Measurement of the axial length of cataract eyes by laser Doppler interferometry.

PURPOSE: To examine the applicability of the recently developed laser Doppler interferometry technique for measuring the axial length of cataract eyes in a realistic clinical situation. To determine the performance of the instrument as a function of cataract grade. To compare the results to those of ultrasound methods. METHODS: A total of 196 cataract eyes of 100 patients were examined. The axial eye length was determined by laser Doppler interferometry and by two different ultrasound techniques, the applanation technique and the immersion technique. The cataract grade was determined by a commercial instrument that measures backscattered light. RESULTS: Laser Doppler interferometry worked very well except in the cases of the highest cataract grades (4% of the eyes of this study were not measurable because of a too-high lens density). Only 3.5% of the other eyes were not measurable because of fixation problems of the patients. The precision of laser Doppler interferometry is not influenced by the cataract grade (except the highest grade). The standard deviation of the geometric eye length is approximately 20 microns. Linear regression analysis revealed a very good correlation of laser Doppler interferometry and ultrasonic measurements, but a systematic difference was found. The eye lengths measured by laser Doppler interferometry were about 0.18 mm longer than those measured by the immersion technique and about 0.47 mm longer than those measured by the applanation technique. CONCLUSION: These differences are attributed to the laser Doppler interferometry results including the retinal thickness and indentation of the cornea by the applanation technique. The main advantages of the laser Doppler interferometry technique are high precision, high accuracy, and more comfort for the patient because it is a noncontact method, anesthesia is unnecessary, and the risk of corneal infection is avoided.

Receptor binding of norgestimate--a new orally active synthetic progestational compound.

Binding of the new progestagen, norgestimate (D-(-)-13 beta-ethyl-17 beta-acetoxy-17-ethinyl-4-gonen-3-one-oxime), and its metabolites (levonorgestrel-3-oxime, levonorgestrel-17-acetate and levonorgestrel) to the progesterone receptor was investigated by competition experiments using cytosol from human myometrial tissue. Compared to R5020, a highly potent synthetic ligand for progesterone receptor analysis, the L-isomer of norgestimate shows only a weak specific behaviour with regard to binding to the progesterone receptor from uterine cytosol with an RBA value of 0.8%, whereas the D-isomer of this compound is characterized by a lack of binding activity to the progesterone receptor. Levonorgestrel-3-oxime, one of the possible metabolites of norgestimate, binds to the progesterone receptor with an RBA value of 8%, whereas levonorgestrel-17-acetate, the other possible metabolite of norgestimate, binds with a binding affinity of 110% which is in the same order of magnitude as levonorgestrel itself. The competition experiments suggest that norgestimate is a prodrug and that the metabolites, levonorgestrel and levonorgestrel-17-acetate, which actively bind to the progesterone receptor, must first be formed from the parent drug via metabolic processes in vivo. These are the actual biologically active compounds which are responsible for the gestagenic potency.

Dihydrospirorenone (ZK30595): a novel synthetic progestagen--characterization of binding to different receptor proteins.

Dihydrospirorenone (6 beta, 7 beta, 15 beta, 16 beta-dimethylen-3-oxo-17a-pregn-4-en-21, 17-carbolacton) is a new type of progestin with no other agonistic activities, in particular no estrogenic, androgenic or glucocorticoid activity. Dihydrospirorenone is a potent aldosterone antagonist 8 times as potent as spironolactone and antiandrogenic (0.3 times cyproterone acetate). The aim of the present study is to make a broad characterization of dihydrospirorenone at the receptor level and to discuss the results in comparison to those of established progestins. Kinetic studies of 3H-dihydrospirorenone uptake show a rapid increase in the amount of specific binding during the first three hours. The dissociation of 3H-dihydrospirorenone from the cytoplasmic human uterine progesterone receptor, measured by displacement of labeled steroid with dextran-coated charcoal treatment at 4 degrees C at various times, showed a monophasic or one-component, first order dissociation curve like progesterone. Sucrose density gradient centrifugation of the 3H-dihydrospirorenone-labeled myometrial cytosol showed that the dihydrospirorenone binding components sedimented in the 4S and 8S region which is typical for the progesterone receptor under low salt conditions. The high binding affinity of dihydrospirorenone to the binding sites of the mineralocorticoid receptor of rat kidney with an RBA value of 230% compared to aldosterone is remarkable. This reflects the strong antimineralocorticoid activity of this compound which was evaluated in adrenalectomized rats. Furthermore, competitive studies indicated that dihydrospirorenone also displays high affinity for the androgen and some affinity for the glucocorticoid receptors but no measurable affinity for the estrogen receptor.

