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Urine specific gravity measurements: comparison of a new reagent strip method with existing methodologies, as applied to the water concentration/dilution tests.

A study was carried out to compare a new, unique chemical strip test for the measurement of urine specific gravity with the traditional methods of hydrometry and refractometry, during water concentration/dilution tests in 12 normal subjects. The pattern of specific gravity results during these serial measurements was virtually identical for the strip and for the comparative methods. Comparison of the strip results with those obtained by both hydrometry and refractometry produced acceptable linear regression data. In addition, a close relationship was demonstrated between the strip specific gravity and urine osmolality. The findings show that the strip method is an acceptable alternative to traditional methods as an indicator of urine concentration.

Adult

Projection-simulated ametropia. A model for teaching subjective refraction.

Teaching subjective refraction techniques ("refractometry") is enhanced by having the student experience the effect of each lens change "through the eyes of the patient." A projector, positioned behind the phoropter, projects a visual acuity chart through the phoropter onto the far wall. "Unknown" refractive errors are simulated by attaching trial lenses to the lens barrel of the projector, and the student progressively clears the projected image as he learns the standard steps of subjective refractometry. The focus sensitivity of each lens change can be varied, if desired, by stopping down the projection system. This simulation, which uses equipment readily available in most ophthalmology offices, has proven useful for both demonstration and practice.

Education, Medical

Study of urinary specific gravity by reagent strip method.

A newly developed reagent strip method for determination of urinary specific gravity is described. This reagent strip (Chemical S.G., Ames Co.) contains polyelectrolyte which is sensitive to the cation in solution. The urinary specific gravity is mainly decided by cation (Na+ and K+) concentrations in urine. As a result, the reagent strip gave measurements that corresponded to those measured by refractometry and gravimetry of NaC1 solution. Urea and glucose gave increased values by the refractometer, whereas the reagent strip did not cause changes. The values of urinary specific gravity obtained from reagent strip showed not only good correlations (r = 0.79-0.93) with the refractometry and gravimetry but also urinary osmolarity. This reagent strip method proved to be a most convenient tool for routine urinalysis because of the good reproducibility and accuracy considering its easy handling.

Contrast Media

[Changes in clinical refraction in future pilots during the course of training].

Changes in clinical refraction parameters were followed up in 343 students of school of pilots in the town of Burguruslan in the course of vocational training. Skiascopy and laser refractometry methods were used. Dynamic follow-up of clinical refraction showed its increase in 24.3% of the examinees both with emmetropia and hypertropia or myopia. The most noticeable increase of refraction was observed during the first two years of training, which may be explained by intensive exercise of the eyes because of theoretical studies and flights. The results evidence practical advantages of laser refractometry in comparison with skiascopy.

Aerospace Medicine

Concentration and composition of plasma proteins in wild mammals.

Total protein concentrations from a variety of wild mammalian species were measured by the Lowry and Biuret techniques and compared with results obtained or derived by refractometry. Values obtained with the Lowry method were the highest and certain derived refractometry results, the lowest. It is suggested that the Biuret method provides the most acceptable values but that care should be exercised in using the correct standard. The percentage composition of plasma from 22 species of wild mammals is also presented and the observations discussed with relation to methodology and A/G ratio.

Animal Population Groups

Thermodynamic characterization of hog kidney D-amino acid oxidase apoenzyme in concentrated guanidine hydrochloride solution. Preferential interaction with the solvent components and the molecular weight of the monomeric unit.

