[Retinal damage by perfluorocarbon liquids - a question of specific gravity? Intraocular pressure peaks and shearing forces]. / Retinale Schäden durch flüssige Perfluorkarbone - eine Frage des spezifischen Gewichts? Intraokulare Druckspitzen und Scherkräfte.

BACKGROUND: Perfluorocarbon liquids (PFCL) cause retinal damage when used as long-term ocular endotamponades. Whether these changes are related to the mechanical or to the chemical properties of PFCL is unclear. The purpose of this study was to evaluate pressure spikes or shearing forces during endotamponade with PFCL and standardised eye movements. MATERIAL AND METHODS: Part 1: In an eye model the resulting pressure forces of 6 PFCL were measured at four different sites during standardised eye movements. Part 2: Shearing forces were determined in a plexiglass eye model and the resulting tangential forces at the PFCL-retina interface were calculated. Part 3: Rabbit eyes were vitrectomised and filled with light and heavy fluorocarbons for 6 weeks. Subsequently, the retina were examined histologically and by immunohistochemistry. RESULTS: With increasing filling of the eye model, the maximum of the pressure peaks moved from the inferior wall of the eye model to the lateral eye walls. For perfluorodecalin (PFD) the highest pressure peak was 407 Pa with a 75 % filling of the vitreous cavity. The lowest pressure peak was 314 Pa with a 50 % filling of hexafluoropropene oxide. Shearing forces for standardised accelerations were dependent on viscosity and ranged between 0.87 mN/m(2) (perfluorohexyloctane) and 8055 mN/m(2) (hexafluoropropene oxide). Part 3: Histological and immunohistochemical analyses did not reveal pressure-related damage or any difference between the effects of the different tamponades in vivo. CONCLUSION: In comparison with physiological dynamic and static pressure peaks, the measured mechanical forces induced by intraocular PFCL tamponades are low. Specific gravity and mechanical damage by intraocular PFCL as a cause of retinal damage seem unlikely. Animal studies underline these findings.

[The influence of a new surface treatment of silicone intracoular lenses with fluoralkylsitan on the adherence of endophthalmitic bacteria in vitro]]. / Effekt einer neuen Oberflächenbehandlung von Intraokularlinsen aus Silikon durch ein Fluoralkylsilan auf das Adhärenzverhalten Endophthalmitis-relevanter Bakterien in vitro.

INTRODUCTION: Dynasilan is a fluoroalkylsilan which is able to bind to surface active molecules of intraocular lenses (IOLs), thereby offering a new option for surface modification of silicone lenses. The purpose of this in vitro study was to investigate the influence of this new surface treatment on the adherence of two typical endophthalmitis-inducing bacteria ( Staphylococcus epidermidis, Propionibacterium acnes). MATERIALS AND METHODS: A total of 14 Dynasilan-treated and 14 untreated silicone lenses were incubated at 37 degrees C for 24 h in brain heart infusion broth (10(8) CFU/ml) either with Staphylococcus epidermidis or with Propionibacterium acnes for 1 h. Subsequently, the adherent bacteria were resuspended using ultrasonification at 35 kHz for 3 x 45 s. After a dilution series and incubation at 37 degrees C for 24 h or 3 days the colonies were counted. RESULTS: On untreated IOLs incubated with Staphylococcus epidermidis the average number of bacteria was 3.6 x 10(7)/ml, and on treated IOLs the number of counted colonies was reduced to 1.09 x 10(7)/ml. Incubated with Propionibacterium acnes the average number of adherent bacteria on untreated IOLs was 4.75 x 10(4)/ml and on modified IOLs the number was reduced to 2.94 x 10(4)/ml. CONCLUSION: Dynasilan surface treatment may reduce the adherence of Staphylococcus epidermidis and Propionibacterium acnes on silicone intraocular lenses. Further studies regarding the stability of this treatment, its biocompatibility and influence on lens epithelial cell adhesion are in progress.

