Nonadrenergic noncholinergic vasodilation in bovine ciliary artery involves CGRP and neurogenic nitric oxide.

PURPOSE: Characterization of the nonadrenergic noncholinergic (NANC) vasodilator innervation in the anterior segment in the bovine eye. METHODS: The neurogenic tetrodotoxin-sensitive response to electrical field stimulation (EFS) of the intraocular segment of the bovine long posterior ciliary artery supplying the ciliary body was recorded using isolated ring segments of this artery mounted on an isometric myograph. After adrenergic and cholinergic receptor blockade (with phentolamine, propranolol, and atropine), the preconstricted vessels were subjected to EFS by passing constant current pulses (0.3 msec, 35 mA, 0.5 to 32 Hz) between two electrodes on either side of the vessel segments. RESULTS: EFS resulted in 60% relaxation of the active tone in 40 vessels. Treatment with capsaicin reduced the NANC response by 16 +/- 2% (P < 0.001) and inhibition of the NO synthase with 1 x 10(-4) M L-NOARG reduced the NANC response by 83 +/- 10% (P < 0.001). Desensitization of the vessels to substance P had no effect. The CGRP(8-37) fragment (1 x 10(-6) M) in the presence of 1 x 10(-4) M L-NOARG reversibly and competitively inhibited the NANC response. L-arginine partly antagonized the inhibition induced by L-NOARG. About 60% of the L-NOARG-sensitive component of the NANC response was inhibited by methylene blue. Combined incubation with capsaicin and L-NOARG nearly abolished the NANC response. The L-NOARG-sensitive/capsaicin-resistant relaxation was present in endothelium denuded vessels. The responses to EFS were blocked by TTX. CONCLUSIONS: The neurogenic NANC vasodilator response in the intraocular part of the bovine long posterior ciliary artery supplying the ciliary body is endothelium independent and consists of two components: a capsaicin-sensitive component mediated by CGRP released from sensory nerve endings and a larger L-NOARG sensitive component mediated by a direct "nitroxidergic" neurotransmission. The size of the nitroxidergic NANC response indicates that it has a physiological relevance in vivo.

Human retinal pigment epithelial cell-induced apoptosis in activated T cells.

PURPOSE: The immune privilege of the eye has been thought to be dependent on physical barriers and absence of lymphatic vessels. However, the immune privilege may also involve active immunologic processes, as recent studies have indicated. The purpose of the present study was to investigate whether human retinal pigment epithelial (RPE) cells can induce apoptosis in activated T cells. METHODS: Fas ligand (FasL) expression was detected by flow cytometry and immunohistochemistry. Cultured RPE cells were cocultured with T-cell lines and peripheral blood lymphocytes for 6 hours to 2 days. Induction of apoptosis was detected by 7-amino-actinomycin D and annexin V staining. RESULTS: Retinal pigment epithelial cells expressed FasL and induced apoptosis in activated Fas+ T cells. Blocking of Fas-FasL interaction with antibody strongly inhibited RPE-mediated T-cell apoptosis. Retinal pigment epithelial cells induced apoptosis in several activated T-cell populations and T-cell lines, including T-cell antigen receptor (TCR)-CD3-negative T-cell lines. In contrast, RPE cells induced little or no apoptosis in resting peripheral T cells. Major histocompatibility complex (MHC) class II monoclonal antibodies, which block alloactivation, had no inhibitory effect on RPE-mediated T-cell apoptotic responses in MHC class II-specific CD4+ T-cell lines. CONCLUSIONS: Retinal pigment epithelial cells express FasL and induce TCR-independent apoptosis in activated human T cells through Fas-FasL interaction. Retinal pigment epithelial cells may constitute an immunologic functional barrier against potentially harmful T cells.

Optic nerve oxygen tension: effects of intraocular pressure and dorzolamide.

AIM: To investigate the influence of acute changes in intraocular pressure on the oxygen tension in the vicinity of the optic nerve head under control conditions and after intravenous administration of 500 mg of the carbonic anhydrase inhibitor dorzolamide. METHODS: Domestic pigs were used as experimental animals. Oxygen tension was measured by means of a polarographic electrode in the vitreous 0.5 mm anterior to the optic disc. This entity is called the optic nerve oxygen tension. Intraocular pressure was controlled by a hypodermic needle inserted into the anterior chamber and connected to a saline reservoir. RESULTS: When the intraocular pressure was clamped at 20 cm H2O optic nerve oxygen tension was 20 (5) mm Hg (n=8). Intravenous administration of dorzolamide caused an increase in optic nerve oxygen tension of 43 (8)% (n=6). Both before and after administration of dorzolamide optic nerve oxygen tension was unaffected by changes in intraocular pressure, as long as this pressure remained below 60 cm H2O. At intraocular pressures of 60 cm H(2)O and below, dorzolamide significantly increased optic nerve oxygen tension. CONCLUSION: Intravenous administration of 500 mg dorzolamide increases the oxygen tension at the optic nerve head during acute increases in intraocular pressure.

