Biomedical subjects

S J Ryan

Publications and source records attributed to S J Ryan.

At least 91 records · Page 5Linked to original sources

PubMed1994

Effect of TGF-beta on interferon-gamma-induced HLA-DR expression in human retinal pigment epithelial cells.

PURPOSE: Retinal pigment epithelial (RPE) cells express human leukocyte antigen (HLA)-DR (class II) antigens when stimulated with interferon gamma (IFN-gamma) and may be capable of local antigen presentation. The authors examined the effect of transforming growth factor-beta (TGF-beta), a cytokine normally found in the eye, on the expression of these immunoregulatory molecules in vitro and attempted to determine the mechanism by which this cytokine acts. METHODS: Human RPE cells were cultured in the presence of IFN-gamma and then stained immunohistochemically for HLA-DR antigens. TGF-beta 1 or TGF-beta 2 was added simultaneously with IFN-gamma or after 3 days of IFN-gamma treatment. In parallel experiments, RPE cells were pretreated with 4-phorbol-12 myristate-13 acetate (PMA), staurosporine, or calphostin C before stimulation with IFN-gamma or TGF-beta. Quantitative analysis was performed by fluorescence-activated cell sorting. RESULTS: IFN-gamma induced HLA-DR expression on RPE cells. Both TGF-beta 1 and TGF-beta 2 were able to inhibit this effect. These inhibitory effects of TGF-beta were augmented by pretreatment with either PMA or calphostin C. Pretreatment of the cells with PMA before stimulation with IFN-gamma downregulated HLA-DR expression. Staurosporine pretreatment suppressed HLA-DR expression by IFN-gamma-stimulated RPE cells, but this was not additive with TGF-beta. CONCLUSIONS: The authors conclude that TGF-beta 1 and TGF-beta 2 strongly inhibit the IFN-gamma-induced upregulation of class II antigens on human RPE cells. The modulation of these IFN-gamma and TGF-beta effects by calphostin C, staurosporine, and PMA treatment suggests involvement of the protein kinase C pathway.

Cells, Cultured↗

PubMed1994

Intercellular gap formation induced by thrombin in confluent cultured bovine retinal pigment epithelial cells.

PURPOSE: Thrombin is formed at the site of intraocular hemorrhage and may be important in the development of progressive retinal damage. The authors observed that thrombin-treated bovine retinal pigment epithelial (RPE) cell cultures develop intercellular gaps and initiated this study to examine in detail the effects of thrombin on RPE cell morphology, adhesion, and cytoskeleton. METHODS: Confluent cultures of bovine RPE cells were incubated for various times (0 to 24 hours) with alpha-thrombin (0.1 to 100 U/ml) or enzymatically inactive thrombin. Intercellular gaps were quantitated by light microscopy in ten representative fields (magnification X400) as number of gaps per field (gaps/f). RPE cytoskeleton was studied using immunofluorescent staining for vinculin and F-actin. The mechanism of thrombin-induced RPE cell gap formation was studied by preincubation with specific drugs, including a protein kinase inhibitor (staurosporine), protein kinase C inhibitors (H-7 and calphostin C), cyclic adenosine monophosphate (cAMP) inducer (forskolin), and cytoskeleton-disrupting agents (cytochalasin B or colchicine). RESULTS: Intercellular gaps (20 to 80 microns in diameter) were markedly increased in number in thrombin-treated cultures in a dose-dependent and time-dependent manner and were associated with an alteration in the distribution of F-actin and vinculin. Whereas control cultures showed 3.3 +/- 2.4 gaps/f, incubation with 8 U/ml of alpha-thrombin for 3 hours resulted in 44.8 +/- 15.3 gaps/f. These changes were most prominent shortly after the 3-hour coincubation, but the cultures did return to their original confluent state within 24 hours. Cultures treated with an enzymatically inactive thrombin showed fewer intercellular gaps than those treated with enzymatically active thrombin but had significantly more intercellular gaps than control cultures. Thrombin-induced intercellular gap formation was blocked by preincubation with forskolin (14.6 +/- 7.1 gaps/f), staurosporine (10.2 +/- 5.0 gaps/f), or H-7 (24.5 +/- 9.8 gaps/f). CONCLUSIONS: Exposure to an enzymatically active thrombin results in formation of intercellular gaps between cultured RPE cells. Inhibition of this phenomenon by protein kinase inhibitors and by a cAMP inducer suggests that this effect is mediated, at least in part, through protein kinase C- and cAMP-dependent pathways. Thrombin generation associated with intraocular hemorrhage may thus result in direct damage to the RPE monolayer, possibly via the same pathway(s).

