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Biomedical subjects

V M Elner

Publications and source records attributed to V M Elner.

At least 73 records · Page 4Linked to original sources

Human and monkey corneal endothelium expression of low-density lipoprotein receptors.

Receptors for low-density lipoprotein are necessary for high-affinity uptake of lipid and protein essential to cell structure and function. Distinct receptors for acetoacetylated low-density lipoprotein internalize oxidized or enzymatically modified low-density lipoprotein and extracellular matrix components. We identified low-density lipoprotein receptors on cultured human and monkey corneal endothelial cells by the avid incorporation of fluorescently labeled low-density lipoprotein that was competitively inhibited by excess unlabeled low-density lipoprotein but not by unlabeled acetoacetylated low-density lipoprotein. Specific uptake of labeled low-density lipoprotein was greatest in nonconfluent, growing cells and increased after low-density lipoprotein deprivation. Intact endothelial monolayers of whole human cornea also incorporated low-density lipoprotein but not acetoacetylated low-density lipoprotein. After scratch injury of human corneas, spreading endothelium adjacent to areas of cell loss internalized more fluorescent low-density lipoprotein than cells distant from the injury. Blood-aqueous barrier breakdown occurring in ocular diseases and after surgical and nonsurgical trauma may allow leakage of circulating low-density lipoprotein, which provides a rich supply of lipid and protein for endothelial use. Efficient, receptor-mediated, low-density lipoprotein uptake may facilitate repair of damaged corneal endothelial membranes and regeneration of intact, functional cell monolayers.

Acetoacetates↗

The effects of subconjunctival mitomycin-C on glaucoma filtration surgery in rabbits.

A prospective, randomized, masked, placebo-controlled study was performed to determine whether a single, intraoperative subconjunctival application of a 0.5-mg/mL solution of mitomycin-C enhances the success of full-thickness filtration surgery in rabbits. Compared with control eyes, mitomycin-C-treated eyes showed significant increases in bleb duration from 8.1 +/- 2.4 to 68.0 +/- 20.8 days and in intraocular pressure reduction from 6.0 +/- 3.0 to 63.6 +/- 21.5 days. Histopathologic evaluation confirmed the inhibitory effects of mitomycin-C on fibrovascular, fibrocellular, and collagenous organization of the filtering blebs that resulted in their preservation. Transient, superficial, corneal vascularization anterior to the bleb occurred in all mitomycin-C-treated eyes. No other clinical or pathologic signs of undesirable side effects were noted. We studied the effectiveness and safety of a single intraoperative application of mitomycin-C in prolonging the success of filtration surgery in rabbits and its potential for similar use in humans.

Administration, Topical↗

Expression of modified low-density lipoprotein receptors by trabecular meshwork cells.

We examined the incorporation of fluoresceinated low-density lipoprotein (LDL) and acetylated or acetoacetylated low-density lipoprotein (A-LDL or AA-LDL) by a number of ocular cells in culture. All the cells investigated, including bovine, monkey, human trabecular meshwork cells, human corneal endothelial cells, human corneal stromal cells and human scleral cells, took up fluorescently labeled LDL. The bovine, monkey and human trabecular meshwork cells showed the strongest fluorescence reactions. In addition, we found that the trabecular meshwork cells became fluorescent after incubations with labeled A-LDL or AA-LDL. They were the only cell type examined that possessed this capacity. The fluorescence intensity was markedly diminished by adding to the incubation solution either fucoidin, a competitive inhibitor of modified LDL uptake, unlabeled A-LDL or AA-LDL. The trabecular meshwork cells in situ also became brightly labeled after exposure to fluoresceinated native LDL, A-LDL or AA-LDL. The uptake of modified LDL separated the trabecular meshwork cells from other types of ocular cells, which may be used to aid identification of trabecular meshwork cells in culture as well as in situ. This property also suggested that trabecular meshwork cells may have some functional similarities to macrophages.

Acetylation↗

Intercellular adhesion molecule-1 in human corneal endothelium. Modulation and function.

