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Potassium fluxes and ouabain binding in growing, density-inhibited and Rous sarcoma virus-transformed chicken embryo cells.

Potassium fluxes, ouabain binding, and Na+ and K+ intracellular concentrations were determined for cultures of growing normal, density-inhibited and Rous sarcoma virus-transformed chicken embryo fibroblasts. No significant differences in K+ influx or ouabain binding were detected between growing normal cells and Rous sarcoma virus-transformed cells; however, ouabain binding and ouabain-sensitive K+ influx were 1.5- to 1.8-fold lower in density-inhibited cells. Thus, potassium influx in this system can be classified as a growth-related, but not transformation-specific change. As determined by both flame photometry and radioisotopic (42K) equilibration, growing normal and density-inhibited cells had similar potassium contents, whereas transformed cells exhibited 1.4-fold higher potassium levels. Sodium ion levels, as measured by flame photometry, were also 2- to 4.5-fold higher in transformed than normal or density-inhibited cells. Complementary studies of potassium efflux showed a 1.3- to 1.5-fold higher rate (based on the percentage of pool exiting the cell) in growing normal versus density-inhibited or transformed fibroblasts. Because of the larger potassium pool in transformed cells, efflux based on absolute number of potassium ions is similar in normal and transformed chicken embryo fibroblasts.

Animals↗

NMR measurement of 39K detectability and relaxation constants in rat tissue.

Differences in the NMR detectability of 39K in various excised rat tissues (liver, brain, kidney, muscle, and testes) have been observed. The lowest NMR detectability occurs for liver (61 +/- 3% of potassium as measured by flame photometry) and highest for erythrocytes (100 +/- 7%). These differences in detectability correlate with differences in the measured 39K NMR relaxation constants in the same tissues. 39K detectabilities were also found to correlate inversely with the mitochondrial content of the tissues. Mitochondria prepared from liver showed greatly reduced 39K NMR detectability when compared with the tissue from which it was derived, 31.6 +/- 9% of potassium measured by flame photometry compared to 61 +/- 3%. The detectability of potassium in mitochondria was too low to enable the measurement of relaxation constants. This study indicates that differences in tissue structure, particularly mitochondrial content are important in determining 39K detectability and measured relaxation rates.

Animals↗

Genome size determination in peronosporales (Oomycota) by Feulgen image analysis.

Genome size was determined, by nuclear Feulgen staining and image analysis, in 46 accessions of 31 species of Peronosporales (Oomycota), including important plant pathogens such as Bremia lactucae, Plasmopara viticola, Pseudoperonospora cubensis, and Pseudoperonospora humuli. The 1C DNA contents ranged from 0.046 (45. 6 Mb) to 0.163 pg (159.9 Mb). This is 0.041- to 0.144-fold that of Glycine max (soybean, 1C = 1.134 pg), which was used as an internal standard for genome size determination. The linearity of Feulgen absorbance photometry method over this range was demonstrated by calibration of Aspergillus species (1C = 31-38 Mb) against Glycine, which revealed differences of less than 6% compared to the published CHEF data. The low coefficients of variation (usually between 5 and 10%), repeatability of the results, and compatibility with CHEF data prove the resolution power of Feulgen image analysis. The applicability and limitations of Feulgen photometry are discussed in relation to other methods of genome size determination (CHEF gel electrophoresis, reassociation kinetics, genomic reconstruction) that have been previously applied to Oomycota.

Aspergillus↗

Posterior sub-Tenon's steroid injections for the treatment of posterior ocular inflammation: indications, efficacy and side effects.

