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Proton-dependent dissociation equilibrium of hemoglobin. 1. A 700-nanometer light-scattering study on horse methemoglobin in the pH range 4.8 to 7.2.

The effect of proton concentration upon the subunit dissociation of horse methemoglobin has been investigated at two ionic strengths by light scattering photometry at 700 nm. Differential refractometry revealed a slight but systematic decrease of the specific refractive index increment with decreasing protein concentration for solutions in dialytic equilibrium with the solvent. In the pH range 4.8-7.2 the dissociation can be described by a simple equilibrium between tetramers and dimers. The dissociation constant Kd of the met derivative is found to be very similar to those of the O2- and CO-ligated states. From the slope of a plot of log Kd vs. pH, the number of protons bound is n = 1.3 +/- 0.1 resulting from an increase in the pK values of two groups upon dissociation. These two groups must be identical because the dissociation is symmetrical.

Animals↗

Refractive change during hyperbaric oxygen therapy. A clinical trial including ultrasound oculometry.

PURPOSE: To record changes in refraction and refractive parameters associated with a standard hyperbaric oxygen treatment protocol consisting of a 95 min session at > 95% oxygen at 2.5 atmospheres (ATA) given daily Monday to Friday, to a total of 30 sessions. PATIENTS AND METHODS: Seventeen of the 26 patients included were able to attend for ophthalmic assessment at time zero (to) and after 20 treatments (t20). Thirteen patients also had a post-treatment follow-up. Most patients were being treated for osteoradionecrosis after radiotherapy of ENT cancers, and drop-outs for the eye exam were common. Refraction was determined subjectively and by refractometry, before and after tropicamide 1% eyedrops. Refractive parameters were assessed by keratometry and by A-scan axial ultrasound measurement. RESULTS: Results are given for the 17 patients with to and t20 assessments. The induced refractive change ranged from 0 to 1.5 D. When observed, refractive changes were myopic in nature. The shift averaged 0.58 D according to the refractometer and 0.49 D as subjectively assessed, with corresponding median change values being 0.62 and 0.39 D. The differences between 0 D and these values, although small, were statistically highly significant. CONCLUSIONS: The refractive changes associated with hyperbaric oxygen therapy were smaller than the literature had led us to expect. No significant change in axial eye length measurements was found, and keratometry readings reflected only minimal change, although this was statistically significant on a 0.05 level. Therefore it is most likely that lens changes, whether in internal refractive indices or curvatures, accounted for the transitory shift towards more myopic/less hyperopic values.

Adult↗

The autoregulatory role of EsaR, a quorum-sensing regulator in Pantoea stewartii ssp. stewartii: evidence for a repressor function.

Capsular polysaccharide synthesis and virulence in the plant pathogenic bacterium Pantoea stewartii ssp. stewartii requires the quorum-sensing regulatory proteins, EsaR and EsaI, and the diffusible inducer N-(3-oxo-hexanoyl)-L-homoserine lactone. Prior mutational studies suggested that EsaR might function as a repressor of quorum sensing in the control of capsular polysaccharide synthesis. Further, a lux box-like palindromic sequence coinciding with the putative -10 element of the esaR promoter suggested a possible negative autoregulatory role for EsaR. This report presents genetic evidence that EsaR represses the esaR gene under inducer-limiting conditions, and that addition of inducer promotes rapid, dose-dependent derepression. DNA mobility-shift assays and analyses by surface plasmon resonance refractometry show that EsaR binds target DNAs in a ligand-free state, and that inducer alters the binding characteristics of EsaR. Physical measurements indicate that the EsaR protein binds N-(3-oxo-hexanoyl)-L-homoserine lactone, in a 1:1 protein:ligand ratio, and that inducer binding enhances the thermal stability of the EsaR protein. These combined genetic and biochemical data establish that EsaR regulates its own expression by signal-independent repression and signal-dependent derepression. Additionally, we provide evidence that EsaR does not govern the expression of the linked esaI gene, thus EsaR has no role in controlling coinducer synthesis.

