Biomedical subjects

C J Murphy

Publications and source records attributed to C J Murphy.

At least 37 records · Page 2Linked to original sources

PubMed2000

Time-resolved spectral observations of cadmium-enriched cadmium sulfide nanoparticles and the effects of DNA oligomer binding.

We measured the steady-state and time-resolved fluorescence spectral properties of cadmium-enriched nanoparticles (CdS-Cd2+). These particles displayed two emission maxima, at 460 and 580 nm. The emission spectra were independent of excitation wavelength. Surprisingly, the intensity decays were strongly dependent on the observation wavelength, with longer decay times being observed at longer wavelengths. The mean lifetime increased from 150 to 370 ns as the emission wavelength was increased from 460 to 650 nm. The wavelength-dependent lifetimes were used to construct the time-resolved emission spectra, which showed a growth of the long-wavelength emission at longer times, and decay-associated spectra, which showed the longer wavelength emission associated with the longer decay time. These nanoparticles displayed anisotropy values as high as 0.35, depending on the excitation and emission wavelengths. Such high anisotropies are unexpected for presumably spherical nanoparticles. The anisotropy decayed with two correlation times near 5 and 370 ns, with the larger value probably due to overall rotational diffusion of the nanoparticles. Addition of a 32-base pair oligomer selectively quenched the 460-nm emission, with less quenching being observed at longer wavelengths. The time-resolved intensity decays were minimally affected by the DNA, suggesting a static quenching mechanism. The wavelength-selected quenching shown by the nanoparticles may make them useful for DNA analysis.

Biopolymers↗

PubMed2000

Adhesion and proliferation of corneal epithelial cells on self-assembled monolayers.

The effect of surface chemistry on the proliferation and adhesion of SV-40 human corneal epithelial cells was investigated. The surface chemistry of substrates was controlled by the deposition of self-assembled monolayers (SAMs) terminated with the following functional groups: -CF3, -CH3, -CO(2)H, and -NH(2). SAMs of alkanethiols on gold and of alkylsiloxanes on SiOx were included in the study. Comparisons are made between different types and functionalities of SAMs and between SAM-covered substrates and tissue culture polystyrene. Adhesion assays were performed after incubation of the cells for 1 h in 10% fetal bovine serum and in serum-free conditions. The cellular response was found to be a function of surface chemistry and the presence of exogenous proteins. The number of cells that adhered to most of the SAMs in 10% serum and in serum-free conditions was not significantly different from the number of cells that adhered to TCPS. Proliferation assays were carried out in 10% serum and in 0.5% serum. Cell behavior was influenced by surface chemistry but did not deviate significantly from the behavior on TCPS for most of the SAMs. Serum level did not play a major role in cell proliferation. Our data establish the expected behaviors for a corneal epithelial cell line under defined conditions on specific surfaces.

Adhesiveness↗

PubMed2000

Nanoscale topography of the basement membrane underlying the corneal epithelium of the rhesus macaque.

This paper quantitatively defines the nanoscale topography of the basement membrane underlying the anterior corneal epithelium of the macaque. Excised corneal buttons from macaques were placed in 2.5 mM ethylenediaminetetraacetate (EDTA) for 2.5 h, after which the epithelium was carefully removed to expose the underlying basement membrane. The integrity of the remaining basement membrane was verified using fluorescent microscopy in conjunction with antibody staining directed against laminin and collagen type IV as well as transmission electron microscopy. Characterization of the surface of the basement membrane was performed using transmission electron microscopy, high-resolution, low-voltage scanning electron microscopy, and atomic force microscopy. Quantitative data were obtained with all three imaging techniques and compared. The basement membrane has a complex topography consisting of tightly cross-linked fibers intermingled with pores. The mean elevation of features measured by transmission electron microscopy, scanning electron microscopy, and atomic force microscopy was 149 +/- 60 nm, 191 +/- 72 nm, and 147 +/- 73 nm, respectively. Mean fiber diameter as measured by SEM was 77 +/- 44 nm and pore diameter was 72 +/- 40 nm, with pores occupying approximately 15% of the total surface area. Similar feature types and dimensions were also found for Matrigel, a commercially available basement membrane-like complex, supporting that a minimum of artifact was introduced by corneal preparative procedures to remove the overlying epithelium. Topographic features amplified the surface area over which cell-substratum interactions occur by an estimated 400%. The three-dimensional structure of the basement membrane exhibits a rich complex topography of individual features, consisting of pores and fibers with dimensions ranging from 30 to 400 nm. These nanoscale substratum features may modulate fundamental cell behaviors such as adhesion, migration, proliferation, and differentiation.