PURPOSE: Posterior sub-Tenon's steroid injections (PSTSI) are a standard drug delivery method used for the treatment of chronic uveitis of the posterior segment. The aim of this study was to analyse the indications, efficacy and complications of PSTSI in the treatment of chronic uveitis. METHODS: During the period 1990-1994, 53 (9.5%) of 558 patients (58 eyes) followed up in the uveitis clinic received a total of 162 PSTSI of triamcinolone acetonide 40 mg in the superior quadrants. Indications for treatment were vision inferior or equal to 0.7 and/or intolerable visual disturbance. Only patients in whom PSTSI were the only treatment parameter changed were analysed. Among the main parameters analysed were visual acuity, aqueous laser flare photometry, intraocular pressure (IOP) and complications. RESULTS: Anatomical location of uveitis was as follows: anterior HLA-B27-related uveitis with CME (1 patient/1 eye), intermediate uveitis (28/32), posterior uveitis (10/10) and panuveitis (14/15). Mean duration of follow-up was 448+/-57 days. Visual acuity improved significantly from 0.40+/-0.03 to 0.79+/-0.07, with 59.4% of eyes having a gain of 2-5 Snellen lines and 18.7% a gain of >5 lines. Mean aqueous flare photometry decreased significantly from 29.6+/-3.5 to 13.6+/-2.2 photons/ms. Mean IOP increased significantly from 13.6+/-0.5 to 18.5+/-0.8 mm Hg with a rise of pressure >8 mm Hg in 23 cases (36%), transient in 16 cases, but chronic in 6 cases, needing filtering surgery. Partial superior ptosis was seen in two cases and cataract progressed in seven cases. CONCLUSION: PSTSI are very effective in restoring visual acuity in chronic uveitis of the posterior segment, without systemic complications, but at the expense of intraocular hypertension, a complication that was found more frequently than expected.

Adolescent↗

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↗

Leukocyte isotopically exchangeable intracellular sodium fractions in lean and overweight hypertensives.

Leukocyte intracellular sodium, as measured by flame photometry, is increased in essential hypertension, especially when associated with a body mass index greater than 27 kg.m-2. A triple isotope method for measuring the isotopically exchangeable pool of intracellular sodium was used to assess if this pool was increased in hypertension. No significant differences in the isotopically exchangeable intracellular sodium concentration were found between lean and overweight hypertensives compared with normotensive controls. Lean hypertensives with systolic blood pressures below the median had significantly lower exchangeable intracellular sodium concentrations than lean normotensives, whereas those with systolic blood pressures above the median had raised exchangeable intracellular sodium concentrations. The obese hypertensives did not show this trend. The exchangeable intracellular sodium concentration was correlated to systolic (r = .53, P less than .001) and diastolic (r = 0.39, P less than .01) blood pressure in hypertensives. We conclude that the increase in total cellular sodium content (as measured by flame photometry) in hypertensives described in previous studies is not associated with any increase in the isotopically exchangeable pool of intracellular Na+, except in those lean hypertensives with systolic blood pressures above the median. By implication, there may be an increased slowly exchangeable pool of intracellular Na+ in leukocytes from most hypertensives.

Adult↗

Increment threshold and purity discrimination spectral sensitivities of X-chromosome-linked color-defective observers.

The goal of the study was to evaluate spectral opponency in nine X-chromosome-linked color-defective observers. The tasks included increment threshold spectral sensitivity on an achromatic background, heterochromatic flicker photometry, and colorimetric purity discrimination. With a task of heterochromatic flicker photometry, the anomalous trichromatic observers showed spectral sensitivity of the corresponding dichromat. The increment threshold spectral sensitivity and colorimetric purity discrimination data were analyzed using the concept of standard cone photopigment spectral sensitivities for normal and defective vision, and a model that postulates one cone-additive and two cone-antagonistic systems. The model incorporated a shift of the peak spectral sensitivity of the long-wavelength-sensitive (LWS) pigment (for protan observers) or of the middle-wavelength-sensitive (MWS) pigment (for deutan observers). Two dichromats and two anomalous trichromats did not show clear evidence of LWS vs MWS cone antagonism. Five anomalous trichromats showed such cone antagonism. Molecular genetic analysis of the opsin genes is presented for eight of the observers.

Color Vision Defects↗

Clinical features of Chinese patients with Fuchs' syndrome.

