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[Focusing strength and a subjective method for determining refraction].

Three hundred and seventy-four eyes were examined by a subjective method, by using trial lenses in natural accommodation. This yielded the following distribution of clinical refraction: hypermetropia (43.6%), emmetropia (30.8%), myopia (12.8%), and astigmatism (12.8%). The classical subjective method of refractometry is based on the schematic eye having no focusing strength. The currently available data by a number of authors show that the focusing strength of more than 0.5 diopters is typical of the human eye, which affects the results of refractometry. When the collecting glass +0.25 D or +0.5 D is applied, the eye with a visual acuity of 1.0 or more retains the level of vision due to the holding the focusing strength across the retina rather than the accommodation strength. By taking in account the correction, the distribution in the study group was as follows: emmetropia (73.0%), myopia (12.8%), astigmatism (12.8%), and hypermetropia (1.4%).

Accommodation, Ocular↗

[Analysis of solutions for parenteral use under field conditions].

Methods for determination of relative density, osmolarity, then refractometry, polarymetry and conductometry as simple methods suitable for work under field conditions have been applied for the quality control of the following solutions for infusion and injections: glucose, mannitol, sodium chloride, sodium hydrogencarbonate, magnesium sulphate and potassium chloride. The results obtained in the analysis of preparations have shown the advantage of refractometry.

Infusions, Parenteral↗

Influence of edge sharpness on the accommodation of the human eye.

Using a subjective method of laser refractometry, the accommodation behavior of six subjects toward real targets with various degrees of blur, dependent on the viewing distance, were investigated monocularly and binocularly. Despite the sharpness of the target, the nominal value is only reached incompletely with a lag of accommodation, because accommodation tends to be as minimal as possible. Increasing blurring leads to a drift of accommodation towards the resting position of accommodation. The resulting underaccommodation causes further physiological blurring of the retinal image of the targets. The approach of the targets to the resting position of accommodation leads to a higher precision of adjustment and less drift. Subjective components (accommodative laziness, cognitive demand) influence the accommodation and increase the phenomenons discovered.

Accommodation, Ocular↗

Active and monomeric human red cell glucose transporter after high performance molecular-sieve chromatography in the presence of octyl glucoside and phosphatidylserine or phosphatidylcholine.

The human red cell glucose transporter (Glut 1) was purified by ion-exchange chromatography in the presence of octyl glucoside. The state of association of the protein was studied, and the transport activity was determined after exchange of copurified membrane lipids for phosphatidylserine (PS) or phosphatidylcholine (PC). The purpose was to analyze the Glut 1 preparation for homogeneity and activity prior to attempts at crystallization. Analyses by high performance molecular-sieve chromatography showed that the Glut 1 was monomeric immediately after the ion-exchange purification: the Mr of the Glut 1 polypeptide was estimated to be 49,000 +/- 6000 by TSKgel G3000SW chromatography monitored by low-angle laser light-scattering photometry, differential refractometry and UV photometry. This required determination of the absorption coefficient of the Glut 1, which was measured to be 1.13 +/- 0.03 ml mg-1 cm-1 at 280 nm, referring to the polypeptide concentration. The Mr value is consistent with the cDNA-deduced Mr 54,117 of the very similar HepG2 glucose transporter polypeptide. At 2 degrees C, pH 7 and an ionic strength of 0.06 M, the Glut 1 associated gradually during three days to form oligomers. These formed much more rapidly at room temperature or at high ionic strength. Freshly prepared Glut 1 retained high activity after separation from membrane lipids on a TSKgel G3000SW column in the presence of 40 mM octyl glucoside and 1 mM PS or PC. In contrast, most of the activity was lost when the membrane lipids were separated from the protein in the absence of eluent lipids. The presence of a phospholipid was thus essential for retention of high activity of the Glut 1 in octyl glucoside and PC was nearly as effective as PS.

