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PubMed1992

Selective laser-induced inactivation of proteins (SLIP) by labelling with chromophores.

Coupling a fluorochrome (e.g. fluorescein-isothiocyanate, FITC) to a molecule can enhance specific laser light absorption, thus leading to alteration or even destruction of the molecule itself. Therefore antibodies were labelled with FITC (absorption maximum 480 nm) and irradiated with laser light (488 nm) under various conditions. Inactivation of antibodies could only be achieved at the absorption maximum of FITC (as measured by direct and indirect immunofluorescence). Positive linear correlation exists between the amount of destruction and both exposure time and energy. Similar destructive effects were obtained when FITC-labelled peroxidase was irradiated. In these cases, enzyme activities measured by absorption photometry also showed a positive correlation to the total amount of energy transferred. Non-labelled proteins were not affected by irradiation. So we conclude that labelling of proteins with fluorochromes provides a highly specific means of selection of target molecules to be destroyed or inactivated. The method is based upon the laws of linear optics and is different from photodynamic or photochemical actions. The destructions observed are most likely caused by thermally induced changes of the molecules' tertiary structure.

Antibodies↗
PubMed1982

Metabolism of arachidonic acid in leukocytes: isolation of a 5,15-dihydroxy-eicosatetraenoic acid.

A novel metabolite of arachidonic acid was isolated from incubations of peripheral blood leukocytes with the fatty acid and the ionophore A23187. The compound was purified by high performance liquid chromatography and identified by ultraviolet photometry and gas chromatography-mass spectrometry as a 5,15-dihydroxy-6,8,11,13-eicosatetraenoic acid. The compound was also isolated from incubations of human leukocytes with the 15S-hydroperoxy-5,8,11,13(Z,Z,Z,E)-eicosatetraenoic acid, suggesting a double dioxygenation mechanism in the formation of this new metabolite of arachidonic acid. A 5S,15S-dihydroxy-6,8,11,13(E,Z,Z,E)-eicosatetraenoic acid was obtained from incubations of 5S-hydroxy-6,8,11,14(E,Z,Z,Z)-eicosatetraenoic acid with the soybean lipoxygenase and reduction with stannous chloride, and was used as reference compound.

Animals↗
PubMed1986

In vivo photometric analysis of hemoglobin.

Since virtually all the oxygen carried by blood at normal hematocrit is reversibly bound to red blood cell hemoglobin, the distribution of oxygen within the microcirculation can be determined from measurements of hemoglobin concentration and hemoglobin oxygen saturation in vessels of the network. Photometric methods that rely on light absorption and scattering properties of blood are described. Criteria for selecting the wavelengths needed to analyze hemoglobin in the microcirculation are specified. Two theoretical descriptions of light absorption and scattering, multiple scattering theory and photon diffusion theory, are applied to the problem. Practical approaches to the determination of hemoglobin concentration and oxygen saturation in the microcirculation follow from these theoretical formulations. Technical aspects of microscope photometry including light sources, microscopy, and detection systems are described with special emphasis on the problem of glare. The importance of in vitro as well as in vivo calibrations is stressed, and several recent applications of a working system are discussed. Current problems as well as future developments of this methodology are delineated as a guide to future work in this area.

Animals↗
PubMed1994

Interstitial and tissue cations and electrical potential after experimental spinal cord injury.

Interstitial and tissue cations and electrical potential were studied in an experimental model of spinal cord contusion injury in anaesthetised cats. Measurements of interstitial ion activity in the grey matter at the injury site (with ion-selective electrodes), showed a decrease of sodium and calcium, an increase of potassium, a small acidification and a negative shift in the electrical potential 5 min after injury. The interstitial ionic changes were completely reversible within 90 min following injury. Measurements of the ion content in a tissue sample from the injury site (flame photometry) showed an increase of sodium and calcium and a decrease of potassium 5 min after injury. The magnitude of the post-injury sodium change was much larger than the potassium change, both for interstitial and tissue measurements. Treatment of the animals with the calcium entry blocker flunarizine before the injury did not influence the magnitude of post-injury interstitial calcium decrease but significantly increased the rate of subsequent recovery. Pre-injury flunarizine treatment also significantly increased the recovery rate of the electrical potential. The experiments suggest the occurrence of a net ionic shift towards the intracellular space, which may contribute to oedema formation in the very early post-injury period. The post-injury decrease of interstitial calcium activity is probably not mediated by flunarizine-sensitive calcium entry mechanisms; such mechanisms may, however, be involved in the subsequent recovery period for interstitial calcium activity. Calcium ions may be involved in the recovery process of the negative electrical potential after injury.

