bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bacteriuria testing by the ATP method as an integral part in the diagnosis and therapy of urinary tract infection (UTI)
Authors: Lundin A. , Hallander H. , Kallner A. , Lundin U. Karnell , Österberg E. 
Published: May 2005

AbstractRapid tests for bacteriuria have the highest value, if the test result is available while the patient is with the doctor. At the bacteriological laboratory rapid testing of samples obtained by mail may be cost‐effective but is of little clinical value. In a previous study performed at a health care centre using conventional urine culture as a reference the ATP test came out as the most reliable one among several rapid bacteriuria tests. The present study was performed to see how the ATP test could be fitted into the routine of the health care centre. Female patients with UTI symptoms were asked to deliver a urine sample to the health care centre laboratory and to wait for the result before seeing the doctor. After having the symptoms confirmed the doctor based the diagnosis on the ATP value. A low ATP value ruled out UTI and a high ATP value confirmed UTI. In patients with an intermediary ATP value (10–50 nmol/I) a positive nitrite test was used to confirm UTI. Only those patients with intermediary ATP values and negative nitrite test had to wait for conventional urine culture. Thus in most patients the decision on antibiotic therapy or not was based on clinical symptoms and ATP results only. Antibiotics (trimethoprim) were given as single dose or as a conventional 7‐day regime in a double‐blind comparison. The correlation between the ATP method and conventional culture was good. Although results of the present study are promising the ATP test as performed is too complicated to become widely accepted at health care centres. However, the dipstick version of the ATP test at present being developed will make the method ideally suited for rapid bacteriuria testing at health care centres and similar doctor's surgery situations.
Determination of adenosine triphosphate in human semen to estimate the fertilizing potential and to quantify sperm antibodies
Authors: Comhaire Frank H. , Vermeulen Lutgart , Monsieur Lieve , Hinting Aucky 
Published: May 2005

AbstractThe measurement of the ATP content of fresh semen is as accurate as the estimation of sperm motility by conventional methods in discriminating between semen of fertile versus subfertile men. The ATP content of frozen thawed donor semen is correlated with the probability of conception per cycle of insemination. Exact quantification of cytotoxic sperm antibodies in serum is possible with the adenosine‐triphosphate‐release‐cytotoxicity test, since measurement is free of the bias of microscopic examination. The procedure has been simplified by testing only one serum dilution and calculating the ‘sperm toxicity index’.
Luciferase of Luciola mingrelica fireflies. Kinetics and regulation mechanism
Authors: Ugarova N. N. 
Published: May 2005

AbstractBiochemical properties, spectral parameters of bioluminescence and reaction kinetics for Luciola mingrelica firefly luciferase are described and analysed. The kinetic scheme of the enzymatic process is proposed and discussed. Allosteric regulation of luciferase activity by ATP and its analogues is considered and binding Mg2+ to luciferase shown to increase its activity. Regulation mechanism of luciferase activity by phospholipids is analysed and choline‐containing phospholipids shown to be specific luciferase activators. Some properties of firefly luciferae and the luciferase synthesized during firefly mRNA translation in frog oocytes are compared.
New approaches to the preparation and application of firefly luciferase
Authors: Filippova N. Yu. , Dukhovich A. F. , Ugarova N. N. 
Published: May 2005

AbstractAllowing for the lipid nature of firefly luciferase we have developed a new method for obtaining high‐activity and high‐stability enzyme preparations for bioluminescent microassay. The method includes the step of differential centrifugation in presence of stabilizing additives which entails a partial purification of the enzyme and its essential stabilization likely due to the fact that luciferase retains its lipid environment which plays an important role in catalysis. The resultant luciferase preparation is stable in solution at 4 °C for 2‐3 months and allows the detection of down to 10−11M ATP.A new method has been offered for luciferase immobilization on film carriers precoated with a phospholipid layer. By sorption of the enzyme on such carriers, the samples of immobilized luciferase have been obtained suitable for constructing chemiluminescent biosensors, in the form of luciferase‐containing films. There are many‐fold applications for detection of ATP micro‐quantities.
Chemiluminescent assay of co‐factors
Authors: Tsuji Akio , Maeda Masako , Arakawa Hidetoshi 
Published: May 2005