The endothelium lining the posterior corneal surface performs physiologic pump functions essential to corneal clarity and integrity. A hallmark of keratitis, anterior ocular inflammation, and corneal allograft rejection is leukocyte adherence to the corneal endothelium (CE) forming keratitic precipitates. To elucidate mechanisms governing cornea-leukocyte interactions, cultured human CE cells and intact corneas were examined for expression of intercellular adhesion molecule-1 (ICAM-1), which binds the lymphocyte function-associated antigen-1 (LFA-1) on all leukocytes and enhances delayed-type hypersensitivity mediated by class II major histocompatibility complex antigens. Immunohistochemistry on culture CE cells using monoclonal anti-ICAM-1 antibody yield positive staining that increased after exposure to interleukin-1-beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (gamma-IFN). Standard leukocyte adherence assays demonstrated ICAM-1-mediated CE-neutrophil binding, which was specifically blocked by antibody to ICAM-1 or antibodies to LFA-1 on neutrophils. In whole human corneas, gamma-IFN increased CE and stromal keratocyte ICAM-1 immunoreactivity and enhanced CE-neutrophil adherence. As in CE cell cultures, antibody to ICAM-1 effectively blocked neutrophil binding to the CE cells of whole corneas. These results are the first to demonstrate ICAM-1 in ocular tissue. They indicate that CE cells express functional ICAM-1, which may be modulated by inflammatory cytokines, ICAM-1 provides mechanisms for keratitic precipitate formation, regulation of corneal leukocyte trafficking and the generation of immune responses that may be crucial to allograft rejection.

Cell Adhesion Molecules↗

Human corneal interleukin-8. IL-1 and TNF-induced gene expression and secretion.

Corneal leukocytic infiltration is an important component of numerous ocular diseases, but specific corneal-derived leukocyte chemotaxins have not been identified. In this study, the authors identified interleukin-8 (IL-8), a known neutrophil and lymphocyte chemotaxin, to be an important chemotaxin produced by human corneal tissue. In situ hybridization and immunohistochemistry of corneas exposed to human recombinant (r) interleukin-1-beta (rIL-1 beta) or tumor necrosis factor-alpha (rTNF-alpha) revealed significant increases in corneal endothelial and stromal cell IL-8 mRNA (P less than 0.001) and marked increases in cell-associated immunoreactive IL-8 compared with unstimulated controls. ELISA assays revealed four- to eight-fold increases in corneal IL-8 secretion after 24-hour exposures to either cytokine over that obtained with unstimulated corneas (P = 0.01). In neutrophil chemotactic bioassays, significant increases in functional IL-8 were detected in media conditioned by corneas exposed to rIL-1 beta or rTNF-alpha for 24 hours (P less than 0.001). Preincubation of these corneal media with anti-IL-8 antibody significantly reduced neutrophil chemotaxis by more than 80%. These results suggest that the cornea is an active participant in ocular inflammation and raise the possibility that agents used in experimental corneal pocket models may produce indirect effects by inducing corneal secretion of other factors, such as IL-8.

Chemotaxis, Leukocyte↗

Monocyte chemotactic protein gene expression by cytokine-treated human retinal pigment epithelial cells.

Inflammation involving the retina and choroid is a common clinical problem, but the mechanisms that elicit and maintain ocular inflammation remain poorly understood. Interposed between the sensory retina and the systemic blood circulation within the choroid is the neural-derived retinal pigment epithelium (RPE), which forms part of the blood-retina barrier. The RPE is actively phagocytic and shares several features with mononuclear phagocytes of bone marrow origin, including the production of a neutrophil chemotactic factor, interleukin 8, after stimulation with interleukin 1 beta (IL-1 beta) or tumor necrosis factor alpha (TNF-alpha). Because monocyte-derived macrophages are present in retinal lesions of many common and blinding diseases, we monitored human RPE cells or monocyte chemotactic protein (MCP) mRNA expression and activity following cytokine stimulation. Cultured human RPE cells were left unstimulated or exposed to recombinant human IL-1 beta, TNF-alpha, or lipopolysaccharide. MCP mRNA expression in RPE cells and biologically active MCP in RPE cell supernatants were present 1 hour after stimulation and maintained for 24 hours. Conditioned media from RPE cells stimulated with 20 ng/ml of IL-1 beta or TNF-alpha for 24 hours contained biologically active monocyte chemotactic activity that rose rapidly from baseline levels over 4 hours and plateaued over the subsequent 20 hours. RPE chemotactic activity was dose dependent using concentrations of these cytokines ranging from 20 pg/ml to 20 ng/ml of 4-hour assays. Time- and concentration-dependent expression of RPE cell MCP mRNA was also found in the same cultures. Peak MCP mRNA expression occurred after 8 hours of stimulation with IL-1 beta or TNF-alpha. Maximal steady-state MCP mRNA expression occurred at 20 ng/ml for IL-1 beta. Immunohistochemical staining using specific anti-MCP antibodies resulted in distinctive RPE cell staining, confirming the presence of MCP in human RPE cells. These findings demonstrate that cytokine-stimulated RPE cells may evoke or augment mononuclear phagocyte-mediated ocular inflammation by synthesizing MCP.

Cells, Cultured↗

Ocular and associated systemic findings in suspected child abuse. A necropsy study.