Chromatography, Gel↗

Determination of cholesterol and cortisone absorption in polyurethane. I. Methodology using size-exclusion chromatography and dual detection.

A size-exclusion chromatographic method is described for measuring the absorption of the steroid-based lipids cholesterol and cortisone into Pellethane 2363, a polyurethane used in biomedical implants. The method uses refractometry and ultraviolet diode-array detection, with tetrahydrofuran as the mobile phase. Using an injection volume of 150 microliters, the lower limit of accurate measurement for cholesterol (refractive index detection) was 6 micrograms/ml with a lower limit of detection, based on a 2:1 signal-to-noise ratio, of 0.15 micrograms (1 microgram/ml). For cortisone (ultraviolet detection), the lower accurate limit was 0.6 micrograms/ml with a lower limit of 0.015 micrograms (0.1 micrograms/ml). The results show that after 44 h, 2037 micrograms/g cholesterol and 3131 micrograms/g cortisone were absorbed by the polyurethane. The method eliminates extensive sample manipulation and is sensitive to low levels of lipid in the presence of a high-molecular-mass synthetic polymer.

Absorption↗

A new surface plasmon resonance sensor for high-throughput screening applications.

We report a new high-throughput surface plasmon resonance (SPR) sensor based on combination of SPR imaging with polarization contrast and a spatially patterned multilayer SPR structure. We demonstrate that this approach offers numerous advantageous features including high-contrast SPR images suitable for automated computer analysis, minimum crosstalk between neighboring sensing channels and inherent compensation for light level fluctuations. Applications of a laboratory prototype of the high-throughput SPR sensor with 108 sensing channels for refractometry and biosensing are described. In refractometric experiments, the noise-limited refractive index resolution of the system has been established to be 3 x 10(-6) refractive index unit (RIU). Experimental data on detection of human choriogonadotropin (hCG) suggest that in conjunction with monoclonal antibodies against hCG, the reported SPR imaging sensor is capable of detecting hCG at concentrations lower than 500 ng/ml.

Biosensing Techniques↗

Counter-propagating optical trapping system for size and refractive index measurement of microparticles.

We propose and demonstrate a novel approach to measure the size and refractive index of microparticles based on two beam optical trapping, where forward scattered light is detected to give information about the particle. The counter-propagating optical trap measurement (COTM) system exploits the capability of optical traps to measure pico-Newton forces for microparticles' refractive index and size characterization. Different from the current best technique for microparticles' refractive index measurement, refractometry, a bulk technique requiring changing the fluid composition of the sample, our optical trap technique works with any transparent fluid and enables single particle analysis without the use of biological markers. A ray-optics model is used to explore the physical operation of the COTM system, predict system performance and aid system design. Experiments demonstrate the accuracy of refractive index measurement of Deltan=0.013 and size measurement of 3% of diameter with 2% standard deviation. Present performance is instrumentation limited, and a potential improvement by more than two orders of magnitude can be expected in the future. With further development in parallelism and miniaturization, the system offers advantages for cell manipulation and bioanalysis compatible with lab-on-a-chip systems.

Biopolymers↗

Tubulin dipole moment, dielectric constant and quantum behavior: computer simulations, experimental results and suggestions.

We used computer simulation to calculate the electric dipole moments of the alpha- and beta-tubulin monomers and dimer and found those to be |p(alpha)| = 552D, |p(beta)| = 1193D and |p(alphabeta)| = 1740D, respectively. Independent surface plasmon resonance (SPR) and refractometry measurements of the high-frequency dielectric constant and polarizability strongly corroborated our previous SPR-derived results, giving Deltan/Deltac approximately 1.800 x 10(-3)ml/mg. The refractive index of tubulin was measured to be n(tub) approximately 2.90 and the high-frequency tubulin dielectric constant k(tub) approximately 8.41, while the high-frequency polarizability was found to be alpha(tub) approximately 2.1 x 10(-33)C m(2)/V. Methods for the experimental determination of the low-frequency p are explored, as well as ways to test the often conjectured quantum coherence and entanglement properties of tubulin. Biobits, bioqubits and other applications to bioelectronics are discussed.