PURPOSE: To characterize the clinical features of Chinese patients with Fuchs' syndrome. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: One hundred eighteen eyes of 104 consecutive patients with Fuchs' syndrome initially examined between January 1999 and March 2005. METHODS: The history and clinical findings of all consecutive Fuchs' patients attending the Zhongshan Ophthalmic Center were reviewed. Auxiliary examinations, including laser flare-cell photometry, ultrasound biomicroscopy (UBM), fundus fluorescein angiography (FFA), and serologic tests for Toxoplasma gondii, were performed in certain cases. MAIN OUTCOME MEASURES: Patients' demographics, clinical presentation, and auxiliary examination findings. RESULTS: One hundred four patients (49 male, 55 female) were included in this study. Unilateral involvement was noted in 90 patients (86.5%). The most common symptom was blurred or decreased vision (86%). Stellate and medium-sized keratic precipitates (KPs) were noted in 108 eyes (91.5%). A mild anterior chamber (AC) reaction was observed in all the affected eyes. Heterochromia was observed in only 15 affected eyes, although there were varying degrees of iris depigmentation in all patients. Iris nodules, mostly Koeppe, were present in 28.0% of the affected eyes. Complicated cataract, vitreous opacity, and secondary glaucoma were observed in 84 of 118 eyes (70.7%), 31 eyes of 42 eyes (73.8%), and 24 of 118 eyes (23.1%), respectively. The mean laser flare photometry value (6.4+/-2.3 photon counts per millisecond) and the cell number in the AC (1.5+/-1.2 cells per 0.5 mm3) in 25 patients were both significantly higher than those in 25 healthy controls (5.3+/-2.3 photon counts per millisecond and 0.8+/-0.6 cells per 0.5 mm3) (P<0.05). Ultrasound biomicroscopy revealed exudates adjacent to the ciliary body in 18 of 24 patients (75%). Serological tests failed to confirm an association of Fuchs' syndrome with toxoplasmosis. Retinal capillary leakage in the midperipheral fundus and disc staining at the late stage were observed in most of the eyes examined by FFA. CONCLUSION: Fuchs' syndrome in Chinese patients is characterized by a mild uveitis with characteristic KPs, varying degrees of iris depigmentation, and, occasionally, heterochromia. Exudates adjacent to the ciliary body and subclinical retinal and optic nerve involvement were common in the patients who were studied by UBM and FFA.

Adolescent↗

Randomized clinical trial of cryotherapy versus laser photocoagulation for retinopexy in conventional retinal detachment surgery.

PURPOSE: To investigate whether the method of retinopexy influences the visual recovery rate and the breakdown of the blood-ocular barrier after conventional retinal detachment surgery. METHODS: Forty-eight patients (48 eyes) with primary rhegmatogenous retinal detachment entered into the study. All eyes were phakic, had an attached macula, and were scheduled for conventional scleral buckling surgery. Patients were randomly assigned to have either laser or cryotherapy for retinopexy. All visual acuity and flare measurements were performed by a masked observer. The interventional procedure was cryopexy at the time of scleral buckling surgery or postoperative (4 weeks) laser photocoagulation. Visual acuity testing with ETDRS chart and aqueous flare measurement with laser flare photometry were performed by a masked observer at standard intervals: preoperatively and 1 day, 7 days, 4 weeks, and 10 weeks postoperatively. Analysis of covariance by multiple linear regression was used for statistical evaluation. RESULTS: Postoperative flare values from patients receiving cryotherapy were significantly higher at each measurement point in time (P < or =.001). The visual recovery was slower in the patients receiving cryotherapy (1 week, P =.003; 4 weeks, P =.03; 10 weeks, P =.081). CONCLUSION: Laser flare photometry proved sufficiently sensitive to quantify an increase in aqueous flare after limited external retinal cryotherapy. Postoperative flare, as a measure of blood-ocular barrier breakdown, was significantly higher and visual recovery slower in the cryotherapy group. Visual acuity after 10 weeks was not significantly different between both groups.