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1993

Relaxing retinotomy with silicone oil or long-acting gas in eyes with severe proliferative vitreoretinopathy. Silicone Study Report 5. The Silicone Study Group.

In the Silicone Study, 117 of 404 eyes (29%) with severe proliferative vitreoretinopathy (> or = C-3, full-thickness retinal folds in three or more quadrants) enrolled in the study were treated with vitrectomy, underwent a relaxing retinotomy, and were randomly assigned to treatment with long-acting gas or silicone oil. Forty-six eyes (20%) had undergone no previous vitrectomy (group 1); 71 eyes (42%) had undergone previous vitrectomy (group 2) with intraocular gas tamponade (P < .001). Group 1 eyes not undergoing retinotomy had better anatomic (six months) and visual (six and 24 months) outcomes and less hypotony (six months) than eyes that did regardless of tamponade (P < .05). For eyes undergoing retinotomy, silicone oil decreased the likelihood of hypotony (six months, P < .05). These differences were not found in group 2 eyes. We conclude that eyes undergoing a vitreous operation for the first time for the treatment of proliferative vitreoretinopathy can in most instances be successfully treated by conventional techniques without the need for relaxing retinotomy. Retinotomy may be required more often in patients undergoing repeat vitreous surgery for proliferative vitreoretinopathy, in which case both silicone oil and long-acting perflouropropane gas appear to be equally effective.

Eye Diseases↗

PubMed1993

Differential permeance of retina and retinal pigment epithelium to water: implications for retinal adhesion.

Water movement from the vitreous to the choroid would tend to maintain retinal apposition, if the retina were less permeable than the retinal pigment epithelium (RPE). We used a rabbit model, to test the relative permeability of retina and RPE to tritiated water. The retina was found less permeable than the RPE. This result suggests the presence of a hydrostatic pressure gradient across the retina which could enhance apposition of retina to the RPE.

Animals↗

PubMed1993

Transplantation of cultured human retinal pigment epithelium into rabbit subretina.

Transplantation of normal retinal pigment epithelium (RPE) into a diseased eye holds promise for treatment of several blinding disorders. Previous studies have involved immunosuppression and implantation of freshly isolated cells. We report here the successful transplantation of cultured human RPE cells into rabbits that were not immunosuppressed. A modified pars plana transvitreal technique was used for RPE transplantation. The cultured RPE cells, loaded with carbon as a marker, were transplanted into the denuded Bruch's membrane of albino rabbits. The animals were followed for from 1 week to 3 months. On histologic examination at 2 months, no infiltrating lymphocytes were found in the vitreous cavity or choroid, even though Bruch's membrane was damaged. At about 3 months there were some macrophages in the subretina of transplanted eyes, indicating that an immunoreaction does occur eventually. Electron microscopy of the transplanted RPE showed apical-basal polarity and gap junctions. Restored function was attested to by the presence of phagosomes and phagocytosed outer segments in the transplanted cells. Our findings suggest that there is a weak, delayed immunoreaction to human RPE cells transplanted beneath the retina of the rabbit; however, functional recovery of the transplanted cells occurs before this immune response develops.

Adult↗

PubMed1993

Traction retinal detachment. XLIX Edward Jackson Memorial Lecture.

This review had two main objectives. The first goal was to attempt to define a final common pathway for traction retinal detachment. To illustrate this, I considered the following three common clinical entities: penetrating ocular trauma, proliferative vitreoretinopathy, and proliferative diabetic retinopathy. My second goal was to emphasize the role of research in developing a rational approach to treatment. To illustrate the second goal, I reviewed the contributions from three realms of research--laboratory, animal, and clinical research--examining their interactions and considering their roles in our current understanding and management of these clinical entities.

