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Breakdown of the blood-aqueous barrier after argon laser panretinal photocoagulation for proliferative diabetic retinopathy.

BACKGROUND: Breakdown of the blood-aqueous barrier (BAB) after panretinal photocoagulation (PRP) was measured with a laser flare photometer over a study period of 8 weeks. METHODS: Twenty-five eyes of 25 patients who had no previous photocoagulation and required such treatment for proliferative diabetic retinopathy (PDR) were included in the trial. They received 2000 burns (0.1-second exposure, 200 mu m spot) via a panfunduscope and 500 burns (0.1-second exposure, 500-mu m spot) with a Goldmann lens. Power levels were adjusted to produce a mild blanching of the retina. Only an argon green laser (514 nm) was used. Laser photometry was performed on both eyes at 3, 24, 48, 72, 96, and 168 hours and 8 weeks after laser treatment. RESULTS: Including all of the eyes treated, there was a significant increase in flare value of 3, 24, and 48 hours compared with baseline (Student's t test) but not at 72, 96, and 168 hours or at 8 weeks. Peak values occurred at 24 hours. When blue and brown irides were analyzed separately, there was a significant increase in flare for blue irides compared with baseline levels at 3 and 24 hours, whereas for brown irides the increased flare was sustained at 3, 24, 48, 72, and 96 hours (Student's paired t test). In addition, when the increase in flare value from baseline was compared between blue and brown irides (pooled Student's test), there was a sustained increase at 24, 48, 72, and 96 hours for brown irides compared with blue. Clinically significant uveitis, posterior synechiae, or peripheral anterior synechiae did not develop in any of the patients. CONCLUSIONS: Breakdown of the BAB may occur after PRP, particularly in more heavily pigmented irides. The time course of this suggests that the phenomenon is related directly to laser effects in the anterior segment, although other factors may contribute.

Anterior Eye Segment↗

Digestive vacuolar pH of intact intraerythrocytic P. falciparum either sensitive or resistant to chloroquine.

We present the first single cell-level analysis of digestive vacuolar pH for representative chloroquine resistant (strain Dd2) versus sensitive (strain HB3) malarial parasites. Human red blood cells harboring intact intraerythrocytic parasites were attached to glass substrate, continuously perfused with appropriate buffer, and pH was analyzed via single cell imaging and photometry techniques. We find that digestive vacuolar pH (pH(vac)) is near 5.6 for HB3 parasites. Surprisingly, we also find that pH(vac) of Dd2 is more acidic relative to HB3. Notably, in vitro pH titration of hematin confirms a very steep transition between soluble heme (capable of binding chloroquine) and insoluble heme (not capable of binding chloroquine, but still capable of polymerization to hemozoin) with a distinct midpoint at pH 5.6. We suggest the similarity between the hematin pH titration midpoint and the measured value of HB3 pH(vac) is not coincidental, and that decreased pH(vac) for Dd2 titrates limited initial drug target (i.e. soluble heme) to lower concentration. That is, changes in pH(vac) for drug resistant Dd2 relative to drug sensitive HB3 are consistent with lowering drug target levels, but not directly lowering vacuolar concentrations of drug via the predictions of weak base partitioning theory. Regardless, lowering either would of course decrease the efficiency of drug/target interaction and hence the net cellular accumulation of drug over time, as is typically observed for resistant parasites. These observations contrast sharply with the common expectation that decreased chloroquine accumulation in drug resistant malarial parasites is likely linked to elevated pH(vac,) but nonetheless illustrate important differences in vacuolar ion transport for drug resistant malarial parasites. In the accompanying paper (Ursos, L. et al., following paper this issue) we describe how pH(vac) is affected by exposure to chloroquine and verapamil for HB3 versus Dd2.

Acridine Orange↗

The diagnosis of endometrial carcinoma by means of jet wash and DNA flow-through cytophotometry.

