摘要:

AbstractWhole blood luminol‐enhanced chemiluminescence (CL) was studied usingN‐formyl‐methionyl‐leucyl‐phenylalanine (FMLP) or opsonized zymosan particles as stimuli. The peak and the integral responses of CL were recorded. In the FMLP‐induced CL the initial activation (1‐minute values) was also studied, because it coincides with the extracellular production of oxy radicals. Correction factors based on neutrophil count and on haemoglobin concentration were found to decrease dispersion and shape the distributions of the CL responses close to normal in a study of 50 healthy adults. One‐minute values were significantly lower in women than in men but there were no significant differences for peak or integral values between sexes. Depressed reaction is in accordance with the previous findings that phagocytic oxy radical production is depressed in female plasma. Thus, our results suggest that 1‐minute value is a variable more sensitive than peak or integral value o

ISSN:0884-3996    

DOI:10.1002/bio.1170050302

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractThe effects of gliadin and glyc‐gli on leukocyte chemiluminescence response were assessedin vitro. A dose‐dependent increase in chemiluminescence response of neutrophils stimulated by zymosan was observed by using gliadin at concentrations ranging between 1 and 20 μg. By increasing glyc‐gli concentration, a bimodal response was observed with an enhancement up to 50 μg/ml, followed by suppressive effects, which were again dose‐dependent. The possible implications of these findings in human pathology are

ISSN:0884-3996    

DOI:10.1002/bio.1170050303

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractHuman as well as canine and rat polymorphonuclear cells (PMN) were separated from whole blood by centrifugation. Two‐step discontinuous Percoll gradients with distinct different densities were used. The chemiluminescence properties of the isolated PMN and of phagocytes in small quantities of whole blood were compared in luminol‐enhanced assays after stimulation with various agents: non‐opsonized zymosan (3.5 g/I), phorbol myristate acetate (PMA, 2.8 × 10−6mol/I), calcium ionophore A 23187 (10−5mol/l) andN‐formylmethionyl‐leucyl‐phenylalanine (FMLP, 3.5 × 10−6mol/l). The isolated cells of the three species responded to all of the various stimuli. Species‐related sensitivity could be ordered: human>canine>rat. Response to the various agents in the human cells can be ranked: PMA ⩾ A 23187>zymosan>FMLP; for the dog: A 23187>PMA>zymosan>FMLP; and for the rat: zymosan ⩾ PMA>FMLP ⩾ A 23187. Time course and peak maximum response were different upon stimulation in the absence and presence of autologous plasma. Distinct soluble stimuli resulted in maximum responses below the baseline in the whole blood assays with canine (FMLP) and rat

ISSN:0884-3996    

DOI:10.1002/bio.1170050304

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractCytochalasin B (CB) is known to interfere reversibly with the cytoplasmic contractile filamental network of mammalian cells. The role of the microfilament system in the mechanism of the reactive oxygen intermediates release of polymorphonuclear leukocytes (PMNL) was studied for different kinds of stimuli. PMNL from fresh human blood were treated with CB and stimulated by adherence on plastic surfaces, by opsonized zymosan, by phorbol myristate acetate and byN‐formylmethionyl‐phenylalaline. The production of reactive oxygen species were monitored by simultaneous detection of native, luminol‐independent, luminescence (NL) and luminol‐dependent luminescence (LDL) using a method of spectral discrimination. Different influences of CB on NL with respect to LDL as well stimuli‐dependent influences of CB on the luminescence response of PMNL were observed. Especially phagocytosis‐associated activation of PMNL was strongly inhibited by CB, whereas LDL was reduced to a much greater extent in comparison with NL. A firm involvement of the microfilament system is indicated, but it depends on the kind of stimu

ISSN:0884-3996    

DOI:10.1002/bio.1170050305

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractAn enzyme‐linked immunosorbent assay (ELISA) is described for the detection of a solubleListeria monocytogenesserogroup 4 antigen in cerebrospinal fluid samples (CSFs). In the ELISA an anti‐Listeriamonoclonal antibody, immobilized onto assay wells, was used to capture antigen from CSFs. the captured antigen was then reacted with a fluorescein isothiocyanate (FITC) conjugate of the same anti‐Listeriaantibody, which was detected with a horseradish peroxidase conjugate of a monoclonal antibody to FITC. The presence of antigen was detected by an enhanced chemiluminescence assay using a camera luminometer.Antigen was detected in the CSFs taken from five out of seven patients with culture provenL. monocytogenesserogroup 4 central nervous system infections, and in none of the CSFs taken from 25 other pat