Diabetic Retinopathy↗

PubMed1993

Postoperative intraocular pressure abnormalities in the Silicone Study. Silicone Study Report 4.

BACKGROUND: Chronically abnormal intraocular pressure (IOP) may follow surgery for proliferative vitreoretinopathy (PVR), using either long-acting gas or silicone oil tamponade. Its prevalence and clinical significance are unclear. METHODS: In the Silicone Study, 241 eyes with severe (> or = C-3) PVR were treated with vitrectomy, randomized to perfluoropropane gas (C3F8) or silicone oil, and followed for 6 months or longer. Chronic IOP abnormalities, based on findings at two consecutive or any three postoperative visits, were defined as (1) low IOP (hypotony), 5 mmHg or less, or (2) elevated IOP, more than 25 mmHg. RESULTS: Eleven (5%) eyes had chronically elevated IOP and 58 (24%) had chronic hypotony. Chronically elevated IOP was more prevalent in eyes randomized to silicone oil than in those randomized to C3F8 gas (8% versus 2%; P < 0.05). Chronic hypotony was (1) more prevalent in eyes randomized to C3F8 gas than in those randomized to silicone oil (31% versus 18%; P < 0.05); (2) more prevalent in eyes with anatomic failure (48% versus 16%; P < 0.01); and (3) correlated with poor postoperative vision (P < 0.0001), corneal opacity (P < 0.001), and retinal detachment (P < 0.001). Factors prognostic of chronic hyotony included preoperative hypotony (P < 0.01), diffuse contraction of the retina anterior to the equator (P < 0.01), rubeosis (P = 0.02), and large retinal breaks (P = 0.02). In a multivariate analysis, diffuse contraction of the retina anterior to the equator remained an independent factor prognostic of chronic hypotony (odds ratio = 4.2), regardless of whether the retina was attached postoperatively. CONCLUSION: Intraocular pressure abnormalities are a common postoperative complication in eyes with PVR, and may occur with either C3F8 gas or with silicone oil. The presence of diffuse contraction of the retina anterior to the equator should alert the vitrectomy surgeon that the eye is likely to be hypotonus postoperatively.

Chronic Disease↗

PubMed1993

Vitrectomy with silicone oil or perfluoropropane gas in eyes with severe proliferative vitreoretinopathy. Silicone Study Report 3.

Between 1985 and 1990, 340 eyes with rhegmatogenous retinal detachment (RRD) and severe (stage C3 or worse) proliferative vitreoretinopathy (PVR) were treated with vitrectomy and randomly selected to receive perfluoropropane gas or silicone oil; 183 eyes had undergone no prior vitrectomy (group 1), and 157 eyes had undergone prior vitrectomy with intraocular gas tamponade (group 2). No differences were found between eyes in the two groups in achieving visual acuity of 5/200 or better (44% vs. 39%), macular reattachment (78% vs. 77%), or complete retinal reattachment (67% for both groups). In group 1, 74 eyes achieved complete retinal reattachment after only one operation; 41 additional eyes achieved reattachment after a second surgical procedure. In group 2, these numbers were 74 and 26, respectively. Eyes treated successfully after more than one operation were less likely to regain a visual acuity of 5/200 or better than those successfully treated with one operation (P < 0.01). There was no difference in hypotony between groups, but keratopathy was more frequent in eyes in group 2 (P < 0.05). The results suggest that differences in outcomes between group 1 and group 2 eyes are not as great as previously believed.

Adult↗

PubMed1993

Ageing and blood pressure regulation: dose-response relationships for angiotensin, blood pressure, atrial natriuretic peptide and aldosterone in normal subjects of varying ages.

1. Infusion of increasing doses of angiotensin II (AII) in normal subjects sequentially increased blood pressure, aldosterone and atrial natriuretic peptide (ANP) levels. 2. The slope of ANP response to AII was positively correlated with basal ANP and with the slope of blood pressure response to AII (pressor slope) but not with age. 3. This is consistent with the response of ANP to AII being mediated partly by the rise in blood pressure, independent of ageing. 4. As expected in a selected normotensive population, there was no correlation between basal blood pressure and age, but pressor slope was positively correlated with age. 5. Thus, dose-response relationships may be an index of age-induced alterations in pressure regulatory mechanisms.