AbstractBioluminescent methods are widely used for the assay of the co‐factors, NADH and ATP. Although the bioluminescent method is highly sensitive, the enzymes used are unstable and expensive. Therefore a chemiluminescent method would be valuable in clinical routine assay.We have developed a chemiluminescent method for the assay of NADH using the 1‐methoxy‐5‐methylphenazinium methyl sulphate (1‐MPMS)/isoluminol(IL)/microperox‐idase(m‐POD) system.In order to increase the sensitivity of this method, enzymatic cycling system was coupled to the chemiluminescent assay of NADH.Alcohol dehydrogenase and malate dehydrogenase were used as the cycling enzyme. The standard curve was obtained in the range from 3 × 10−14 to 5 × 10−12mol/assay. The detection limit of NADH was 30fmol/assay which was comparable to that of the bioluminescent method using bacterial luciferase.Two chemiluminescent methods for the assay of ATP have been developed. Method 1 is the system using hexokinase/G6PDH and 1‐PMS/IL/m‐POD, and method 2 is the system based on the enzymatic cycling reaction of ATP using hexokinase/pyruvate kinase. Method 2 is 1000/fold more sensitive than the method 1. The detection limit of ATP was 10 fmol/assay.
On the mechanism of peroxyoxalate chemiluminescence. Quenched chemiluminescence as a detection method in HPLC
Authors: Gooijer C. , van Zoonen P. , Velthorst N. H. , Frei R. W. 
Published: May 2005

AbstractSeveral analytes such as the inorganic anions bromide, iodide, sulphite and nitrite and organic compounds as substituted anilines and sulphur compounds cause quenching of peroxyoxalate chemiluminescence. A detection method for liquid chromatography based on the quenching phenomenon has been developed. It makes use of an immobilized luminophore, i.e. 3‐aminofluoranthene covalently bound via an alkyl‐spacer on controlled pore glass, packed in the detector cell.The mechanism behind the quenching has been elucidated by investigating the roles of luminophores (both in the liquid and in solid state) and oxalates in peroxylate CL with respect to quenchers. Most probably the quencher destroys the radical ion pair produced after electron transfer in the last stage of the CIEEL reaction scheme, thus preventing the formation of electronically excited luminophore.
Effects of serine protease inhibitors on oxyradical burst from phagocytizing neutrophils—analysis by chemiluminescence counting and its microscopic imaging
Authors: Suematsu Makoto , Houzawa Shigenari , Miura Soichiro , Nagata Hiroshi , Kitahora Tetsuji , Morishita Tetsuo , Oshio Chikara , Tsuchiya Masaharu 
Published: May 2005

AbstractReaction difference of oxyradical generation and luminol‐dependent photoemission of zymosan‐ and phorbol ester‐treated neutrophils were investigated using a conventional photomultiplier and ultrasensitive photonic imaging technique. Zymosan‐treated cells released a concentrated photonic burst corresponding to the cellular distribution. In contrast, phorbol ester‐treated cells produced a negligible level of photoemission, and the additional application of Ca2+ ionophore enhanced the photonic burst, which was gradually spread out into extracellular space. Serine protease inhibitors did not attenuate PMA‐induced chemiluminescence but did attenuate zymosan‐induced chemiluminescence. This suggests the involvement of serine protease in the respiratory burst of phagocytizing neutrophils.
Design of luminescence photobiosensors
Authors: Blum Loïc J. , Gautier Sabine M. , Coulet Pierre R. 
Published: May 2005

AbstractThe potential of immobilized enzyme membranes in biosensors has been explored in our group for several years. Although part of our work has been mainly devoted to electrochemical transducers and oxidases for the design of enzyme electrodes, the demand for ultrasensitive and highly selective sensors led us to consider the use of luminescent enzyme systems associated to optical transduction. When considering the need for operational and reliable biosensors in biotechnology, immobilization and stability of the sensing element still remain, in most cases, an unavoidable problem. We recently proposed a very fast and reliable procedure for preparing enzymatic membranes from Pall (Biodyne Immunoaffinity membranes) supplied in a pre‐activated form. Both the firefly and bacterial systems as well as peroxidase for the chemiluminescent determination of various analytes, could be bound to such a support.Based on this approach, a fibre‐optic sensor with immobilized enzymes has been designed which permits bio‐ or chemiluminescent analysis of ATP, NADH or H2O2 respectively. With the NADH‐based system, other analytes could be detected using coupled dehydrogenases. This device appears very promising and includes the convenience of both the luminescence sensitivity as well as the handling of the biosensor design.
Stable‐light‐emitting Escherichia coli as a biosensor
Authors: Korpela Matti , Mäntsälä Pekka , Lilius Esa‐Matti , Karp Matti 
Published: May 2005