Bacterial Proteins↗

[Effect of pupil size on longitudinal focal distribution after photorefractive keratectomy ].

BACKGROUND: Optical aberrations induced by the transition zone of a myopic photorefractive keratectomy (PRK) may degrade the image quality. The subjectively evaluated visual acuity and contrast sensitivity may be reduced significantly especially with mydriasis. The purpose of this study was to calculate the image forming properties of the eye using raytracing techniques of a spot light source based on topographic height data of a scanning slit videokeratoscope. METHODS: Refractive surfaces were calculated from raw height data sets of the anterior and posterior corneal surface measurements (Orbscan, Orbtec, USA). The characteristics of the residual refractive surfaces were derived from Navarro's eye model. The focal distance was calculated using the exact raytracing calculation. Point spread function was determined at the focal plane and at 5 (delta = 0.2 mm) parafocal planes in front of and behind the focal distance in accordance to the pupil diameter (2, 3 and 6 mm). The algorithm was applied to 11 selected eyes (6 left, 5 right) of 8 patients before and after PRK with myopia (n = 6) and myopic astigmatism (n = 5). RESULTS: Before PRK the focal distance did not decrease significantly with increasing pupil size (2 to 6 mm), but decreased significantly after PRK (-0.41 +/- 0.12 mm, p = 0.02). Preoperatively, the variance of the point spread function did not differ significantly between a 2 mm (0.046 +/- 0.015 mm) and a 6 mm (0.055 +/- 0.021 mm) pupil size but was affected significantly by the pupil size postoperatively (2 mm: 0.049 +/- 0.018 mm, 6 mm: 0.096 +/- 0.045 mm, p = 0.04). CONCLUSIONS: Raytracing of corneal topography height data based on refined eye models with the option of calculation of the focus has the potential of tracing the optical resolution of the eye as a function of the pupil size. Due to of the altered surface geometry of the cornea after myopic PRK, both the variance of the point-spread function and the depth of focus increase with pupil size. This may be an explanation for impaired subjective refractometry and reduced contrast sensitivity of the patient after conventional keratorefractive surgery.

Adult↗

[Clinical application of a new method for the objective estimation of minimum visual acuity].

PURPOSE: Recently, a new method for the objective estimation of the minimum visual acuity (OEM) by means of suppression of the optokinetic nystagmus has been presented (Klin Monatsbl Augenheilkd 1998;212:196-202). This study reports on the clinical use of this method. METHODS: In 120 individuals referred to our clinic either to procure an ophthalmological expert opinion or for differential diagnosis of an unclear visual impairment, an OEM was performed. The result of the OEM was compared to the clinical findings (history, biomicroscopy of anterior and posterior segments, objective refractometry, pupillary responses, binocular alignment, motility, binocular vision, colour vision, applanation tonometry, electrophysiology, fluorescein angiography, neurologic, radiologic, psychiatric findings, reproducibility of visual acuity and visual field statements under different conditions, comparison of subjective and objective visual field data, statistical prove of false visual acuity and visual field statements) which were critically interpreted concerning the credibility of the subject's statements. RESULTS: As a result of the clinical examinations, the stated visual acuity of 62 individuals was credible. In one of these individuals, the OEM pointed to a slightly (1dB) better visual acuity. The statements of 7 individuals could not be categorized clinically. The OEM pointed to a better acuity in 3 cases. The stated vision of the remaining 51 individuals was not credible. In 38 of these cases, the OEM pointed to a better acuity, or false statements could be proven by the OEM. CONCLUSION: A significant rate (75%) of the individuals whose statements were not credible was detected by the computer-aided, nystagmographic method of OEM. The method allows an estimation of the actual minimum visual acuity and yields evidence of false statements concerning the detection acuity.

Biofeedback, Psychology↗

[Sensitivity of refraction measurement with the infrared videorefractometer VRB 200].