Animals↗

PubMed2000

Nanoscale topography of the corneal epithelial basement membrane and Descemet's membrane of the human.

PURPOSE: Quantitatively define and compare the nanoscale topography of the corneal epithelial basement membrane (anterior basement membrane) and Descemet's membrane (posterior basement membrane) of the human. METHODS: Human corneas not suitable for transplantation were obtained from the Wisconsin Eye Bank. The corneas were placed in 2.5 mM EDTA for 2.5 h or 30 min. for removal of the epithelium or endothelium, respectively. After removal of the overlying cells, specimens were fixed in 2% glutaraldehyde and either examined in this state by atomic force microscopy only or dehydrated through an ethanol series and prepared for transmission electron microscopy (TEM), scanning electron microscopy (SEM), and atomic force microscopy (AFM). RESULTS: The subepithelial and subendothelial basement membrane surfaces have a similar appearance that consists of an interwoven meshwork of fibers and pores. Topographic feature sizes were found to be in the nanometer size range with the epithelial basement membrane features larger and less densely packed than Descemet's membrane features. The topographic features are fractile in nature and increase surface area for cell contact. CONCLUSION: With the use of the TEM, SEM, and AFM, a detailed description of the surface topography of corneal epithelial basement membrane and Descemet's membrane of the human cornea are provided. The significance of differences in corneal basement membrane topography may reflect differences in function of the overlying cells or may be related to differences in cell migration and turnover patterns between the epithelium and endothelium.

Aged↗

PubMed2000

Effect of topical administration of epidermal growth factor on healing of corneal epithelial defects in horses.

UNLABELLED: OBJECTIVE-To characterize healing of corneal epithelial defects in horses and to evaluate the ability of epidermal growth factor (EGF) to modulate rate of corneal epithelial healing in horses. SAMPLE POPULATION: 20 eyes in 12 adult horses. PROCEDURE: Corneal epithelial wounds were created by mechanically debriding the limbus. Corneal healing was recorded for 3 treatment groups: 50 microg of EGF/ml (n = 5 eyes), 5 microg of EGF/ml (7), and PBS solution (8). Corneal healing was recorded once daily after instillation of fluorescein stain by use of photography and calculating the area of the wound, using imaging software. RESULTS: After corneal debridement, re-epithelialization was rapid and progressed in a linear fashion for the first 5 to 7 days after surgery in all groups. After that period, rates of healing decreased. A profound increase in the degree of inflammation, neovascularization, melanosis, and scarring was observed in eyes treated with the high dose of EGF (50 microg/ml), but there was not a statistical difference in mean healing time or in mean decrease in radius during the linear phase between the control and either EGF treatment groups. However, for all 8 horses in which both eyes were debrided, the first eye healed significantly faster than the second eye, regardless of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Beneficial effects of topical administration of a high dose of EGF for acceleration of healing of corneal defects in eyes of horses are outweighed by the intensity of the associated inflammatory response.

Administration, Topical↗

PubMed2000

Iridology.

Explore the source record for details and available documents.

Diagnostic Techniques, Ophthalmological↗

PubMed1999

Usefulness of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals.

OBJECTIVE: To determine the diagnostic value of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis (IUK). DESIGN: Prospective study. ANIMALS: 48 animals (26 dogs, 13 horses, 7 cats, 1 bird, and 1 llama) with corneal ulcers. PROCEDURE: Scrapings from corneal ulcers were examined cytologically. Corneal swab specimens were submitted for microbial culture. Animals were grouped according to whether they had been receiving antimicrobials at the time of admission. RESULTS: Of the 38 animals receiving antimicrobials, 19 had positive results for IUK on cytologic evaluation, 20 on microbial culture, and 26 on cytologic evaluation, microbial culture, or both. Of the 10 animals not receiving antimicrobials at the time of admission, 7 had positive results for IUK on cytologic evaluation, and 9 had positive results on microbial culture. In this group of 10 animals, additional animals with IUK were not identified on the basis of cytologic evaluation alone. When all 48 animals were considered irrespective of antimicrobial treatment, 26 and 29 had positive results for IUK on cytologic evaluation and microbial culture, respectively, whereas IUK was confirmed in 35 animals on the basis of cytologic evaluation, microbial culture results, or both. CONCLUSIONS AND CLINICAL RELEVANCE: Microbial culture and cytologic evaluation of corneal specimens maximizes identification of IUK, especially in animals receiving antimicrobial treatment. Because of serious consequences of untreated IUK, we recommend that both diagnostic tests be used to tailor treatment and reduce risk of vision impairment in animals.

