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Red cell aggregation in blood flow. I. New methods of quantification.

The rheological behavior of normal and pathological red cell aggregates in viscometric flow (artificial flow in cone plate chamber) is studied by direct microscopy, (rheoscopy) viscometry and photometry. Marked differences between normal and pathological blood are measured in the microrheological properties of red cell aggregates; only discreet differences are measured by blood viscometry (macrorheology). Both in normal and abnormal blood, red cell aggregation is a reversible process in the presence of adequate shear forces; their respective influences on apparent blood viscosity at low rates of shear are complex functions of shear rate, shear time, hematocrit and plasma viscosities. Pathological red cell aggregation (RCA) forms more rapidly and extensively than normal RCA. The pathological aggregates frequently have a tendency to grow at low rates of shear and they are highly shear resistant.

Adolescent↗

Photometric oculometry: a slit-lamp technique for determination of intraocular distances.

The optics of slit-lamp fluorophotometry were analyzed. This analysis forms the basis of the formulation of a slit-lamp technique called photometric oculometry, which makes it possible to measure intraocular distances. A comparison between ultrasonography and photometric oculometry in the determination of the ocular axial length was carried out. Measurements in 11 eyes showed statistical agreement, with a mean of 22.90 mm, SD +/- 1.815 (ultrasound), and a mean of 23.17 mm, SD +/- 1.667 (photometry). The principle involved in photometric oculometry is of special importance for the calculation of the blood-retinal barrier permeability to fluorescein by vitreous fluorophotometry, since the method allows the volume of the vitreous body to be estimated directly from the slit-lamp fluorophotometer recording.

Eye↗

Kinetics of cellular proliferation in regenerating mouse liver pretreated with the alkylating drug dipin.

The effect of prior exposure to the alkylating drug 1,4-Bis[N,N'-di(ethylene)-phosphamide] piperazine (Dipin) on the cell cycle progression in a regenerating mouse liver is reported, using cytological, autoradiographical and DNA-cytophotometrical methods. In Dipin-treated animals 3H-TdR pulse labelled nuclei appeared 22 h after the partial hepatectomy, followed by a rapid rise in labelling index to a very high level between 43-54 h, and then by a gradual decline to 72 h. Four injections of 3H-thymidine at 16 h intervals resulted in 89% labelling of hepatocytes at 72 h. The DNA-synthesis time was assessed by DNA photometry combined with 3H-TdR autoradiography on the same nucleus. The DNA-content in Dipin-pretreated nuclei was doubled after 26 h, approximately three times longer than in the controls. Mitoses were completely absent during the first 2-4 days. They were observed subsequently for up to 2 months but with varying frequency. The frequency of binucleated hepatocytes remained constant during liver regeneration. Hence, pretreatment of resting liver with Dipin altered the kinetics of cell proliferation following partial hepatectomy. Delay in division resulted primarily from an increase in the DNA-synthesis time and a prolonged block of the transition from G2 to mitosis. No changes in the duration of the prereplicative period and in the size of the proliferative pool were noticed. The increase of the mitotic cycle time led to synchronization of the cell population. About a half of the hepatocytes were seen in the S phase by the end of the second day and by the end of the third day about 80% of the cells had passed into the G2 phase.

Alkylating Agents↗

Pharmacokinetic evaluation of co-administration of nefazodone and lithium in healthy subjects.

OBJECTIVES: To evaluate the possible pharmacokinetic interaction between nefazodone and lithium. METHODS: Twelve healthy volunteers received nefazodone 200 mg b.i.d. for 5 days. A 4-day washout phase followed from day 6 to day 9. From day 10 to day 20, escalating doses of lithium 250 mg b.i.d. to 500 mg b.i.d. were given; the daily dose of 1000 mg was obtained on day 13. From day 16 to day 20, nefazodone 200 mg b.i.d. was added to the lithium dosing regimen. Venous blood sampling was performed on days 5, 15 and 20 for 0- to 48-h-pharmacokinetic analysis. Nefazodone and its metabolites, hydroxynefazodone, mCPP and triazoledione were assayed by high-performance liquid chromatography (HPLC). Lithium was assayed by flame photometry. RESULTS: Co-administration of nefazodone did not modify pharmacokinetic parameters of lithium at steady-state. Comparison of the area under the plasma or serum concentration-versus-time curve calculated from 0-12 h (AUC0-12) of nefazodone and hydroxynefazodone revealed no significant differences when nefazodone was administered alone or with lithium. The mean maximum peak plasma concentration Cmax and AUC0-12 of meta-chlorophenyl-piperazine (mCPP) were significantly reduced by 27% (P < 0.001) and 16% (P < 0.001) with the co-administration. The mean Cmax and AUC0-12 of triazoledione were reduced by 23% (P < 0.005) and 16% (P < 0.01) by the co-administration. CONCLUSION: Since there were no clinically significant changes in the pharmacokinetics of the parent compounds or metabolites, and the combination was well tolerated, no dosage adjustments of nefazodone or lithium are necessary when they are co-administered.