AIM OF THE STUDY: This study investigates the sensitivity of a computerised videorefractor (VRB 200) for refractive errors, especially amblyopiogenic, in babies and infants. PATIENTS AND METHODS: Videopictures from 312 eyes of 156 babies and infants were taken, evaluated and compared with the current measurements (skiaskopy or refractometry) of the university eye hospital Erlangen-Nürnberg. RESULTS: 255 (81.7%) of 312 eyes were evaluable. The VRB 200 achieves a sensitivity of 96.1% in hyperopic eyes (219 of 228 eyes) and 94.7% in myopic eyes respectively (18 of 19 eyes). Of 8 emmetropic eyes 6 were recognized correctly negative. Astigmatism could be estimated coarsely quantitatively in power and qualitativ in the axis with a sensitivity of 80% (92 of 115 eyes) and a specificity of 86% (121 of 140). Anisometropia over 2 dpt was recognized in 5 of 6 patients. A general statement concerning the specificity is not possible with this study, because it is not a mass screening procedure. CONCLUSION: This videorefractor is able to recognize semi-quantitatively ambylopiogenic refractive errors with high sensitivity.

Amblyopia↗

[How accurate is video refraction measurement in infants in comparison with skiascopy? How much do the values change within a half a year?].

BACKGROUND: Isotropic photorefraction (Atkinson et al., 1981) is a method used to screen the refraction in infants. The aim of this study is to analyze its limitations as a measuring instrument. MATERIALS AND METHODS: 215 children aged between 5 and 12 months were examined in cycloplegia. The results were subsequently compared (1) to the results of streak retinoscopy also performed at the same time and (2) to photorefraction performed a second time, 6 months later (with 145 children). RESULTS: Spherical refraction: The mean spherical values differed by 0.70 dpt through photorefraction methods, compared to retinoscopy. The standard deviation (SD) was 0.67 dpt. The values of the photorefraction differed therefore in 95% of the children within a range of 2.7 dpt (or +/- 2 SD). Compared to the second photorefractive measurement 6 months later, the average refraction showed a small myopic shift (-0.45 dpt). Astigmatism: The errors of measurement were more pronounced here. Extreme differences of up to 9 dpt were found in single cases. The mean difference, compared to retinoscopy, was 0.73 dpt. The standard deviation (SD) was 0.90 dpt. In 95% of the children, the values of photo-refraction differed therefore by a range of 3.6 dpt (or +/- 2 SD). Compared to the second photorefractive measurement 6 months later, the average refraction showed a mean change of 0.73 dpt. CONCLUSIONS: Our photorefractive measurements showed errors in the range of 1-2 dpt for the spherical refraction, and in the range of 2-3 dpt for astigmatism. In 5% of the children examined, even greater aberrations were found. As long as the aim of photorefraction is restricted to disclosing high refractive errors, this method is judged suitable for the refractive screening of infants. For more precise refraction, and for the prescription of spectacles, we believe that is should be supplemented by a second examination technique, such as retinoscopy or automated refractometry.

Astigmatism↗

[Eliminating reflection of intraocular lenses].

Reflections on the refractive surfaces of intraocular lenses can be a nuisance when performing retinoscopy or refractometry. Some patients with IOLs also see reflections which disturb their vision. This article examines the conditions for coating IOLs. It also explains why conventional coating methods cannot be used for IOLs.

Humans↗

[Liindner's method of cylinder retinoscopy without theory].

Brief description of Lindner's cylinder retinoscopy for practical application: The main criteria of this accurate method for checking refraction are the direction, shape, speed and brightness of the retinoscopic phenomena moving in the pupil of the examined eye. To determine astigmatism the use of plus cylinders is recommended. Difficulties may be due to the optical aberration at the periphery of a wide pupil, to a scissors phenomenon, and to optical irregularities of the senile lens, corneal opacities etc. In many cases retinoscopy can be performed well without using mydriatics and cycloplegics, respectively. Children must not be overstrained. Generally, final subjective checking is crucial for the prescription of corrective glasses. Mastery of Lindner's cylinder retinoscopy enables the eye specialist to keep refractometry with simple aids, as an essential part of the ophthalmological diagnosis, in his own hands, against the increasing claim of optometrists and opticians.