We reviewed complete ocular and systemic necropsy findings of 10 consecutive children who died of suspected child abuse. All 10 children had evidence at necropsy of blunt head trauma, although external signs of blunt trauma occasionally were covert. Ocular injuries were observed in 7 of the 10 cases and when present always included retinal, vitreous, and subdural optic nerve hemorrhages. In 5 cases, intrascleral hemorrhage from the circle of Zinn occurred at the sclera-optic nerve junction. In 4 cases, traumatic retinoschisis or tractional retinal folds were present. Anterior segment findings were uniformly consistent with blunt trauma. Hemosiderin, indicating old hemorrhage, was present in 3 cases. Intracranial hemorrhage, present in all cases with abnormal ocular findings, was always accompanied by signs of direct head trauma, such as subgaleal hemorrhage, skull fracture, cerebral contusion, or external contusions, which are sometimes subtle or hidden beneath the hair.

Child Abuse↗

Neutrophil chemotactic factor (IL-8) gene expression by cytokine-treated retinal pigment epithelial cells.

The neural-derived retinal pigment epithelium (RPE) underlies the sensory retina and is central to both retinal homeostasis and many common retinal diseases. Retinal pigment epithelium cells are actively phagocytic and share several features with macrophages that have recently been shown to produce a neutrophil chemotactic factor (NCF), also known as interleukin-8, after cytokine stimulation. Because RPE cell responses to cytokines are largely unknown, human RPE cell NCF production was monitored after interleukin-1-beta (IL-1 beta), tumor necrosis factor-alpha, or lipopolysaccharide stimulation. RPE NCF mRNA expression and RPE production of biologically active NCF was time and concentration dependent. Maximal NCF mRNA expression occurred at 20 ng/ml for IL-1 beta. Messenger RNA expression in RPE cells and biologically active NCF in RPE cell supernatants were found 1 hour after stimulation and were maintained for 24 hours. These findings demonstrate that cytokine-stimulated RPE cells may evoke or augment neutrophil-mediated inflammation by synthesizing NCF, a cytokine that may be important in ocular disease mechanisms.

Biological Assay↗

Monkey trabecular meshwork cells in culture: growth, morphologic, and biochemical characteristics.

We established tissue cultures of trabecular meshwork cells from cynomolgus monkey eyes. The cultures were initiated within 4 h of enucleation on Falcon Primaria flasks. Using medium containing 10% fetal bovine serum and 5% calf serum, trabecular meshwork cells could be grown for up to eight passages without additional growth factors. The growth pattern and cell morphology were distinct from those seen in fibroblastic or endothelial cultures derived from neighboring tissues. Ultrastructurally, our cells showed the characteristics of trabecular meshwork cells, exhibiting prominent basement membranes, intercellular junctions, pinocytotic vesicles, microvillous projections, and branched cell extensions. These cells were grown mostly as monolayers. However, they also appeared to form multi-layered arrays in densely confluent areas when plated at a high density. The extracellular matrix material was surrounded by cells and cell processes, simulating in vivo trabecular beam formation. Radiolabeling experiments demonstrated that our trabecular meshwork cells had the capacity to produce collagen. These results indicated that our cultured cells retain many in vivo characteristics and may be used for various biologic studies of trabecular meshwork.

Animals↗

Intraocular neurilemmoma arising from the posterior ciliary nerve in neurofibromatosis. Pathologic findings.

A classic neurilemmoma was found in the eye of a 31-year-old man with known neurofibromatosis. The pathologic features of this rare intraocular tumor were examined histologically, immunohistochemically, and ultrastructurally. Contiguity of the neurilemmoma with its nerve of origin, the long posterior ciliary nerve, was clearly demonstrated. Expansion of the nerve by proliferating Schwann's cells was present.

Adult↗

Anterior proliferative vitreoretinopathy. Clinicopathologic, light microscopic, and ultrastructural findings.

Proliferative vitreoretinopathy (PVR) involving the posterior and equatorial retina is an established clinicopathologic entity. Clinically, a similar process, anterior PVR (APVR), results in anterior dragging of the peripheral retina by membranes which connect to the ciliary body or iris and cause circumferentially and radially fixed retinal folds. The pathology of APVR, however, has not been reported. The authors describe pathologic findings in 28 cases of APVR and ultrastructural pathologic findings in 6 surgical APVR specimens. Anterior PVR was frequently associated with retinal detachment (RD) repair (96%) and trauma (38%). Residual vitreous at the vitreous base virtually always provided a scaffold for membranes containing proliferating cells and deposited extracellular matrix. Major components of APVR membranes were fibrovascular tissue (71%), pigment epithelial cells (43%), fibrous and corneal stromal ingrowth (32%), and glial proliferation (18%). Because of its anterior location, APVR membranes also incorporated ciliary epithelium and corneal endothelium. Contraction of APVR membranes caused anterior retinal displacement and detachment in anatomic configurations corresponding to narrow and wide peripheral troughs. The authors' findings indicate that APVR is a distinctive clinicopathologic entity which may complicate rhegmatogenous RD and its repair.