Animal Diseases↗

PubMed1999

Combined cycloablation and gonioimplantation for treatment of glaucoma in dogs: 18 cases (1992-1998).

OBJECTIVE: To evaluate a combined cycloablative and gonioimplantation technique for treatment of glaucoma in dogs. DESIGN: Retrospective study. ANIMALS: 18 adult dogs with glaucoma. PROCEDURE: Medical records of dogs that received a valved gonioimplant and a cyclodestructive procedure (cyclocryoablation or diode laser cyclophotocoagulation) during a 6-year period were reviewed. Retention of vision and intraocular pressure control were assessed, as well as number and nature of complications. RESULTS: 19 eyes of 18 dogs received a valved gonioimplant and either cyclocryoablation (n = 12) or diode laser cyclophotocoagulation (7). At > or = 1 year after surgery, 11 of 19 eyes had vision and 14 of 19 eyes had intraocular pressure < 25 mm Hg. Two dogs (2 eyes) were lost to follow-up 3 and 6 months after surgery. Despite the alternative route for aqueous humor flow created by the gonioimplant, 7 eyes had increased intraocular pressure (27 to 61 mm Hg) < 24 hours after surgery. Other complications included excessive intraocular fibrin, focal retinal detachment, corneal ulcer, retinal hemorrhage, cataract, and implant migration. CONCLUSIONS AND CLINICAL RELEVANCE: Combined cycloablation and gonioimplantation appears to be a promising technique for retention of vision and control of intraocular pressure in dogs with glaucoma.

Animals↗

PubMed1999

Effects of synthetic micro- and nano-structured surfaces on cell behavior.

Topographical cues, independent of biochemistry, generated by the extracellular matrix may have significant effects upon cellular behavior. Studies have documented that substratum topography has direct effects on the ability of cells to orient themselves, migrate, and produce organized cytoskeletal arrangements. Basement membranes are composed of extracellular matrix proteins and found throughout the vertebrate body, serving as substrata for overlying cellular structures. The topography of basement membranes is a complex meshwork of pores, fibers, ridges, and other features of nanometer sized dimensions. Synthetic surfaces with topographical features have been shown to influence cell behavior. These facts lead to the hypothesis that the topography of the basement membrane plays an important role in regulating cellular behavior in a manner distinct from that of the chemistry of the basement membrane. This paper describes the topography of the basement membrane and reviews the fabrication of synthetic micro- and nano-structured surfaces and the effects of such textured surfaces on cell behavior.

Animals↗

PubMed1999

Glutamine metabolism by lymphocytes, macrophages, and neutrophils: its importance in health and disease.

Many aspects of the cell biology of lymphocytes, macrophages, and neutrophils have been studied extensively. Our recent work on these cells has investigated how fuel metabolism, especially glutamine metabolism, is related to the specific function of these cells in the inflammatory response. The high rate of glutamine utilization and its metabolism in such immune cells has raised the question of why glutamine is responsible for these functions. The macrophage has access to a variety of metabolic fuels both in vivo and in vitro. The quantitatively important role of glutamine in the processes of free radical and cytokine production has been established in our laboratories. Our current understanding of the rate of utilization and the pathway of metabolism of glutamine by cells of the immune system raises some intriguing questions concerning therapeutic manipulation of utilization of this amino acid, specifically the phagocytic and secretory capacities of cells of the defense system can be beneficially altered.

Journal Article↗

PubMed1999

Accommodation in the cuttlefish (Sepia officinalis).

We have studied natural accommodation in the eye of six specimens of cuttlefish (Sepia officinalis) as they were fed with fish and shrimp. Using infrared photoretinoscopy, we observed (1) that the resting refractive state of the cuttlefish was emmetropic or slightly hyperopic, (2) that accommodation took place only a fraction of a second before a strike and (3) that accommodation focused selectively only in the frontal visual field while no change in refraction could be measured in the lateral field of view. Accommodation was bilateral and amounted to approximately 5 diopters (the reciprocal of the focal length expressed in meters). Simultaneously, the eyes converged. It appears that, as in most teleost fishes, accommodation in the cuttlefish involves a movement of the crystalline lens perpendicular to the axis of the eye. In histological sections, we observed the position and arrangement of the ciliary muscles, confirming earlier anatomical descriptions, and developed a model of how accommodation could be achieved.