Adult↗

Compensation for spectacle lenses involves changes in proteoglycan synthesis in both the sclera and choroid.

PURPOSE: It has been demonstrated that chick eye growth compensates for defocus imposed by spectacle lenses: the eye elongates in response to hyperopic defocus imposed by negative lenses and slows its elongation in response to myopic defocus imposed by positive lenses. We ask whether the synthesis of scleral extracellular matrix, specifically glycosaminoglycans, changes in parallel with the changes in ocular elongation. In addition, there is a choroidal component to compensation for spectacle lenses; the choroid thickens in response to myopic defocus and thins in response to hyperopic defocus. We ask whether choroidal glycosaminoglycan synthesis changes in parallel with changes in choroidal thickness. METHODS: Chicks wore either a +15 diopter (D) or -15 D spectacle lens over one eye, or they wore one lens of each power over each eye for 5 days. At the end of this period, we measured refractive errors and ocular dimensions by refractometry and A-scan ultrasonography, respectively. Pieces of the scleras and choroids from these eyes were put into culture and the synthesis of glycosaminoglycans was assessed by measuring the incorporation of radioactive inorganic sulfur. RESULTS: We here report that the compensatory modulation of the length of the eye involves changes in the synthesis of glycosaminoglycans in the sclera, with synthesis increasing in eyes wearing -15 D spectacles lenses and decreasing in eyes wearing +15 D lenses. In addition, changes in the synthesis of glycosaminoglycans in the choroid are correlated with changes in choroidal thickness: eyes wearing +15 D lenses develop thicker choroids and these choroids synthesize more glycosaminoglycans than choroids from eyes wearing -15 D lenses. CONCLUSIONS: Changes in scleral glycosaminoglycan synthesis accompany lens-induced changes in the length of the eye. Furthermore, changes in the thickness of the choroid are also associated with changes in the synthesis of glycosaminoglycans. These results are consistent with the regulation of the growth of the eye being bidirectional, and with the retina being able to sense the sign of defocus.

Adaptation, Physiological↗

Changes of posterior corneal astigmatism and tilt after myopic laser in situ keratomileusis.

PURPOSE: The purpose of this study was to assess the changes of posterior corneal astigmatism and tilt after laser in situ keratomileusis (LASIK) and to correlate these changes with the amount of correction and the residual stromal bed thickness. METHODS: This prospective nonrandomized (self-controlled) comparative trial included 57 eyes from 14 females and 15 males, whose mean age (+/- standard deviation [SD]) at the time of surgery was 33 +/- 9 years (range, 19-53), with a spherical equivalent (SEQ) of -1.00 to -15.50 (mean, -5.07 +/- 2.81) diopters (D). All LASIK procedures were accomplished with the Keratom II Coherent-Schwind excimer laser and the Moria Model One microkeratome (150-microm head). Subjective refractometry, Orbscan slit-scanning corneal topography analysis, and pachymetry were performed before and 3 months after LASIK for myopia (n= 35; -1.00 to -15.50 D [mean, -4.75 +/- 3.07]) or myopic astigmatism (n= 22; sphere, 0.00 to -9.75 D [mean, -4.75 +/- 2.36]; cylinder, -0.75 to -3.50 D [-1.68 +/- 0.86]). Intended ablation depth ranged from 12 to 108 (mean, 48 +/- 22) microm. Topographic raw data were decomposed into a set of Zernike polynomials as published in detail previously, and parameters for detection of asymmetric mechanical deformation of the cornea were derived. Posterior corneal astigmatism and tilt before and after LASIK were compared, and changes in these variables were correlated with the SEQ change (DeltaSEQ) and the residual corneal bed thickness (RBT). RESULTS: The RBT after LASIK ranged from 186 to 373 (mean, 280 +/- 42) microm. Overall, astigmatism (0.19 +/- 0.07 D/0.22 +/- 0.13 D; p= 0.80) and tilt (3.58 +/- 0.35 degrees /3.65 +/- 0.48 degrees; p= 0.61) did not change significantly by 3 months after LASIK. In eyes with RBT < or =250 microm, the average change in astigmatism (0.05 +/- 0.11 versus 0.01 +/- 0.13 D; p= 0.46) and tilt (0.21 +/- 0.45 degrees versus 0.04 +/- 0.55 degrees; p= 0.30) was not greater than in eyes with RBT > 250 microm. Change in astigmatism (p= 0.19) and tilt (p= 0.56) did not correlate with the RBT during LASIK. CONCLUSIONS: Zernike decomposition of topographic height data discloses that no significant asymmetric mechanical deformation of the posterior corneal curvature occurs after myopic LASIK. Further studies with long-term follow-up are needed to clarify whether this symmetry of the posterior corneal surface can indeed be preserved over time after LASIK if the RBT is < 250 microm.

