulus amebocyte lysate (LAL) microtiter test was used. It can be concluded that LPS and lipid A do not pass from either side through the filter system used when saline was recirculated for more than 10 h on both sides of the membrane.
Subject(s)
Blood , Endotoxins/blood , Membranes, Artificial , Renal Dialysis/adverse effects , Ultrafiltration/instrumentation , Humans , Lipid A/blood , Lipopolysaccharides/blood , Permeability , Polymers , SulfonesABSTRACT
An experimental model was used in mice in which septicemia develops following invasion of the animals' own intestinal flora after cecal ligation and puncture. Pretreatment with 1 microgram of endotoxin administered 24 hours before surgery significantly reduced the rate of lethality. Bacteria were counted and differentiated in cardiac blood at various times throughout a 48-hour period after induction of septicemia in mice, with and without pretreatment. Endotoxin levels and plasma-related interference of the Limulus-amebocyte-lysate (LAL)-endotoxin reaction also were determined as were hematologic and metabolic parameters. A shift from mixed gram-positive and gram-negative to predominantly gram-negative bacteria occurred in both groups. In pretreated mice, a reduction in aerobic bacterial counts and reduced hyperglycemia were seen in the initial phase; and a decrease in anaerobic and aerobic bacteria and in endotoxin levels were observed at the end of the experiment. This appears to be related to endotoxin-induced increased resistance against the consequences of intraabdominal sepsis. These studies also indicate that the measured amount of circulating endotoxin does not necessarily correlate to the severeness of infection. Individual differences in plasma-related interference with the LAL-endotoxin reaction also emphasize the requirement for sample-internal standardization in order to reliably quantify endotoxin in plasma.
Subject(s)
Endotoxins/pharmacology , Escherichia coli , Sepsis/physiopathology , Shock, Septic/physiopathology , Animals , Bacteriological Techniques , Blood Glucose/metabolism , Cecum/microbiology , Disease Models, Animal , Endotoxins/blood , Feces/microbiology , Female , Lactates/blood , Lactic Acid , Limulus Test , MiceABSTRACT
A turbidometric, automated limulus amoebocyte lysate (LAL) microtiter test has been developed based on the evaluation of the LAL-endotoxin reaction kinetics. The maximal increase in optical density of each reaction mixture within 1 min is recorded. With this method an endotoxin standard curve is achieved which is linear over a concentration range of six decades. With presently available LAL methods sample-related inhibition or enhancement of the LAL endotoxin reaction may be overlooked and lead to false results. The quality of interfering factors can be characterized with our methods by spiking serial dilutions of the sample with constant endotoxin concentrations. The additional introduction of an internal standardization in our system allows the determination of endotoxin with simultaneous detection of quality and quantity of sample-induced interference. This procedure is based on a mathematic model which describes interference-caused alterations of the reaction revealed by addition of endotoxin in increasing concentrations. In comparison to the LAL tube test and the turbidometric determination at a given time the advantages of the developed method are demonstrated using three different samples (gelatin solution, adenine-HCl solution and a concentrate of coagulation factors (PPSB)). These are paradigmaticly selected because and enhancement of the LAL endotoxin reaction.