Adolescent↗

Comparison of 0.1% dexamethasone phosphate eye gel (Dexagel) and 1% prednisolone acetate eye suspension in the treatment of post-operative inflammation after cataract surgery.

UNLABELLED: A prospective, multi-centre, clinical parallel group study was conducted to assess the efficacy and safety of a new 0.1% dexamethasone phosphate eye gel (Group 1, n= 117) compared to 1% prednisolone acetate eye suspension (Group 2I, n=119) in a total of 236 patients (safety population), aged 39-92 years, following cataract surgery. Both drugs were given four times a day for 14 days starting 24+/-4 h after surgery. Criteria for evaluation were the reduction in anterior chamber flare and inflammation severity score (primary efficacy criteria) as well as different secondary efficacy and safety evaluation criteria. Laser photometry (LFM-500, Kowa), slit lamp assessment and the examination of other objective and subjective symptoms of ocular discomfort were performed between the last preoperative and 14th post-operative day. There were no statistically significant differences between the treatment groups concerning primary and secondary efficacy criteria. The mean reduction in anterior chamber flare from day 1 to day 14 post-operatively was 8.34+/-20.80 photons/ms with 0.1% dexamethasone eye gel and 5.72+/-16.70 photons/ms with 1% prednisolone eye suspension. The mean reduction of inflammation severity score was 1.8+/-1.3 points in Group 1 and 2.0+/-1.1 points in Group 2. Intra-ocular pressure did not increase after treatment with 0.1% dexamethasone phosphate eye gel. CONCLUSION: the results of the study underline the protective effect of topically applied 0.1% dexamethasone phosphate eye gel on the blood-aqueous barrier. This drug is an effective and safe steroidal antiinflammatory agent for topical use following cataract surgery and intraocular lens implantation.

Administration, Topical↗

Alkaline haematin D-575, a new tool for the determination of haemoglobin as an alternative to the cyanhaemiglobin method. II. Standardisation of the method using pure chlorohaemin.

Chlorohaemin with high purity (greater than 99%), a stable and well-defined compound, can be used as a primary standard for the standardisation of a haemoglobin assay based on alkaline haematin D-575 [6]. Dissolved in a solution of 25 g Triton X-100 per litre of 0.1 mol/l NaOH ('AHD solution'), the millimolar absorbance coefficient of the end product (alkaline haematin D-575) is 6.960 +/- 0.046 [l X mmol-1 X cm-1] at 575 nm. Within the range of haemoglobin concentrations of 5 to 25 g/100 ml there is a strong linear relation between chlorohaemin concentration and absorbance with a deviation of less than or equal to 2% from the theoretical values. As compared to the conventional cyanhaemiglobin standard solutions, standardisation with pure chlorohaemin is the method of choice because of the simplicity of the preparation of standard solutions, which can be done in every laboratory, and the stability of both the solid compound chlorohaemin and its solutions in alkaline Triton X-100. For the first time a real standard for quality control in haemoglobinometry is recommended: a concentrated solution which behaves like blood, i.e. the simulation of all steps in haemoglobin determination (dilution and photometry) is possible.

Heme↗

Enzymatic determination of sodium and chloride in sweat.

