Photometry in goldfish by electrophysiological recording: comparision of criterion response method with heterochromatic flicker photometry.
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Adaptation, Ocular↗
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Ektachem slides for direct potentiometric determination of sodium in plasma: effect of natremia, blood pH, and type of electrolyte reference fluid on concordance with flame photometry and other potentiometric methods.
With electrolyte reference fluid (ERF)00, results from Kodak Ektachem slides for the direct potentiometric assay of sodium in plasma were significantly correlated with results from flame photometry, but also appeared to be systematically higher, especially in hypernatremic patients. Indirect potentiometry with the Technicon RA-1000 yielded intermediate values. In 23 hypernatremic patients with greater than or equal to 6 mmol/L difference in sodium between Ektachem ERF00 and flame photometry, a clinical survey disclosed the frequent association of large between-method differences with renal failure, diabetes mellitus, and gastrointestinal disease. However, there was no correlation between differences in sodium on the one hand and anion gaps or (lipo)protein concentrations on the other, nor did in vitro addition studies implicate metabolites that often accumulate in the above-mentioned disorders. Unlike indirect methods, sodium measurements by direct potentiometry on Ektachem and Corning were influenced by in vitro changes of pH between 7.0 and 7.9. However, in a group of patients that included many acidotic individuals, between-method differences in sodium appeared not significantly correlated with in vivo blood pH. Use of the equitransferant ERF04 on Ektachem strongly diminishes the systematic differences with flame photometry, reduces the pH-dependency of the results to that of the direct Corning method, and brings the mean analytical recovery of sodium to below 95% (instead of 115% previously) without affecting the ability of Ektachem slides to avoid spuriously low results in the presence of increased (monoclonal) protein concentrations.
Blood↗
A new test for endothelial viability. The Janus green photometry technique.
A new in vitro technique was developed to measure, by simple photometry, the amount of Janus green extracted with absolute alcohol from isolated corneas after vital staining. The amount of stain corresponded to the percentage of damaged endothelial cells and was a reliable parameter of endothelial viability. More rapid and easier to perform than the usual cell counting of devitalized cells, this technique can be used to study the endothelial damage induced, for example, by experimental surgery, intraocular solutions, intraocular lens material, corneas storage solutions. As an example of application, results of the endothelial status after medium-term preservation of pig corneas (n = 40) in a modified McCarey-Kaufman medium at 4 degrees C were determined. The mean +/- endothelial damage was 8.5% +/- 1.8% (mean +/- SD) after 4 days, 29.5% +/- 3.7% after 7 days, 45.5% +/- 5.5% after 10 days, 69.3% +/- 3.7% after 14 days, and 100% after 20 days of storage in McCarey-Kaufman medium.
Animals↗
Use of a wedge cuvette in thin layer photometry and its application to oximetry.
A wedge cuvette was constructed by fixing 2 glass plates at a known angle with a spacer at one end. This resulted in a thin layer with thickness varying from 0 to 250 micrometer. By measuring the intensity of a beam of light through the thin layer as a function of distance along the wedge (and thus layer thickness), the absorption coefficient at the light wavelength used could be obtained without a separate measurement of I0, the reference light intensity. In addition, the difficult problem of determining accurate layer thickness as encoutered in conventional thin layer photometry has been avoided. Tests of the wedge cuvette method with Evans Blue and Malachite Green serial dilutions as well as with haemoglobin solutions at several oxygen saturations demonstrate that accuracy of the order of 1% can be obtained. Application of the wedge cuvette in experiments on oxygen uptake by layers of haemoglobin solution are discussed.
Hemoglobins↗
The relationship between the course of psoriasis and transepidermal water loss, photoelectric plethysmography and reflex photometry.
The transepidermal water loss (TWL), photoelectric plethysmography and reflex photometry were parallelly registered on the involved hands and forearms of psoriatics. The TWL values showed a fairly good correlation with the clinical course of the disease: they increased during the active phase and decreased after return to normal. The vascular tests did not show a strict parallelism with the clinical state. On the basis of the findings, the temporal relations between epidermal and vascular factors during the different phases of the psoriatic process are discussed. The TWL can be used to predict a relapse occurring on the area investigated but not on a distant area.
Erythema↗
Nystatin-induced potassium efflux from Saccharomyces cerevisiae measured by flame photometry: a potential bioassay approach.