AbstractWe have studied possibilities for constructing Escherichia coli strains capable of producing stable light. Light production in E. coli is achieved by cloning the genes encoding bacterial luciferase from Vibrio harveyi. To gain the advantage of sensitive detection of light we transferred the genes under the control of a strong, regulatable promoter system.Stabilization of light produced by E. coli clones was accomplished by finding the optimal plasmid construction and growth conditions as well as suitable measuring buffers. The adjustment of the luciferase synthesis for bioluminescence measurements to a high but not harmful level gives healthy cells and stable luciferase. Cultivation at 30 °C in an uninduced state was found to be the most important factor in getting stable‐light production. The overall cell metabolism being unstressed gives us the possibility of monitoring cell physiology and factors affecting it via bioluminescence reactions in vivo. To make the results easy to interpret the light emission has to be stable during a measurement period of one to several hours. In the case of the original light‐producing bacteria, Vibrio and Photobacterium strains it has not thus far been possible to find conditions where light emission would be stable for several hours. Based on our findings an automated biosensor system can be developed to monitor the effects of biologically active compounds against stable‐light‐producing bacteria.
A chemiluminescent immunoassay for the direct measurement of urinary 5α‐androstane‐3α, 17β‐diol‐glucuronide in urine
Authors: Pinzani P. , Franceschetti F. , Magini A. , Orlandini A. , Pazzagli M. 
Published: May 2005

AbstractWe described a chemiluminescent immunoassay (CIA) for 5α‐androstane‐3α, 17β‐diol‐glucuronide (3α‐diol‐G) in human diluted urine. This method allowed the direct measurement in 1μl of urine avoiding the hydrolysis and extraction steps for sample pretreatment commonly used in routine methods. The hapten 3α‐diol‐G was synthesized by a Koenigs–Knorr reaction. The immunogenic complex, 3α‐diol‐G conjugated to bovine serum albumin (BSA), was employed to induce the formation of specific antibodies in New Zealand rabbits. In addition, the required chemiluminescent (CL) tracer was prepared. The characteristics of the antibody was determined as regard to specificity and sensitivity and the precision of the assay methods established. In 22 hirsute women affected by policystic ovarian syndrome we found 3α‐diol‐G values significantly (p < 0.01) higher (146.28 ± 73.77μg/g of creatinine; mean ± SD) than those observed in normal women (72.1 ± 32.58 μg/g of creatinine; mean ± SD).
Magic lite design and development
Authors: Woodhead J. Stuart , Weeks Ian 
Published: May 2005

AbstractChemiluminescence immunoassays have now achieved a recognized place in the diagnostic laboratory. The advantages of this non‐isotopic technology derive from the use of acridinium esters which can be used to label antigens and antibodies to high specific activities, as well as from optimized immunochemistry. The availability of simple, reliable instrumentation for chemiluminescence measurement together with a range of assay kits offers a logical alternative to traditional radioimmunoassay.
Extended standard curve stability on the CCD magic lite immunoassay system using a two‐point adjustment
Authors: Lynch Michael J. 
Published: May 2005

AbstractCiba Corning Diagnostics has developed a two‐point adjustment algorithm for use with the Magic Lite System which allows for extended stability of a full 7–10 point calibration curve over the life of a kit. This adjustment is accomplished by using a master calibration curve established during manufacturing along with two calibrators for each assay.Most conventional non‐automated immunoassays contain anywhere from 6 to 10 calibrators which are included with each run of an assay.Eliminating the need to run full standard curves and using the two‐point adjustment algorithm results in significant savings in both labour and reagent usage.
Isoluminol as a marker in direct chemiluminescence immunoassays for steroid hormones
Authors: de Boever J. , Kohen F. , Leyseele D. , Vandekerckhove D. 
Published: May 2005

