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The magnitude of longitudinal chromatic aberration of the human eye between 458 and 633 nm.

The longitudinal chromatic aberration (LCA) of the human eye was determined between wavelengths 458 and 633 nm for ten observers by incorporating Argon and Helium-Neon Lasers into a Badal optometer system such that tonic (or "dark focus") resting positions of accommodation could be measured under darkroom conditions. A mean chromatic range of 1.87 D was found between 488 and 633 nm. The range increased slightly to 1.91 D when the experiment was repeated under cycloplegia on a subset of three subjects. Additional wavelengths (458 and 476 nm) for a further subset (four subjects) increased the range to 2.65 D. This magnitude of LCA would support recent predictions based on Abbe and Pulfrich refractometry analyses of the dispersion of the human crystalline lens and ocular media.

Accommodation, Ocular↗

Solution characterization of starch nicotinates with different degrees of esterification.

A series of starch-nicotinic acid copolymers with a degree of esterification ranging from about 15% to about 90% was prepared, and the stability in solution of two representative samples of the series was tested. The variation of some physico-chemical characteristics in the series was examined by solubility tests, viscometry and refractometry, and found to be not simply correlatable to the nicotinylation degree.

Esters↗

Estimation of molecular weights of membrane proteins in the presence of SDS by low-angle laser light scattering combined with high-performance porous silica gel chromatography. Confirmation of the trimer structure of porin of the E. coli outer membrane.

An assessment study was carried out to evaluate the performance of the low-angle laser light-scattering technique combined with high-performance porous silica gel chromatography in the presence of sodium dodecyl sulfate and precision differential refractometry. It was found that the combined technique is highly promising as a reliable method for determining the molecular weight of a membrane protein solubilized by the surfactant. As a test, molecular weights of porin forming the permeability channel of the outer membrane of E. coli B in an oligomeric form were measured before and after heat treatment, which is known to cause dissociation. The results obtained indicate that the porin oligomer is a trimer with stoichiometric composition.

Chromatography, High Pressure Liquid↗

Determination of molecular weight of membrane proteins by the use of low-angle laser light scattering combined with high-performance gel chromatography in the presence of a non-ionic surfactant.

An assessment study was carried out to evaluate the performance of the low-angle laser light scattering technique combined with high-performance gel chromatography in the presence of a nonionic surfactant, octaethyleneglycol n-dodecyl ether, precision differential refractometry and ultraviolet photometry. It was found that the combined technique is highly promising as a method for the determination of the molecular weight of a membrane protein solubilized by the surfactant. For trial, molecular weights of the following membrane proteins of Escherichia coli, both solubilized in oligomeric forms, were measured; porin that forms the transmembrane diffusion pore in the outer membrane, and lambda-receptor protein that facilitates the diffusion of maltose-maltodextrins across the outer membrane. The result obtained indicates that both porin and lambda-receptor protein exist as trimers in the surfactant solution.

Bacterial Outer Membrane Proteins↗

A comparison of three methods for determining the concentration of rat urine.

The concentrations of 78 rat urines were compared using osmometry, refractometry and test strips for specific gravity. Test strip specific gravity values are a guide to urine concentration; where small changes of urine concentration are expected, the test strips should not be a replacement for more accurate methods such as osmometry.

Animals↗

A field lab method to determine urine concentration in small mammals.

1. The concentrations of 136 urine samples from four species of small mammals were compared using osmometry, refractometry and a colorimetric test for urea concentration. 2. To obtain a wide range of concentrations (430-3950 mOsm/kg), urine samples were collected under normal and dehydration conditions. 3. Regression analyses of paired values indicate that measurements of total solids concentration (refractometric method) permit evaluations of urine osmolality and estimations of the concentration of urea with a high degree of confidence.

Animals↗

Dilatometric, refractometric and viscometric study of lysozyme-cation interaction.

The interaction between hen egg-white lysozyme and Cu(II) or Co(II) cations has been studied by dilatometry, equilibrium dialysis-differential refractometry and viscometry at different metal cation concentrations. Delta V isotherms in copper and cobalt solutions have been obtained from dilatometry. Preferential adsorption parameters and specific viscosity have been determined from refractometric and viscosimetric measurements. It has been observed that this interaction produces structural alterations in lysozyme. The magnitude of these conformational changes depends on the metal ion and protein concentration. The results obtained using the three techniques are in good agreement.

Animals↗

Insulin association in neutral solutions studied by light scattering.