Binding of oral contraceptive progestogens to serum proteins and cytoplasmic receptor.

Some progesterones widely used in oral contraceptives are characterized at the level of high-affinity receptor binding as well as binding to sex hormone-binding globulin and corticosteroid-binding globulin. With regard to binding to sex hormone-binding globulin, gestodene, levonorgestrel, and to a lesser extent 3-ketodesogestrel (which is only formed from the prodrug desogestrel in the body), show a behavior that is manifested in the relatively high affinity to sex hormone-binding globulin, whereas desogestrel and norgestimate do not display any measurable affinity for this specific steroid-binding serum protein. Furthermore, levonorgestrel and gestodene dissociate very much more slowly from the binding sites of sex hormone-binding globulin than 3-ketodesogestrel. A natural affinity of all these synthetic progestogens tested for corticosteroid-binding globulin could not be established. Gestodene, levonorgestrel, and 3-ketodesogestrel bind to the progesterone, glucocorticoid, and androgen receptor with high affinity, apart from slight differences, whereas estrogen receptor affinity could not be demonstrated in any of the progestogens investigated. In relation to aldosterone, the relative binding affinity values of gestodene, levonorgestrel, and the natural progestogen progesterone are relatively high, whereas 3-ketodesogestrel does not display any measurable affinity for this receptor species.

Lack of binding of gestodene to estrogen receptor in human breast cancer tissue.

Competition studies with progesterone and estradiol receptors of human myometrial tissue as well as of mammary cancer tissue showed that gestodene bound with high affinity to the progesterone receptor, as did other synthetic and natural progestogens. However, gestodene did not bind to the estradiol receptor. The relative binding affinities of all tested synthetic and natural ligands showed no organ-specific differences and no differences between neoplastically transformed and normal tissues.

An enzyme-linked immunosorbent assay (ELISA) for human corticosteroid-binding globulin.

A specific and sensitive enzyme-linked immunosorbent assay (ELISA) of the "sandwich-type" for human corticosteroid-binding globulin was developed. A polyclonal rabbit anti-CBG antibody is immobilised to a microtitre plate. Following incubation of standards and samples a second monospecific rabbit anti-CBG antibody, labelled with alkaline phosphatase, is added. After colour development the microtitre plate is read at 405 nm wavelength. The assay shows good agreement to CBG binding capacity assay and commercially available RIA.

Gestodene: a novel synthetic progestin--characterization of binding to receptor and serum proteins.