AbstractWe have used isoluminol steroid hormone conjugates in competitive heterogeneous chemiluminescence immunoassays (ClA) for estradiol, progesterone and estriol in body fluids. The assays did not require prior extraction of the steroid hormone with an organic solvent. Polyclonal or monoclonal antibodies, covalently coupled to polyacrylamide microspheres or adsorbed onto the plastic surface of microtritation plates via an intermediate second antibody, were used as solid‐phase. The latter solid‐phase system allows rapid processing of incubation mixtures. Sensitivie and reliable direct CIAs for estradiol in serum, progesterone in serum and in saliva and for estriol in saliva have been developed and their clinical utility has been assessed. Sensitivities ranged from 0.54 to 0.04nmol/l, and precision (% CV) ranged from 10% to 16%.
A sensitive and rapid luminescence sandwich assay for antibodies to hepatitis‐B surface antigen (Anti‐HBsAg)
Authors: Missler Ulrich , Wood William Graham 
Published: May 2005

AbstractA chemiluminescent assay for hepatitis‐B surface antigen is described which used an isoluminol derivative as the label. The assay is precise intra‐assay CV, 1.96‐2.45%; inter‐assay CV, 5.26‐8.11% and has a lower detection limit for hepatitis‐B surface antigen of 1.3U/I.
Optimization of an HPLC peroxyoxalate chemiluminescence detection system for some dansyl amino acids
Authors: Baeyens W. , Bruggeman J. , Dewaele C. , Lin B. , Imai K. 
Published: May 2005

AbstractBis(2,4,6‐trichlorophenyl) oxalate (TCPO)‐hydrogen‐peroxide‐generated chemiluminescence (CL) of four dansyl amino acids has been used as a model system for the optimization of a detection system in reversed‐phase high‐performance liquid chromatography. Dansylated alanine, glutamic acid, methionine, and norleucine were subjected to peroxyoxalate induced CL in a static system and in a flow system under various conditions with respect to TCPO (ethyl acetate) and hydrogen peroxide (acetone) concentrations, solvent composition and flow, using a two‐pump or a one‐pump post‐column reagent system. From the CL‐decay curve, the influence on the emission signal from the total flow rate in the detector was investigated. Special attention was focused on the mixing of the LC eluate and the reagent in order to combine an efficient collection of the emitted light using a 74μI flow cell (originally 10μI in the fluorescence detector) with minimal extra column band broadening. Therefore, a capillary fused‐silica tubing of about 100μm i.d. was inserted against the end‐frit of the column and brought through a mixing tee, in which the solutions of TCPO and hydrogen peroxide were added. The column end tubing ended in the flow cell and the LC eluate and the reagents were mixed when entering the flow‐cell. Average detection limits (S/N=2) of 200fmol injected dansylated amino acid could be reached. A comparison is made between the use of TCPO and DNPO (bis (2, 4‐dinitrophenyl) oxalate).
Luminol‐amplified chemiluminescence activity in human monocytes: A comparison with the activity induced in granulocytes
Authors: Johansson Agneta , Dahlgren Claes 
Published: May 2005

AbstractThe characteristics of the luminol‐amplified chemiluminescence (CL) induced in mononuclear phagocytes interacting with PMA, FMLP or ionomycin were determined. Azide reduced the CL activity by more than 80%, while superoxide dismutase and catalase had minor effect on the monocyte CL response. The sensitivity of the monocyte CL response, to the addition of extra peroxidase differed depending on the stimulus used. Furthermore, no direct correlation was obtained between the CL response and superoxide anion or hydrogen peroxide production. In comparison with the response in granulocytes, minor quantitative differences were observed. The mechanism for the light‐generating reaction, seems to be the same in both cell types.
Enhancement of granulocyte luminescence by murine macrophage secretory products: Suggestive evidence for the release of a new activator
Authors: Willems J. , Leclercq G. , Joniau M. 
Published: May 2005