Molecular weights and weight distributions of sulfated, Zn-free, and 2Zn insulins have been measured at pH 7.3 as a function of concentration from 0.1 to 2 mg/ml by use of a combination of light scattering, refractometry, and size-exclusion chromatography. Results show that sulfated insulin is monomeric over the studied concentration range. Weight average molecular weights between those of a monomer and a hexamer were found for both zinc-free and 2Zn insulins. Zinc stabilizes the hexamer, and the dimer-hexamer equilibrium constant is approx. 400-times higher in the presence of zinc than in its absence. An average hydrodynamic radius of 5.6 nm, close to the crystallographic size of the insulin hexamer, was determined from dynamic light scattering of 2Zn insulin solutions.

Journal Article↗

Impact of phototherapeutic keratectomy on the outcome of subsequent penetrating keratoplasty in patients with stromal corneal dystrophies.

PURPOSE: To examine the impact of previous phototherapeutic keratectomy (PTK) on the outcome of subsequent penetrating keratoplasty (PK) in patients with stromal corneal dystrophies. DESIGN: Retrospective, cross-sectional, clinical single-center study. METHODS PATIENT POPULATION: Fifteen patients (21 eyes) age 39.9 +/- 11.4 years. INCLUSION CRITERIA: Primary homologous PK performed in phakic patients with granular or macular dystrophy; no use of combined surgical procedures; defined graft size and technique. The study group comprised eight eyes of five patients, PK performed 3.7 +/- 2.3 years after PTK. The control group (no previous PTK) comprised 13 eyes of 10 patients. In both groups, 38% had granular and 62% had macular dystrophy. Intervention Procedures: Phototherapeutic keratectomy was performed using a 193 nm excimer laser. All PKs were also performed using this laser, with trephination using a metal mask. Subjective refractometry (trial lenses), standard keratometry (Zeiss ophthalmometer), and corneal topography (Tomey TMS-1) were performed preoperatively, 6 months after PK, and after first and second suture removal (1.1 +/- 0.2 years; 1.6 +/- 0.2 years). MAIN OUTCOME MEASURES: Keratometric, topographic net astigmatism, and refractive cylinder; keratometric and topographic central power; best-corrected visual acuity (BCVA); surface regularity index (SRI), surface asymmetry index (SAI), potential visual acuity (PVA). RESULTS: Refractive power and astigmatism, BCVA, and PVA values did not differ significantly between the two groups at any time-point; SRI tended to be better in the study group after first suture removal (P =.05). CONCLUSION: Preceding PTK does not appear to impair the outcome of subsequent penetrating keratoplasty in stromal corneal dystrophy patients.

Adult↗

Repeat keratoplasty for correction of high or irregular postkeratoplasty astigmatism in clear corneal grafts.

PURPOSE: To evaluate the functional results of repeat penetrating keratoplasty in clear corneal grafts with high/irregular postkeratoplasty astigmatism. DESIGN: Retrospective, longitudinal, single-center, consecutive clinical case series. METHODS: We studied 17 eyes (16 keratoconus, 1 Fuchs' dystrophy) of 16 patients (age, 54.9 +/- 12.6 years). They were treated with repeat PK, performed using the 193-nm Zeiss-Meditec MEL-60 excimer laser using round metal masks (diameter, 7.5-8.0 mm), and employing double running sutures. main outcome measures: Subjective refractometry, standard keratometry, and corneal topography (Tomey TMS-1) were used to assess best-corrected visual acuity (BCVA), spherical equivalent (SEQ), keratometric and topographic central corneal power (CP), refractive, keratometric and topographic astigmatism, surface regularity index (SRI), surface asymmetry index (SAI), and potential visual acuity (PVA) preoperatively, before and after first suture removal (1.1 year), and after second suture removal (1.8 years). RESULTS: Visual acuity improved significantly (BCVA from 0.2-0.5, P = .04 or better) for all postoperative measurements. CP decreased significantly, but SEQ did not change. All measures of astigmatism and SRI and SAI values showed postoperative improvement with sutures in place; however, astigmatism increased significantly after second suture removal. CONCLUSIONS: With all-sutures-in, BCVA and astigmatism improve significantly after repeat PK for high/irregular astigmatism. However, to present significant increase in astigmatism, final suture removal should be postponed as long as possible in such eyes.

Adult↗

Spontaneous long-term changes of corneal power and astigmatism after suture removal after penetrating keratoplasty using a regression model.