Blood-Retinal Barrier↗

Transfection of mu MDR 1 inhibits Na(+)-independent Cl-/-HCO3 exchange in Chinese hamster ovary cells.

We have used single-cell photometry to measure intracellular pH (pHi) for several MDR cell lines constructed by stably transfecting LR73 chinese hamster ovary fibroblasts with mutant and wild type murine MDR 1 genes. In addition, plasma membrane electrical potential (delta psi) has been measured for the same cells by the K+/valinomycin null point titration method using the ratiometric styryl probe di-4-ANEPPS. Both the untransfected, parental cell line and a cell line expressing substantial mutant MDR 1 protein (K432R/K1074R) that is unable to confer the MDR phenotype are found to have delta psi > or = -40 (+/- 5) mV and pHi < or = 7.16 (+/- 0.03) units. In contrast, MDR cell lines constructed by transfecting wild type mu MDR 1 cDNA are found to exhibit delta psi from 15 to 19 mV lower and pHi from 0.13 to 0.34 units higher. A cell line that overexpresses crippled MDR protein (S941F) that is not resistant to colchicine or doxorubicin, but which is resistant to vinblastine [Gros, P., Dhir, R., Croop, J., & Talbot, F. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 7289-7293], exhibits elevated pHi and slightly elevated delta psi, relative to LR73. Northern and western blot analyses confirm the substantial overexpression of the mu MDR genes and proteins in these lines, as well as the mild overexpression of endogenous hamster p-GP mRNA in some lines. In general agreement with previous studies that examined myeloma cells overexpressing hu MDR 1 protein [Roepe, P.D., Wei, L.-Y., Cruz, J., & Carlson, D. (1993) Biochemistry 32, 11042-11056] we find that overexpression of wild type mu MDR 1 protein inhibits Cl(-)- and -HCO3-dependent pHi homeostasis. Via single-cell photometry studies we now conclude that this is due to inhibition of Na(+)-independent Cl-/-HCO3 exchange (strict anion exchange or AE). As concluded previously for other MDR cells, decreased AE activity is not due to decreased expression of the exchanger; in fact, again similar to previous work [Roepe et al. (1993) Biochemistry 32, 11042-11056], we find increased levels of AE mRNA in some MDR cell lines. Models that may explain these data that are also consistent with the known physiology of cells that endogenously express MDR protein are suggested. These data are consistent with a model for MDR protein function wherein overexpression of the protein decreases delta psi and/or elevates pHi via Cl(-)- and -HCO3-dependent mechanisms.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

[Dose-effect relation in therapy of cytostatic-induced leukopenia with lithium carbonate].

Lithium is used increasingly for attenuation of chemotherapy induced leukopenia in infants and adolescents. Thus, pharmacological data are needed. The purpose of this study was to give recommendations for oral dosage yielding efficient serum levels. Furthermore, two methods of serum level determination were compared. Flame emission photometry using microsamples gave results comparable to atomic absorption photometry. The correlation of oral dose and serum level was investigated in a phase I study. A wide range of serum levels related to an identical oral dose was found. Therefore, frequent serum level determinations are needed to realize efficacy and to avoid toxicity. The correlation between increasing serum levels and leucocyte count was investigated in the first part of a phase II study. No significance was found; i.e. a linear progressing dose-effect relation does not exist. The time course of leukocyte count was analysed for two different serum level groups in the second part of the phase II study. A significant correlation was found in the group with a serum level greater 0,7 mmol/l.

Adult↗

The dust coma of Comet P/Giacobini-Zinner in the infrared.

