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Analysis of violet-excited fluorochromes by flow cytometry using a violet laser diode.

BACKGROUND: Low power violet laser diodes (VLDs) have been evaluated as potential replacements for water-cooled argon-ion and krypton-ion ultraviolet and violet lasers for DNA content analysis using the Hoechst dyes and 4,6-diamidino-2-phenylindole (Shapiro HMN, Perlmutter NG: Cytometry 44:133-136, 2001). In this study, we used a VLD to excite a variety of violet-excited fluorescent molecules important in biomedical analysis, including the fluorochromes Cascade Blue and Pacific Blue, the expressible fluorescent protein cyan fluorescent protein (CFP), and the fluorogenic alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazoline (ELF-97; for endogenous AP detection and cell surface labeling with AP-conjugated antibodies). METHODS: Comparisons were made between VLD excitation and a krypton-ion laser emitting at 407 nm (both at higher power levels and with the beam attenuated at levels approximating the VLD) on the same FACSVantage SE stream-in-air flow cytometer. We evaluated a Power Technology 408-nm VLD (30 mW) equipped with circularization optics (18 mW maximum output, set to 15 mW) and a Coherent I-302C krypton-ion laser emitting at power levels ranging from 15 to 75 mW. RESULTS: Cascade Blue, Pacific Blue, and CFP showed comparable signal-to-noise ratios and levels of sensitivity with VLD excitation versus the krypton-ion laser at high and VLD-matched power outputs. Multicolor fluorescent protein analysis with 488-nm excitation of green fluorescent protein and DsRed and VLD excitation of CFP was therefore feasible and was demonstrated. Similar levels of excitation efficiency between krypton-ion and VLD sources also were observed for ELF-97 detection. CONCLUSIONS: These evaluations confirmed that VLDs may be cost- and maintenance-effective replacements for water-cooled gas lasers for applications requiring violet excitation in addition to DNA binding dyes.

Alkaline Phosphatase↗

Temporal relation of changes in regional coronary flow and myocardial lactate and nucleoside metabolism during pacing-induced ischemia.

The temporal relation between myocardial lactate and hypoxanthine metabolism and regional changes in krypton-81m perfusion during pacing-induced ischemia was studied in 17 patients with coronary artery disease (CAD). During incremental atrial pacing, lactate production and hypoxanthine release occurred early and simultaneously, accompanied by ST-segment changes, but before angina and only few minutes after a significant (17%) reduction in krypton-81m perfusion in areas with more than 90% luminal diameter reduction. During maximal pacing heart rates, krypton-81m distribution decreased to 68 +/- 7% of control in areas with more than 90% diameter reduction and to 80 +/- 4% in 70 to 90% reduction (both p less than 0.05 vs control). Maximal lactate production occurred 15 seconds after pacing (extraction -15 +/- 7% vs 16 +/- 2% during control, p less than 0.05) and peak hypoxanthine release 1 minute after pacing (delta arteriovenous -2.64 +/- 0.8 microM vs 0.08 +/- 0.21 microM during control, p less than 0.05). Krypton-81m perfusion decreased in 20 of the 21 CAD areas. Angina, ST-segment changes, hemodynamic alterations and lactate production occurred in 15, 14, 9 and 15 patients, respectively. In contrast, hypoxanthine release was found in all cases. After pacing, lactate production and all general indexes of ischemia persisted for only 2 to 3 minutes. In contrast, krypton-81m perfusion was still significantly reduced 5 minutes after pacing and was only accompanied by hypoxanthine release (delta arteriovenous -1.41 +/- 0.6 microM, p less than 0.05 vs control).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Inhalation lung imaging with radioactive aerosols and gases.

Krypton lung-imaging is considered the ventilation procedure of choice when it is available. Aerosol and krypton wash-in images both reveal regional abnormalities of ventilation, but the aerosol images also disclose evidence of major airways disease. Xenon wash-out images are probably the most sensitive indicators of regional abnormalities of ventilation and/or small airways obstruction, whereas xenon wash-in images are far less sensitive and may be frequently misinterpreted. The Tc-DTPA aerosol inhalation procedure is perferred over xenon and krypton ventilation imaging for routine use immediately after a perfusion examination in pulmonary embolism suspects. This combined method provides the referring physician with the necessary diagnostic information quickly and with around-the-clock availability. In our opinion, this new aerosol procedure deserves wider application in the diagnosis and management of pulmonary disease. Lung imaging procedures, performed after th inhalation of 99mTechnetium labeled aerosols, 133Xenon and and 81mKrypton gases, were used to visualize the sites of airway obstruction and regional abnormalities of ventilatory function in normal volunteers, patients with obstructive airway disease and pulmonary embolism suspects. This chapter presents intercomparisons of these three methods regarding their functional significance, diagnostic merits, and limitations. A new nebulizer-radioaerosol delivery system is described. Test agent kits are inexpensive and readily available for on site assembly. Currently, we consider radioaerosol imaging in multiple views as more informative and suitable for routine use than xenon methods to detect regional abnormalities of the airways and ventilatory function. The krypton procedure is preferred in pulmonary embolism suspects because it requires far less patient cooperation than the xenon and aerosol methods and the lung images disclose regional ventilatory impairment quickly and accurately. However, krypton gas is cyclotron-produced and not yet commercially available.