New substances for intraocular tamponades: perfluorocarbon liquids, hydrofluorocarbon liquids and hydrofluorocarbon-oligomers in vitreoretinal surgery.

UNLABELLED: Perfluorocarbon liquids (PFCLs) and heavy fluorocarbon liquids (HFCLs) are being increasingly used as soft tools during vitreoretinal surgery. However, since long-term intraocular tolerance is still unsatisfactory, at present complete removal at the end of surgery is recommended. With the aim to improve long-term intraocular compatibility and to enlarge the spectrum of clinical applications, modified HFCLs have been developed. HFCL-oligomers with a higher viscosity represent the latest perspective. All three groups of fluorocarbon liquids will be compared with respect to their physical and chemical properties, experimental and clinical results, and prospects for clinical applications. Common features of PFCLs, HFCLs and HFCL-oligomers are biological inertness, specific gravity higher than water, immiscibility with water or blood, and a high gas binding capacity. In PFCLs such as decalin, octane, or phenanthrene. All carbon atoms of the carbon backbone are completely fluorinated. In experimental and clinical use, emulsification, vascular changes and structural alterations of the retina have been described. By only partial replacement of hydrogen atoms by fluorine, the specific gravity of HFCLs is reduced, whereas lipophilic properties increase. Thus HFCLs are potential solvents for intraocular silicone oil remnants. However. after long-term application, side-effects are similar to those observed with PFCLs. Substances of this group, such as F6H6, F6H8, 044, and 062 are used intraoperatively and are currently being investigated for clinical long-term application. With the aim to avoid emulsification and to improve intraocular tolerance, we have developed HFCL-oligomers consisting of 2-4 HFCL molecules with increased viscosity. The oligomers were tolerated well in rabbit eyes for up to 4 months. In contrast to PFCLs or monomers, they did not emulsify nor show vascular alterations. ERGs returned to normal after removal of the oligomer from the eye. Histology of the retina showed mild alterations. CONCLUSION: according to physical properties, experimental intraocular compatibility and stability against emulsification, HFCL-oligomers are promising candidates for improved long-term tamponade of the lower retina. At present, indications for an application in human eyes have to be determined in clinical trials.

Combined use of partially fluorinated alkanes, perfluorocarbon liquids and silicone oil: an experimental study.

BACKGROUND: Partially fluorinated alkanes (FALKs) are a new class of substances which can be used in vitreoretinal surgery as an intraoperative tool and as a long-term tamponade. The aim of this in vitro study was (1) to investigate the solubility of FALKs in silicone oil during direct exchange, (2) to study their combined use and solubility in PFCLs, (3) to evaluate their lipophilic properties and (4) to investigate the possibility of preparing "heavy silicone oil". METHODS: (1) Four different FALKs (F6H6, F6H8, O44 and O62) were directly exchanged with silicone oil (5,000 mPas). The dissolved amount of fluorocarbons in the removed silicone oil was determined by gas chromatography and by gravimetry. Furthermore, the diffusion phenomena during the exchange process were studied. (2) The behaviour of FALKs in PFCLs was investigated and the solubility of the resulting mixtures in silicone oil was measured. (3) The solubility of FALKs and PFCLs in native olive oil was analysed. (4) Different FALKs were added to silicone oil and measurements of the resulting specific gravity and the viscosity were performed. RESULTS: (1) FALKs dissolved in silicone oil up to the following values: F6H6=45 m%, F6H8=54 m%, 044=100 m%, O62=18 m%. (2) FALKs dissolved in PFCL, thereby changing the physicochemical properties of PFCL depending on the type of FALK and ratio used. (3) The lipophilic properties of FALKs and PFCLs could be characterized by their dissolution in native olive oil (F6H8=23.4 m%, 044=16.7 m%, F6H6=12.3 m%, 062=5.3 m%, PFD=1.1 m%, PFO=0.6 m%). (4) It was possible to prepare "heavy silicone oil" e.g. by adding 30 vol% F6H8, resulting in a specific gravity of 1.08 g/ml, or by adding 80 vol% 044, resulting in a specific gravity of 1.25 g/ml, but decreasing the viscosity of the mixtures dramatically. CONCLUSION: (1) If FALKs are used as an intraoperative tool, a direct exchange with silicone oil should be avoided owing to their capacity to dissolve in silicone oil, resulting in a mixture with unpredictable properties. (2) A combined use with PFCLs and silicone oil is possible, if the right ratio is chosen. (3) The solubility of FALKs in native olive oil may be an indicator for their tissue penetration and may render feasible their use as a long-term tamponade. (3) "Heavy silicone oil" preparation using FALKs is possible, but the mixture needs further evaluation in terms of emulsification, mobilization of oligosiloxanes, tissue penetration and long-term stability.

