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Detection of low serum immunoglobulin concentrations in clinically ill calves.

The sensitivity, specificity, and accuracy of classification of 4 tests for failure of passive transfer (FPT) were examined in clinically ill neonatal calves. Comparisons were made with serum IgG1 concentrations determined by radial immunodiffusion. Serum samples were obtained from 27 clinically ill calves < or = 21 days of age. The results of 4 commonly used assays, the sodium sulfite turbidity test, the zinc sulfate turbidity test, refractometry, and the serum gamma-glutamyl transferase (GGT) activity test, were compared with radial immunodiffusion determinations of serum IgG1 concentration. Serum GGT activity using a 50 IU/L threshold resulted in correct classification of the highest percentage of calves (93%) with regard to their passive transfer status. The sodium sulfite test with a 1+ end point and refractometry using a 5.5 g/dL end point resulted in correct classification of 85% of the calves studied. When using the sodium sulfite test, the 2+ and 3+ test end points had lower specificity, 0.58 and 0.00, respectively, than the 1+ end point. This loss in specificity resulted in misclassification of calves with adequate serum immunoglobulin concentrations as having FPT. The zinc sulfate turbidity test was inadequately specific (0.33) and resulted in misclassification of 33% of calves.

Animals↗

The density of the cell sap and endoplasm of Nitellopsis and Chara.

We measured the densities of the cell sap, endoplasm and cell wall of Nitellopsis obtusa and Chara corallina using interference microscopy, refractometry, immersion refractometry, equilibrium sedimentation and chemical microanalysis techniques. These values are important for the determination of many rheological properties of the cytoplasm as well as for understanding buoyancy regulation, dispersal mechanisms and how cells respond to gravity. The average densities of the cell sap, endoplasm and cell wall are 1,006.9, 1,016.7 and 1,371 kg m-3 for Nitellopsis and 1,005.0, 1,013.9, and 1,355.3 kg m-3 for Chara.

Cell Wall↗

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↗

The effects of diaper brands, urine volume, and time on specific gravity measurement.

Nurses use urine specific gravity to assess fluid volume status in pediatric patients. Specific gravity of five leading brands of disposable diapers was measured to determine the effects of diaper brand, urine volume, time elapsed, and method of specific gravity measurement comparing the two methods of refractometry and N-Multistix SG (Ames Division, Miles Inc., Elkhart, IN). Immediately after a baby voided 20 mL of urine into any disposable diaper, the specific gravity by refractometer accurately compared with the control standard. Measurement accuracy by N-Multistix SG was only assured if Pampers (Procter & Gamble, Cincinnati, OH) were used. However, both methods were inaccurate at 4 hours with 20 mL of urine. Immediately after a baby voided 40 mL of urine into any disposable diaper, both methods were accurate when compared with the control. At 4 hours, only Pampers and Chux (Procter & Gamble, Cincinnati, OH) were accurate by refractometry, and Pampers alone was accurate by N-Multistix SG.

Humans↗

Accommodation amplitude after lens refilling with injectable silicone by sealing the capsule with a plug in primates.

OBJECTIVE: To restore accommodation in primate eyes by refilling the lens capsule with injectable silicone compounds. MATERIALS AND METHODS: Eight eyes of 8 monkeys (Macaca fascicularis) were treated by the lens refilling procedure. To prevent leakage of the injected liquid silicone before it polymerized in the capsule in vivo, a silicone plug for sealing the capsular opening was developed. After endocapsular phacoemulsification following an upper minicircular capsulorhexis, the plug was introduced into the capsulorhexis opening. A silicone mixture was injected into the capsular bag through the delivery tube of the plug. Automated refractometry was performed 1 week and 3 months after surgery. Accommodation amplitude was determined as the difference between the refractions before and 1 hour after topical application of 4% pilocarpine chloride. RESULTS: Five of 8 eyes could be refilled. In 4 of 5 eyes, refraction could be measured. Accommodation amplitude ranged from 1.0 to 4.5 diopters, with a mean of 2.3 +/- 1.3 diopters (8.0 +/- 2.0 preoperative values). At 3-month examination, thick posterior capsule opacification precluded refractometry in all eyes. CONCLUSIONS: The lens refilling procedure with the use of a silicone plug for sealing the capsular opening was feasible in primate eyes. The accommodation amplitude attained was a small fraction of the value before surgery. This may result from the loss of so-called intracapsular accommodation, ie, active participation of lens fiber cells in accommodation. However, since the obtained accommodation may be sufficient for near vision after cataract surgery, this lens refilling procedure warrants further study. Elucidation of the mechanism of intracapsular accommodation may also be necessary.