Adult↗

Lysosomal enzyme activities in cultured trabecular-meshwork cells.

We examined the lysosomal enzyme levels in cultured trabecular meshwork cells. Histochemical studies revealed that bovine trabecular meshwork cells stained prominently for acid lipase, acid phosphatase, and acid esterase activities. Following phagocytic challenges, such as incubations with latex microspheres or zymosan particles, lysosomal enzyme staining was visibly enhanced. The uptake of foreign particles by trabecular-meshwork cells was evident. Data from biochemical assays for acid lipase, cholesteryl esterase, and acid esterase confirmed the histochemical findings. These results indicate that trabecular meshwork cells are actively phagocytic in culture and that lysosomal enzyme levels in these cells may be modulated by phagocytic challenge.

Acid Phosphatase↗

Cultured trabecular-meshwork cells: immunohistochemical and lectin-binding characteristics.

We examined the extracellular matrix formation and lectin-binding properties of cultured trabecular-meshwork cells established from cynomolgus monkey and bovine eyes. Using an avidin-biotin complex method, we found that the extracellular matrix in both the monkey and bovine cultures stained intensely with antibodies to fibronectin, type IV collagen, and laminin. These materials were especially prominent when the cultured monkey trabecular-meshwork cells aggregated in clusters, manifesting their close anatomic relationship to trabecular beams. Cell-surface and intracellular lectin-binding properties were studied using eight biotinylated lectins: concanavalin A (Con A), peanut agglutinin (PNA), Ricinus communis agglutinin (RCA), Dolichos biflorus agglutinin (DBA), soy bean agglutinin (SBA), Phaseolus vulgaris (PHA), wheat germ agglutinin (WGA), and Ulex europaeus agglutinin I (UEA I). Both monkey and bovine trabecular-meshwork cells showed positive cell surface and intracellular binding to Con A, WGA, and PHA. Only moderate cell-surface staining was observed with RCA, and no visible staining occurred with PNA, DBA, SBA, and UEA-1.

Animals↗

Neoplastic angioendotheliomatosis. A variant of malignant lymphoma immunohistochemical and ultrastructural observations of three cases.

Neoplastic angioendotheliomatosis (NAE) is a rare, fatal disease characterized by widespread intravascular proliferations of malignant cells of putative endothelial origin. Clinically, dermatologic and bizarre neurological manifestations predominate, but review of the reported cases of NAE reveals ophthalmic involvement to be frequent. To our knowledge, no reports of NAE have appeared in the ophthalmic literature. We describe three cases of NAE with the ocular manifestations of visual loss, cells in the vitreous, retinal artery occlusion, retinal vascular and pigment epithelial alterations, nystagmus, and cortical blindness. Autopsies (including eyes and central nervous system) revealed pancorporal involvement by intravascular anaplastic cells in each patient. In two patients massive extravascular involvement was also present. The tumor cells lacked ultrastructural features of endothelial cells and failed to stain for factor-VIII-related antigen. Common leukocyte antigen, a maker for hematopoietic cells, particularly lymphocytes, was detected on tumor cells in all cases, indicating that NAE is probably an extranodal lymphoma. The dramatic response of the central nervous system lesions to radiotherapy in one case supports this contention. It is suggested that this disorder be treated as a malignant lymphoma.

Aged↗

Sarcomas arising after radiotherapy for peptic ulcer disease.

Therapeutic gastric irradiation has been used to reduce peptic juice secretion in patients with peptic ulcer disease. Between 1937 and 1968 a total of 2049 patients received such therapy at the University of Chicago. Three of these patients are known to have developed sarcomas in the field of radiation. Two gastric leiomyosarcomas of the stomach were diagnosed 26 and 14 years after treatment and a malignant fibrous histiocytoma of the anterior chest wall was removed six years after gastric irradiation. Of 743 peptic ulcer patients treated without irradiation and constituted as a control group for the study of therapeutic gastric radiation, none is known to have developed sarcoma. As the incidence of sarcoma in these patient groups is known only from the tumor registry of the University of Chicago, other cases of sarcoma may exist in the groups. While an increased incidence of sarcoma has not been proven to occur in patients who received therapeutic gastric irradiation for peptic ulcer disease, the possibility of such a risk should be borne in mind by physicians caring for such patients.

Adult↗