Adult↗

A simple field test for assessing salt balance in heat-stressed miners.

A simple colorimetric test for urinary chloride (Fantus test) corrected for urine specific gravity (measured by refractometry) enabled 24-hour urinary sodium excretion rates to be predicted accurately from a single spot sample of urine. In 29 acclimatized miners working in an environment in excess of 40 degrees C, salt-conserving mechanisms, involving renin and aldosterone, were not active until the urinary sodium excretion rate fell below 50 mmol/24 h. A predicted sodium excretion rate greater than 50 mmol/24 h indicates that dietary sodium supplementation is not necessary.

Adult↗

Laser-hole boring into overdense plasmas measured with soft X-Ray laser probing

A laser self-focused channel formation into overdense plasmas was observed using a soft x-ray laser probe system with a grid image refractometry (GIR) technique. 1.053 &mgr;m laser light with a 100 ps pulse duration was focused onto a preformed plasma at an intensity of 2x10(17) W/cm (2). Cross sections of the channel were obtained which show a 30 &mgr;m diameter in overdense plasmas. The channel width in the overdense region was kept narrow as a result of self-focusing. Conically diverging density ridges were also observed along the channel, indicating a Mach cone created by a shock wave due to the supersonic propagation of the channel front.

Journal Article↗

On the Stiles-Crawford effect and ocular imagery.

To study the modifying effect on spherical aberration of the retinal directional sensitivity (the Stiles-Crawford effect), ocular imagery was simulated by computer ray-tracing in a phakic and a pseudophakic model eye. The Stiles-Crawford effect was found to reduce the distance between the paraxial focus and the effective least blur focus. The modifying effect on spherical aberration was found to be insignificant for pupil sizes below 4 mm, and to increase in magnitude above this size. For an 8 mm pupil, the effective focus was found to move 0.3 mm and 0.2 mm toward the paraxial focus for the phakic and the pseudophakic eye, respectively. This corresponded to a shift of 1.0 D and 0.6 D in the spectacle plane, respectively. The modifying effect of the retinal directional sensitivity should be taken into account in the theory of ocular refraction and in objective keratometry and refractometry.

Cataract↗

Analysis of transtracheal aspirates and pleural fluid from clinically healthy llamas (Llama glama).

Seventeen clinically healthy adult llamas were used to study the characteristics of transtracheal aspirates (TTA) and pleural fluid samples. Results of complete blood counts, fibrinogen determination and thoracic radiographs were within normal limits prior to sampling. Cytologic evaluation of TTA revealed the majority of cells were vacuolated macrophages (60-100%), with 0-40% neutrophils, and fewer lymphocytes (0-1%), eosinophils (0-3%), and ciliated respiratory epithelial cells (0-10%). In TTA from 10 of 17 llamas, neither aerobic nor anaerobic bacteria were isolated. Bacteria isolated in pure culture from TTA were similar to isolates found in clinically healthy animals of other species, and included Acinetobacter sp., Staphylococcus sp. and Bacillus sp. Results (mean +/- SD) of pleural fluid analyses were: total nucleated cell count 576 +/- 361/microliter; specific gravity 1.0133 +/- 0.002; glucose concentration 135.1 +/- 9.02 mg/dL; and lactate concentration 2.95 +/- 1.34 mg/dL. Pleural fluid total protein concentrations determined by refractometry ranged from < 2.5 to 3.5 g/dL. The refractive index ranged from 1.3396 to 0.0013. In pleural fluid, small lymphocytes were the predominant cell type.