We present 1-20 micrometers photometry of P/Giacobini-Zinner obtained at the NASA Infrared Telescope Facility, during 1985 June-September (r = 1.57-1.03 AU). A broad, weak 10 micrometers silicate emission feature was detected on August 26.6; a similar weak emission feature could have been hidden in the broadband photometry on other dates. The total scattering and emitting cross section of dust in the inner coma was similar to that in other short-period comets, but a factor of 10 (r = 1.56 AU) to 100 (r = 1.03 AU) lower than the amount of dust in Comet Halley. The thermal emission continuum can be fit with models weighted toward either small or large absorbing grains. The dust production rate near perihelion was approximately 10(5) g/s (small-grain model) to approximately 10(6) g/s (large-grain model). The corresponding dust/gas mass ratio on August 26 was approximately 0.1-1. A silicate-rich heterogeneous grain model with an excess of large particles is compatible with the observed spectrum of Giacobini-Zinner on August 26. Thus, weak or absent silicate emission does not necessarily imply an absence of silicates in the dust, although the abundance of silicate particles < or = 1 micrometer radius must have been lower than in Comet Halley.

Astronomy↗

The Optical Gravitational Lensing Experiment: Red Clump Stars as a Distance Indicator.

We present relation of the mean I-band brightness of red clump stars on metallicity. Red clump stars were proposed to be a very attractive standard candle for distance determination. The calibration is based on 284 nearby red giant stars whose high-quality spectra made it possible to determine accurate individual metal abundances. High-quality parallaxes (sigmapi&solm0;pi<10%) and photometry of these very bright stars come from Hipparcos measurements. Metallicity of the sample covers a large range: -0.6 dex<&sqbl0;Fe&solm0;H&sqbr0;<+0.2 dex. We find a weak dependence of the mean I-band brightness on metallicity ( approximately 0.13 mag dex-1). What is more important, the range of metallicity of the Hipparcos sample partially overlaps with metallicity of field giants in the LMC, thus making it possible to determine the distance to the LMC by almost direct comparison of brightness of the local Hipparcos red clump giants with that of LMC stars. Photometry of field red clump giants in nine low-extinction fields of the LMC halo collected during the OGLE II microlensing survey compared with the Hipparcos red clump stars data yields the distance modulus to the LMC: &parl0;m-M&parr0;LMC=18.24+/-0.08 mag.

Journal Article↗

Brightness contrast perception in the mesopic region.

Subjective evaluations of brightness contrast were obtained for Munsell chips of varying hues, lightness values and sizes against a white background illuminated in turn by four white illuminants at four background luminances in the mesopic region. Chip lightness was the strongest variable, resulting in a family of monotonic functions relating brightness contrast ratings to chip lightness. At the lowest light levels a nearly linear relationship was found between chip lightness and subjective ratings. As light levels increased, the relationship became curvilinear such that at the lowest contrast, chromatic brightness became important for subjective ratings. Within the level of precision for this experiment, chip hue and size as well as the spectral power distribution of the illuminants did not significantly impact the overall brightness contrast ratings. The subjective ratings of brightness contrast were analysed as a function of the luminance contrast of the stimuli as determined by conventional photometry and by three models of mesopic vision. The response functions relating subjective rating to the stimulus characterized in terms of these models were similar to those obtained with conventional photometry.

Adult↗

Evaluation and assessment of new disposable strip for determination of plasma potassium concentration.

Overall laboratory reproducibility for the Ames Seralyzer and plasma potassium test strips was less than 4% within the plasma reference range; and linearity extended from 2.1 to 10.2 mmol/l. Neither bilirubin nor lipaemia interfered in the analysis and selectivity for potassium over sodium was 450:1. Laboratory comparison with automated flame photometry and an indirect ion selective electrode showed a Seralyzer bias of +0.10 (+/- 25D 0.26) and +0.11 (0.27) mmol/l. Against flame photometry, comparison of analyses by six doctors from a coronary care unit and four nurses from a renal dialysis unit showed smaller positive biases (0.06 and 0.02 mmol/l) but substantially larger error ranges (0.43 and 0.55 mmol/l, respectively). Some 49% of the nurses' and 37% of the doctors' results differed from laboratory values by more than 0.2 mmol/l. Potassium test strip results proved acceptably comparable and reproducible when produced by an experienced analyst, but greater variability in clinical hands rendered the system unsuitable for precise monitoring of potassium concentration.

