ABSTRACT
An amplified flow immunoassay (AFIA) was developed for cocaine, which combines a noncompetitive immunoenzymometric assay (IEMA) with an on-line detection of the enzyme label alkaline phosphatase (ALP) by a substrate-recycling biosensor. In the IEMA, the analyte cocaine first binds to a labeled polyclonal anti-cocaine antibody. Then, the excess labeled antibody is separated on an affinity column that contains a perfusion chromatography carrier modified by immobilized cocaine. The unbound complexes of the analyte cocaine with the ALP-labeled antibody are detected postcolumn. The detector senses phenol produced by ALP from phenyl phosphate. As detector, an amperometric substrate-recycling biosensor was used, which consists of a Clark-type oxygen electrode covered by tyrosinase and pyrroloquinoline quinone-dependent glucose dehydrogenase. The lower limit of detection is 380 pM (38 fmol) for cocaine. The sampling rate is 26/h. Cocaine could be detected from "real samples" with an imprecision of +/- 10% (n = 3) and with a recovery of 49 +/- 3% for various concentrations. AFIA is generally important as a new approach for the fast detection of picomolar concentrations of haptens.
Subject(s)
Cocaine/analysis , Immunoassay/methods , Alkaline Phosphatase/analysis , Antibodies , Automation , Biosensing Techniques , Immunoassay/instrumentation , Phenols/analysisABSTRACT
Patient death is a stressful experience for the patient, family, and the healthcare team. Nurses-often have only informal resources for coping with the sadness and grief they might experience. Realizing the need for nursing grief support, a group of staff nurses from the intensive care unit formed a grief support group. Using information from the literature and critical incident stress debriefing, the group developed support interventions to aid intensive care unit staff after patient death.
Subject(s)
Attitude to Death , Burnout, Professional/prevention & control , Critical Care/psychology , Grief , Nursing Staff, Hospital/psychology , Self-Help Groups/organization & administration , Adaptation, Psychological , Burnout, Professional/psychology , Communication , Humans , Program Development , Social SupportABSTRACT
A bienzyme substrate-recycling biosensor in a flow injection analysis system is described for the sensitive measurement of alkaline phosphatase (ALP) and applied to the fast readout of a competitive immunoassay for the widely used pesticide 2,4-dichlorophenoxyacetic acid (2,4-D). The phenol-indicating biosensor consists of a Clark-type electrode covered by a membrane with coentrapped tyrosinase and quinoprotein glucose dehydrogenase. ALP dephosphorylates phenyl phosphate to phenol (K(m) = 36 microM) outside the flow system. Phenol is oxidized in the sensor membrane by the oxygen-consuming tyrosinase via catechol to o-quinone. The quinone is reconverted to catechol by glucose dehydrogenase. This substrate cycling results in a 350-fold amplified sensor response to phenol. The oxygen consumption of the enzyme couple in the presence of phenol is monitored as a decrease in current. A total of 3.2 fM ALP (320 zmol/ 100 microL) has been detected after a 57.5 min incubation with phenyl phosphate. All involved reagents are stable over the time of measurement. The sensor does not produce any measurable blank signals. The immunoassay detects 0.1 microgram/L 2,4-D, the maximum concentration for pesticides allowed in drinking water by European Community regulations. The applicability of this biosensor for fast immunoassay readout is demonstrated by a 2 min incubation. By comparison, a standard photometric method (p-nitrophenyl phosphate) requires overnight incubation.
Subject(s)
2,4-Dichlorophenoxyacetic Acid/analysis , Alkaline Phosphatase/analysis , Biosensing Techniques , Herbicides/analysis , Calibration , Electrochemistry , ImmunoassayABSTRACT
The amplification cycle consisting of NADH independent oligosaccharide dehydrogenase (ODH) and laccase has been recently reported to be highly sensitive to several catecholamines and p-aminophenol. A competitive immunoassay for 2,4-dichlorophenoxyacetic acid has been developed by combining this amplification cycle with beta-galactosidase as enzyme label resulting in p-aminophenol as product. The combination of enzymatic amplification cycles with a competitive immunoassay yields a highly sensitive measurement of 2,4-dichlorophenoxyacetic acid. Using a monoclonal antibody the linear range of the assay was between 0.02 and 100 ng/l and the c(50) was found at 0.2 ng/l; the detection limit was at 5 pg/l (25 fmol/l) corresponding to 5 amol.
Subject(s)
Dental Abutments , Dental Porcelain , Dental Veneers , Denture Design , Denture, Partial, Removable , Aged , Bicuspid/pathology , Cuspid/pathology , Dental Clasps , Dental Porcelain/chemistry , Dental Prosthesis Design , Denture Retention , Female , Follow-Up Studies , Glass Ionomer Cements/chemistry , Humans , Maleates/chemistry , Resin Cements/chemistryABSTRACT
Examined the degree of interspouse similarity in three samples from previous studies. Two of these samples were matched on age, education, and SES. The third sample represented couples who were not matched on these variables. Contrary to prior assumptions, the matched groups differed from each other nearly as much as they did from the unmatched group. This has implications for the ability to generalize from interspousal data even when comparison groups are matched on several traditional variables.