Flow-through cytophotometric determination of nuclear DNA content on jet wash material from the endometrium was employed in the diagnosis of endometrial carcinoma. One hundred and thirty cases were studied. In comparison to results from cytodiagnosis, flow-through photometry yielded a false negative rate of 31.6% and a false positive rate of 42.2%. The false negative findings resulted in part from the small relative frequency of atypical cells in a mixed population of normal and atypical cells in certain cases. Besides this, we often found carcinomas with a diploid DNA stem line, which could not be distinguished cytophotometrically from normal corpus endometrium (Sandritter, 1952; Atkin et al., 1959; Hustin, 1976). The flow-through photometrically false positive findings may have resulted either from cell aggregates or from a nuclear DNA content elevated over the diploid value in proliferating cells (D. Wagner et al., 1968; D. Wagner and Richard, 1968). The observed false negative and false positive rates demonstrate that flow-through photometric determination of nuclear DNA content is unsuitable for the diagnosis of endometrial carcinoma.

Biopsy↗

Microionization constants: novel approach for the determination of the zwitterionic equilibrium of hydroxyphenylalkylamines by photometric titration.

The record of the formation of the phenolate and the zwitterionic form in the course of titration by photometry makes it possible to estimate the tautomeric equilibrium, K(Z), between the zwitterionic and the uncharged form of an ampholyte, provided that (1) the absorptivity of the phenolate and the zwitterionic form are identical and (2) the absorptivities of both forms are distinct from the absorptivities of the protonated and the uncharged form. The relation between the absorbance and K(Z), the degree of titration and the degree of overlapping of the basic and the acid ionization constant is given.

Algorithms↗

Corneal endothelial toxicity of different lidocaine concentrations.

PURPOSE: To examine the potential damaging effect on the corneal endothelium of unpreserved lidocaine in concentrations of 1%, 5%, and 10%. SETTINGS: Department of Ophthalmology, Charité Medical Faculty, Humboldt University, Berlin, Germany. METHODS: Experimental porcine corneas (n = 18) were exposed to 100 microL of unpreserved lidocaine hydrochloride at concentrations of 1%, 5%, and 10% for 60 minutes. Additional corneas (n = 6) were treated with lidocaine hydrochloride 1% for 30 minutes to simulate clinical conditions. Balanced salt solution (BSS((R))) served as a control to evaluate corneal endothelial cell damage using Janus Green photometry. Morphology, damage pattern, and changes in the ultrastructural appearance of corneal endothelial cells were examined by light and scanning electron microscopy. RESULTS: Lidocaine 1% used for 30 or 60 minutes did not cause significantly more corneal endothelial damage (mean 3.00% +/- 0.76% [SD] and 3.26% +/- 1.00%, respectively) than in the control group (mean 3.32% +/- 0. 86%) (P >.01). Significant corneal endothelial cell loss was observed with lidocaine 5% (mean 10.7% +/- 6.4%) (P <.001) and lidocaine 10% (42.3% +/- 17.0%) (P <.001). CONCLUSION: Experimental exposure of corneal endothelial cells to higher concentrations of lidocaine resulted in significant cell loss, indicating that the 1% concentration only should be used clinically.

Anesthesia, Local↗

Spectral sensitivity of cones in an ungulate.

Ungulates have been classified as having arrhythmic eyes in the sense that they contain features appropriate both to diurnal and nocturnal life. The former is typically associated with multiple classes of cones and a color-vision capacity. To see if an arrhythmic animal has these features, the number of cone classes was determined and the spectra of these cones were measured in a common ungulate, the domestic pig (Sus scrofa). Examination with electroretinogram (ERG) flicker photometry revealed the presence of two classes of cones in the pig's eye having average maximum sensitivity (lambda max) at 439 nm and 556 nm, respectively. This ungulate thus has the requisite retinal basis for dichromatic color vision.

Animals↗

L and M cone proportions in polymorphic New World monkeys.