Adolescent↗

Laser energy and dye fluorescence transmission through blood in vitro.

PURPOSE: Because of the potential usefulness of evaluating and treating choroidal neovascularization obscured by blood, we designed this study to quantify the transmission of dye fluorescence and laser energy through blood. METHODS: Blood preparations anticoagulated with ethylenediaminetetraacetic acid with hematocrits of 0% (plasma), 46%, and 99% were placed in open cuvettes with path lengths of 100, 200, or 500 microns and were exposed for one minute to either 100% oxygen or 100% carbon dioxide. Each cuvette was then sealed. Photographs of the cuvettes of blood in front of a flask of fluorescein or indocyanine green solution were decoded and used to calculate the percent transmission of fluorescence through blood. Cuvettes of blood were also placed in the path of argon, krypton, and diode lasers for energy transmission measurements. RESULTS: Plasma transmission of fluorescein and indocyanine green fluorescence and argon, krypton, and diode laser energy was 89% to 100% for all samples tested. Transmission of fluorescein fluorescence and argon laser energy through 99% hematocrit samples were both less than 5%. Transmission of indocyanine green fluorescence through 100-, 200-, and 500-microns-thick cuvettes filled with 99% hematocrit blood was 57%, 34%, and 4%. Transmission of krypton laser energy was 50%, 25%, and 6%; and transmission of diode laser energy was 60%, 35%, and 12% through 99% hematocrit blood. Intermediate transmission values were obtained for 46% hematocrit samples. CONCLUSIONS: Krypton and, to a slightly greater degree, diode laser energy penetrate a thin film of blood. Indocyanine green fluorescence also penetrates a thin film of blood. If a layer of blood appears thinner than 500 microns, then indocyanine green angiography may be useful in imaging underlying pathologic features. If a lesion can be imaged with indocyanine green, then it can probably be treated with a krypton or diode laser.

Blood↗

Functional outcome of branch retinal vein occlusion after photocoagulation with various laser wavelengths--an automated perimetric analysis.

This study highlights the comparative efficacy of three different laser wavelengths--argon green, argon blue-green, and krypton red--for sector photocoagulation in fresh cases of branch retinal vein occlusion. The mean Logarithm of minimal angle of resolution (LogMAR) improved in the argon green and argon blue-green groups but decreased in the krypton red group. Statistical analysis of the automated visual fields in these eyes showed a mean change in corrected pattern standard deviation value, with increase in all three groups--minimal increase in the argon blue-green group and maximal in the krypton red group, suggesting maximum visual field defect in the krypton laser group. To conclude, the green and the blue-green wavelengths gave results that were decidedly superior to those given by krypton red: further, argon blue-green had an edge over the green wavelength.

Fluorescein Angiography↗

Partition coefficient ratios and tumour perfusion studied with 85mKr and 133Xe.

The relative partition coefficients of krypton and xenon, and the regional blood flow in 27 superficial malignant tumour nodules in 22 patients with diagnosed tumours were measured using the 85mKr- and 133Xe-clearance method. In order to minimize the effect of biological variables on the measurements the radionuclides were injected simultaneously into the tumour. The distribution of the radiotracers was assumed to be in equilibrium at the beginning of the experiment. The blood perfusion was calculated by fitting a two-exponential function to the measuring points. The mean value of the perfusion rate calculated from the xenon results was 13 +/- 10 ml/(100 g.min) [range 3 to 38 ml/(100 g.min)] and from the krypton results 19 +/- 11 ml/(100 g.min) [range 5 to 45 ml/(100 g.min)]. These values were obtained, if the partition coefficients are equal to one. The equations obtained by using compartmental analysis were used for the calculation of the relative partition coefficient of krypton and xenon. The partition coefficient of krypton was found to be slightly smaller than that of xenon, which may be due to its smaller molecular weight.