Journal Article↗

Plasma biochemistry reference values of wild bonnethead sharks, Sphyrna tiburo.

BACKGROUND: Elasmobranchs (sharks, skates, and rays) are of commercial, sport, research, and exhibit importance, however, blood chemistry reference values have been determined for few of these species. OBJECTIVE: The purpose of this study was to establish plasma biochemistry and PCV reference values for wild bonnethead sharks (Sphyrna tiburo). METHODS: Heparinized blood samples were collected from 24 bonnethead sharks at the time of capture in trawl nets off the coast of South Carolina and Georgia. Weight, length, PCV, total solids (TS, by refractometry), and plasma biochemical analyses were done using standard techniques. Wilcoxon rank-sum and Kendall tau b tests were used to compare values by animal size, boat and sex; 1-way ANOVA was used to compare TS and total protein (TP) concentrations. RESULTS: Median (quartile; minimum-maximum) values were as follows: PCV 22% (22%, 26%; 17-28%), TS 6.3 (6.0, 6.8; 5.8-7.5) g/dL, total protein 2.9 (2.7, 3.4; 2.2-4.3) g/dL, albumin 0.4 (0.4, 0.4; 0.3-0.5) g/dL, globulins 2.6 (2.3, 3.0;1.9-3.8) g/dL, sodium 282 (279, 285;273-292) mmol/L, potassium 7.3 (6.4, 7.9; 5.7-9.2) mmol/L, chloride 290 (285, 296; 277-304) mmol/L, total CO2 3 (2, 4; 0-5) mmol/L, calcium 16.8 (16.2, 17.4;15.8-18.2) mg/dL, phosphorus 8.8 (7.5, 10.0; 5.9-12.7) mg/dL, urea nitrogen 1004 (986, 1028; 944-1068) mg/dL, creatinine <0.1 mg/dL, glucose 184 (165, 191; 155-218) mg/dL, aspartate aminotransferase 42 (33, 66; 15-132) U/L, lactate dehydrogenase <5 U/L, creatine kinase 82 (47, 233; 18-725) U/L, and osmolality 1094 (1078, 1111; 1056-1139) mOsm/kg. No differences based on sex were detected. TS and TP values were related by the fitted line TS = (1.006 X TP) + 3.318. CONCLUSION: Values reported here will be useful for evaluating the health status of bonnetheads in wild and captive research conditions and in exhibits.

Animals↗

Some relations between dissolution rates and physical parameters of a drug in aqueous micellar solutions of a non-ionic surfactant.

Dissolution rates of salicylic acid from a constant surface area into a series of aqueous micellar polysorbate 20 solutions at pH 1-0 to 4-0 have been measured using two different methods; a stirred beaker and a rotating disc technique. The micellar molecular weight of polysorbate 20 has been obtained from light scattering and differential refractometry data and used with other independently determined physical data to calculate diffusion coefficients of the diffusing species. Linear multiple regression analysis was used to assess the dependence of drug dissolution rate on the diffusion coefficient and the viscosity of the dissolution medium.

Colloids↗

Sweet-sensitive protein from bovine taste buds: isolation and assay.

Using refractometry and ultraviolet-difference spectroscopy to indicate interaction between proteins and coinpounds of low molecular weight, we found a protein fraction in bovine tongue extracts that coinplexes sugars and saccharin. The strengths of the coinzplexes parallel the degrees of sweetness of the compounds, and the effects of pH upon formation of complexes parallel the effects of pH upon sensitivity of taste buds to sweet compounds in vivo.

Animals↗