OBJECTIVE: To develop methods based on enzyme activation for the analysis of sweat sodium and chloride using beta-galactosidase and alpha-amylase, respectively. METHODS: Both were monitored kinetically on the Cobas Fara centrifugal analyzer. The sweat, collected with the Macroduct system, was diluted no more than five-fold for the volumes obtained of 16 to 80 mu L, median 32.5 mu L. The sodium assay utilized a sodium-binding cryptand to maximize linearity. RESULTS: Between-run coefficients of variation (%) at 10, 20, and 50 mmol/L were 3.6, 4.5, and 1.3 for sodium and 7.1, 6.1, and 6.0 for chloride, respectively. The sodium method showed excellent agreement with flame photometry (y = 0.997x + 0.742; r = 0.998), and chloride with a mercuric thiocyanate method (y = 0.995x + 0.485; r = 0.996), giving equivalent discrimination between patients with and without cystic fibrosis. CONCLUSIONS: The methods enable the rapid analysis on the same analyzer of both sodium and chloride in a single dilution of sweat collections of low volume.

Bridged Bicyclo Compounds, Heterocyclic↗

Remodeling of the myoblast membrane accompanies development.

We have prepared a library of cloned hybridomas that produce monoclonal antibodies reactive with the surface of E63 rat myoblasts. Using immunofluorescence analysis of antigens on single cells we have studied the expression of determinants at distinct stages of development. Conditions were established for quantitative photometry and were used to confirm the diversity in stage-specific expression that accompanies development. The remodeling of the myoblast membrane also involves stage-specific and transient changes in topography and aggregation of many antigens, and the period surrounding fusion is one of particular activity. The localization of antigens on the upper and attached surfaces of myogenic cells was often distinct, and quantitative and spatial stage-specific reorganizations of antigens differed with respect to these two surfaces. This polarity represents an additional level of complexity in the continuous remodeling of the muscle cell membrane. Comparisons of quantitative and topographic analyses of antigens on E63 cells with Rat-1 fibroblasts and developmentally defective (fu-) myoblasts indicate that the outer membranes of these nonmyogenic cells are distinct from differentiating myoblasts. One determinant, H36, is absent on Rat-1 cells and on all fu- lines tested, and undergoes interesting stage-specific changes in expression and topography.

Animals↗

Carbachol has opposite effects to glucose in raising the sodium content of pancreatic islets.

Integrating flame photometry was used for measuring sodium in single pancreatic islets from ob/ob mice. Exposure to 100 microM carbachol resulted in a 25-40% increase in sodium without any effect on potassium during incubation with 0-5 mM glucose in media deficient or not in Ca2+. This action of carbachol was abolished by 10 microM atropine or by raising the glucose concentration to 20 mM. A minor increase of the steady state content of sodium occurred in the presence of 200 microM ATP or 10 nM tetradecanoylphorbol 13-acetate (TPA). Carbachol differed from TPA in markedly stimulating sodium accumulation after ouabain inhibition of the Na/K pump. The results indicate that muscarinic receptor activation has opposite effects to glucose in inducing a rise of the islet content of sodium. It is suggested that the cholinergic control of the endocrine pancreas involves entry of Na+ in addition to the Na+ entry mediated by protein kinase C activation of Na+/H+ countertransport.

Adenosine Triphosphate↗

Sulfonylureas mimic glucose in stimulating the uptake of Na+ in pancreatic islets exposed to ouabain.

The increase of sodium in response to ouabain inhibition of the Na+/K+ pump was measured in beta-cell-rich pancreatic islets from ob/ob mice using integrating flame photometry. D-Glucose promoted the uptake of sodium when added at a concentration of 6 mM or above. The hypoglycemic sulfonylurea compound, tolbutamide, mimicked the action of D-glucose in stimulating the sodium uptake at concentrations of 10 microM or above. There was no stimulation beyond that obtained with 20 mM glucose during exposure to 100 microM tolbutamide. Other test substances also affected sodium uptake in a way reflecting known effects on insulin release. Accordingly, the sodium uptake was stimulated with glibenclamide, glipizide, HB 699 (4-[2-(5-chloro-2-methoxybenzamido)ethyl]benzoic acid) and high K+. Sulphonamides (sulfamethazole, sulfadiazine and sulfadoxine) had practically no effect when added at a concentration of 1 mM. Sodium uptake in response to glucose and tolbutamide was antagonized by 400 microM diazoxide or 4 microM tetrodotoxin. It is concluded that both glucose and hypoglycemic sulfonylureas stimulate Na+ entry into the pancreatic beta-cells, a process presumably involving depolarization.