AbstractThe incubation of macropages (MΦ) in the presence of lipopolysaccharides (LPS) usually results in the release of a variety of immunoregulatory cytokines such as interleukins (IL), tumour necrosis factor (TNF) and colony stimulating factors (CSF). We recently observed that conditioned media (CM) from LPS‐treated murine MΦ lines probably contain another protein endowed with granulocyte stimulatory activity. This cytokine, which has an apparent MW of about 55 kDa enhances the PMA‐induced luminescence of granulocytes and also stimulates their degranulation as measured by lactoferrin release. In contrast to IL1 and IL6 this factor is destroyed by brief treatment at pH 2, but is stable for 60 minutes at 65°C. Unlike CSF, its activity is unchanged by reducing agents such as beta‐mercaptoethanol. Furthermore, pretreatment of the MΦ with dexamethasone, in order to reduce the release of IL1 and TNF, hardly reduces the effect on granulocyte activation. Finally, treatment with a neutralizing polyclonal anti‐murine TNF antiserum only partly abolishes its activity.These results show that, in addition to the already well‐described cytokines, LPS‐treated murine MΦ lines most probably secrete another granulocyte activator.
Luminol chemiluminescence: Chemistry, excitation, emitter
Authors: Merényi Gábor , Lind Johan , Eriksen Trygve E. 
Published: May 2005

AbstractThe mechanism of luminol chemiluminescence is a special case of nucleophilic addition to carbonyl compounds. The breakdown of the key intermediate, an alpha hydroxy hydroperoxide, produces a peracid ortho to an acyl diazene group. After intramolecular addition of the peracid, the energy from nitrogen expulsion is utilized in the formation of an anti‐aromatic endoperoxide. Rupture along the O,O bond leaves a substantial part of the ensuing phthalate in its excited state. The emitter is shown to be a mono‐protonated phthalate unaccessible by photoexcitation. The dark reaction is a concerted decomposion of the alpha hydroxy hydroperodixe to yield ground‐state phthalate.
Fibre‐optic biosensor based on luminescence and immobilized enzymes: Microdetermination of sorbitol, ethanol and oxaloacetate
Authors: Gautier Sabine M. , Blum Loïc J. , Coulet Plerre R. 
Published: May 2005

AbstractWe have investigated highly selective and ultrasensitive biosensors based on luminescent enzyme systems linked to optical transducers. A fibre‐optic sensor with immobilized enzymes was designed; the solid‐phase bioreagent was maintained in close contact with the tip of a glass fibre bundle connected to the photomultiplier tube of a luminometer. A bacterial luminescence fibre‐optic sensor was used for the microdetermination of NADH. Various NAD(P)‐dependent enzymes, sorbitol dehydrogenase, alcohol dehydrogenase and malate dehydrogenase, were co‐immobilized on preactivated polyamide membranes with the bacterial system and used for the microdetermination of sorbitol, ethanol and oxaloacetate at the nanomolar level with a good precision.
The luminescent bacteria toxicity test: Its potential as anin vitroalternative
Authors: Bulich Anthony A. , Tung Ker‐Kong , Scheibner Grace 
Published: May 2005

AbstractDuring the past several years, the use of animals for toxicity testing has come under critical surveillance. For ethical and economic reasons, various techniques have been developed and proposed as potential alternatives for some of the whole animal toxicity assays. One assay proposed as an alternative to animal testing is the luminescent bacteria toxicity test (LBT), provided under the trade name of Microtox®. The sensitivity and specificity of the LBT was compared with two commonly used toxicity tests–‐the L‐929 Minimal Eùgle's Medium (MEM) elution cytotoxicity test and the Draize test. Cytotoxicity and LBT test data from 709 medical device and biomaterial extracts were compared using a positive/negative ranking system which provided a measurement of false positive and false negative results. These data were compiled from nine separate laboratories producing or using a wide variety of biomaterials and medical device products. The LBT was more sensitive than the tissue culture assay and displayed few false negatives. LBT EC50values were compared with eye irritancy categories for a group of 34 chemicals and 27 personal care products. As with tissue culture, the LBT was more sensitive and produced minimal false negatives. The data from this study indicate the LBT has potential as a rapid, simple method to screen biomaterials and personal care products for toxicity and irritancy.
From Luc and Phot genes to the hospital bed
Authors: Campbell Anthony K. , Sala‐Newby Graciela , Aston Paul J. , Kalsheker Noor , Kishi Yoshito , Shimomura Osamu 
Published: May 2005

AbstractThe Luc gene from the firefly Photinus pyralis has been isolated by cloning it in pcDV1 PL plasmid primer and Honjo linker and the Phot gene isolated from Aequorea victoria using the polymerase chain reaction. A method has been established using SP6 RNA polymerase for transcribing and translating bioluminescent genes in vitro. It should now be possible to engineer these genes to measure intracellular ATP and the covalent modification of proteins in single, live cells, providing unique insights into the molecular basis of disease.