A potential bioassay method for nystatin has been developed based on the ability of this antibiotic to bring out dose-related potassium ion efflux from susceptible cells of Saccharomyces cerevisiae, such extracellular potassium ion concentration being measured by flame photometry.
Nystatin↗
Multiangle light scattering flow photometry of cultured human fibroblasts: comparison of normal cells with a mutant line containing cytoplasmic inclusions.
Multi-angle light scattering flow photometry was used to study the light scattering properties of normal cultured fibroblasts and a mutant fibroblast line containing cytoplasmic lysosomal inclusions. The effect of glutaraldehyde fixation on the light scattering properties of the cells was also examined and correlated with their ultrastructure. Normal fibroblasts showed uniform organelle distribution with few vacuoles or dense bodies in the cytoplasm while the mutant line showed abnormal cytoplasmic inclusions of varying morphology, density and lucency. As predicted by light scattering theory, the mutant cells containing the cytoplasmic inclusions scattered more light at large angles (greater than theta = 1.85 degrees) than did the normal cells. Glutaraldehyde fixation decreased light scattering at small angles (less than theta = 1.85 degrees), increased light scattering at larger angles (greater than theta = 1.85 degrees) in both normal and mutant cells and enhanced resolution of the light scattering signatures. The mutant line scattered 2-3 times more light at a wide angle (greater than theta = 12.74 degrees) than did the normal cells. These data suggest that abnormal lysosomal storage inclusion bodies in the cytoplasm of the cells can be detected by differential light scattering methods.
Cell Line↗
Quantification of islet cell antibodies by microscope photometry and antibody titration compared.
Islet cell antibodies can predict eventual insulin-dependent diabetes mellitus. In the standard method used by the Immunology of Diabetes Workshop (IDW), predictive power depends on antibody titer as determined by testing multiple serum dilutions. Here I report use of a microscope-based photometry system (MPS) to test whether islet fluorescence intensities from undiluted sera can predict IDW results. MPS testing of 120 undiluted IDW specimens correctly identified 27 of 31 positive and 84 of 89 negative specimens. The relationship between IDW consensus values and MPS readings of undiluted sera is: IDW = 2.45 MPS - 1.44 (r = 0.67, P less than 0.001). Thus, corrected islet intensities from undiluted sera correlated well with IDW values. However, the ability of MPS to predict high IDW consensus values is limited, which may reflect the variable antigenic content of pancreas specimens; moreover, background correction as currently applied is incomplete; nonspecific immunofluorescence appears to emanate from within the islet.
Autoantibodies↗
[Use of methods of derivative conductometry and photometry for assessing the structure of a cellular suspension].
Experimental proofs for the possibility of studying the dynamical structure of rotatory moving erythrocyte suspension with the methods of derivative conductometry and photometry are presented. The amplitude of the recorded impulses was proportional to the cell concentration and the angle velocity, and strongly depended on the shape of the suspended cells.
Cell Movement↗
[Comparison of ion-selective electrodes and flame photometry for the determination of serum Na+ and K+ for clinical purposes (author's transl)].
The ORION SPACE-STAT (SS-30) and the TECHNICON STAT/ION were used to investigate quality control of Na+- and K+-determinations in test sera (n=8) and in the plasma of 100 patients. The flame photometer IL 543 was used as a reference apparatus. The ion-selective electrode instruments, SS-30 and STAT/ION, gave results very similar to those of flame photometry. The values from the SS-30 were higher by an average of 3% to 5% compared with the values from the other instruments. An explanation for this bias may be the fact that the SS-30 (unlike the STAT/ION and the IL 543) measures electrolyte activities or concentrations in undiluted plasma water. On the basis of these results the routine use of ion-selective electrodes for Na+- and K+-determination can be recommended for clinical laboratories.
Electrodes↗
[Optimal measurement technic for the vasoconstrictor effect of corticosteroids. Comparative studies with reflection photometry and flow measurement].
In 20 healthy male test subjects, a non-specific ointment base was applied to the back of the hand over 16 hours. Two days later, the same ointment base with 0.25% triamcinolone acetonide was once more applied to the back of the hand over 16 hours. The light reflection of the skin was measured by reflection photometry and the heat transport coefficient was measured fluvographically under the condition of the tourniquet test. The results show that the vasoconstrictor effect of triamcinolone acetonide is best detected by the reflection photometric measurements. In fluvographic measurement, the heat transport coefficient is most affected in maximal reactive hyperemia when the tourniquet has been removed. Application of the base led to a reduction of the blood flow and the water content of the skin after 16 hours, whereas the water content of the skin was unaffected in the presence of triamcinolone acetonide.