Platyrrhine monkeys typically have only a single X-chromosome opsin gene. Alleles of this gene code for multiple versions of middle- to long-wavelength cone photopigments. X-chromosome inactivation provides heterozygous females with a retinal mosaic of cones containing either of two types of M and L pigment, thus establishing the photopigment basis for trichromatic color vision. This study examined the proportions of L and M cones created by this process. For that purpose, electroretinogram flicker photometry was used to obtain complete spectral sensitivity functions from 60 heterozygous female monkeys drawn from seven genera of platyrrhine monkeys. To obtain estimates of cone proportions, these functions were subsequently fit with linear combinations of L and M cone fundamentals that were derived from similar recordings made on conspecific animals having only one type of M/L pigment. Consistent with a random X-chromosome inactivation process, the average L:M cone weighting across the sample was close to unity. At the same time, there were significant individual variations in L:M cone proportions. The genesis of this variation and its implications for seeing are discussed.

Animals↗

Visual evoked potentials to red-green stimulation in schoolchildren.

The aim is to study chromatic visual evoked potentials (VEP) to isoluminant red-green (R-G) stimulus in schoolchildren. Sixty children (7-19 years) with normal color vision were examined, 30 binocularly and 30 monocularly. The isoluminant point was determined for each child subjectively by using heterochromatic flicker photometry, and objectively from recordings. The stimulus was a 7 degrees circle composed of horizontal sinusoidal gratings, with spatial frequency 2 cycles/degrees and 90% contrast, presented in onset-offset mode. VEP were recorded from Oz (mid-occipital) position. Age-dependent waveform changes and changes of the positive and negative wave were studied to both binocular and monocular R-G stimulation. Age-dependent waveform changes were observed to binocular and monocular R-G stimulation. In younger children the positive wave was prominent, whereas in older children also the negative wave became more evident. The latency of the positive wave decreased linearly with age to R-G binocular stimulation. To monocular stimulation no significant changes of the latency were observed. The amplitude of the positive wave dropped exponentially with age to binocular and monocular stimulation. The latency of the negative wave increased linearly with age to binocular and monocular stimulation, whereas the amplitude did not show age-dependent changes. These findings suggest that the chromatic VEP response undergoes evident age-dependent changes during the school-age period.

Adolescent↗

Photosynthetic water oxidation at high O2 backpressure monitored by delayed chlorophyll fluorescence.

The atmospheric dioxygen is produced by photosynthetic organisms. This light-driven process culminates in what appears as one step: a four-electron abstraction from two water molecules bound to the Mn4Ca complex of photosystem II. Recently, an intermediate of the O2-producing reaction sequence was stabilized by elevated oxygen backpressure and detected by UV flash photometry [Clausen, J., and Junge, W. (2004) Nature 430, 480]. We scrutinized its properties by delayed chlorophyll fluorescence measurements. Half-suppression of oxygen evolution was observed at a similar O2 pressure of 2.3 bar, as previously, now with photosystem II membrane particles from spinach, without artificial electron acceptors, and at a high signal-to-noise ratio. The data are tentatively interpreted as the stabilization of a 2-fold oxidized state of the catalytic center (S2*) with bound peroxide and its slow conversion into the normal S2 state by the release of peroxide.

Chlorophyll↗

Ground-based detectability of terrestrial and Jovian extrasolar planets: observations of CM Draconis at Lick Observatory.