Computer Simulation↗

Changes in the refractive index of lens fibre membranes during maturation--impact on lens transparency.

PURPOSE: Local variations in refractive index are the physical cause of light scattering in a material or tissue and also induce phase changes of propagating light waves. The goal of this study was to analyse local differences in refractive index by phase contrast microscopy of sections of human lenses. METHODS: Refractive index was estimated by immersion refractometry. Cryo-sections of quick-frozen human donor lenses were embedded in a graded series of bovine serum albumin solutions, and in immersion oil, ranging in refractive index from 1.34 to 1.52. RESULTS: Fibre membranes in the lens cortex prove to have a refractive index considerably above that of fibre cytoplasm at the same location. Fibre membranes in the lens nucleus have a refractive index approximately the same as that of fibre cytoplasm at the same location. CONCLUSION: In the lens cortex, transparency is obtained by a high spatial order of the lens fibre lattice to compensate for the light scattering caused by differences in refractive index between fibre membranes and cytoplasm. In the lens nucleus, high spatial order is less important, because the minor differences in the refractive index between fibre membranes and fibre cytoplasm lead only to minimal scattering.

Cryopreservation↗

Direct determination of amino acids and carbohydrates by high-performance capillary electrophoresis with refractometric detection.

This is an initial report to propose a novel approach in high-performance capillary electrophoresis (HPCE) for the direct detection of compounds without natural absorbance in the UV and visible spectral range, such as amino acids and carbohydrates. A refractometry detector with the 2 nl cell (Applied Systems, Minsk, Belarus) was employed to identify amino acids and carbohydrates without derivatization. The first results are provided on separation of seven free amino acids in the phosphate running buffer and three free carbohydrates in the borate-sodium dodecyl sulfate running buffer and detection by refractometer. Fused capillaries of 50 or 75 microm internal diameter and separation voltage (10-23 kV) were applied. Detection limits ranged typically from 10 to 100 fmol and the response was linear over two orders of magnitude for most of the amino acids and carbohydrates. The HPCE system demonstrated good long-term stability and reproducibility with a relative standard deviation, less than 5% for the migration time (n=10).

Amino Acids↗

Polymer characterization by size-exclusion chromatography with multiple detection.

Size-exclusion chromatography with coupled multiangle light scattering and differential refractometry detectors has been used to obtain molecular mass and radius of gyration distributions of polydisperse polymer samples. From these data the scaling law between dimensions and the absolute molecular mass is obtained with just one sample of each polymer. Three different kinds of polymers are presented: polystyrene which serves as reference polymer, polyphosphazenes which behave abnormally in solution and poly(ethylene oxide) which is soluble in water. Since the relationship between dimensions and molecular mass depends on the extent of interactions between chain segments and solvent molecules, the scaling law provides information about the solution properties of the polymer.

Chromatography, Gel↗

Influence of chain architecture on the mechanochemical degradation of macromolecules.

We detail here studies into the nature of mechanochemical degradation of macromolecules, effected by means of ultrasonic (US) irradiation. Specifically, we have investigated the effect of long-chain branching (LCB), in the star configuration, on the degradation mechanism of polystyrene dissolved in DMAc/LiCl. The information obtained from size-exclusion chromatography with triple detection (refractometry, viscometry, multi-angle light scattering) shows that the degradation mechanism of stars is radically different from that of linear polymers. Whereas in the latter, from a macromolecular standpoint, it is merely necessary for the molar mass to be greater than some limiting value (M(lim)), in the former both molar mass and structural factors affect ultrasonic degradation. We have examined the effects of arm number and of arm molar mass on star degradation, and propose the concept of a spanning molar mass (M(span) approximately 2M(arm)) such that, even in the event that M(arm) M(lim). This mechanism has been extended to other types of architecture (e.g., H-branched, dendritic), where it is proposed that a continuous path must exist with M(path)>M(lim) for degradation to occur. Examination of the different radii afforded by viscometric and light-scattering detection gives insight into the solution thermodynamics and conformation of the stars with differing arm number and molar mass and of the effects of insonation on macromolecular structure.