PURPOSE: To assess the diagnosis-based spontaneous long-term changes in corneal power and refraction with a regression model in the all-sutures-out time period following non-mechanical penetrating keratoplasty (PK). DESIGN: Retrospective non-randomized clinical trial. METHODS: setting: Clinical practice. study population: 147 eyes [47 Fuchs dystrophy (FD); 100 keratoconus (KC)] were studied after suture removal in this retrospective longitudinal study. main outcome measures: Zeiss keratometry [equivalent power (KEQ) and astigmatism (KAST)], corneal topography analysis [equivalent power (TEQ) and astigmatism (TAST)], and subjective refractometry [spherical equivalent (SEQ) and refractive cylinder (RAST)] were assessed in at least three up to 16 ophthalmologic examinations in the all-sutures-out time period. observation procedure: The time course of each target variable was analyzed in a longitudinal manner (time interval > or = 12 months) separately for each patient with a linear regression model. RESULTS: Post-keratoplasty follow-up ranged from 31 months to 10.3 years. In the linear regression model, the annual change in FD/KC showed an increase/a decrease in KEQ (0.29 +/- 0.50/-0.63 +/- 0.46 diopters, P = .02) and an increase/a decrease in TEQ (0.37 +/- 0.54/-0.69 +/- 0.49 diopters, P = .04) corresponding to a decrease/an increase in SEQ (-0.31 +/- 0.47/0.63 +/- 0.43 diopters, P = .02). KAST/TAST/RAST showed a minimal annual decrease (-0.06 +/- 0.41/-0.05 +/- 0.45/-0.06 +/- 0.41 diopters) in FD but an increase in KC (0.46 +/- 0.41/0.51 +/- 0.43/0.46 +/- 0.38 diopters) (P = .05/0.06/0.12). CONCLUSIONS: In the follow-up after post-keratoplasty suture removal, patients with FD/KC tend to develop a spontaneous myopic shift (steepening of the cornea)/hyperopic shift (flattening of the cornea). In contrast with those with FD, patients with KC should be counseled on the fact that astigmatism may increase again over time after suture removal.

Astigmatism↗

Changes in corneal power and refraction due to sequential suture removal following nonmechanical penetrating keratoplasty in eyes with keratoconus.

PURPOSE: To assess the changes in corneal power and refraction due to sequential suture removal after penetrating keratoplasty (PK). DESIGN: Retrospective consecutive case series. METHODS: setting: Clinical practice. study population: We studied 67 phakic keratoconus eyes (central excimer laser trephination, primary keratoplasty, graft/recipient diameter 8.1/8.0 mm; double running suture) in this longitudinal study. main outcome measures: Zeiss keratometry (equivalent power (KEQ), astigmatism (KAST)), corneal topography (equivalent power (TEQ), astigmatism (TAST)) and subjective refractometry (spherical equivalent (SEQ), refractive cylinder (RAST)) were assessed with sutures in place (interval 1), with one suture out (interval 2), and with all sutures out (interval 3). observation procedure: Corneal power and refraction was decomposed into vector components and the changes were derived between time stages. RESULTS: The mean follow-up period was 3.9+/-1.7 years. At interval 1, the axes of KAST/TAST/RAST were almost randomly distributed. At interval 2, the with/against the rule component of KAST/TAST/RAST decreased slightly and the oblique component increased significantly, so that the axes tended to have a preferred oblique direction. At interval 3, the with/against the rule component of KAST/TAST/RAST increased slightly and the oblique component decreased significantly, so that the with/against the rule component exceeded the oblique component by approximately 23%/28%/25%. Median KEQ/TEQ/SEQ changed by 0.64/0.62/-1.11 diopters (interval 1 to interval 2) and by -0.85/-0.90/1.56 diopters (interval 2 to interval 3). CONCLUSIONS: As a result of removal of the first running suture, corneal astigmatism as well as the refractive cylinder tend to oblique axes. As a result of removal of the second running suture, the final corneal astigmatism and refractive cylinder tend to orientation axes with/against the rule.

Astigmatism↗

Ethanol biosensors based on alcohol oxidase.