Humans↗

Radionuclide assessment of the effects of chest physical therapy on ventilation in cystic fibrosis.

This study assesses the use of krypton 81m scintigraphy as a measurement tool in evaluating the effectiveness of bronchial drainage with percussion and vibration on peripheral ventilation in patients with cystic fibrosis. Ten patients with cystic fibrosis participated. Each patient underwent a krypton 81m ventilation study and traditional pulmonary function tests. Forty-five minutes later, these studies were repeated before and after a chest physical therapy treatment. Each patient acted as his own control. All krypton 81m scintiscans were recorded and analyzed visually and numerically using a digital computer to assess distribution of ventilation. Visual analysis of the scintiscans indicated individual variation in treatment response: in some patients ventilation improved with therapy; in others, no change was noted; still others had changes independent of treatment. Numerical data derived from the scintiscans and pulmonary function tests showed no important differences among the three studies of each patient. Airway abnormalities characteristic of cystic fibrosis, progression of the disease, sputum production, or a combination of these factors may account for the individual variation in response to treatment. Krypton 81m scintigraphy is a reliable measure of regional ventilation and should be useful for assessing the efficacy of chest physical therapy because of the consistent, high quality visual data retrieved.

Adolescent↗

Solution of protein crystallographic structures by high-pressure cryocooling and noble-gas phasing.

Room-pressure flash-cryocooling of protein crystals is the standard way to reduce radiation damage during data collection. Typically, it is necessary to find cryoprotection conditions by trial and error, a process that is not always successful. Recently, a new method, high-pressure cryocooling, was developed that does not require penetrative cryoprotectants and typically yields very high quality diffraction. Since this method involves helium gas as a pressurizing medium, it was of great interest to see whether the method could be extended to diffraction phasing by the incorporation of heavy noble gases such as krypton. A modified Kr-He high-pressure cyrocooling procedure is described wherein crystals are first pressurized with krypton gas to 10 MPa for 1 h. The krypton pressure is then released and the crystals are repressurized with helium over 150 MPa and cooled to liquid-nitrogen temperatures. Porcine pancreas elastase (PPE; 240 residues, 26 kDa) was selected as a test case for this study. Excellent diffraction was achieved by high-pressure cryocooling without penetrating cryoprotectants. A single 0.31 occupied krypton site in a PPE molecule [Bijvoet amplitude ratio (|DeltaF|/F) of 0.53%] was successfully used for SAD phasing at 1.3 A. This method has the potential to greatly simplify obtaining protein structures.

Air Pressure↗

Laser photocoagulation of the choroid through experimental subretinal hemorrhage.

OBJECTIVE: To study the differential abilities of diode infrared, krypton red, and argon blue-green laser energies to penetrate experimental subretinal hemorrhage and coagulate the underlying choroid. METHODS: Autologous, heparinized whole blood was injected beneath the neurosensory retina of pigmented rabbit eyes. After 30 to 60 minutes, confluent patches of moderate or severe diode, krypton, or argon laser burns were applied to adjacent healthy retina and continued into the region of the subretinal hematoma without varying the power setting or focal plane. Histopathologic evaluation of early lesions was performed in a masked fashion, and subretinal hemorrhage thickness was determined with computer-assisted image capture and analysis. RESULTS: Retina overlying treated subretinal hemorrhage showed no ophthalmoscopically visible signs of photocoagulation with diode energy, a faint gray reaction with krypton energy, and an intense white reaction with argon energy. Histopathologic analysis revealed photocoagulative inner choroidal damage beneath a mean (+/- SD) maximum blood thickness of 0.56 +/- 0.14 mm with severe diode burns, 0.42 +/- 0.09 mm with severe krypton burns, and 0.22 +/- 0.04 mm with severe argon burns. CONCLUSIONS: These data demonstrate that laser penetration of subretinal blood increases with longer wavelengths in vivo. Diode infrared laser energy is capable of penetrating subretinal blood to coagulate the choroid in the absence of ophthalmoscopically visible changes in the overlying retina.

Animals↗

Flow cytometric measurement of fluorescence (Förster) resonance energy transfer from cyan fluorescent protein to yellow fluorescent protein using single-laser excitation at 458 nm.