Chromatography, Gel↗

Autorefractometry after laser in situ keratomileusis.

PURPOSE: To correlate cycloplegic subjective refraction with cycloplegic autorefractometry in eyes that have had laser in situ keratomileusis (LASIK). SETTING: Vlemma Eye Institute, Athens, Greece. METHODS: Subjective refraction and autorefractometry under cycloplegia were performed in 73 eyes of 46 patients 1, 6, and 12 months after LASIK to correct myopia or myopic astigmatism. The preoperative subjective refraction and autorefractometry under cycloplegia in the same eyes served as controls. RESULTS: A statistically significant difference between subjective refraction and autorefraction was found in the sphere and cylinder at all postoperative times. No statistically significant difference was found in the axis. There was no statistically significant difference in the control eyes. CONCLUSIONS: Automated refractometry in eyes that had had LASIK was reliable in the axis only. Retreatments after LASIK should always be based on subjective refraction.

Adult↗

Investigation of disaccharide recognition by molecularly imprinted polymers.

The selectivity of carbohydrate-imprinted polymers for several disaccharides, namely cellobiose, maltose, lactose and gentiobiose, is investigated. An ternary ligand-Cu(II)-carbohydrate complex was formed in alkaline solution and captured afterwards in the polymer. The accessibility of the polymer matrix for disaccharides was investigated by HPLC analysis, refractometry and 1H NMR spectroscopy applying excess of the original template during rebinding experiments under saturation conditions in unbuffered, aqueous solution at neutral pH and 20 degrees C. The selective discrimination of the alpha- and beta-glycosidic linkage of cellobiose and maltose is demonstrated. It is further shown, that the disaccharide-imprinted polymers slightly distinguish between the 1,4-beta- and the 1,6-beta-glycosidic linkage of cellobiose and gentiobiose, while cellobiose and lactose are not selectively recognized. Due to the weak apparent binding constant of the functional Cu(II) monomers with the targeted disaccharides at physiological pH, the recognition process is dominated by the shape of the created imprinted cavity under the applied conditions.

Carbohydrate Sequence↗

Use of the refractometer as a tool to monitor dietary formula concentration in gastric juice.

BACKGROUND AND AIMS: Critically ill patients do not always tolerate nasogastric tube feeding. Gastric residual volumes are widely used to evaluate feeding tolerance, but controversy exists about what constitutes the residual volume (diet formula or digestive juice). In this paper, we describe the use of the refractometer as a tool to monitor dietary formula concentration in gastric juice and evaluate gastric juice refractometry as a possible clinical application. METHODS: Brix value (an index of the total solutes in solution) readings for polymeric diet at pH 1, 4, 7 and 8, and at 4 degrees C, 25 degrees C and 37 degrees C, and in fasting gastric juice were determined with a refractometer. RESULTS: We found that distilled water, minerals, and vitamins had low Brix values of 0+/-0, 1.2+/-0.1, and 0.4+/-0.1, respectively. On the other hand, because carbohydrate (17 g/100 ml), protein (5.3 g/100 ml), fat (4.1 g/100 ml), and full-strength polymeric diet had high concentrations of dissolved nutrients, they also had high Brix values (12.1+/-0.6, 6.5+/-0.1, 6.0+/-0.1, and 23.5+/-0.1, respectively). The Brix values of polymeric diet had a linear additive relationship with the diet formula concentration at various pHs, temperatures, and in the gastric juice. CONCLUSION: Brix value measurement can be used to monitor stomach dietary formula concentration. Such information can be obtained at the bedside and used to evaluate feeding-intolerant patients receiving enteral feeding.