Adult↗

PubMed1993

Allotransplantation of rabbit retinal pigment epithelial cells double-labelled with 5-bromodeoxyuridine (BrdU) and natural pigment.

To facilitate identification of transplanted retinal pigment epithelial (RPE) cells, we sought to double-label the cells with 5-bromodeoxyuridine (BrdU) and with natural pigment. The BrdU is not lost during cell division but does require immunohistochemical methods for visualization; the pigment, on the other hand, allows immediate, obvious identification, but is gradually lost with cell division. Together they provide a convenient, long-term double label. Herein we report the successful allotransplantation of double-labelled RPE cells onto Bruch's membrane of albino rabbits. The labelled cells were localized by anti-BrdU antibody and the avidin biotin-alkaline phosphatase complex (ABC-AP) method, and by visible inclusions of pigment. Using this double-label method, allotransplanted RPE cells could be readily and reliably identified on the recipient Bruch's membrane eight months after transplantation. The cells had distinct basal and apical morphology, and were in close contact with the photoreceptor outer segments of the host. This successful allotransplantation raises the possibility that the subretinal space of the rabbit might enjoy some degree of immunologic privilege.

Animals↗

PubMed1992

Vitrectomy for perforating eye injuries from shotgun pellets.

Pars plana vitrectomy and scleral buckling were performed on 22 eyes of 19 patients for treatment of perforating eye injuries from shotgun pellets. We reviewed the intraoperative findings at the time of vitrectomy to determine what factors might influence final visual acuity. The presence of a total retinal detachment at vitrectomy portended a poor prognosis when compared with eyes without total retinal detachment, as only one of ten eyes with total retinal detachment obtained useful vision (P = .002). Preoperative separation of the posterior vitreous was associated with a favorable outcome when compared with the absence of posterior vitreous detachment (P = .035), as ten of 16 eyes with posterior vitreous detachment at the time of vitrectomy ultimately achieved functional vision. The locations of the exit wounds did not affect visual success in the overall series of patients. However, in the patients who achieved visual success, exit wounds outside the vascular arcades were more likely to be associated with final visual acuities of, or better than, 20/70 than were those within the arcades (P = .022). Other prognostic factors, such as the number of perforations and the use of cryotherapy, were also examined for their effects on final visual outcome. However, these factors did not appear to affect visual outcome statistically.

Adolescent↗

PubMed1992

Maturation of newly-formed subretinal vessels.

The maturation process of newly-formed subretinal vessels was studied ultrastructurally. Primitive new vessels have thick endothelial cells with cytoplasm that contains many ribosomes and rough endoplasmic reticulum. Some immature endothelial cells show budding, and many of the intercellular junctions have small gaps. As the newly-formed vessels mature, the endothelial cells become attenuated and fenestrated, and their junctions become tight. Mature subretinal new vessels closely resemble those of the normal choriocapillaris.

Animals↗

PubMed1992

Retinal pigment epithelium implantation in the rabbit: technique and morphology.

Dysfunction of the retinal pigment epithelium (RPE) is a major component of many degenerative disorders of the retina. We have developed a modified pars plana technique for subretinal implantation of homologous, cultured, carbon-loaded RPE cells in the rabbit. The natural history and morphologic behavior of the implanted RPE cells for up to 6 weeks after transplantation were observed in successful procedures.

Animals↗

PubMed1992

Systemic antibiotic prophylaxis in penetrating ocular injuries. An experimental study.