Animals↗
PubMedDate not supplied

Cadmium binding proteins induced in exposed freshwater crayfish Procambarus clarkii.

This work describes results on the characterization of cadmium binding proteins (Cd-BPs) obtained from cadmium exposed freshwater crayfish Procambarus clarkii. After acclimation to laboratory conditions, induction of Cd-BPs was achieved by water exposure at a concentration of 100 micrograms Cd/L during 2, 15, and 30 d. In accordance with the method followed by Engel and Brouwer, in each case two midgut glands were minced and homogenized in Tris-HC1 buffer with PMSF to prevent protease activity and DTT to maintain reducing conditions. The homogenate was centrifuged, heat treated, applied to a column of Sephadex G-75, and eluted with the same buffer (pH 8.6). Absorbances of the fractions collected were measure at 254 and 280 nm. Cadmium concentrations were determined by flame photometry. In midgut glands of two-d treated crayfish, significant levels of cadmium occurred in the void volume, and no cadmium-binding protein peak was resolved. On the contrary, a cadmium peak was clearly resolved in samples of 15 and 30 d. Cadmium was accumulated in the low molecular weight fractions (about 20,000). These fractions had high ultraviolet absorption at 254 nm and a higher 254/280 ratio.

Animals↗
PubMedDate not supplied

Trypan Blue as a marker of plasma membrane permeability in alloxan-treated mouse islet cells.

Suspensions of pancreatic islet cells from noninbred ob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30:1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant for nuclear binding of the dye was 3.1 X 10(4)l/mol. Albumin partially inhibited the nuclear staining. More than 0.5% Trypan Blue in the medium was toxic per se. In the absence of albumin, 0.5 or 20 mmol/l alloxan, 1 mmol/l N-ethylmaleimide, or 0.1 mmol/l chloromercuribenzene-p-sulphonic acid, but not 20 mmol/l streptozotocin, increased the frequency of islet cells stained with 0.1% Trypan Blue. The absorbance of nuceli was also increased in cells treated with alloxan or N-ethylmaleimide, but not in those treated with chloromercuribenze-p-sulphonic acid. It is concluded that alloxan rapidly increases the permeability of the plasma membrane in mouse beta-cells. This action of alloxan appears to be more acute than any such effect of streptozotocin.

4-Chloromercuribenzenesulfonate↗
PubMed1988

Autoradiographical evidence for increased thyrotropin binding to autonomously functioning thyroid gland tissue.

The aim of this study was to quantify thyrotropin (TSH) binding to individual cells from autonomously functioning thyroid gland tissue (AFTT) in comparison with the corresponding non-autonomous cells. Cryostatic tissue sections from ten thyroid glands with autonomously functioning adenomas were incubated in 125I-labeled TSH. TSH-binding density (TBD) was visualized by means of autoradiography and quantified by means of reflection photometry. Eight out of ten specimens showed a significant (p less than 0.001), up to tenfold increase in TBD of AFTT cells. All ten specimens had a significantly increased number of TSH molecules bound per thyrocyte in comparison both with the corresponding non-autonomous cells and also with tissues from non-functioning thyroid neoplasms (n = 6). The increased TBD of AFTT persisted after transplantation onto nude mice. The paranodular tissue from thyroid glands with scintigraphically compensated adenomas showed in four out of six cases an increased TBD when compared with normal thyroid tissue. It can be concluded that AFTT possesses distinct cytoplasm membrane characteristics as evinced by TSH binding analysis. Increased TSH binding should be considered as a possible cause of metabolic hyperactivity of AFTT cells. The concomitant existence of functional autonomy in the paranodular tissue might lead to a false supposition of a compensated autonomous adenoma as revealed by scintigraphical evidence.

Adenoma↗
PubMed1997

Renal response to infusion of dopamine precursors in anaesthetized rats.