Use of O44 as a solvent for silicone oil adhesions on intraocular lenses.

PURPOSE: To describe a solvent that removes silicone oil adhesions on intraocular lenses (IOLs). METHOD: The solvent O44 is a partially fluorinated alcane that dissolves silicone oil. Silicone oil adhesions on silicone and poly(methyl methacrylate) IOLs were treated with O44. The extent of silicone oil adhesions and the effectiveness of O44 were studied by gross microscopy. RESULTS: The solvent O44 removed silicone oil adhesions from both IOL types. CONCLUSION: The substance O44 may be a successful intraoperative tool to remove silicone oil IOL adhesions, avoiding IOL explantation.

[O44--a solvent for silicone oil adhesions on intraocular lenses]. / O44--ein Lösungsmittel für Silikonöladhäsionen auf Intraokularlinsen.

AIM: To examine the efficiency of O44, a partial fluorinated octane, as a solvent for silicone oil adhesions on intraocular lenses. MATERIALS AND METHODS: After placing silicone- and PMMA-lenses in silicone oil, the adhesions were treated with O44. The extent of silicone oil adhesions and the effectiveness of O44 were studied by gross microscopy by scanning electrone microscopy (SEM) and combined energy dispersive spectrometry (EDX). Furthermore an explanted silicone lens with oil adhesions was treated with O44 and examined. To simulate the effect of adhesion proteins PMMA- and silicone lenses were placed in silicone oil and human plasma. RESULTS: With EDX it was possible to prove that O44 is able to remove silicone oil adhesions from PMMA-lenses. The removal from an explanted silicone lens by O44 could be demonstrated by light microscopy. Silicone oil adhesions on intraocular lenses (IOL's) in vitro showed a different oil coverage than the IOL in vivo. Silicone lenses in vitro were often covered by a homogenous oil film and therefore the demonstration of the silicone adhesions and of the efficacy of O44 was difficult. The adhesions of IOL's placed additional in human plasma did not show any differences. CONCLUSIONS: O44 may be a successful intraoperative tool to remove silicone oil adhesions avoiding explantation of silicone oil contaminated IOL's. Silicone oil adhesions in vivo seem to be influenced by adhesive proteins.

Photomutagenesis test development: I. 8-Methoxypsoralen, chlorpromazine and sunscreen compounds in bacterial and yeast assays.

Two in vitro genotoxicity tests have been adapted to the evaluation of photomutagenic activity of test compounds. The study was initiated to obtain an experimental basis relating to newly proposed guidelines of the EC which request the screening of UV-absorbing compounds, for example, those employed in sunscreen preparations, for their photomutagenic potential. The well established photomutagens 8-methoxypsoralen and chlorpromazine were used to define relevant test protocols. The compounds were evaluated with the Ames test and the Saccharomyces cerevisiae D7 test for gene conversion. The influence of various parameters such as UV light sources, spectral composition, UV sensitivity of the test systems, absorbance by test materials and different exposure conditions is indicated. Two exemplary screening experiments with cosmetic ingredients are presented. Both test systems can be employed for the evaluation of compounds for photomutagenic activity although the standard excision-deficient strains of S. typhimurium pose problems because of their high UV sensitivity. The present experience in this complex field suggests that rigid test protocols and a restrictive test battery would be inadequate.