Critical Care↗

Wave-front analysis as screening technique for amblyogenic ametropia with and without cycloplegia.

INTRODUCTION: For many years, attempts have been made to find an easy, efficient and inexpensive method to screen children for amblyogenic ametropia. Wave-front analysis is a new way to determine the refractive state of the eye from a distance. This technique could be a useful tool for infant screening. PURPOSE: The purpose of the study was the evaluation of the efficacy of a commercially available wave-front analyzing autorefractometer (SureSight, software version 2.0, Welch Allyn, Skaneateles Falls, NY 13153, U.S.A.) in detecting amblyogenic ametropia in patients with and without cycloplegia. METHODS: 256 eyes (-28.25 D to +7.88 D spherical equivalent) of 128 patients (1-81 years) were examined with the wave-front autorefractometer under cycloplegia. Prior to this investigation, 108 eyes (-21.38 D to 75 D) of 54 of these patients (1-76 years) were refracted without cycloplegia. The readings of the wave-front autorefractometer were compared with the results of retinoscopy under cycloplegia. RESULTS: Without cycloplegia, the sensitivity in detecting any amblyogenic ametropia such as anisometropia, astigmatism, myopia or hyperopia was 94%, while the specificity was 63%. Following cycloplegia, the sensitivity decreased to 87% and the specificity increased to 80%. CONCLUSION: Wave-front analyzing refractometry is highly applicable for infant screening. At the present state of development, the efficacy in detecting amblyogenic ametropia is similar to that of other screening techniques and instruments that operate from distance. Cycloplegia enhances the sensitivity in detecting hyperopia and decreases the sensitivity in detecting astigmatism.

Adolescent↗

Nectar-secreting floral stomata in Maxillaria anceps Ames & C. Schweinf. (Orchidaceae).

BACKGROUND AND AIMS: Although it was generally assumed that Maxillaria spp. do not produce nectar, in recent years, nectar has been reported for a number of these orchids. Nevertheless, our current understanding of nectary structure and nectar secretion in Maxillaria is based solely on M. coccinea (Jacq.) L.O. Williams ex Hodge, which, since it shows many features characteristic of ornithophilous flowers, is atypical of this largely entomophilous genus. The aim of the present paper is to describe, for the first time, nectar secretion in a presumed entomophilous species of Maxillaria. METHODS: The structure of the nectary of M. anceps Ames & C. Schweinf., nectar composition and the process of nectar secretion were investigated using light microscopy, scanning electron microscopy, transmission electron microscopy, histochemistry, refractometry and high performance liquid chromatography. KEY RESULTS AND CONCLUSIONS: Nectar appears as droplets that are exuded by modified stomata borne upon the labellar callus and collects upon the labellum and at the base of the column-foot. Although such stomata are known to occur in a number of angiosperm families, this is the first time for them to be observed in orchids. The callus consists largely of parenchyma with raphides and is supplied by eight to ten collateral bundles. This tissue, together with the single-layered epidermis, seemingly contains terpenoids. During the bud stage, the callus cells contain an organelle complement consistent with secretory cells whereas by day 4 of anthesis, much of the cell is occupied by a vacuole. The nectar is sucrose-dominant but also contains low concentrations of glucose, fructose, free amino acids and possibly terpenoids. The high sugar concentration (approx. 66 %) is consistent with melittophily and may indicate that, like the majority of Maxillaria spp., M. anceps is visited by stingless bees (Meliponini).

Chromatography, High Pressure Liquid↗

An automatic interference refractometer.

Interference refractometry is a well-established method of determining the concentration of anaesthetic agents in gas mixtures. A photo-electric detector system has been fitted to a small portable, direct-reading interference refractometer to display automatically the concentration of the agent being measured. The design of the instrument is described and two possible methods of information display are suggested.

Anesthetics↗