Animals↗

Quantitative immunoautoradiography at the cellular level. I. Design of a microphotometric method to quantitate membrane antigens on single cells using 125I-labeled antibodies.

Iodine-125 has become a commonly-used radioisotope, especially for immunoautoradiographic investigations. Microphotometry of grain density, a well-established method in autoradiography with tritium and carbon-14, was applied to nucleated cells with 125I-labeled membranes. Geometric and absorption factors of radiation were investigated in order to find suitable conditions for quantitative evaluation. A preparatory device is given and a set-up of appropriate measuring conditions is presented. With these prerequisites the reflected-light bright-field photometry of immunoautoradiographs permits to determine automatically the content of surface antigens of single cells. Measurement examples were demonstrated.

Animals↗

Highly sensitive amperometric enzyme immunoassay for alpha-fetoprotein in human serum.

A sandwich amperometric enzyme immunoassay with flow injection for alpha-fetoprotein in human serum has been developed with alkaline phosphatase as the enzyme label. p-Hydroxyphenyl phosphate was used as the substrate for alkaline phosphatase. The hydrolysis product, hydroquinone, was detected by oxidative amperometry in a flow injection system. The amperometric wall jet detector was fitted with a glassy carbon working electrode held at 350 mV vs. Ag/AgCl. The detection limit of hydroquinone in 30 mM borate buffer pH 9.5 was 1.2 x 10(-10) M (linearity range: 10(-9)-5.12 x 10(-6 M). A detection limit for free alkaline phosphatase of 1.2 x 10(-15) M (linearity range: 10(-15)-10(-13) M), or about 36 000 molecules, was observed (same borate buffer and incubations of 10 min at 25 degrees C). These conditions were maintained for the amperometric alpha-fetoprotein immunoassay. For comparison purposes, a photometric detection system was set up, with p-nitrophenyl phosphate as enzyme substrate and the same pair of antibodies and incubation conditions. The detection limit for alpha-fetoprotein obtained by amperometry, 0.07 ng/ml (linearity range = 5-500 ng/ml), was 14 times lower than by photometry. The amperometric enzyme immunoassay correlates well with a commercial colorimetric immunoassay (r = 0.986, slope = 0.967, n = 240).

Alkaline Phosphatase↗

Temporal induction of blackness--II. Spectral efficiency and tests of additivity.

The perception of blackness was investigated by measuring spectral efficiency and field additivity under conditions of temporal induction. For both purposes, observers foveally viewed a 0.75 deg, inducing stimulus for 5 sec followed immediately (or 200 msec later) by a broadband (5500 K) reference stimulus of the same size and spatial location. For spectral efficiency measurements, the inducing field was a monochromatic light between 400 and 700 nm (10 nm steps), while the additivity studies involved various wavelength mixtures. The psychophysical task was to increase the radiance of the inducing field until the reference stimulus just turned completely black. For two observers the spectral efficiency function of temporally-induced blackness more closely resembled their heterochromatic flicker photometry (HFP) function than their direct brightness-matching function. Their brightness functions were characterized by an inflection at about 580 nm which is generally ascribed to subtractive cone interactions, but their blackness-induction and HFP functions did not show this inflection. The brightness function of the third observer did not show an inflection at 580 nm, thereby making it difficult to differentiate between her three spectral efficiency functions. Overall, the subtle differences between the various spectral efficiency functions made it difficult to determine whether blackness induction was more similar to HFP or brightness matching. The results from the additivity tests of blackness induction, HFP and direct brightness matching removed this ambiguity from the spectral efficiency findings. Blackness induction and HFP were shown to be additive, whereas the results from brightness matching showed clear additivity failures of the cancellation type. These data support the view that the perception of blackness is mediated by neural mechanisms that additively combine the inputs of middle- and long-wave photoreceptors.

Adult↗

The minimum motion technique applied to determine isoluminance in psychophysical experiments with monkeys.