This paper describes the physical characterization of the monomeric unit of hog kidney D-amino acid oxidase apoenzyme in 6 M guanidine hydrochloride (GuHCl) solution by means of differential refractometry, densimetry, light scattering, equilibrium sedimentation, and high-speed gel filtration chromatography. In 6 M GuHCl solution, the oxidase interacts preferentially with GuHCl: the values of the preferential interaction parameter are 0.11 +/- 0.03 (S.D.) g/g of protein by densimetry and 0.14 +/- 0.04 g/g of protein by refractometry. The volume change, delta V, of the oxidase on transfer from the native to the denatured state is -350 ml/mol. The molecular weight of the monomeric apoenzyme is 39,600 +/- 1,700 by light scattering and 38,000 +/- 1,200 by high-speed equilibrium sedimentation. The values of the molecular weight estimated by the empirical methods, i.e., sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and high-speed gel filtration chromatography in 6 M GuHCl, agree well with those obtained by the thermodynamic methods mentioned above. These results confirm definitely that the complex of the apoenzyme with SDS normally behaves in the same manner as those of standard proteins in SDS-gel electrophoresis. This is also supported in this study by the analysis of the electrophoretic data at several gel concentrations by Ferguson plots. The molecular weight of quasi-D-amino acid oxidase apoenzyme was also examined by the empirical methods.

Animals

Subunit constitution of carbonic anhydrase from Chlamydomonas reinhardtii.

Carbonic anhydrase purified from the cell surface of Chlamydomonas reinhardtii was inactivated by treatment with dithiothreitol. This treatment caused dissociation of the holoenzyme into 35-kDa (A) and 4-kDa (B) subunits as revealed by SDS/PAGE. The 35-kDa subunit was further separated into two components A1 (35 kDa) and A2 (36.5 kDa) by SDS/PAGE using a gradient gel. These two components have the same amino acid sequence up to at least the 10th amino acid from the N-terminus. The molecular masses were estimated at 76 kDa and 35 kDa for the holoenzyme and the large subunit, respectively, and the molar ratio of the former to the latter at 1:2, by using the techniques of low-angle laser light-scattering photometry and precision differential refractometry combined with gel-filtration HPLC. The molar ratio of the 35-kDa/4-kDa subunits was estimated at 1:1 the gel-filtration HPLC monitored with precision differential refractometry. Atomic-absorption spectrophotometry revealed that the holoenzyme contains two atoms of zinc. These results suggest that the holoenzyme is a heterotetramer composed of two large subunits (A1 and A2) and two small subunits (B).

Amino Acid Sequence

[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

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

Structure of the nucleoid in cells of Streptococcus faecalis.

The structure of the nucleoid of Streptococcus faecalis (ATCC 9790) was examined and compared in the unfixed and fixed states by immersive refractometry and electron microscopy. It appears from these studies that the nucleoid structure is much more centralized in unfixed chloramphenicol-treated (stationary-phase) cells than it is in cells in the exponential phase of growth. The more dispersed configuration of the exponential-phase nucleoid could be preserved by fixation in glutaraldehyde, but not in Formalin or in osmium tetroxide. One important factor in explaining these differences in preservation is that glutaraldehyde (but not Formalin or osmium tetroxide) can rapidly cross-link the amino groups of macromolecules in cells. It was also observed that osmium tetroxide resulted in a preferential breakdown of nascent ribonucleic acid. These results are interpreted as indicating that glutaraldehyde is able to stabilize the exponential-phase nucleoid before it assumes the more central appearance seen in osmium tetroxide- and Formalin-fixed cells. These results are discussed in terms of the proposed organization of the exponential-phase nucleoid in unfixed cells.

Cell Nucleus

The clinical refractometer: a useful tool for the determination of specific gravity and osmolality in canine urine.

For the determination of specific gravity of canine urine the urinometer is often used. An alternative method is based on refractometry. Both methods were compared and a table for converting refractometer values to urinometer values is given. The refractometer was also compared with an osmometer. A linear relation was found between the readings of both instruments. The results clearly indicate that the refractometric method is very suitable for the small animal practitioner. The instrument appears to be a good alternative for both the urinometer and the osmometer.

Animals

A human model of allergic conjunctivitis.

Using the method of refractometry to measure protein concentration in tears, a simple model was developed to evaluate the allergic response in humans. Timothy grass pollen was instilled into the cul de sac of a human subject, and the protein content of the subject's tears was sampled every 15 minutes for three hours. Once the time course of this experiment was predictable, various eye medications were instilled into the subject's eye 30 minutes after allergic challenge. It was noted that 1.0% and 0.1% prednisolone, each combined with 0.12% phenylephrine hydrochloride, as well as 1% epinephryl borate eye drops, produced an almost immediate return to normal of tear protein levels, whereas 1.0% medrysone, 0.1% fluorometholone, 1.0% prednisolone, and 0.12% prednisolone had less pronounced effects on reduction of elevated tear protein concentration.