The detection of terrestrial-sized extrasolar planets from the ground has been thought to be virtually impossible due to atmospheric scintillation limits. However, we show that this is not the case especially selected (but nevertheless main sequence) stars, namely small eclipsing binaries. For the smallest of these systems, CM Draconis, several months to a few years of photometric observations with 1-m-class telescopes will be sufficient to detect the transits of any short-period planets of sizes > or = 1.5 Earth radii (RE), using cross-correlation analysis with moderately good photometry. Somewhat larger telescopes will be needed to extend this detectability to terrestrial planets in larger eclipsing binary systems. (We arbitrarily define "terrestrial planets" herein as those whose disc areas are closer to that of Earth's than Neptune's i.e., less than about 2.78 RE.) As a "spin-off" of such observations, we will also be able to detect the presence of Jovian-mass planets without transits using the timing of the eclipse minima. Eclipse minima will drift in time as the binary system is offset by a sufficiently massive planet (i.e., one Jupiter mass) about the binary/giant-planet barycenter, causing a periodic variation in the light travel time to the observer. We present here an outline of present observations taking place at the University of California Lick Observatory using the Crossley 0.9-m telescope in collaboration with other observatories (in South Korea, Crete, France, Canary Islands, and New York) to detect or constrain the existence of terrestrial planets around main sequence eclipsing binary star systems, starting with CM Draconis. We demonstrate the applicability of photometric data to the general detection of gas giant planets via eclipse minima timings in many other small-mass eclipsing binary systems as well.

Astronomy↗

Ultrasound biomicroscopy examination of intraocular lens haptic position after phacoemulsification with continuous curvilinear capsulorhexis and extracapsular cataract extraction with linear capsulotomy.

PURPOSE: Intraocular lens (IOL) haptic position in 35 eyes that had undergone cataract surgery was examined with ultrasound biomicroscopy (UBM). METHODS: In a prospective randomized study the patients were operated by phacoemulsification using continuous curvilinear capsulorhexis (CCC) (group I) or by extracapsular cataract extraction (ECCE) using linear capsulotomy (group II). Ultrasound biomicroscopy was used to localize both haptics of the implanted intraocular lenses and to measure anterior chamber depth (ACD), iris thickness and anterior chamber angle. The inflammatory reaction in the anterior chamber was assessed with laser flare photometry. Slit lamp examination was performed. RESULTS: Both IOL haptics were found in the lens capsule in all 18 eyes in group I. In group II one of the haptics was located out of the capsule in 7 of 17 eyes (41%). The difference is statistically significant (p=0.01). Postoperatively mean ACD measured with the UBM was 4.06+/-0.30 mm in group I and 3.64+/-0.24 mm in group II (p=0.00025). CONCLUSION: The UBM examinations indicate that phacoemulsification with continuous curvilinear capsulorhexis is a more reliable technique than ECCE with linear capsulotomy to achieve implantation of the intraocular lens haptics in the capsular bag.

Aged↗

Characterization of extrasolar terrestrial planets from diurnal photometric variability.

The detection of massive planets orbiting nearby stars has become almost routine, but current techniques are as yet unable to detect terrestrial planets with masses comparable to the Earth's. Future space-based observatories to detect Earth-like planets are being planned. Terrestrial planets orbiting in the habitable zones of stars-where planetary surface conditions are compatible with the presence of liquid water-are of enormous interest because they might have global environments similar to Earth's and even harbour life. The light scattered by such a planet will vary in intensity and colour as the planet rotates; the resulting light curve will contain information about the planet's surface and atmospheric properties. Here we report a model that predicts features that should be discernible in the light curve obtained by low-precision photometry. For extrasolar planets similar to Earth, we expect daily flux variations of up to hundreds of per cent, depending sensitively on ice and cloud cover as well as seasonal variations. This suggests that the meteorological variability, composition of the surface (for example, ocean versus land fraction) and rotation period of an Earth-like planet could be derived from photometric observations. Even signatures of Earth-like plant life could be constrained or possibly, with further study, even uniquely determined.

Earth, Planet↗

Protection of corneal endothelium from irrigation damage: a comparison of sodium hyaluronate and hydroxypropylmethylcellulose.