Isoluminance ratios for red/green, red/blue and blue/green sinusoidal gratings were determined in macaque monkeys using the minimum motion heterochromatic photometry technique (Anstis & Cavanagh, 1983), in which the motion of specially constructed test grating reverses at the point of isoluminance. The point of luminance equality between the two colors was determined using (1) overt responses of monkeys trained to discriminate motion direction and (2) reversals in the direction of the nystagmic eye movements. Both measures yielded essentially identical isoluminance points. Both the spatial and the temporal frequency of the grating patterns were found to influence the isoluminance settings in all animals tested. The amount of blue equating a preset red or green value was found to increase consistently with increased spatial frequency and with increasing eccentricity. These results show the necessity of spatial and temporal frequency dependent adjustment of visual patterns used in psychophysical and physiological experiments dealing with the processing of color. Furthermore, eye movement measurements appear to be sufficiently in order to determine isoluminance in foveally presented moving patterns with nonhuman primates.

Animals↗

Genetic studies of variation in Rayleigh and photometric matches in normal trichromats.

The inheritance of Rayleigh match midpoints and photometric matches (551-667 nm) was studied in observers with normal color vision. An analysis was performed to evaluate whether the measured interobserver variations in these two traits were consistent with single gene allelic variation, polygenic variation, or environmental factors. A bipartite 2 deg field and a computerized tracking method were used to obtain Rayleigh matches; a new photometric technique, termed heterochromatic modulation photometry (HMP), was used to obtain photometric matches. Data were collected from 72 nonrelated males to determine distribution characteristics for the normal population. The distributions were analyzed for evidence of multimodality and the results indicated that the distributions for Rayleigh match midpoints and HMP matches obtained in this study were unimodal and symmetrical. Data from 52 observers from 5 families were used to study the transmission of the two traits in pedigrees. Statistical analysis of the pedigree data suggested that the major source of variations for Rayleigh match midpoints and for HMP matches were each due to allelic variation at single gene loci; that is, each may be determined by a single gene. Results were inconclusive as to whether variation in the two traits could be determined by the same gene.

Adolescent↗

The contribution of chromatic and achromatic valence to spectral saturation.

The spectral efficiency of the achromatic and opponent chromatic channels was measured in three subjects by use of heterochromatic flicker photometry and hue cancellation, respectively. Heterochromatic brightness matching was also used for measuring achromatic spectral efficiency. These data were then used to predict spectral saturation based on Hurvich and Jameson's (1957; Psychological Review, 64, 384-404) opponent colors model. A standard color-naming procedure and a saturation matching technique were used for measures of spectral saturation. The ratio of saturation of short-wave to long-wave lights was found to be less than that predicted by the linear valence model. Allowing for nonlinearity at the opponent site of the yellow-blue channel plus a desaturating signal from the rods provided a good fit between data and theory.

Adult↗

Brightness matching and flicker photometric data obtained over the full mesopic range.

Three normal observers made visual matches from 0.03 to 100 td on a 10 degree central field at 445, 560 and 630 nm, using two different methods: heterochromatic flicker photometry and direct comparison brightness matching. In a relative sensitivity vs illuminance plot, the brightness matches underwent smooth changes, while the flicker curve exhibited a step-like transition around 1 td, between two separate branches. A reverse Purkinje shift was found on the high mesopic branch at 630 nm, possibly due to cone-cone interactions, and on the low mesopic branch at 445 nm, with questionable origin. At 630 nm and 0.03 td, both methods yielded matches that were not truly scotopic but might not have addressed the same mechanisms.

Adult↗

Confusion points and constant-luminance planes for trichromats, protanopes and deuteranopes.

The confusion points of dichromats are derived from the constant-luminance planes of trichromats, protanopes and deuteranopes experimentally defined by heterochromatic-flicker photometry: (1) the zero-luminance planes of the observers considered in this experiment intersect almost exactly in a line that crosses the plane of the chromaticity diagram in the tritanopic-confusion point and confirm that the short-wavelength sensitive cones can be considered to have no contribution to luminance; (2) protanopic- and deuteranopic-confusion points are taken as being defined by the intersection of the tangent line to the long-wavelength region of the spectrum locus and the zero-luminance plane for protanopes and deuteranopes, respectively.

Adaptation, Ocular↗