Bcl-2, Bax, and c-Fos expression correlates to RPE cell apoptosis induced by UV-light and daunorubicin.

PURPOSE: The aim of this study was to determine the role of Bcl-2, Bcl-X L, Bax, and c-Fos in regulation of apoptosis, induced by ultraviolet-light A (UV-A) and daunorubicin (DNR), in retinal pigment epithelium (RPE) cells grown on bovine extracellular matrix (ECM)-coated or uncoated plastic dishes. METHODS: Apoptosis in confluent RPE cells cultured on ECM-coated or uncoated dishes was induced by UV-A or DNR. Apoptosis was detected by 7-amino-actinomycin D labeling followed by flow cytometry and by terminal deoxy-transferase mediated X-dUTP nick end labeling (TUNEL). Cellular expression of Bcl-2, Bcl-X L, Bax, and c-Fos was determined by the use of antibodies and flow cytometry, Western blot analysis, and immunocytochemical staining. RESULTS: Both UV-A and DNR induce apoptosis in human RPE cells in vitro. Human fetal RPE cells grown on ECM-coated dishes were significantly more resistant to UV-A or DNR induced apoptosis than cells grown on uncoated dishes. RPE cells grown on ECM-coated dishes expressed higher Bcl-2 levels and lower Bax levels compared to cells grown on uncoated dishes. However, Bcl-X L and c-Fos levels were comparable in the two cultures. After UV-A or DNR treatment, Bcl-2, Bcl-X L, Bax, and c-Fos levels were differently regulated in cells grown on ECM-coated dishes compared to cells grown on uncoated dishes. CONCLUSION: A significant protection against apoptosis of RPE cells grown on ECM compared to cells grown on uncoated plastic dishes was found after exposure to UV-A or DNR. This protection was found to be proportionally correlated to the anti-apoptotic protein Bcl-2 and inversely correlated to the expression of Bax. Furthermore a sustained induction and expression of c-Fos was found to correlate to a higher percentage of apoptotic cells of RPE cells grown on plastic. These findings demonstrate that ECM is of great importance for RPE cell survival during noxious stimuli and points out the essential role for a healthy Bruch's Membrane (BM) for RPE survival.

[AIDS--knowledge, behavior and attitude at the University of Aarhus in 1990. A questionnaire study]. / AIDS--viden, adfaerd og holdning på Aarhus Universitet i 1990. En spørgeskemaundersøgelse.

To describe knowledge, attitude and behaviour regarding AIDS among students and employees at the University of Aarhus an anonymous self-administered questionnaire survey was conducted in April 1990. Of the 2169 and 850 questionnaires to students and employees, respectively, 1548 (71.4%) and 568 (67%) were returned. The study group had an excellent knowledge about AIDS and the transmission of HIV-virus. However, when defining "risk behaviour" in relation to transmission of HIV-virus among heterosexuals as "> or = 2 sexual partners within the past year without using condoms" 27% of the male and 20% of the female students showed risk behaviour. Among employees the rate was highest (23%) among the male scientific staff than among the remainder (17%). It is concluded that risk behaviour was not related to age and occurred in spite of the fact that 42% of the students and 31% of the employees, who showed risk behaviour were also aware of a personal risk of being infected. There is still a need for information about AIDS, and future campaigns should be directed towards all age groups as risk behaviour is not only a phenomenon among young people.

The anterior lens capsule used as support material in RPE cell-transplantation.

PURPOSE: To investigate the use of an ocular basement membrane as support material for transplanted porcine RPE cells. METHODS: Porcine RPE cells were grown on bovine corneal extracellular matrix (ECM), isolated bovine- and porcine lens capsules, and tissue culture plastic. Cell density, and cell morphology were studied by phase contrast microscopy and transmission electron microscopy. RESULTS: RPE cells grown on porcine anterior lens capsule and on ECM obtained better morphology and higher final cell density than cells grown on plastic and on bovine anterior lens capsule. It was possible to transplant the porcine anterior lens capsule to the subretinal space in pigs. Within two weeks of observation, the lens capsule was well tolerated in the subretinal space. CONCLUSION: The anterior lens capsule seems to be promising as support material for use in RPE cell-transplantation.