The detection and quantification of ethanol with high sensitivity, selectivity and accuracy is required in many different areas. A variety of methods and strategies have been reported for the determination of this analyte including gas chromatography, liquid chromatography, refractometry and spectrophotometry, among other. The use of the enzyme alcohol oxidase (AOX) on the analysis of ethanol in complex samples allows a considerable enhancement in specificity. This paper reviews the state of the art on ethanol determination based on AOX sensors, using either electrochemical electrodes or immobilised enzyme reactors. Almost all AOX-based ethanol sensors developed so far are based on the monitoring of O2 consumption or H2O2 formation. This has been mostly achieved using amperometric electrodes set at appropriate potentials namely, -600 mV for O2 monitoring or +600 mV for H2O2 monitoring. Mediated and non-mediated bienzymatic systems have also been assembled using AOX coupled to horseradish peroxidase (HRP). Different types of electrodes have been proposed for the detection of ethanol, namely, membrane electrode, carbon paste electrodes, screen-printed electrodes and self-assembled monolayers. Another approach to work with this sensitive enzyme is to use high amounts of AOX in order to create an enzyme reservoir, a strategy which can be implemented using immobilised enzyme reactors. These reactors can be combined with a colorimetric detection in a flow-injection analysis system or with electrochemical transducers.

Alcohol Oxidoreductases↗

Development of a digital video-microscopy technique to study lactose crystallisation kinetics in situ.

Polarised light microscopy was employed non-invasively to monitor lactose crystallisation from non-seeded supersaturated solutions in real time. Images were continuously recorded, processed and characterised by image analysis, and the results were compared with those obtained by refractometry. Three crystallisation temperatures (10, 20 and 30 degrees C) and three different levels of initial relative supersaturation (C/C(s)=1.95; 2.34; 3.15) were investigated. Induction times using the imaging technique proved to be substantially lower than those determined using refractive index. Lactose crystals were isolated digitally to determine geometrical parameters of interest, such as perimeter, diameter, area, roundness and Feret mean, and to derive crystal growth rates. Mean growth rates obtained for single crystals were fitted to a combined mass transfer model (R(2)=0.9766). The model allowed the effects of temperature and supersaturation on crystallisation rate to be clearly identified. It also suggested that, in this set of experiments, surface integration seemed to be the rate controlling step. It is believed that a similar experimental set-up could be implemented in a real food system to characterise a particular process where crystallisation control is of interest and where traditional techniques are difficult to implement.

Crystallization↗

Determination of lignin by size exclusion chromatography using multi angle laser light scattering.

A method was developed using high-performance size exclusion liquid chromatography (HPSEC) with multi-angle laser light scattering (MALLS), quasi-elastic light scattering (QELS), interferometric refractometry (RI) and UV detection to characterize and monitor lignin. The combination proved very effective at tracking changes in molecular conformation of lignin molecules over time; i.e. changes in molecular weight distribution, radius of gyration, and hydrodynamic radius. Until this study, UV detection (280 nm) had been the primary lignin determination method for chromatography. Three different HPLC columns were used to study the effects of pH, flow conditions, and mobile phase compositions (dimethyl sulphoxide, water, 0.1M NaOH, and lithium bromide) on the chromatography of lignin. Since light scattering accuracy is highly dependent on solute concentration, both the UV and RI detectors were calibrated for use as concentration detectors. Shodex Asahipak GS-320 HQ column with 0.1M NaOH (pH 12.0) run at 0.5 ml/min was found to give the highest separation and most consistent recovery. The study also revealed that the lignin aggregated at pH below 8.5. This aggregation was detected only by MALLS and was not observed on UV or RI detectors. It is very important to take this loss in apparent concentration due to aggregation into consideration before collecting reliable depolymerization data.

Calibration↗

Emmetropisation under continuous but non-constant light in chicks.

It has been suggested that ambient lighting at night influences eye growth and might play a causal role in human myopia. To test this hypothesis, we reared newly hatched chicks under 12 hr light-dark or light-dim cycles with a light phase intensity of 1500 microW/cm(2) and variable dim phase intensities between 0.01 and 500 microW/cm(2). Other chicks were reared under constant light conditions with intensities between 1 and 1500 microW/cm(2). After three weeks, the chicks were examined by refractometry, ultrasound and caliper measurements of enucleated eyes. To relate ocular parameters with a retinal neurotransmitter likely involved in eye growth control, retinal and vitreal levels of dopamine and its principal metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), were measured by high performance liquid chromatography with electrochemical detection in the light, dark and dim phases. Diurnal fluctuations in axial length and choroidal thickness also were measured twice daily by partial coherence interferometry (PCI) in chicks under light-dark and the two brightest light-dim conditions. The eyes of chicks reared under most light-dim conditions had refractions and ocular dimensions comparable to those reared under light-dark conditions. At dim phase light intensities of 10 microW/cm(2) and above, the day-night changes in retinal dopamine metabolism were not observed. The daily fluctuations of axial length and choroidal thickness were altered with rearing under the two brightest dim light intensities, compared to the light-dark condition. Rearing under constant light with intensities ranging between 1 and 1500 microW/cm(2) produced a shallow anterior chamber and other eye alterations previously described for constant light rearing even though rearing under continuous light that fluctuated between these same intensities generally permitted normal eye growth. Thus, continuous but fluctuating light exerts different developmental effects on the eye than constant non-fluctuating light. Light-dim rearing may be more relevant to daily human light exposures than other laboratory lighting conditions and may provide an opportunity to study developmental interactions of visual quality (e.g. blur, defocus, etc.) and features of the light-dark cycle under conditions that perturb daily rhythms in dopamine metabolism and ocular dimensions. Such studies also could provide mechanistic insights into whether and how daily rhythms in retinal dopamine metabolism, axial length or choroidal thickness modulate refractive development.