Gestodene, 17 alpha-ethinyl-13-ethyl-17 beta-hydroxy-4,15-gonadien-3-one, is a new orally active progestational agent, which is available for clinical use in oral contraceptives. The aim of the present study is to make a broad characterization of gestodene at the receptor level and to discuss the results in comparison to those of established progestogens. Kinetic studies of 3H-gestodene uptake show a rapid increase in the amount of specific binding during the first three hours. After saturation, the amount of specifically bound 3H-gestodene remained almost constant up to 24 hours at 4 degrees C. The dissociation of 3H-gestodene from the cytoplasmic myometrial progestone receptor, measured by displacement of labeled steroid with dextran-coated charcoal treatment at 4 degrees C at various times, showed a biphasic or two-component first order dissociation curve. As anticipated, sucrose gradient centrifugation analysis of the 3H-gestodene-labeled cytosol of human myometrial tissue showed that the gestodene binding components sedimented in the 4S and 8S region. A 200-fold molar excess of nonradioactive gestodene reduced only the 8S binding of 3H-gestodene. 4S binding of 3H-gestodene was not reduced, which indicate the existence of a second high capacity binding component. In biological test systems, such as the Clauberg test or Kaufmann test, gestodene has proved to be a very effective progestogen. Among nortestosterone derivatives it is one of the most potent and resembles progesterone biologically in its progestogenic effects. This biologically identical gestagenic activity of gestodene and progesterone is reflected by a very similar behavior in vitro in terms of binding to progesterone receptors of human uterus cytosol. Furthermore, competitive studies indicated that gestodene like other synthetic progestagens also displays some affinity for androgen and glucocorticoid receptors but no measurable affinity for the estrogen receptor. Remarkable is the high binding affinity of gestodene to the binding sites of the mineralocorticoid receptor of rat kidney with a RBA value of 350% compared to aldosterone.

Tissue reaction after silicone and poly(methyl methacrylate) intraocular lens implantation: a light and electron microscopy study in a rabbit model.

We describe the histopathologic results of extracapsular lens extraction and silicone and poly(methyl methacrylate) (PMMA) intraocular lens (IOL) implantation in 36 rabbit eyes. Phase-contrast microscopy was used to examine precipitates on IOL surfaces and posterior capsules. Semithin and ultrathin sections were taken from the central cornea, anterior uvea, capsular bag, and retina near the posterior pole. The follow-up was one to 16 weeks. Silicone IOLs did not cause significantly less precipitation than PMMA IOLs. Precipitates consisted of spindle-shaped fibroblast-like cells, various forms of inflammatory cells and multinucleated giant cells, single melanophages, and irregularly arranged birefringent collagen fibers. Corneal endothelial edema was slightly more prominent in PMMA IOL implanted eyes. Significant retinal edema in the posterior pole area was not observed with either of the two lens types. Severe precipitation in the form of large clusters of pigment cells and inflammatory reactions seemed to depend on mechanical trauma (iris capture and lens dislocation) and individual animal reactions, but not on the lens type used.

3H-ZK 98,734, a new 11 beta-aryl substituted antigestagen: binding characteristics to receptor and serum proteins.

Recently, in the laboratories of Schering in Germany a competitive progesterone antagonist, ZK 98,734, was synthetized, which is characterized by a similar antigestagenic activity as RU 38,486, synthezised by Roussel-Uclaf in France, as assessed by inhibition of nidation tests in rats and guinea pigs. However, this compound has a substantially lower antiglucocorticoid activity measured in cell culture systems than RU 38,486. The purpose of this study was to present a comparison of biochemical and physical properties of the complexes formed by the human uterine progesterone receptor with 3H-ZK 98,734 on one hand and with other well-established progestins on the other hand. ZK 98,734 competed in the same order of magnitude as progesterone or RU 38,486 for the 3H-R5020 binding site of progestin receptor, whereas R5020, Org 2058 or progesterone were unable to compete against 3H-ZK 98,734. This apparent contradiction could be explained by means of FPLC-chromatography and sucrose density centrifugation technique. FPLC-chromatography with an anion exchange column (Mono Q, Pharmacia, Uppsala, Sweden) showed that 3H-ZK 98,734 forms at least two stable complexes with uterine cytosol, on one hand with serum albumin, which presents almost 90% of bound radioactivity, and on the other hand with the two native progestin receptor forms, corresponding to 4S and 8S receptor forms in sucrose density gradient analysis. Competition experiments in liver cytosol of adrenalectomized rats with increasing concentrations of unlabeled ligands other than dexamethasone showed that ZK 98,734, RU 38,486 and cortisol displaced 3H-dexamethasone efficiently from the binding sites in the cytosol. Furthermore, results concerning the specificity of 3H-cortisol binding to serum proteins in diluted pregnant serum demonstrated that ZK 98,734 did not compete with 3H-cortisol for serum binding.

Evaluation of the first 60 cases of poly HEMA posterior chamber lenses implanted in the sulcus.