PURPOSE: To compare the endothelial protection of sodium hyaluronate and hydroxypropylmethylcellulose against endothelial damage induced by irrigation. METHODS: An in vitro assay with freshly excised porcine eyes was developed using the Janus green photometry technique. Irrigation and aspiration technique was standardised. Forty pairs of porcine eyes were used. One randomly chosen eye was filled with sodium hyaluronate (SH) and the other with hydroxypropylmethylcellulose (HPMC). Irrigation and aspiration was carried out with balanced salt solution for 5 min. Twenty additional pairs of porcine eyes served as controls. Student's t-test was used for statistical analysis. RESULTS: Both viscoelastic agents protected the endothelium as compared with controls. The endothelial protection, determined with the Janus green photometric technique, was significantly greater with HPMC than with SH. CONCLUSIONS: Viscoelastic agents are effective in protecting the endothelium from irrigation damage in porcine eyes in vitro. HPMC provided greater protection than SH in this particular model.

Animals↗

Angiotensin II-mediated calcium signals and mitogenesis in human prostate stromal cell line hPCPs.

Western blots and immunocytochemistry were used to detect angiotensin 1 (AT(1)) and angiotensin 2 (AT(2)) receptors in human primary cultures of the prostate stromal compartment (hPCPs). Immunohistochemistry was performed on human prostate tissue-embedded paraffin. In addition, pharmacological tools were applied in combination with photometry experiments to characterize the physiological activity of AT(1) and AT(2) receptors in hPCPs cell culture. A proliferation assay was used to describe the mitogenic activity of angiotensin II (Ang II) on hPCPs cells. Only the AT(1) receptor was detected in Western blot analysis. Immunocytochemistry of hPCPs cells showed that the AT(1) receptor is present in both the smooth muscle type and the fibroblastic type. In the stromal compartment of human prostate tissue, immunoreaction with antibodies against the AT(1) receptor was detectable.Fura-2-loaded hPCPs cells showed an instantaneous and linear rise in free intracellular calcium ion concentration ([Ca(2+)](i)) after local perfusion with Ang II in concentrations of 10 nM. Removing of external calcium or emptying intracellular calcium stores before Ang II application diminished or abolished this [Ca(2+)](i) response. The response to Ang II was also diminished when hPCPs cells were perfused with the AT(1) receptor inhibitor losartan prior to Ang II application. No inhibition of the [Ca(2+)](i) increase was detectable after perfusion with PD 123319, a specific inhibitor of the AT(2) receptor.hPCPs cells were stimulated with Ang II in various concentrations over a period of 2 days. The subsequently performed proliferation assay revealed a mitogenic effect of Ang II on hPCPs in concentrations starting at 10 nM. This effect could be inhibited by losartan.

Angiotensin II↗

The role of bound potassium ions in the hydrolysis of low concentrations of adenosine triphosphate by preparations of membrane fragments from ox brain cerebral cortex.

1. The intrinsic Na(+), K(+), Mg(2+) and Ca(2+) contents of a preparation of membrane fragments from ox brain were determined by emission flame photometry. 2. Centrifugal washing of the preparation with imidazole-buffered EDTA solutions decreased the bound Na(+) from 90+/-20 to 24+/-12, the bound K(+) from 27+/-3 to 7+/-2, the bound Mg(2+) from 20+/-2 to 3+/-1 and the bound calcium from 8+/-1 to <1nmol/mg of protein. 3. The activities of the Na(+)+K(+)+Mg(2+)-stimulated adenosine triphosphatase and the Na(+)-dependent reaction forming bound phosphate were compared in the unwashed and washed preparations at an ATP concentration of 2.5mum (ATP/protein ratio 12.5pmol/mug). 4. The Na(+)-dependent hydrolysis of ATP as well as the plateau concentration of bound phosphate and the rate of dephosphorylation were decreased in the washed preparation. The time-course of formation and decline of bound phosphate was fully restored by the addition of 2.5mum-magnesium chloride and 2mum-potassium chloride. Addition of 2.5mum-magnesium chloride alone fully restored the plateau concentration of bound phosphate, but the rate of dephosphorylation was only slightly increased. Na(+)-dependent ATP hydrolysis was partly restored with 2.5mum-magnesium chloride; addition of K(+) in the range 2-10mum-potassium chloride then further restored hydrolysis but not to the control rate. 5. Pretreatment of the washed preparation at 0 degrees C with 0.5nmol of K(+)/mg of protein so that the final added K(+) in the reaction mixture was 0.1mum restored the Na(+)-dependent hydrolysis of ATP and the time-course of the reaction forming bound phosphate. 6. The binding of [(42)K]potassium chloride by the washed membrane preparation was examined. Binding in a solution containing 10nmol of K(+)/mg of protein was linear over a period of 20min and was inhibited by Na(+). Half-maximal inhibition of (42)K(+)-binding required a 100-fold excess of sodium chloride. 7. It was concluded (a) that a significant fraction of the apparent Na(+)-dependent hydrolysis of ATP observed in the unwashed preparation is due to activation by bound K(+) and Mg(2+) of the Na(+)+K(+)+Mg(2+)-stimulated adenosine triphosphatase system and (b) that the enzyme system is able to bind K(+) from a solution of 0.5mum-potassium chloride.