Photomutagenesis test development: II. 8-Methoxypsoralen, chlorpromazine and sunscreen compounds in chromosomal aberration assays using CHO cells.

Chromosomal changes were analysed in Chinese hamster ovary (CHO) cells treated with 8-methoxypsoralen (8-MOP) or chlorpromazine (CPZ) and irradiated with either a UVA fluorescent tube (emission spectrum ranging from 350 to 400 nm) or a xenon burner (continuous emission spectrum simulating ambient sunlight). In the dark neither 8-MOP nor CPZ was genotoxic by itself. If these compounds were used in combination with UV irradiation the rate of chromosome aberrations was significantly increased. The magnitude of the clastogenic response was dependent on compound concentration and UV dose. The spectral composition also played an important role. Care must be taken to account for spectral changes caused, e.g., by passage of the light through the plastic lid of the container. The possible clastogenicity of two sunscreens was tested with two protocols: (1) cells attached to the culture dish were treated in presence of the sunscreen in the medium or (2) cells were irradiated through a layer of sunscreen solution as a filter. With this a clear UVB-absorbing effect and a decreased frequency of UVAB-induced chromosome aberration was evident with the UVB-absorbing compound Parsol HS but was absent, as expected, with the UVA-absorbing compound Parsol 1789. The presence of the sunscreens in the irradiated cell sample did not cause a significant increase in UV-induced chromosome aberrations.

Normal chromosomal aberration frequencies in peripheral lymphocytes of healthy human volunteers exposed to a maximum daily dose of paracetamol in a double blind trial.

Paracetamol (acetaminophen) has been examined for mutagenic potential in numerous studies: gene mutation tests consistently gave negative results while in vitro chromosomal aberration tests showed equally consistently positive effects. In vivo studies for chromosome breaking activity gave clearly negative, equivocal or weakly positive results. In particular two reports have indicated that human volunteers taking a maximum daily dose of paracetamol (3 x 1000 mg over 8 h) exhibited significantly elevated frequencies of chromatid breaks in their peripheral lymphocytes 24 h later. In the one study evaluating the time course, levels returned to normal between 3 and 7 days later. We performed a carefully controlled double-blind study in which volunteers were pre-screened for normal liver function, they all were non-smoking and their diet and environmental exposures were controlled during the study. Cell-cycle kinetics were monitored and paralleled and a placebo group was included. Although a larger number of cells than in the other studies was analysed we were unable to reproduce their findings. No significant increases in structural chromosome aberrations (CA) were found either when the paracetamol group (male, female or both) post-dosing values were compared with pre-dosing values, or when treated groups at any sampling time were compared with the placebo groups. There was not even any evidence that individuals responded to the clastogenic potential of paracetamol or that a group response may have been masked by non-responders. In conjunction with the recently published results of the NTP bioassay, showing no carcinogenic activity in mice and no carcinogenic activity in rats except an increase of mononuclear cell leukaemia in female rats which is of doubtful relevance, the study presented here argues that paracetamol does not pose an unacceptable (if any) genotoxic/carcinogenic risk to man.

Direct analysis of the clastogenic effect of formaldehyde in unstimulated human lymphocytes by means of the premature chromosome condensation technique.

The cytogenetic effect of formaldehyde (FA) on unstimulated human lymphocytes was studied by means of conventional chromosome analysis and the premature chromosome condensation (PCC) technique. In first post-treatment metaphases no significantly increased yields of chromosomal changes could be observed. The analysis of PCCs, however, showed high yields of chromosome fragments. Bleomycin (BLM) used as positive control was also highly clastogenic in PCCs and resulted in significantly increased yields of chromosome-type aberrations. As recently argued, a premitotic selection against heavily damaged cells could be an explanation for the discrepancy between the chromosome findings in metaphase and PCC analysis after FA treatment. In addition, a differential effectiveness may exist in unstimulated lymphocytes to convert multiple fragmentation into chromatid- or chromosome-type aberrations through S-phase-dependent or S-phase-independent mechanisms.