Flexible lenses allow folding and inserting through a small incision. Using Faulkner's forceps, 60 hydrogel lenses were implanted in the sulcus through a 3.5 mm pocket incision following phacoemulsification. Mean follow-up was six months. The lens was suitable for the procedure when it was properly handled. When it was not, untimely unfolding or tears occurred, necessitating prolonged maneuvering in the eye or sometimes replacement of the lens which caused increased trauma. Though functional results were comparable to those achieved with polymethylmethacrylate lenses, all eyes achieving a visual acuity of 20/40 or better, morphological results were less satisfactory. Because of hydrogel's lack of adhesiveness and the inability to estimate the individual sulcus diameter, pseudophakodonesis was a common observation. The lens touching the posterior iris surface caused pigment dispersion (37%), sometimes leading to secondary pigment glaucoma. For this reason, we now favor capsular bag fixation.

Preparation of electrophoretic variants of corticosteroid-binding globulin (CBG) using liquid liquid partition chromatography.

Human corticosteroid-binding globulin (CBG) was purified to homogeneity by application of three different chromatographic methods. After fractionation of pregnancy serum with ammonium sulfate the 80%-pellet was used for affinity chromatography based on tresyl activated Sepharose (Pharmacia, Uppsala, Sweden). The affinity eluate was injected into a Mono Q anion exchange column (Pharmacia). Fractions containing CBG were finally purified by liquid liquid chromatography on LiParGel 750 (Merck, Darmstadt, F.R.G.). The purified protein was characterized by IEF and PAGE. This paper describes a method for the chromatographic separation of the two variants of CBG without a loss of binding activity towards steroids for each of the two characteristic bands of this protein.

Influence of indomethacin on PGE2 in rabbit aqueous humor after YAG laser photodisruption of the iris.

In an experimental study with rabbits, the influence of indomethacin on the postoperative PGE2 level in the aqueous humor was investigated, following YAG laser traumatization of the iris. Indomethacin is a drug with an inhibitory effect on the synthesis of prostaglandins. Using albino rabbits, the right eye was treated with indomethacin eye drops three times daily for 3 days. On the 4th day, high-energy YAG laser was applied to the iris of both eyes (Q-switched Nd: YAG Laser, Model Cilco Lasertek PV 135; ten photodisruptive lesions, with 50 mJ to the midperiphery of the iris). Subsequently, the rabbits were subdivided into three groups. In group 1 the aqueous humor was removed from both eyes 12 h postoperatively; in group 2 the aqueous humor was tapped 36 h after the intervention; for group 3, it was 60 h afterwards. The results from 15 rabbits were evaluated. Local indomethacin treatment was continued until tapping of the aqueous humor. As a control, another group was used with 3 rabbits without treatment. Twelve hours after YAG laser treatment there was still a clearly significant difference in the PGE2 concentrations between the eyes that had received indomethacin and the untreated eyes; 36 h postoperatively, the difference was no longer statistically significant, and after 60 h the PGE2 concentrations of the treated and untreated eyes were the same.

An enzyme-linked immunosorbent assay (ELISA) for human sex hormone-binding globulin.

An enzyme-linked immunosorbent assay (ELISA) of the "sandwich-type" for sex hormone binding globulin (SHBG) has been developed. A rabbit anti-SHBG antibody (RAb) is immobilized to the microtitre plate. After incubation with standards and samples a second monospecific rabbit anti-SHBG antibody, labelled with alkaline phosphatase is added (RAb). Following further washing substrate is added, colour developed and the plate read at 405 nm wavelength on a standard ELISA plate reader. The assay is not influenced by the presence of steroids at the binding site, and shows good agreement to SHBG binding capacity assay and commercially available IRMA. Its sensitivity, specificity and precision allows its use in the routine laboratory. The SHBG ELISA has been used to measure SHBG concentrations in sera of normal men, women, pregnant women, and women receiving high-dose medroxyprogesterone acetate as a treatment of metastatic breast cancer.