Adenosine Triphosphatases↗

Changes in sodium transport during the human menstrual cycle and pregnancy.

1. We have studied the transport of Na+ and K+ by erythrocytes during the follicular and luteal phases of the human menstrual cycle, and in pregnant compared with non-pregnant women. Venous blood was drawn from 10 healthy young women (not taking any medication or hormones) 1-2 days after menstruation and from the same women 7-9 days after ovulation. For the pregnancy part of the study, blood was drawn from eight other normotensive non-pregnant women and from eight age-matched normotensive pregnant women (36-43 weeks gestation). 2. Intracellular erythrocyte and plasma Na+ and K+ concentrations were measured by flame photometry. The increase in the intracellular Na+ concentration during a 1 h 37 degrees C incubation of fresh whole blood with 0.2 mmol/l ouabain (compared with no ouabain) was measured to determine the rate of active Na+ efflux. The Na(+)-K+ pump rate constant was calculated by dividing the active Na+ efflux rate by the intracellular Na+ concentration. 3. In fresh blood, the intracellular erythrocyte Na+ concentration (P < 0.002) and the plasma K+ concentration (P < 0.01) were both lower in pregnant than in non-pregnant women. The Na(+)-K+ pump rate constant was higher (P < 0.02) during the luteal phase than during the follicular phase, and in pregnant compared with non-pregnant women. 4. We conclude that short-acting hormones in the plasma most probably account for the changes in the Na(+)-K+ pump rate constant during the menstrual cycle.

Adult↗

[Determination of oxygen content: a comparison between the new Oxystat-Photometer and the Lex-O2-Con].

The Oxystat-photometer (Fa. Andos) is a new instrument for measuring of oxygen content and haemoglobin of blood. Using specific agents oxygen and haemoglobin can be registered in small cuvettes. The principle of measurement is photometry. Additionally O2-saturation can be calculated. In 139 experiments we checked the reliability of the Oxystat-photometer in comparison to the Lex-O2-Con (Lexington Instruments Corporation). High significance could be demonstrated (r = 0.96). There was a range from the control value below 5%. Following investigations using calculation, results could be confirmed.

Adult↗

[Sodium content of liquid antacid preparations].

The sodium content of 13 liquid antacids was determined using two independent analytical methods (flame photometry). With a neutralisation buffering capacity of 564 mmol/d (pH 3.5) the possible body sodium load was calculated by evaluating the sodium content in the supernatant of the antacid suspensions, ranging from 0.56 to 568 mmol/d (method A) and from 0.42 to 468 mmol/d (method B). Therefore, in patients who are on a sodium restriction of 50 mmol/d this critical intake may be easily exceeded by the intake of certain antacids.

Antacids↗