Accommodation, Ocular↗

Two-beam optical traps: refractive index and size measurements of microscale objects.

A counter-propagating optical trap measurement (COTM) system is proposed and analyzed based on the ray-optics model. In this system, refractive index and size of trapped objects can be estimated by using forward scattered light from the two-beam laser trap with resolution Delta n = 0.013 for the refractive index measurements and 3.3% for the size measurements, which is comparable with current bulk techniques, such as refractometry, and flow cytometry. The unique advantage of the COTM system over conventional approaches lies in its capability of marker-free single-particle characterization in whatever transparent buffer required by living cell, eliminating the necessity of changing the fluid composition of the sample in refractometry, and of tagging target with toxic fluorescence dyes in flow cytometry. Noise analysis predicts a potential improvement in the system resolution by more than two orders of magnitude. This non-invasive and sterile tool complements lab-on-a-chips with single cell manipulation and analysis in living friendly ambient.

Biopolymers↗

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↗

Characterization and analysis of thermal denaturation of antibodies by size exclusion high-performance liquid chromatography with quadruple detection.

Size exclusion chromatography (SEC) coupled with online light scattering, viscometry, refractometry, and UV-visible spectroscopy provides a very powerful tool for studying protein size, shape, and aggregation. This technique can be used to determine the molecular weight of the component peaks independent of the retention times in the SEC column and simultaneously measure the hydrodynamic radius and polydispersity of the protein. We applied this technology by coupling an Agilent Chemstation high-performance liquid chromatography system with a diode array UV-visible detector and a Viscotek 300 EZ Pro triple detector (combination of a light scattering detector, refractometer, and differential pressure viscometer) to characterize and compare the molecular properties of a number of monoclonal antibodies. Our studies reveal that different monoclonal immunoglobulin Gs (IgGs) and chimeric IgGs show slightly different retention times and therefore different molecular weights in gel filtration analysis. However, when they are analyzed by light scattering, refractometry, and viscometry, different IgGs have comparable molecular weight, molecular homogeneity (polydispersity), and size. Gel filtration coupled with UV or refractive index detection suggests that antibodies purified and formulated for preclinical and clinical development are more than 95% monomer with little or no detectable soluble aggregates. Light scattering measurements showed the presence of trace amounts of soluble aggregate in all the IgG preparations. The different IgG molecules showed different susceptibility to heat and pH. One of the murine antibodies was considerably less stable than the others at 55 degrees C. The application of this powerful technology for the characterization of monoclonal antibodies of therapeutic potential is discussed.

Animals↗

The influence of anticoagulants on the measurement of total protein concentration in equine peritoneal fluid.

The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The solution of K3EDTA in distilled water only caused TP overestimation at high K3EDTA concentrations (>5 micromol/ml). By contrast, lithium heparin did not influence the refractometric values of TP. Neither anticoagulant modified TP values when measured by the biuret method. In conclusion, the use of K3EDTA as anticoagulant may result in a significant overestimation of TP values of peritoneal fluid samples measured by refractometry.

Animals↗

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↗

Comparison of refractometer and biuret methods for total protein measurement in body cavity fluids.

Most hand-held medical refractometers have internal scales that limit protein measurement to results >/=2.5 g/dL. Tables for conversion of refraction (r) to protein concentration for values as low as 0.1 g/dL were published in the 1960s, but their accuracy for use on body fluids has not been established. The purpose of this study was to assess the reliability of body cavity fluid protein determination by refractometry. We compared the protein concentration of 25 body cavity fluids as determined by 2 Goldberg type hand-held refractometers with results obtained by the biuret method. Published charts converting refraction (r) to protein concentration were used to determine protein concentration in samples with protein <2.5 g/dL. Higher protein values were read directly from the instruments. The range of comparison was limited to >/=0.6 g/dL, the lowest concentration of the biuret method's standard curve. Twenty-one peritoneal fluid, 2 pleural fluid and 2 pericardial fluid samples from 16 horses, 5 cattle, 3 dogs, 2 llamas and 1 cat were tested. The results obtained by the two refractometers were closely and linearly related to biuret results (P<.001), with slopes by linear regression analysis close to 1, and correlation coefficients >0.977. Based on this study, the range for quantification of body cavity fluid protein concentration by refractometry can be extended below 2.5 g/dL, allowing for quantitative assessment of most clinical samples.