In the present study the renal response to intravenous infusion of the catecholamine precursors L-dihydroxyphenylalanine (L-DOPA) or L-tyrosine was investigated in thiopentone sodium-anaesthetized Sprague-Dawley rats. Glomerular filtration rate (GFR) was assessed by renal clearance of inulin, urinary concentration of dopamine (U(DA)V) by HPLC and sodium excretion (U(Na)V) by flame photometry. We found that basal U(DA)V was 6.5 +/- 0.5 pmol/min per 100 g body weight (mean +/- SEM). Intravenous infusion of L-tyrosine at 0. 1-3.0 micromol/min dose dependently enhanced U(DA)V (17 +/- 3 to 144 +/- 14 pmol/min respectively) with higher doses of L-tyrosine resulting in no further increase in U(DA)V. Compared with L-tyrosine administration significantly lower doses of L-DOPA (0.07 to 35 nmol/min) caused increases in U(DA)V which were orders of magnitude higher (18 +/- 1 to 7800 +/- 470 pmol/min, respectively) and did not show saturation characteristics. GFR did not change in response to L-tyrosine or L-DOPA infusion. No variations in urinary flow rate or in U(Na)V could be observed which were significantly correlated to changes in U(DA)V. In contrast, intravenous infusion of dopamine at a dose of 6 nmol/min significantly increased GFR by 35 +/- 6.2% and urinary flow rate by over 2-fold. Immunohistochemistry with light microscopy revealed no tyrosine hydroxylase in the kidney. Therefore, dopamine synthesis in the tubular cells mainly depends on the renal supply of L-DOPA. The unchanged GFR and U(Na)V in spite of large variations of U(DA)V argue against the hypothesis that intratubular dopamine plays a functional role in the regulation of hemodynamics or sodium transport in the kidney. Renal dopamine excretion may rather represent an effective pathway for the elimination of catecholamine precursors from the plasma.

Adrenal Glands↗
PubMed1999

Green Fluorescent Protein As a Cell-Labeling Tool and a Reporter of Gene Expression in Transgenic Rainbow Trout.

: Green fluorescent protein (GFP) has been used as an indicator of transgene expression in living cells and organisms. For testing the utility of GFP in rainbow trout, we microinjected fertilized eggs with four types of supercoiled constructs containing two variants of GFP complementary DNA (S65T and EGFP), driven by two ubiquitous regulatory elements, human cytomegalovirus immediate early enhancer-promoter (CMV) and Xenopus laevis elongation factor 1alpha enhancer-promoter (EF1). Green fluorescence was first observed at 3 days postfertilization, when the embryo was in the mid-blastula stage. Fluorescence could be detected mosaically in various types of embryonic cells and tissues of swim-up fry. Both the percentage of fluorescent cells and the fluorescence intensity of GFP-expressing cells on blastoderms, measured with a microscopic photometry system, were highest in CMV-EGFP-microinjected embryos. We conclude that GFP is capable of producing detectable fluorescence in rainbow trout, and can be a powerful tool as a cell marker and reporter gene for cold-water fish, and that analysis of GFP expression in living cells is useful for characterizing the activity of cis-elements in vivo.

Journal Article↗
PubMed2003

On the in vitro biocompatibility of Elgiloy, a co-based alloy, compared to two titanium alloys.

AIM: The aim of the present investigation was to contribute to an understanding of the effects of surface topography and chemical composition on the corrosion behavior and thus the biocompatibility of Elgiloy (RMO, Denver, CO, USA), a common Co-based alloy. MATERIAL AND METHODS: The results are compared with those obtained for a binary NiTi alloy, Neo Sentalloy (GAC, Central Islip, NY, USA) and a beta-III-Ti alloy, TMA (Ormco, Glendora, CA, USA). In the present study, the surface topography and the chemical composition of two different grades of Elgiloy, Blue Elgiloy (soft) and Yellow Elgiloy (ductile), were examined by means of scanning electron microscopy (SEM) and energy-dispersive spectroscopy analysis (EDS). Their corrosion behavior in half-strength Ringer solution and in an artificial saliva solution according to Barrett [1] was investigated using potentiodynamic corrosion testing (PDC). The photometry-based PAN method was used to quantify the released Ni and Co ions. The in vitro biocompatibility of the two grades of Elgiloy was tested in three different cell cultures: in L929, a commercially available mouse fibroblast cell line, and in primary human epithelial cells and fibroblasts. RESULTS: The results of the corrosion testing showed satisfactorily high pitting corrosion potentials but lower repassivation potentials and a strong increase in current density once pitting had occurred. The photometric results revealed the release of Ni and Co ions in both tested electrolytes. The tested native surfaces exhibited numerous grinding and polishing grooves, inclusions and inhomogeneities of the microstructure. After corrosion testing the same surfaces displayed numerous signs of corrosion, especially in areas with microstructural inhomogeneities. In vitro biocompatibility testing showed a substantially reduced dehydrogenase activity in the presence of Elgiloy. The reduced quality of surface finish resulting from the manufacturing process led in the case of the tested Elgiloy types to decreased corrosion resistance with consequently reduced in vitro biocompatibility. CONCLUSIONS: In this context it is also conceivable that patients with a proven allergy to nickel, cobalt or chromium may react sensitively to the deployment of this alloy, at least in the surface quality tested by us. From this aspect, the introduction of a binding standard for the surface quality of materials used in orthodontic appliances is urgently recommended.