3,4-Dihydroxyphenylacetic Acid↗

The heat-sensitive Escherichia coli grpE280 phenotype: impaired interaction of GrpE(G122D) with DnaK.

GrpE is the nucleotide-exchange factor of the DnaK chaperone system. Escherichia coli cells with the classical temperature-sensitive grpE280 phenotype do not grow under heat-shock conditions and have been found to carry the G122D point mutation in GrpE. To date, the molecular mechanism of this defect has not been investigated in detail. Here, we examined the structural and functional properties of isolated GrpE(G122D) in vitro. Similar to wild-type GrpE, GrpE(G122D) is an elongated dimer in solution. Compared to wild-type GrpE, GrpE(G122D) catalyzed the ADP/ATP exchange in DnaK only marginally and did not compete with wild-type GrpE in interacting with DnaK. In the presence of ADP, GrpE(G122D) in contrast to wild-type GrpE, did not form a complex with DnaK detectable by size-exclusion chromatography with on-line static light-scattering and differential refractometry. Apparently, GrpE(G122D) in the presence of ADP binds to DnaK only with much lower affinity than wild-type GrpE. GrpE(G122D) could not substitute for wild-type GrpE in the refolding of denatured proteins by the DnaK/DnaJ/GrpE chaperone system. In the crystal structure of a (Delta1-33)GrpE(G122D).DnaK-ATPase complex, which as yet is the only available structure of a GrpE variant, Asp122 does not interact directly with neighboring residues of GrpE or DnaK. The far-UV circular dichroism spectra of mutant and wild-type GrpE proved slightly different. Possibly, a discrete change in conformation impairs the formation of the complex with DnaK and renders GrpE(G122D) virtually inactive as a nucleotide exchange factor. In view of the drastically reduced ADP/ATP-exchange activity of GrpE(G122D), the heat sensitivity of grpE280 cells might be explained by the ensuing slowing of the chaperone cycle and the increased sequestering of target proteins by high-affinity, ADP-liganded DnaK, both effects being incompatible with efficient chaperone action required for cell growth.

Adenosine Diphosphate↗

The sensitivities and specificities of total plasma protein and plasma fibrinogen for the diagnosis of traumatic reticuloperitonitis in cattle.

We performed a prospective clinical study to select cut-off points for total plasma protein (TPP) and plasma fibrinogen (PF) to differentiate between traumatic reticuloperitonitis (TRP) and other gastrointestinal diseases with similar clinical signs, and to estimate the dependence and accuracy of TPP and PF when used in series or in parallel. TPP and PF were estimated preoperatively by refractometry in 93 cattle with TRP and 65 cattle with gastrointestinal diseases such as vagus indigestion (n = 16), liver abscesses (n = 17), omasal impaction (n = 19) and Johne's disease (n = 13). Three different cut-off points were selected by two-graph receiver-operating characteristic (TG-ROC) analysis for TPP and PF. Conditional covariances were calculated as a measure of dependence between sensitivities and specificities of TPP and PF. The cut-off points of 7.22, 7.78 and 8.82g/dl for TPP and 622, 691 and 766mg/dl for PF were suggested by TG-ROC based on different requirements of test performances. There was moderate negative dependence between sensitivities of TPP and PF at the 8.82g/dl and 766mg/dl cut-off points, and mild negative dependence between their specificities at the 7.78g/dl and 691mg/dl cut-off points, respectively. Acceptable accuracy (98 or 86% specificity with 62 or 88% sensitivity, respectively) was obtained with serial interpretation of the tests.

Animals↗