Adrenal Cortex Hormones↗
Measurement of plasma levels of tricylic psychoactive drugs and their metabolites by UV reflectance photometry of thin layer chromatograms.
A method has been developed for the determination of perazine, clozapine, imipramine and amitriptyline and their demethylated metabolites in plasma. Other metabolites measured were perazine sulfoxide and the N-oxides of clozapine and perazine, the latter two following their reduction to the parent drugs with ascorbic acid. 10-Hydroxynortriptyline was identified as an amitriptyline metabolite in plasma. The general procedure included extraction of alkalinized plasma samples (3 - 6 g) with benzene or toluene and thin layer chromatography of the extracts, followed by reflectance photometry of the plates at appropriate wave lengths in ultraviolet light. Spots of questionable identity were further characterized by two-dimensional chromatography and by colour reactions. Therecoveries of compounds added in therapeutic concentrations were between 70 and 98 %. The limits of detectability were 5 - 10 ng/g plasma.
Amitriptyline↗
Minimum enzyme unit for Na+/K+-ATPase is the alpha beta-protomer. Determination by low-angle laser light scattering photometry coupled with high-performance gel chromatography for substantially simultaneous measurement of ATPase activity and molecular weight.
The oligomeric state of canine renal NA+/K+ -ATPase solubilized by octaethylene glycol n-dodecyl ether (C12E8) was studied by means of low-angle laser light scattering photometry coupled with high-performance gel chromatography (HPGC). At around 0 degree C the solubilized enzyme was separated into the (alpha beta)2-diprotomeric and alpha beta-protomeric protein components with Mr values of 302,000 +/- 10,000 and 156,000 +/- 4,000, respectively, in approximately equal quantities. As the temperature of chromatography was increased toward 20 degrees C, the two protein components converged into a single major component. The Mr of this component depended on the monovalent cation included in the elution buffer, and was 255,000 or 300,000 in the presence of 0.1 M NaCl or 0.1 M KCl, respectively. A computer simulation technique showed that the solubilized enzyme was in a dissociation-association equilibrium of 2 protomers = diprotomer at 20 degrees C, and the difference in apparent Mr of the solubilized enzyme between the two species of monovalent cation was interpreted by an association constant (Ka) in the presence of 0.1 M KCl that was about 50-fold larger than in the presence of 0.1 M NaCl. In order to measure ATPase activity and Mr of the solubilized enzyme simultaneously, a TSKgel G3000SW column had been equilibrated and was eluted with an elution buffer containing 0.30 mg/ml C12E8 and 60 microgram/ml phosphatidylserine (bovine brain) as well as the ligands necessary for the enzyme to exhibit the activity at pH 7.0 and 20 degrees C. The solubilized enzyme was always eluted as a single protein component irrespective of the the amount of the protein applied to the column, ranging between 240 and 10 microgram. The Mr of the protein component, however, decreased from 214,000 and 158,000 with the decrease of the protein amount. The specific ATPase activity, however, remained constant at a level of 64 +/- 4% of that of the membrane-bound enzyme even in the range of protein concentration sufficiently low as to allow the enzyme to exist only in the protomeric form. Thus, the alpha beta-protomer is concluded to be the minimum functional unit for the ATPase activity. The value of Ka obtained from the concentration-dependent dissociation curve was 5 . 10(5) M-1 for the enzyme turning over, and 1.1 . 10(7) M-1 for the enzyme inhibited with ouabain. It was discussed, based on the values of Ka obtained, that the enzyme would exist as the diprotomer or the higher oligomer in the membrane.
Animals↗
Interfacing gradient elution ion-exchange chromatography and low-angle laser light-scattering photometry for analysis of proteins.