BACKGROUND: Use of distinct green fluorescent protein (GFP) variants permits the study of protein-protein interactions and colocalization in viable transfected cells by fluorescence (Förster) resonance energy transfer (FRET). Flow cytometry is a sensitive method to detect FRET. However, the typical dual-laser methods used in flow cytometric FRET assays are not generally applicable because they require a specialized krypton ultraviolet (UV) laser. The purpose of this work was to develop a flow cytometric method to detect FRET between cyan fluorescent protein (CFP; donor) and yellow fluorescent protein (YFP; acceptor) by using the 458-nm excitation from a single tunable argon-ion laser. METHODS: FUSE-binding protein (FBP) interacting repressor (FIR) and FBP are c-myc transcription factors and are known to interact physically. To examine their interaction within viable cells, FIR and the binding motif of FBP, the FBP central domain (FBPcd), were fused with CFP and YFP, respectively, and this pair of fluorescently-tagged proteins was used to detect FRET in vivo. Cells transfected with expression plasmids encoding a CFP-FIR fusion protein and YFP as a negative control, a CFP-YFP fusion protein as a positive control, or CFP-FIR and YFP-FBPcd fusion proteins were examined for FRET after excitation with a 458-nm line from a tunable argon-ion laser. FRET was measured as the ratio of YFP:CFP emission or as YFP emission at 564-606 nm. Conventional FRET using the 413-nm UV line from a krypton laser was examined for comparison. Fluorescence signals were separated with a customized optical filter configuration using 530-nm shortpass, 500-nm longpass, and 560-nm shortpass dichroics in addition to 488/30 nm (CFP), 530/30 nm (YFP), and 585/42 nm (FRET) bandpass filters. Further, a laser-scanning confocal microscopic photobleach technique was used to document that FRET occurred by showing that the intensity of donor CFP fluorescence increased after its acceptor YFP was photobleached. Steady-state spectrofluorometry was used to confirm and validate the results detected by flow cytometry. RESULTS: Upon excitation with the 458-nm line of the argon-ion laser, the enhancement of the acceptor YFP signal and the decrease of the CFP signal were easily detected in cells transfected with the CFP-YFP construct or CFP-FIR and YFP-FBPcd. Similarly, FRET was detected under these conditions when the YFP emission was assessed at 564-606 nm. A strong correlation was observed between the increase in the YFP:CFP ratio and the YFP emission detected at 564-606 nm, consistent with the conclusion that FRET was detected comparably by both methods. A conventional flow cytometric krypton UV-laser technique was also used to confirm that FRET occurred with the CFP-YFP fusion protein and from CFP-FIR --> YFP-FBPcd. FRET also was confirmed by a confocal photobleaching technique, in which donor CFP intensity was enhanced after its acceptor YFP was photobleached. The flow cytometric and confocal microscopic results were confirmed by spectrofluorometry. CONCLUSION: These results demonstrated the feasibility of flow cytometric detection of FRET signals from CFP to YFP by excitation with the 458-nm line from the tunable argon-ion laser. The method was as efficient as excitation with the krypton UV laser and therefore should make FRET a more generally available flow cytometric technique.

Argon↗

Understanding the origin of visual percepts elicited by electrical stimulation of the human retina.

BACKGROUND: The success of a retinal prosthesis for patients with outer retinal degeneration (ORD) depends on the ability to electrically stimulate retinal cells other than photoreceptors. Experiments were undertaken in human volunteers to ascertain whether electrical stimulation of cells other than photoreceptors will result in the perception of light. METHODS: In two subjects, two areas of laser damage (argon green and krypton red) were created in an eye scheduled for exenteration due to recurrent cancer near the eye. In the operating room prior to exenteration, under local anesthesia, a hand-held stimulating device was inserted via the pars plana and positioned over the damaged areas and normal retina. Subjects' psychophysical responses to electrical stimulation were recorded. RESULTS: In both subjects, electrical stimulation produced the following perceptions. Normal retina: dark oval (subject 1), dark half-moon (subject 2); krypton red laser-treated retina: small, white light (both subjects); argon green laser treated retina: thin thread (subject 1), thin hook (subject 2). Histologic evaluation of the krypton red-treated retina showed damage confined to the outer retinal layers, while the argon green-treated area evinced damage to both the outer and the inner nuclear layers. CONCLUSION: The perception produced by electrical stimulation was dependent on the retinal cells present. Electrical stimulation of the krypton red-ablated area best simulated the electrically elicited visual perceptions of our blind, ORD patients, suggesting that the site of stimulation in blind patients is the inner retinal neurons.

Aged↗

Size versus polarizability in protein-ligand interactions: binding of noble gases within engineered cavities in phage T4 lysozyme.