Chromosome aberrations in human lymphocytes induced by fission neutrons.

The dose-response relationships of dicentrics and excess acentrics were analysed after exposure of human lymphocytes to a mixed fission neutron-gamma-ray beam. From the analysis of exclusively first division cells a linear-quadratic relation was obtained for dicentrics with the ratio of linear and quadratic components, zeta, equal to 2.76 Gy. Over the range of doses studied (0.04-1.97 Gy) intratrack events therefore predominated. This also applied to acentrics which were linearly related to dose. At the lowest level of observed effect and dose, r.b.e. values with respect to 60Co gamma-rays of up to about 11 were derived for dicentrics and acentrics. With increasing neutron dose the r.b.e. decreased.

Cytogenetic effects of an alkyl-lysophospholipid derivative in human peripheral lymphocytes exposed in vitro and in vivo.

Chromosome analyses were carried out in human lymphocytes exposed to an antitumor alkyl-lysophospholipid (ALP) in vitro and in vivo. There was no evidence for a significant clastogenic effect or incidence of an increased sister-chromatid exchange (SCE) after short-term and long-term treatment of unstimulated or stimulated lymphocytes with concentrations of 0.1-50 micrograms ALP per ml. Cell proliferation was delayed after G0 exposure to 50 micrograms/ml. Although no ALP monotherapy was present for 3 cancer patients examined before and during ALP treatment with accumulated doses of 15-516 mg/kg, the cytogenetic data were in accord with the negative experimental results.

Dose-effect relationship for 14.5 MeV (d + T) neutron-induced chromosome aberrations in human lymphocytes irradiated in a man phantom.

Human lymphocytes were irradiated with 14.5 MeV (d + T) neutrons at the positions sternal, retrosternal, liver and gonads of a waterfilled Plexiglas man phantom of 37 degrees C. After BUdR treatment of lymphocyte cultures and FPG-staining of metaphases, exclusively first post-irradiation divisions (M1-cells) were analysed. A regression analysis was carried out by means of a weighted least-squares method. Dose-effect relations of dicentrics and acentrics could be fitted by the linear-quadratic model. A comparison of the dose-effect curves established with either conventional or FPG-staining technique revealed statistically significant differences. The M1-evaluation yielded about 46 per cent more dicentrics and about 37 per cent more acentrics. The consequences of these results are discussed in relation to the application of chromosome analysis in 'biological dosimetry'.

Chromosome changes in lymphocytes after occupational exposure to toluene.

Cytogenetic analyses were carried out in peripheral lymphocytes from 20 male workers exposed only to toluene in a rotogravure plant for more than 16 years. As compared with a group of 24 unexposed controls, significantly higher yields of chromatid breaks, chromatid exchanges and gaps were observed. The number of SCEs was significantly increased in smoking and non-smoking toluene-exposed workers compared with the corresponding control groups.

Chromosome changes in lymphocytes after occupational exposure to pentachlorophenol (PCP).

Chromosome analyses were carried out on peripheral lymphocytes from 22 male workers employed at a pentachlorophenol (PCP) producing factory. As compared with a group of 22 matched controls a small, but significant, increase in the frequency of dicentrics and acentrics was observed. There was no significant increase of sister-chromatid exchange (SCEs) in smoking PCP workers, as compared with smoking controls. Within the control group, smokers had a higher incidence of SCEs than non-smokers.

Analysis of structural chromosome changes and SCE after occupational long-term exposure to electric and magnetic fields from 380 kV-systems.

Chromosome analyses were carried out in lymphocytes of 32 workers occupationally exposed for more than 20 years to 50 Hz alternating electric magnetic fields in 380 kV switchyards. As compared with a control group of 22 workers of similar age and occupation but without field exposure neither the yields of structural chromosome changes nor the SCE-frequencies were increased. The difference of cytogenetic data after occupational exposure to ionizing radiation is discussed.