Molecular weights (MWs) of different proteins were determined by interfacing gradient elution ion-exchange chromatography and low-angle laser light-scattering photometry (IEC-LALLS). A high-performance strong cation-exchange column was used to elute proteins using fast (5 min) and conventional (15-30 min) gradients. The eluted proteins were characterized on-line by determining their MWs using LALLS. The specific refractive index (RI) increment (dn/dc) and the RI of the solvent used over the gradient range were determined off-line and used to calculate the absolute weight-average MWs. Four proteins, ribonuclease A, alpha-chymotrypsinogen A, trypsinogen and beta-lactoglobulin A (beta-LACT) were studied. Accurate MWs were obtained for all the proteins using fast and conventional gradients, except for beta-LACT, which aggregated as a function of the gradient employed. The degree of aggregation of beta-LACT increased as the rapidity of the gradient was increased over a fixed gradient range. This study indicated that it is possible to separate and characterize proteins rapidly using IEC-LALLS.
Chromatography, Ion Exchange↗
Protein-protein interaction of detergent-solubilized Ca2(+)-ATPase during ATP hydrolysis analyzed by low-angle laser light scattering photometry coupled with high-performance gel chromatography.
Protein-protein interaction of detergent-solubilized Ca2(+)-ATPase was examined, employing low-angle laser light scattering photometry coupled with high-performance gel chromatography. When solubilized with octa(ethylene glycol) mono-n-dodecyl ether (C12E8) and chromatographed in the presence of 0.3 mg/ml C12E8, the Ca2(+)-ATPase emerged as a single peak with an intermediate molecular weight between the monomer and the dimer, showing a dissociation-association equilibrium of the two components. In the presence of 50 micrograms/ml phosphatidylcholine and 0.3 mg/ml C12E8 at 0 degrees C, the Ca2(+)-ATPase (0.8 mg) emerged as the two distinct components with molecular weights of 125,000 +/- 2100 (n = 3) and 211 300 +/- 7300 (n = 3), indicating that there was no rapid interconversion between the monomer and the dimer. Under the latter conditions, addition of ATP induced fusion of two components. The apparent molecular weight of the fused peak shifted from the monomer to the dimer as the amount of protein increased. Addition of ADP or adenosine 5'-(beta, gamma-methylene triphosphate), however, did not induce such fusion of the peaks. The ATP-induced fusion of the peaks was not observed either in 5 mM CaCl2, the conditions in which the rate of ATP hydrolysis was extremely slow. Thus, the solubilized Ca2(+)-ATPase underwent a rapid interconversion between the monomer and the dimer during ATP hydrolysis. These results suggest that the protein-protein interaction during ATP hydrolysis is an intrinsic nature of Ca2(+)-ATPase and that such interaction may be important for Ca2+ transport by Ca2(+)-ATPase in the sarcoplasmic reticulum membranes.
Adenosine Triphosphate↗
Function of DR-positive thyrocytes from patients with Graves' disease: quantitative analysis of thyroid peroxidase content by fluorescent photometry.
A considerable number of thyrocytes in patients with autoimmune thyroiditis ectopically express HLA-DR antigen. Furthermore, it has been reported that interferon-gamma-induced DR-positive thyrocytes in vitro secrete less thyroid hormone in response to TSH stimulation compared with DR-negative ones. However, the function of the intrinsically DR-positive thyrocytes is unknown. To evaluate their function, we stained by immunofluorescence for both DR antigen and thyroid peroxidase (TPO) in thyroid epithelial cells from patients with Graves' disease. We also measured the quantity of DR antigen and TPO using fluorescent photometry. The content of TPO was not significantly reduced in DR-positive thyrocytes compared with that in DR-negative thyrocytes. The TPO content is one measure of thyrocyte function. There was no significant difference between DR-positive and DR-negative thyrocytes. In conclusion, the function of DR-positive thyrocytes in vivo was not suppressed compared with that of DR-negative thyrocytes.
Adult↗
[Determination of Fe, Cu and Zn by a mechanised micro-method of flame photometry (author's transl)].
By sample application in measured quantities ("injection method") the quality of serum required for the determination of trace elements can be considerably decreased with essentially no loss in sensitivity. The volume of solution, 50 or 100 microliters, required for a single measurement is added to the nebulizer of the AAS apparatus manually with a bulb pipette, or automatically by a suitable sample changing system. A total of 500 microliters serum is sufficient for the determination of traces of Fe, Cu, and Zn. It was shown that under suitable conditions a total volume of 25 microliters serum is sufficient for the determination of Cu and Zn by flame atomic absorption-atomic emission photometry.
Copper↗
On the use of liquid scintillation vials in ATP-photometry.
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Adenosine Triphosphate↗