ISSN:0884-3996    

DOI:10.1002/bio.1170050306

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractAn enhanced chemiluminescence enzyme‐linked immunosorbent assay has been developed for the detection of soluble antigen in the urine of patients with Legionnaires' disease (LD). In the assay antigen(s) in the urine samples are captured by a rabbit anti‐L. pneumophilaantibody coated onto microtitre strips. A fluorescein‐isothiocyanate (FITC) conjugate of the same antibody is then added which binds to the captured antigen. Any immobilized FITC‐labelled antibody is then detected with a horseradish peroxidase (HRP) conjugate of a monoclonal anti‐FITC antibody. HRP activity is monitored after oxidation of luminol in the presence of H2O2and iodophenol. The resulting luminescence is recorded using a camera luminometer.Urine specimens were available for testing from 31 patients with evidence of ongoingL. pneumophilaserogroup 1 infection. A positive result was obtained in the cases of 12/12 specimens from culture‐proven LD patients, and 16/19 specimens from patients with serological evidence of LD. Thus the sensitivity is estimated to be 28/31 (90%) The specificity was estimated using urine specimens from eight patients with non‐L. pneumophilapneumonias of known aetiology. All eight specimens gave a ne

ISSN:0884-3996    

DOI:10.1002/bio.1170050307

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractA modified purification method for bacterial luciferases and NAD(P)H:FMN oxidoreductases is described which uses FMN‐Sepharose alone or coupled to DEAE ion exchange chromatography for the simultaneous purification of luciferase and the various oxidoreductases fromVibrio harveyi, a bright mutant ofVibrio harveyi, Vibrio fischeri, andPhotobacterium phosphoreum. This purification method is compared with DEAE‐Sepharose CI 6B fractionations from these organisms. Both methods allow the separation of oxidoreductases specific for either NADH or NADPH. The use of FMN‐Sepharose coupled to DEAE‐Sepharose fractionation allows the isolation of highly purified enzymes. Lacking interfering factors, these are very suitable for various analytical applications based on bacterial bioluminescence enzymes. The partially purified enzymes from the affinity column have higher specific activities than those obtained using DEAE‐

ISSN:0884-3996    

DOI:10.1002/bio.1170050308

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractWe evaluated the Amerlite system (Amersham, Bucks, UK) for hCG and FT4. The within‐run imprecision (CV%) for hCG was 4.05 at 19.6 U/I (n= 10), 6.28 at 43.45 U/I (n= 10) and 4.62 at 298.57 U/I (n= 10). The between‐run imprecision (five replicates for ten days) was 4.8%, 15% and 11%, respectively. The system was linear up to 200 U/I. A good correlation between Amerlite hCG and an IRMA assay (Becton Dickinson,r= 0.91), Delfia (Pharmacia,r= 0.91) and an automated ELISA assay on ES 600 (Boehringer,r= 0.92) was observed on 70 samples.Within‐run imprecision for FT4 was 3.8% at 0.7 ng/dl (n= 10), 3.3% at 1 ng/dl (n= 10) and 4.32% at 5.15 ng/dl (n= 10), and between‐run was 5.95%, 4.4% and 8.2%, respectively. The comparison with a commercial direct RIA (Becton Dickinson) showed good correlation (r= 0.90,n= 100 samples). The diagnostic value of the association of thyrotropin and FT4, in comparison with the traditional thyroid tests (T3, T4, thyrotropin, FT4, FT3) has been assessed in various thyroid d

ISSN:0884-3996    

DOI:10.1002/bio.1170050309

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractInflammation is accompanied by leukocyte activation (LA). We decribe a simpleex vivotechnique for studying LA that might help to find new LA inhibitors for the treatment of pathologic events related to LA.Arterial and venous blood samples obtained from six permanently catheterized beagle dogs −60, 0, +15 min and +23 h after i.v. challenge with C 48/80, and also blood samples from six normal beagles, were minimally diluted 1:2.5 with buffer. Total leukocyte counts (LC), and luminol amplified CL, induced by opsonized zymosan (C3‐Z), were estimated. Blood samples from dogs elicited CL responses of almost 1/10 the magnitude of erythrocyte‐free human leukocytes, whereas blood samples from rats reacted three orders of magnitude less. Obviously quenching of CL by accompanying erythrocytes in blood samples from dogs is not important, for CL correlated almost linearly with the CL in differently diluted samples. In arterial, but not in venous samples from catheterized dogs, absolute CL and LC, both were significantly depressed (p<0.05) 15 min after C 48/80 challenge. CL/106leukocytes was augmented twofold. All leukocyte deviations returned to pre‐values 23 h post‐

ISSN:0884-3996    

DOI:10.1002/bio.1170050310

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractSpontaneous and induced chemiluminescence of rat blood plasma following irradiation of the animals with fast neurons was studied. Dynamics of the luminescence reflected the degree of radiation injury and an oscillatory response of blood chemiluminescent effect was observed.

ISSN:0884-3996    

DOI:10.1002/bio.1170050311

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY