Animals ; Fluorescence ; Fluorescence Recovery After Photobleaching ; Fluorescence Resonance Energy Transfer ; Fluorometry ; Green Fluorescent Proteins ; Humans ; Immunohistochemistry

No abstract available.
Cyclosporine* ; Fluorescence Polarization Immunoassay* ; Fluorescence Polarization* ; Fluorescence*

OBJECTIVEThis study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum.

METHODSTo measure the concentration of theophylline (n=122) and evaluate the assay.

RESULTSThe linear range of the CLIA method was 0.51-40 mg/L (Y=1.02X+0.44, r=0.995). The intra and inter CV (coefficient variance) of CLIA were 3.20% and 3.57%, respectively. The average recovery rate was 102.3%. This method was free from interference by brilirubin (<200 micromol/L), hemoglobin (<10 g/L), and triglycerides (<15 mmol/L).

CONCLUSIONThis method is simple, convenient and precise for clinical pharmacokinetics study of theophylline.

Blood Chemical Analysis ; methods ; Female ; Fluorescence Polarization Immunoassay ; methods ; Humans ; Luminescent Measurements ; methods ; Lung Diseases ; blood ; Male ; Middle Aged ; Reproducibility of Results ; Sensitivity and Specificity ; Theophylline ; blood

A stable dark variant separated from photobacterium phosphoreum (A2) was fixed in agar-gel membrane and immobilized onto an exposed end of a fiber-optic linked with bioluminometer. The variant could emit a luminescent signal in the presence of genotoxic agents, such as Mitomycin C (MC). The performance of this whole-cell optical fiber sensor system was examined as a function of several parameters, including gel probe thickness, bacterial cell density, and diameter of the fiber-optic core and working temperature. An optimal response to a model genotoxicant, Mitomycin C, was achieved with agar-bacterial gel membrane: the thickness of gel membrane was about 5 mm; the cell density of bacteria in gel membrane was about 2.0 x 10(7)/ml; the diameter of fiber-optic core was 5.0 mm; the working temperature was 25 degrees C. Under these optimized conditions, the response time was less than 10 h to Mitomycin C, with a lower detection threshold of 0.1 mg/L.
Biosensing Techniques ; Chemiluminescent Measurements ; Fiber Optics ; Luminescent Proteins/*genetics ; Mitomycin/*pharmacology ; Mitomycin/toxicity ; Photobacterium/*genetics ; Transcription, Genetic/drug effects ; Variation (Genetics)

OBJECTIVESTo summarize neonatal screening for phenylketonuria (PKU) and congenital hypothyroidism (CH) in China, to further clarify incidence of the two kinds of diseases in newly-born babies, and to explore issues in neonatal screening and their solutions.

METHODSNeonatal screening for PKU and CH was conducted by 39 neonatal screening centers all over the country, sponsored by the Group of Neonatal Screening, Chinese Society of Child Health Care, Chinese Preventive Medical Association and the Center for Neonatal Screening Quality Control Laboratory, National Center for Clinical Laboratories (NCCL). In each infant a heel prick blood sample was collected at 72 hours postnatal onto standard filter paper. PKU was screened by bacterial inhibition assay and fluorometric method, and CH was screened by TSH measurement by time-resolved fluorescence immunoassay (TRFIA), fluorescence enzyme immunoassay (FEIA) and enzyme immunoassay (EIA).

RESULTSFrom 1985 to 2001 in China, totally of 5 817 280 newborns were screened for PKU, 522 cases of PKU detected with an incidence of 1:11 144, and 5 524 019 newborns were screened for CH, 1 836 cases of CH detected with an incidence of 1:3 009. Annual average number of newborns screened for congenital genetic diseases was increased by 45.5% in recent six years.

CONCLUSIONSNeonatal screening was developed quickly in China in recent years, especially in some developed cities, such as Shanghai with a coverage of 98.3% in 2001. But, its coverage was about only 10% in China as a whole. In development of neonatal screening, it is necessary to attach more importance to quality of screening and increasing coverage of screening, as well as gradual development of new screening techniques for other neonatal preventable diseases, in addition to PKU and CH, and their application, and improvement of level of child health care in China.

China ; epidemiology ; Congenital Hypothyroidism ; Fluorescence Polarization Immunoassay ; Fluorometry ; Humans ; Hypothyroidism ; epidemiology ; Immunoenzyme Techniques ; Infant, Newborn ; Neonatal Screening ; Phenylketonurias ; epidemiology ; Thyrotropin ; blood

A fluorescence polarization immunoassay (FPIA) was developed for the analysis ofaflatoxins (AFs) using an anti-aflatoxin B1 (AFB1) monoclonal antibody and a novel fluorescein-labeled AFB1 tracer. The FPIA showed an IC50 value of 23.33 ng/mL with a limit of detection of 13.12 ng/mL for AFB1. The cross-reactivities of AFB1, AFB2, AFG1, AFG2, AFM1, and AFM2 with the antibody were 100%, 65.7%, 143%, 23.5%, 111.4%, and 2%, respectively. The group-specificity of anti-AFB1mAb indicated that the FPIA could potentially be used in a screening method for the detection of total AFs, albeit not AFG2 and AFM2. The total time required for analyzing 96 samples in one microplate was less than 5 min. This study demonstrates the potential usefulness of the FPIA as a rapid and simple technique for monitoring AFs.
Aflatoxins ; analysis ; Fluorescence Polarization Immunoassay

In this review we summarize the progress made towards understanding the role of protein-protein interactions in the function of various bioluminescence systems of marine organisms, including bacteria, jellyfish and soft corals, with particular focus on methodology used to detect and characterize these interactions. In some bioluminescence systems, protein-protein interactions involve an "accessory protein" whereby a stored substrate is efficiently delivered to the bioluminescent enzyme luciferase. Other types of complexation mediate energy transfer to an "antenna protein" altering the color and quantum yield of a bioluminescence reaction. Spatial structures of the complexes reveal an important role of electrostatic forces in governing the corresponding weak interactions and define the nature of the interaction surfaces. The most reliable structural model is available for the protein-protein complex of the Ca(2+)-regulated photoprotein clytin and green-fluorescent protein (GFP) from the jellyfish Clytia gregaria, solved by means of Xray crystallography, NMR mapping and molecular docking. This provides an example of the potential strategies in studying the transient complexes involved in bioluminescence. It is emphasized that structural studies such as these can provide valuable insight into the detailed mechanism of bioluminescence.
Animals ; Anthozoa ; physiology ; Aquatic Organisms ; physiology ; Bacteria ; metabolism ; Binding Sites ; Calcium ; metabolism ; Crystallography, X-Ray ; Fluorescence Resonance Energy Transfer ; Green Fluorescent Proteins ; metabolism ; Hydrozoa ; physiology ; Imidazoles ; metabolism ; Luciferases ; metabolism ; Luminescent Measurements ; instrumentation ; methods ; Luminescent Proteins ; metabolism ; Models, Molecular ; Protein Binding ; Pteridines ; metabolism ; Pyrazines ; metabolism ; Scyphozoa ; physiology ; Spectrometry, Fluorescence

Digoxin plays a part in healing of congestive heart failure in clinic. Its therapeutic dose is very approximate to toxic dose and even they overlap each other sometimes. There are many influencing factors on blood drug level of digoxin. Pharmacodynamics and pharmacokinetics varies with different individuality. It is indispensable to detecting blood drug level in order to treat disease and prevent intoxication. Integrating with the detecting-methods of blood drug level of digoxin home and broad, characteristic of many methods are summarized from sensitivity, linearity range, cross-reaction and precision. These methods include radio immunoassay, enzyme immunoassay, chemiluminescence immunoassay, fluorescence immunoassay and HPLC-MS-MS. These methods are popular for their specialized ascendancy. The cost of radio immunoassay is low. Enzyme immunoassay has good specificity. Sensitivity and stability of chemiluminescence immunoassay is very excellent. Fluorescence polarization immunoassay is sensitive and convenient. HPLC-MS-MS has high resolution and good specificity. One of the development tendencies is to combine two or more methods in detecting the blood drug level of digoxin which contribute to these methods integrated use.
Chemistry Techniques, Analytical ; methods ; Chromatography, High Pressure Liquid ; Digoxin ; blood ; Enzyme-Linked Immunosorbent Assay ; Fluorescence Polarization Immunoassay ; Fluoroimmunoassay ; Humans ; Radioimmunoassay ; Reproducibility of Results ; Tandem Mass Spectrometry

BACKGROUND: It was purposed to estimate correlation between fluorescence polarization immunoassay (FPIA) and high performance liquid chromatography (HPLC), and precision of individual methods. It was also objected to describe distribution of plasma total homocysteine in Korean adults. METHODS: The subjects were 100 adults admitted to Inha University Hospital during the month of October, 1998. The total plasma homocysteine concentration was measured by FPIA (IMx analyzer, Abbott Laboratories, IL, USA) and by HPLC (ACCLAIM Biogenic Amines Testing System, Bio-Rad Laboratories, CA, USA) using Bio-Rad Homocysteine. RESULTS: Plasma homocysteine levels (mean+/-SD) from Korean healthy adults by FPIA and HPLC were 9.75+/-3.80micromol/L, 7.72+/-3.36micromol/L, respectively. Plasma homocysteine levels according to sex by FPIA were 11.79micromol/L for male, 7.71micromol/L for female, and those by HPLC were 9.47micro mol/L for male, 5.98micromol/L for female, respectively. Intra-assay coefficient variations (CVs) of low, medium, and high concentration by FPIA are 1.83%, 0.47%, and 1.66%, and those by HPLC are 5.53%, 5.37%, and 4.56%, respectively. Inter-assay CVs of low, medium, and high concentration by FPIA are 2.28%, 1.44%, and 1.29%, and by HPLC are 7.23%, 5.54%, and 4.95%, respectively. CONCLUSION: Plasma homocysteine levels from male were significantly higher than female in Korean. Plasma homocysteine levels were increased according to increment of age. FPIA was more convenient, automatic, rapid, and reproducible than HPLC and also excellently correlated with HPLC. It is concluded that FPIA will potentially benefit for quantifying homocysteine in clinical laboratories.
Adult ; Biogenic Amines ; Chromatography, High Pressure Liquid ; Chromatography, Liquid* ; Female ; Fluorescence Polarization Immunoassay* ; Fluorescence Polarization* ; Fluorescence* ; Homocysteine* ; Humans* ; Male ; Plasma*

BACKGROUND: The penetraton in vivo of topically applied substances can be assessed by physiological or pharmacalogical signs or analysed by chemical or histological techniques. In vitro absorption can be commonly quantitated by measuring the passage of a radioisotope-labelled substance across skin that has been mounted in a diffusion chamber. OBJECTIVE: Fluorescence polarization immunoassay technique has made the possible rapid growth of therapeutic drug nonitoring. We applied this methodology in measuring percutaneous absorption in a diffusion chamber. METHODS: We utilized sheets of whole epidermis prepared from the circumcised prepuce. Some epidermal sheets were treated with 2 ml of acetone for 2 minutes, and others not. The epidermal sheet was mounted in a diffusion chamber between the donor compartment for the penetrant and the receptor compartment containing saline. Lidocaine HC1(10 microgram/cm2) in vehicle(propylene glycol:ethanol; 7:3, vol/vol) was applied to the donor compartment for the penetrant. With flow rate of about 3 ml/h all of the receptor phase collected during 2 hours interval were quantitated for 10 hours by the fluorescence polarization immunoassay. RESULTS: Total absorption of lidocaine HC1 in the acetone-untreated group was 2.14+/-0.74% of the applied dose. Total absorption in the acetone-treated group showed no substantial difference (2.09+/-1.25%) compared to those of acetone-untreated group. The amount of lipid extracted from a epiderrnal sheet with acetone was 19+/-2.97%. CONCLUSION: Fluorescence polarization immunoassay may be a useful method in measuring percutaneous absorption in vitro.
Absorption ; Acetone ; Diffusion* ; Epidermis ; Fluorescence Polarization Immunoassay* ; Fluorescence Polarization* ; Fluorescence* ; Histological Techniques ; Humans ; Lidocaine ; Skin ; Skin Absorption* ; Tissue Donors