Animals↗
PubMed1999

Free radical scavengers to prevent reperfusion injury following experimental warm liver ischaemia. Is there a real physiological benefit?

Free radical scavengers have been utilized to prevent the consequences of ischemia, however, results do not seem conclusive. In our study we analyzed the blood flow, function, and histology of rat liver tissue after warm liver ischemia, in order to assess the effect of free radicals in liver reperfusion injury. N-acetyl cysteine (NAC), tocopherol, allopurinol, and superoxide dismutase (SOD), pharmacological agents expected to protect from injury mediated by free radicals, were investigated. Laser Doppler flowmetry and photometry were utilized to measure post-ischemic microcirculatory changes as an expression of ischemia-reperfusion injury in a model of segmental liver ischemia in the rat, with an ischemic time of 45 min. Galactose elimination capacity, ALT and histology were used to assess the functional and morphological consequences of ischemia after 24 h of reperfusion. The overall mean blood flow over 1 hour after reperfusion was of 33.9% (SD 11.2) of the normal, non-ischemic control. NAC (31.2% SD 10.9) did not show any protective effect and in some cases the effect seemed to be negative. Tocopherol (41.7% SD 5.1) marginally improved post ischemic liver tissue blood flow. Treatment with allopurinol did not show any beneficial effects (37.5% SD 14.2). Only animals treated with SOD showed an improvement of the post ischemic liver microcirculation (57.9% SD 14.4)(P < 0.001) and function. Only SOD produced statistically significant differences in galactose elimination capacity, compared with those of the ischemic control group. This moderately protective effect of SOD is encouraging, however, the relevance of all these compounds in a broader pathophysiological setting remains unproven.

Acetylcysteine↗
PubMed2006

Effect of quinupristin/dalfopristin on 3T3 and Eahy926 cells in vitro in comparison to other antimicrobial agents with the potential to induce infusion phlebitis.

Infusion phlebitis is a common clinical problem that is observed with some antimicrobial agents, when being administered intravenously. In this study, cultured murine fibroblasts and immortalised human endothelial cells were exposed to three antibiotics at clinically relevant concentrations to assess their toxic potential in two established cytotoxicity assays. BALB/c 3T3 fibroblasts and Eahy926 endothelial cells were exposed to quinupristin/dalfopristin (QD), erythromycin and levofloxacin at increasing concentrations. For assessment of cytotoxicity the cells were incubated with neutral red (NR) or stained with crystal violet (CV). Measurements were done by photometry. At the concentration range tested QD and erythromycin showed a concentration-dependent cytotoxic effect in both cell cultures. In 3T3 cells the half-maximal effect concentration (EC50) was 20 mg/l for QD and 340 mg/l for erythromycin in the NR uptake test and 12 and 200 mg/l, respectively, in the CV assay. In Eahy926 cells the EC50 was 50 mg/l for QD and 880 mg/l for erythromycin in the NR uptake test and 40 and 750 mg/l, respectively, in the CV assay. No EC50 could be established in both cell types for levofloxacin. Eahy926 cells were less sensitive to cytotoxic stimuli than 3T3 fibroblasts. Cytotoxic effects in both cell cultures occurred in the following order: QD > erythromycin >> levofloxacin. This ranking correlates well with the frequency of local adverse effects observed with the infusion of these antibiotics in patients. Thus, these in vitro assays may serve as an estimate for the prediction of local tolerability of antibiotics when administered parenterally.