To investigate the relative importance of size and polarizability in ligand binding within proteins, we have determined the crystal structures of pseudo wild-type and cavity-containing mutant phage T4 lysozymes in the presence of argon, krypton, and xenon. These proteins provide a representative sample of predominantly apolar cavities of varying size and shape. Even though the volumes of these cavities range up to the equivalent of five xenon atoms, the noble gases bind preferentially at highly localized sites that appear to be defined by constrictions in the walls of the cavities, coupled with the relatively large radii of the noble gases. The cavities within pseudo wild-type and L121A lysozymes each bind only a single atom of noble gas, while the cavities within mutants L133A and F153A have two independent binding sites, and the L99A cavity has three interacting sites. The binding of noble gases within two double mutants was studied to characterize the additivity of binding at such sites. In general, when a cavity in a protein is created by a "large-to-small" substitution, the surrounding residues relax somewhat to reduce the volume of the cavity. The binding of xenon and, to a lesser degree, krypton and argon, tend to expand the volume of the cavity and to return it closer to what it would have been had no relaxation occurred. In nearly all cases, the extent of binding of the noble gases follows the trend xenon>krypton>argon. Pressure titrations of the L99A mutant have confirmed that the crystallographic occupancies accurately reflect fractional saturation of the binding sites. The trend in noble gas affinity can be understood in terms of the effects of size and polarizability on the intermolecular potential. The plasticity of the protein matrix permits repulsion due to increased ligand size to be more than compensated for by attraction due to increased ligand polarizability. These results have implications for the mechanism of general anesthesia, the migration of small ligands within proteins, the detection of water molecules within apolar cavities and the determination of crystallographic phases.

Amino Acid Substitution↗

Lung scan abnormalities in asthma and their correlation with lung function.

We have used asthma as a model of airways disease to test how well an automated, quantitative method of analysis of lung scans correlates with physiological measurements of disturbed lung function and gas exchange. We studies 25 asthmatics (age 16-73) of widely differing severity (forced expiratory volume in 1-s (FEV1) 22%-123% of predicted value), who had airways tests, arterial blood gas analysis, and krypton-technetium lung scans within a short time of each other. In all patients with airways obstruction and in some with normal function during remission, scans showed the typical appearances of multiple defects of ventilation and perfusion. The severity of ventilation defects was computed from the posterior view of the krypton scan compared to an age- and sex-matched normal range to yield an underventilation score. This correlated closely with the severity of airways obstruction as measured by forced expiratory manoeuvres. Ventilation and perfusion defects were usually imperfectly matched; the severity of this was computed using a subtraction method applied to the counts on the posterior krypton and technetium scans. The degree of mismatch was inversely related to the arterial partial pressure of oxygen (r = -0.86). The results suggest that computer scan analysis can provide useful functional information about the lung in airways disease.

Adolescent↗

A simplified method of 85Kr measurement for dating young groundwaters.

The technique of gas extraction from water and gas sample preparation for 85Kr low-level measurements was further developed to improve its applicability to isotope-hydrological "age" determinations of young groundwaters. A water sample of about 200 L is flushed with helium to extract the dissolved gases. Krypton is separated from the extracted gas mixture by repeated adsorption on charcoal, fractionated desorption and subsequent gas chromatography. The detection limit for a 10,000 min counting time in a 10 mL counter containing the krypton (15 microL) and methane as counting gas corresponds to 25 mBq 85Kr per mL (STP) krypton or 2.5% of the mean atmospheric 85Kr activity concentration in the year 1990.

Geological Phenomena↗

Abnormal regional distribution of ventilation in middle-aged smokers: comparison of changes in 81Krm ventilation scans and computed tomography of the lung.