Clinicopathologic findings after in-the-bag implantation of open-loop polymethylmethacrylate and silicone lenses in the rabbit eye.

Two lens models, one all silicone with J-loop haptics, the second with PMMA optics and angled polypropylene C-loop haptics, were implanted in the capsular bag of 36 rabbit eyes following extracapsular lens extraction. Technical ease of implantation, inflammatory response, lens position, and fibrin exudation as well as precipitate formation were examined. Biomicroscopy, light and phase-contrast microscopy were used. Differences in haptic vulnerability, lens position, and fibrin clot formation were found between the two lens types. Lens-specific differences in precipitate formation could not be substantiated.

Optical resolution of silicone and polymethylmethacrylate intraocular lenses.

The optical resolution and contrast of 18 silicone and 18 polymethylmethacrylate (PMMA) intraocular lenses were determined before implantation in and after removal from rabbit eyes. A specially designed microphotographic system that allows documentation and comparison of the image quality of different lens types was used. Silicone intraocular lenses showed a mean axial resolution of 1.25, PMMA lenses showed 1.60, of an optotype analogous resolution target (1.0 = 20/20) at a 3 mm pupil opening preoperatively. At one to 16 weeks postoperatively, both types of IOLs demonstrated nearly unchanged resolution. Contrast, however, decreased to between 60% and 85% of the preoperative 100% standard. This decrease was due to precipitates deposited on the lens surface, to posterior capsule opacification, and to lens decentration.

[Ocular findings in hemodialysis and following kidney transplantation in childhood and adolescence]. / Augenbefunde bei Hämodialyse und nach Nierentransplantation im Kindes- und Jugendalter.

Between August 1980 and January 1987, 23 patients undergoing treatment for chronic renal failure underwent eye examinations. Hemodialysis and subsequent kidney transplants were performed in 18 patients; in two patients a kidney transplant was performed alone, and in three others hemodialysis without transplant. The interval between dialysis and transplantation averaged 23.1 months, the mean follow-up after transplantation 20 months. Patients who underwent hemodialysis alone were followed up for periods of two, three and 85 months. The patients' ages when hemodialysis treatment was first instituted ranged from six to 17 years (average 11.8 years). The mean age at the time kidney transplants were performed was 13.6 years (ranging from one to 17 years). Seventeen patients had conjunctival and corneal infiltrations in the area of the palpebral fissure. In two cases infiltrations were confined to the conjunctiva. Four patients had no pathologic changes, in either the cornea or the conjunctiva. Slitlamp examination revealed subcapsular losses of lens transparency in eight patients; these losses were manifested by delicate punctiform and patchy configurations. In nine cases fundus ophthalmoscopy revealed constricted retinal arteries. Within the period of observation all but one of the patients had unchanged vision. The one exception (cystinosis) had reduced visual acuity due to an accumulation of crystalline inclusions in the cornea.

Application of liquid-liquid partition chromatography in the simultaneous purification of sex-hormone-binding globulin and corticosteroid-binding globulin.

Two human serum proteins, corticosteroid-binding globulin (CBG) and sex-hormone-binding globulin (SHBG), were purified to homogeneity by the application of a combination of three different modes of chromatography. Human pregnancy serum was fractionated with ammonium sulphate. SHBG (50% pellet) and CBG (80% pellet) were then purified by affinity chromatography on tresyl-activated Sepharose with 15-aminopentadecanoic acid (for SHBG) and 1,12-diaminododecane (for CBG) as spacers and 17 zeta-aminoethyl-5 alpha-androstan-3 beta,17-diol (for SHBG) and 17 alpha-hydroxy-4-androsten-3-one-17 beta-carboxylic acid (for CBG) as specific ligands for these two proteins. The eluate was injected into a Mono Q anion-exchange column. Fractions containing SHBG or CBG were finally purified by liquid-liquid chromatography on Lipar-Gel 750. This chromatographic sequence clearly separated SHBG and CBG from other proteins, mainly serum albumin, without a loss of protein or binding activity.