Journal Article↗

The usefulness and limitations of hand-held refractometers in veterinary laboratory medicine: an historical and technical review.

Medical hand-held refractometers have been used in veterinary practice since their development in the 1960s. They have become ubiquitous for the measurement of protein and urine solute concentrations because of their rapidity of analysis, ease of use, and relatively low cost. Refraction of light offers advantages for the determination of solute concentrations because the measurement requires no chemical alteration of the specimen. Numerous authors have reported that the results of protein estimation by refractometry for domestic mammals correlate well with those obtained by the biuret method, although others have reported both higher and lower refractometric results compared with biuret results. Major discrepancies between biuret and refractometric results have been reported for avian samples. Some of the variation in reported results may be due to differences in design by refractometer manufacturers. Another possible source may be variation in the biuret reagent mixture and assay conditions. Refractometers also can be used to calculate serum water concentration. A table that converts index of refraction to serum water concentration can be used to convert electrolyte concentration from mmol/L of serum to mmol/L of serum water, a more accurate indicator of effective electrolyte concentration. Refractometers are especially useful for determining urine specific gravity on veterinary samples because they require relatively small sample volumes. Specific gravity continues to be the most common unit for reporting total solids concentration. Some solutes, such as acetone, may cause false increases in specific gravity by refractometry, as they increase refraction but are less dense than water.

Journal Article↗

In-fiber reflection mode interferometer based on a long-period grating for external refractive-index measurement.

We present two novel schemes for refractometry based on a long-period fiber grating- (LPG-) based Michelson interferometer. These schemes are designed to overcome the measurement dependence of previously demonstrated LPG-based refractometry on the immersion depth. The first utilizes an unshielded LPG and the second, a shielded one. Both schemes were tested over a certain refractive-index range, and the measurement of glucose concentration in water was experimentally demonstrated. In addition, the temperature sensitivity of the two schemes is discussed.

Journal Article↗

Correlation between automated and subjective refraction before and after photorefractive keratectomy.

PURPOSE: To ascertain whether autorefractometry after photorefractive keratectomy (PRK) provides results that are comparable with subjective refractometry. METHODS: The refractions of 159 eyes before and 1 month after PRK were determined with both automated and subjective refractometry, and the results of the two methods were compared. RESULTS: Before PRK, subjective spherical equivalent refraction ranged from -15.25 to +0.25 D (mean -5.30 +/- 2.64 D) and automated spherical equivalent refraction ranged from -16.13 to -0.50 D (mean -6.10 +/- 2.74 D), with good correlation (r2 = 0.965) between the two methods. One month after PRK, subjective spherical equivalent refraction ranged from -1.63 to +3.50 D (mean +0.98 +/- 0.78 D) and automated spherical equivalent refraction ranged from -2.50 to +3.00 D (mean +0.36 +/- 0.96 D) with poor correlation (r2 = 0.401). CONCLUSION: Autorefractometry is less reliable in eyes after PRK, and consequently software changes are recommended for overcoming this problem.

Adult↗

The new National Collegiate Athletic Association wrestling weight certification program and sport-seasonal changes in body composition of college wrestlers.

Changes in body composition of college-age wrestlers (N = 21, 19.0 +/- 0.2 years of age, 8.2 +/- 0.6 years of experience) were studied throughout the season after implementation of the new National Collegiate Athletic Association (NCAA) wrestling weight certification (WWC) program. Wrestlers were divided into 2 groups, starters (n = 10) and nonstarters (n = 11), based on the number of completed matches during the 1998-1999 season. Body density was measured via hydrostatic weighing, with residual volume determined via the nitrogen washout technique. Hydration was assessed by measuring specific gravity of urine (Usg). For body mass and fat-free mass (FFM), a significant difference in the pattern of change was found (p < 0.05), highlighted by a decrease in body mass (-3.0%, -2.5 kg, -5.8 lb) and a maintenance of FFM from preseason to peak season for the starters. The pattern of change over time was significantly different for refractometry Usg (p < 0.01) but not for the test strip Usg (p = 0.54). No significant interaction effects were seen for percentage of body fat or fat mass. Significant time main effects were found for percentage of body fat, body mass, fat mass, FFM, and refractometry Usg. Taken together, the results of this investigation demonstrate that collegiate wrestlers lose body mass but maintain FFM throughout the season, which may in part be associated with the new NCAA WWC program.

Adolescent↗