In 1980 we found that abnormalities in regional distribution of ventilation, as assessed by 81Krm lung scans, were common in middle-aged smokers with normal chest radiographs and mild impairment of overall lung function. In 1984 we repeated 81Krm scans in 16 continuing smokers then aged 50-64 years and with mean forced expiratory volume in one second 93% (20 SD) of predicted values who had previous 81Krm scans performed in 1980. To assess the role of disease of the peripheral airspaces in causing abnormal regional ventilation, we also obtained computed tomograms (CT) of the lungs and measured carbon monoxide transfer of the lungs in these men. Krypton scans in seven men who had normal or minor focal defects of ventilation in 1980 were unchanged in 1984. Scans in seven of the nine men who had abnormal scans in 1980 remained abnormal in 1984 but there was no overall deterioration in the abnormality of ventilation in these men; in men with similar grading in 1980 and 1984 some of the peripheral defects present in 1980 had resolved and some new abnormal areas had appeared. Minor localised abnormalities of CT scans, as assessed visually, were present in eight of the 16 men and were associated with lower values of carbon monoxide transfer coefficient (mean 78% vs 98% predicted in men with normal scans, P less than 0.01) and lung density (mean -894 vs -869 HU in men with normal scans, P less than 0.054) suggesting the CT changes were due to alveolar destruction. Abnormality of the krypton scan was not significantly associated with abnormality of the CT scan or with a reduction in carbon monoxide transfer. The results of the krypton lung scans confirm that non-uniformity of regional ventilation is often present in asymptomatic middle-aged smokers and suggest that this non-uniformity is in part due to temporary occlusion of airways. Abnormality in regional ventilation was not associated with the anatomical changes shown by the CT scan, suggesting that airway narrowing was more important than alveolar destruction in causing regional abnormalities of ventilation in these men.

Forced Expiratory Volume↗

On the difficulty of assessing the specific surface area of magnesium stearate.

The water content of as-received commercial magnesium stearate batches from animal and vegetable sources have been modified by ageing in humid air at room temperature or by vacuum treatment. The complete adsorption-desorption isotherms of nitrogen and krypton vapours by samples of these as received and modified materials have been measured at liquid nitrogen temperature after standardised vacuum degassing. They are greatly affected by the initial water content of the material. In particular: (a) the BET surface area values computed from the adsorption branch vary widely and is increasing with increasing water content; (b) anomalous hysteresis of varying amplitude is observed in all cases except adsorption of krypton on the material with the lowest water content; (c) the hysteresis loops extend down to very low desorption pressure values and cannot be accounted for by capillary condensation. Lastly, the surface area value of a given material computed from nitrogen and krypton adsorption may differ by a factor as high as six. Accordingly, the very significance of BET surface area values obtained from routine adsorption experiments should be regarded as questionable, at least until the mechanisms of adsorption are fully clarified.

Adsorption↗

Effects of helium and other inert gases on Echinosphaerium nucleofilium (protozoa, heliozoa).

The effects of helium, nitrogen, argon and krypton on Echinosphaerium nucleofilum (Heliozoa) have been studied at partial pressures of 10-130 atm. Additional experiments have been carried out with hydrostatic pressure alone. Helium causes shortening of the axopods over the whole range of pressures, and damage to the cell body at pressures of 60-90 atm, both with a maximum at 80 atm. These effects cannot be explained in terms of hydrostatic pressure alone; a 'pressure reversal' effect may be operating, causing the peak at 80 atm. Nitrogen also causes both cell damage and axopod shortening, the severity increasing with increasing pressure. Argon and krypton cause cell damage but no shortening. The order of potency for cell damage is krypton greater than argon greater than nitrogen greater than helium. It is suggested that there may be tuo sites of action, possibly the microtubules (for axopod shortening) and the cell membrane (for cell damage). In appropriate mixtures of helium and argon, both the cell damage usually caused by argon, and the axopod shortening usually caused by helium, are prevented. Possible mechanisms include the effects of hydrostatic pressure on gas solubility coefficients, reversal of the effects of the gases by the increase in total pressure, and competition for sites of action.

Argon↗

A model of subretinal neovascularization in the pigmented rat.

We produced krypton laser photocoagulation lesions of mild to moderate whiteness in the posterior retinas of one eye of 23 pigmented rats, and identical appearing argon laser burns in the fellow eyes. We observed foci of subretinal neovascularization, histopathologically markedly similar to that which occurs in several human retinal diseases, in the krypton laser treated eyes of 6 of the 14 rats that were followed for one to three months after photocoagulation. No such lesions were observed in the argon laser treated fellow eyes, nor in krypton or argon laser treated eyes examined earlier than one month after photocoagulation. The photocoagulation damaged only the choriocapillaris, the retinal pigment epithelium (RPE), and the photoreceptor layer. In the acute lesions, we did not observe ruptures in Bruch's membrane. The neovascularization was surrounded by multiple layers of RPE cells, a histopathologic finding that has also been reported in some human eyes with subretinal neovascularization in age-related macular degeneration. These observations suggest that the RPE cells may be modifying the proliferative behavior of adjacent choroidal capillaries. This model differs from previous models of subretinal neovascularization in primates, and may be useful for additional studies of this important pathological process.

Animals↗