Intraocular concentrations of systemically administered gentamicin and cefazolin in eyes after a standard penetrating ocular injury were measured to determine the effect of eye injury on ocular pharmacokinetics. Twenty pigmented rabbits were divided into two groups. A standard, 8-mm wound was made at the pars plana of one eye in each animal. Group 1 consisted of 10 rabbits that were treated with cefazolin (75 mg/kg), and Group 2 consisted of 10 rabbits that were given gentamicin (2 mg/kg). Fellow eyes, which sustained no trauma, served as control eyes. All groups received intravenous injections every 8 hours for 72 hours, beginning immediately after repair of the wound. After 72 hours, samples were obtained from the anterior chamber, vitreous cavity, and serum. Standardized bioassays for detection of gentamicin or cefazolin were performed. Significant concentrations of intravitreal cefazolin (9.6 micrograms/ml) and gentamicin (0.60 micrograms/ml) were found in the traumatized eyes in comparison to control eyes (P = 0.0001; P = 0.006). Cefazolin exhibited excellent penetration, achieving concentrations well above minimum inhibitory concentration values for most organisms. Gentamicin levels, however, were well below acceptable therapeutic levels. This study suggests that systemically administered cefazolin can achieve significant intravitreal penetration at minimum inhibitory concentrations after penetrating injury.

Animals↗

PubMed1992

In vitro stimulation of retinal pigment epithelium proliferation by taurine.

Taurine is an amino acid that is essential for retinal integrity and function. Although it has been suggested that the ratio of melatonin to taurine in the interphotoreceptor matrix may regulate the phagocytosis of outer segments by retinal pigment epithelial (RPE) cells, the effect of taurine on the RPE has not been studied. Using cultured RPE cells, we found that in vitro taurine specifically stimulated proliferation of human and rabbit RPE, but had only minimal effect on cultured scleral fibroblasts. The RPE proliferation was due to more cells entering into S-phase and thus an increase in DNA synthesis, was not dependent upon cell density, and was most pronounced in the presence of a low concentration of fetal bovine serum.

Animals↗

PubMed1992

Ultrastructures of the glial epiretinal membrane induced by activated macrophages.

A rabbit model of glial epiretinal membrane was established following the injection of activated macrophages into the vitreous. The membrane was composed entirely of cells with glial characteristics, ie, abundant intermediate filaments, microvilli, junctional complexes and basement membranes. The extracellular matrix of the mature membranes contained collagen fibrils of 10 to 15 and 20 to 25 nm in diameter. Fusiform densities were seen adjacent to the cell membrane and cells with indented nuclei were found in thick membranes. These observations demonstrate that glial cells in epiretinal membranes may synthesize collagen and possess myofibroblast-like properties.

Animals↗

PubMed1992

Cultures of human retinal pigment epithelium. Modulation of extracellular matrix.

The extracellular matrix of retinal pigment epithelial (RPE) cells is an important component of the fibroelastic membranes formed in proliferative vitreoretinopathy (PVR). These membranes consist of cells intermingled with fibrillar proteins, such as elastin and collagen. Because RPE cells may be exposed to vitreous and serum-derived factors in conditions that lead to PVR, the effects of vitreous and serum on the deposition of fibrillar proteins by RPE cells were studied using radioactive labeling and indirect immunofluorescence techniques. The RPE cells, modulated by combined vitreous and serum into a fibrocytic-like appearance, deposited less fibroelastic proteins per cell. However, as a result of simultaneous cell proliferation, the total deposition of fibroelastic proteins per culture was increased. Indirect immunofluorescence studies suggest that it is type I collagen that is altered in the matrix of these modulated cell cultures. This information is important in pathologic conditions characterized by intraocular cell proliferation and fibroelastic membrane formation; these phenomena are seen in many vitreoproliferative disorders.

Cell Count↗

PubMed1992

Prediction of visual acuity recovery in cystoid macular edema.

Three consecutive patients participated in a prospective evaluation of pseudophakic cystoid macular edema. The duration of the macular edema ranged from 6 to 8 months. On the initial visit, the best corrected acuity with spectacles was determined and a potential acuity meter reading was obtained; this test suggested potential for visual recovery in two of the three patients. Sub-tenon's injections of methylprednisolone acetate (20 mg) were administered along with topical 1% prednisolone acetate and 1% atropine. One month later, visual acuity was improved by more than 2 Snellen lines in all three of the patients. A visual acuity measurement with the potential acuity meter that is better than the best corrected acuity with spectacles may reflect the presence of intact, but dysfunctional photoreceptors that are capable of restoring visual acuity upon resolution of the edema.

Aged↗