Humans↗

Effects of oxytocin on cation content and electrophysiology of frog skin epithelium.

We measured effects of oxytocin on current-voltage (I-V) relations of frog (Rana catesbeiana) skins impaled with an intracellular microelectrode. In both Cl- and Cl(-)-free (SO4(2-) solutions, oxytocin caused an approximate doubling short-circuit current (Isc) and a depolarization of the cell membrane. Increase in apical membrane slope conductance, chord conductance, and permeability after oxytocin correlated with the increase in amiloride-sensitive Isc. Oxytocin also increased basolateral membrane conductance (gb). In Cl-, the shift in the voltage intercept of the apical membrane I-V relation (Ea) implied increased intracellular Na+ activity (a(Na)c) after oxytocin. In isolated frog skin epithelia, a similar increase in intracellular [Na+] after oxytocin was demonstrated by flame photometry. In SO4(2-), changes caused by oxytocin in both Ea and in flame photometrically determined cell [Na+] were minimal. The voltage intercept of the basolateral membrane I-V relations (Eb) was shifted by oxytocin in both Cl- and SO4(2-) solutions. Assuming that the basolateral membrane is selectively permeable to K+, changes in K+ obtained from Eb were in disagreement with those obtained by flame photometry. These results suggest that 1) the increase in a(Na)c caused by oxytocin is not essential to produce either the increase in gb or Isc and 2) ions other than K+ make an important contribution to basolateral membrane conductance.

Animals↗

A simple method for evaluation of Rb+ transport and Na(+)-K+ pump stoichiometry in adherent cells.

This report describes a method based on flame photometry for the evaluation of transmembrane Rb+ transport and Na(+)-K+ pump stoichiometry in adherent cells. In monolayers of cultured fibroblasts, the rates of 86Rb+, an isotope widely used as a K+ congener in transport studies, and nonradioactive Rb+ influx were equivalent when measured in the absence and presence of the transport inhibitors ouabain and bumetanide. Ouabain- and bumetanide-sensitive Rb+ fluxes were also equal with the two methods. Flame photometry allowed the simultaneous determination of intracellular [Na+] in the same sample in which Rb was measured. The incubation of human fibroblasts with ouabain for 5 min promoted a significant increase in intracellular [Na+]. Under appropriate experimental conditions, the ratio between the rate of ouabain-promoted increase in intracellular [Na+] and ouabain-sensitive Rb+ influx was 1.4, close to the theoretical value of 1.5 corresponding to a Na(+)-K+ pump stoichiometry of 3 Na+ extruded from the cell in exchange for 2 K+.

Biological Transport, Active↗

Electrophysiological and optical changes in slices of rat hippocampus during spreading depression.

Spreading depression (SD) was studied with intracellular and extracellular recordings and with photometry in slices of rat hippocampus. Repetitive electrical stimulation could initiate SD in either normal medium or in low-Ca2+ medium containing Mn2+, especially during transient hypoxia. The extracellular voltage near CA1 pyramidal somata and dendrites became negative by approximately 18 mV during SD. This negativity peaked more slowly in low-Ca2+ medium containing Mn2+. The wave of negativity propagated across the slice in both media at approximately 6 mm/min. Input resistance of pyramidal neurons became undetectable during SD, and differential voltage recording between neurons and adjacent extracellular space demonstrated that transmembrane potential approached zero. Slices became more opaque during SD. Photometry revealed approximately 10% increase in reflectance and a similar decrease in transmittance of white light, which occurred with a time course similar to the extracellularly recorded voltage shift. These data support the hypothesis that SD represents a large increase in membrane permeability associated with substantial movements of water. The persistance of SD in a bathing solution that blocked electrically evoked postsynaptic potentials suggests that the contribution of synaptic transmitter release to the propagation of SD should be reappraised.

Animals↗