Animals↗
PubMed2003

Store operated Ca2+ influx by selective depletion of ryanodine sensitive Ca2+ pools in primary human skeletal muscle cells.

The contraction and relaxation of skeletal muscle is driven by release of Ca2+ from sarcoplasmic reticulum through the ryanodine receptor type 1 and extruding the ion from the cytosol by Ca2+ ATPases. Efficient refilling of the empty Ca2+ stores is essential for repetitive cycles of muscle contraction and relaxation, but not investigated in human skeletal muscle cells. Here we show that under conditions of selective depletion of the ryanodine-sensitive Ca2+ pool Ca2+ influx occurs in differentiated human skeletal muscle cells using the Ca2+ imaging technique. This Ca2+ influx is not due to permeation through the L-type Ca2+ channel and not observed under conditions of inhibited Ca2+ ATPase. The Ca2+ influx was visualised by quenching the intracellular fura2 signal with Mn2+ on single cell level and also using fluorescence photometry of cell suspensions. The Mn2+ influx was inhibited by the Ca2+ channel blockers La(3+) and SKF96356. The delineation of the signalling cascade leading to Ca2+ influx evoked by selective depletion of ryanodine sensitive Ca2+ stores showed that phospholipase C or protein kinase C were not involved. Interestingly, a Mn2+ influx was triggered by the cell-permeant analogue of diacylglycerol and further augmented by the application of RHC80267, a diacylglycerol lipase inhibitor. This signalling pathway could be attributed to the participation of a protein kinase C activity. However, Mn2+ influx evoked by selective depletion of ryanodine sensitive Ca2+ stores was not altered by RHC80267 or protein kinase C inhibitors. Using RT-PCR, correctly spliced mRNA fragments were detected corresponding to human transient receptor potential (TRPC) Ca2+ channels type 1, 3, 4 and 6. These data show that in skeletal muscle at least two independent mechanisms of Ca2+ influx exist. For Ca2+ influx triggered by the selective depletion of ryanodine sensitive Ca2+ stores we propose a phospholipase C independent coupling of ryanodine receptors to voltage insensitive Ca2+ channels.

Caffeine↗
PubMed2000

Influence of column geometry on the ion chromatographic separation of aluminium species.

The dependence of the degree of disintegration and therefore the applicability of ion chromatography for the speciation of aluminium fluoride species was examined for two different column geometries, a standard bore and a microbore column. Besides mathematical calculations, the temperature of the separation column was varied between -5 and 50 degrees C for the observation of a temperature-dependent decomposition of the species. All species were detected by UV photometry after post-column reaction with Tiron. The results showed that the disintegration of the higher coordinated Al fluoride species (AlFn with n >2) could be dramatically reduced utilizing the microbore technique. In contrast to the standard bore technique the column temperature is of minor importance. The agreement between speciation data experimentally determined by microbore chromatography and those calculated using stability constants is quite good. The standard bore technique showed bigger differences between calculated and experimentally determined species distributions.

Journal Article↗
PubMed2001

Ozone measurement with silica cartidges and HRGC-MS analysis.

Ozone in ambient air is collected onto silica gel cartridges impregnated with pentafluorophenylhydrazine (PFPH) and 1,2-di(4-pyridyl)ethylene (DPE), so that the pyridine-4 aldehyde formed by DPE oxidation is converted into the corresponding PFPH derivative (PPH). The latter product is determined by HRGC/MS. Since the ozonolysis reaction proceeds stoichiometrically on the cartridge, there is no need for calibration in the gas phase with a standardized ozone source. When compared with UV photometry analyzers, this active chromatographic method (ACM) demonstrates a very good accuracy (ACM/UV photometer = 0.97) and precision (12.0-14.0%) under both laboratory and field sampling conditions at ozone concentrations of 20-200 microg m(-3) and exposure times of 1-3 h. The sampling performance was found to be insensitive to relative humidity (r.h.) variations in the 25-90% range and any interference effects could not be observed from various agents, except light, which can be eliminated by using an aluminium shelter. The detection limit for ozone achievable with the ACM in air samples collected at 0.5 L min(-1) for 1 h was found better than 0.5 microg m(-3).