Administration, Topical

Outflow facility and its response to pilocarpine decline in aging rhesus monkeys.

Refractive error and total outflow facility were determined by Hartinger coincidence refractometry and two-level constant-pressure perfusion, respectively, in 17 rhesus monkeys, aged 5 to 29 years. Maximum accommodative response to corneal (iontophoretic) carbachol hydrochloride, baseline outflow facility, and the facility response to strong but submaximal intracameral doses of pilocarpine hydrochloride all declined with age. The correlation between accommodative response to carbachol and facility response to pilocarpine was slightly stronger than that between age and facility response. Since the ciliary muscle plays a major role in controlling both outflow facility and accommodation, and since histologic and videographic techniques demonstrate an age-related decline in rhesus ciliary muscle excursion induced by topical pilocarpine or electrical stimulation of the Edinger-Westphal nucleus, the present data support the hypothesis that an age-related decline in ciliary muscle mobility is associated, perhaps causally, with an age-related decline in facility and facility responsiveness to cholinergic drugs.

Accommodation, Ocular

Inhibition of aceclidine-stimulated outflow facility, accommodation and miosis in rhesus monkeys by muscarinic receptor subtype antagonists.

Aceclidine can dissociate accommodative and outflow facility responses in monkeys and humans. We sought to determine if different muscarinic receptor subtypes control outflow facility, accommodative and pupillary responses to aceclidine in the living rhesus monkey eye, as a possible basis for this separation. Each eye was cannulated with one branched and one unbranched needle. Baseline measurements (of outflow facility by two-level constant pressure perfusion; refraction by Hartinger coincidence refractometry and pupil diameter by vernier calipers) were recorded after anterior chamber exchange with (one eye) or without (opposite eye), muscarinic receptor subtype antagonist (pirenzepine, AF-DX 116 or 4-DAMP). The eyes were then exchanged a second time with these solutions plus added aceclidine. The response to aceclidine (the differences from baseline) in the presence or absence of antagonist were compared. The M3 muscarinic receptor subtype antagonist 4-DAMP was the most potent inhibitor of all three responses to aceclidine. The dissociation of accommodative, outflow facility and miotic responses to aceclidine in rhesus monkeys does not appear to be due to differences in the muscarinic receptor subtypes that can currently be distinguished pharmacologically.

Accommodation, Ocular

A-band mass exceeds mass of its filament components by 30-45%.

We have measured filament lattice spacing in fibrils using X-ray diffraction, and find that STEM-determined mass/length values reported for myofilaments should give 13% w/v as the filament protein concentration in the lattice of the AO-band (filament overlap zone) of both insect flight muscle (IFM) and vertebrate skeletal muscle ( VSkM ). This is well below the actual mass concentration of AO-bands as measured by immersion refractometry of detergent-washed rigor myofibrils under the phase microscope. This technique identifies an immersion fluid of suitable refractive index (RI) for matching out all image contrast between background and the selected cross-band. We used RI fluids in which the effective RI matching component is a large-particle solute (Percoll or hemocyanin) excluded by the filament lattice. The measured RI indicates that protein concentration in AO-bands is 16-17% in Lethocerus and other IFM fibrils including CAF-digested fibrils, and is 18-19% in rabbit VSkM fibrils. On IFM fibrils we also measured absolute buoyant density in Percoll as greater than or equal to 1.042; this supports the value for mass concentration as determined by RI. The mass discrepancy between fibrils and filaments does not seem to arise from faults in the methods used. We therefore accept the STEM-determined mass/length values for thick filaments which indicate 4 myosins/crown in IFM and 3 in VSkM , and we believe there is considerable extra nonfilament material (concentration: 30-50 mg/ml) between the filaments in fibrils. In stretched VSkM , it is the H-bands, not the I-bands, which have an excess over filament mass content. The extra mass has not been identified.

Animals