Air Pollutants↗
PubMed2005

Changes in the urine composition during its passage through the ureter. A concept of urothelial function.

Studies in our and other laboratories have negated the general assumption that the urothelium functions as a simple conduit and support the concept of a dynamic urothelium. Based on this concept, we investigated the hypothesis that urine undergoes changes during its passage in the ureter. Urine samples were separately collected from the renal pelvis and ureters of 23 volunteers (15 men, eight women, mean age 37.6 years) through cutaneous ureterostomy performed as a treatment for bladder cancer. The pH was determined using a pH electrode, osmolality by means of micro-osmometry and Na and K by flame photometry. The pH and osmolality of ureteric urine showed no significant differences from renal pelvic urine, whereas the Na and K levels of the ureteric urine were significantly increased (both P<0.05). Gender and age differences were not significant. Renal pelvic urine undergoes changes in Na and K concentrations during its passage through the ureter. These findings presumably indicate that the ureteral urothelium is a functioning epithelium and that the renal pelvic and ureteric urine are not identical.

Adult↗
PubMed2006

Does the composition of voided urine reflect that of the renal pelvis?

Studies have shown that the urothelium has a transport function and that urine composition changes on its way through the urinary tract. In this study, we investigated the hypothesis that the composition of voided urine differs from and does not reflect that of the renal pelvis. Urine samples were obtained from the renal pelvis and voided urine of 18 healthy volunteers (mean age 36.2+/-5.1 SD years, 10 men, 8 women). The pH was determined using a pH electrode, osmolality by means of micro-osmometry and Na and K using flame photometry. In comparison to the urine of the renal pelvis, voided urine showed significant increases in pH, osmolality and Na and K concentrations (P<0.05 for each). There were no significant differences in gender and age. This study has demonstrated that the pH, osmolality, Na and K of voided urine differ significantly from the values in the renal pelvis. Urine composition is thus modified as it passes through the urinary tract, which would support the concept of a dynamic urothelium. The composition of voided urine does not seem to compare to renal pelvic urine. This concept needs to be considered in urine analysis evaluation and its relation to renal function.

Adult↗
PubMed2002

Risk factors for corneal allograft rejection: intermediate results of a prospective normal-risk keratoplasty study.

PURPOSE: To analyze the incidence of and possible risk factors for endothelial corneal allograft rejection in a well-defined population following penetrating normal-risk keratoplasty. METHODS: Since 1996 a longitudinal prospective study has been conducted to analyze the results of normal-risk penetrating keratoplasty. All patients underwent a standardized protocol of follow-up treatment and examinations in our institution. Diagnosis of corneal endothelial rejection was based on slit-lamp biomicroscopy and laser flare photometry. Data were analyzed using a proportional hazard model for censored data (Cox model), and Kaplan-Meier survival curves. The following parameters were analyzed: age, gender, atopic dermatitis, dry eye symptoms of the recipient; surgeon, graft diameter, post-mortem time, storage time and graft preservation method; and duration of postoperative epithelial defects. RESULTS: Between 1996 and May 2001, 397 patients were recruited and followed with a median follow-up of 18 months. Episodes of endothelial graft rejection were observed in 22 patients (5.5%; 18 eyes with acute diffuse episodes and 4 eyes with chronic focal rejection episodes). In addition, 12 eyes (3%) showed isolated small keratic precipitates ("graft rejection suspects"). All but one graft regained clarity after topical and systemic steroid treatment. Most episodes occurred 11-18 months postoperatively. The percentage of grafts without any episode of endothelial allograft rejection was 95% after 12 months, 89% after 18 months, and 86.5% after 24 months. The following factors were associated with graft rejection: atopic dermatitis (P=0.021), clinically manifest tear insufficiency (P=0.007), and short duration of graft storage (P=0.008). No significant correlation was detected for the remainder of the analyzed factors (P>0.05). CONCLUSION: The incidence of episodes of corneal endothelial allograft rejection following normal-risk keratoplasty was 13.5% within the first two postoperative years. However, the frequency of irreversible immunologic graft failure (3 per thousand) was lower than reported in the literature. Patients should be regularly followed up for at least 18 months postoperatively. Patients with underlying atopic dermatitis or dry eyes should receive special ophthalmological care.

Corneal Diseases↗