Adaptation, Physiological↗

PubMed1999

Naturally occurring vitreous chamber-based myopia in the Labrador retriever.

PURPOSE: To investigate whether myopia is present in a breed of domestic dog, the Labrador retriever, and how the ocular components are related to refractive error in this breed. METHODS: Cycloplegic refractive error was measured in 75 Labrador retrievers by retinoscopy. Corneal and crystalline lens radii of curvature were measured in the right eyes of 57 of these dogs using a video-based keratophakometer, with axial ocular dimensions measured using A-scan ultrasonography. RESULTS: Of the 75 dogs tested, 11 (14.7%) were myopic by at least -0.50 D in one eye, and 6 (8.0%) were myopic in both eyes (full range of refractive errors, +3.50 D to -5.00 D). Of the 57 dogs with ocular component measurements, seven (12.3%) were myopic by at least -0.50 D in the right eye. There was a significant negative correlation between refractive error and vitreous chamber depth (Spearman r = -0.42; P < 0.001). Myopic eyes had an elongated vitreous chamber depth (10.87+/-0.34 mm for myopic dogs, 10.02+/-0.40 mm for nonmyopic dogs; P < 0.0001, Kruskal-Wallis test). There was also a significant quadratic association between lens thickness and vitreous chamber depth (P < 0.005; R2 = 0. 11), indicating that thinner lenses occurred at both shorter and longer vitreous chamber depths. CONCLUSIONS: Myopia in the Labrador retriever is analogous to human myopia in that it is caused by an elongated vitreous chamber. Thinner crystalline lenses found at longer vitreous chamber depths may be analogous to lens thinning documented in human ocular development. The Labrador retriever warrants investigation as a potential model of myopia that is naturally occurring rather than experimentally induced.

Animals↗

PubMed1998

The neuronal growth-associated protein GAP-43 interacts with rabaptin-5 and participates in endocytosis.

Structural plasticity of nerve cells is a requirement for activity-dependent changes in the brain. The growth-associated protein GAP-43 is thought to be one determinant of such plasticity, although the molecular mechanism by which it mediates dynamic structural alterations at the synapse is not known. GAP-43 is bound by calmodulin when Ca2+ levels are low, and releases the calmodulin when Ca2+ levels rise, suggesting that calmodulin may act as a negative regulator of GAP-43 during periods of low activity in the neurons. To identify the function of GAP-43 during activity-dependent increases in Ca2+ levels, when it is not bound to calmodulin, we sought proteins with which GAP-43 interacts in the presence of Ca2+. We show here that rabaptin-5, an effector of the small GTPase Rab5 that mediates membrane fusion in endocytosis, is one such protein. We demonstrate that GAP-43 regulates endocytosis and synaptic vesicle recycling. Modulation of endocytosis by GAP-43, in association with rabaptin-5, may constitute a common molecular mechanism by which GAP-43 regulates membrane dynamics during its known roles in activity-dependent neurotransmitter release and neurite outgrowth.

Brain Chemistry↗

PubMed1998

Orbital lymphosarcoma associated with reticuloendotheliosis virus in a peafowl.

Lymphosarcoma associated with infection by avian reticuloendotheliosis virus was diagnosed in an Indian peafowl with exophthalmia and exposure keratitis. Exenteration of the orbit was complicated by a profound oculocardiac reflex and extensive hemorrhage during surgery. Orbital bleeding was controlled by direct pressure, electrocautery, topical administration of bovine thrombin, and application of sterile gelatin sponges and oxidized regenerated cellulose. A blood transfusion was also performed. In addition to describing methods of handling intraoperative complications of orbital exenteration in birds, to the authors' knowledge, this is the first report to describe an association of reticuloendotheliosis virus, which more commonly affects poultry, with lymphosarcoma in an Indian peafowl.

Animals↗

PubMed1998

Effect of intracameral administration of carbachol on the postoperative increase in intraocular pressure in dogs undergoing cataract extraction.

OBJECTIVE: To determine whether intracameral injection of carbachol at the completion of phacoemulsification in dogs would prevent the increase in intraocular pressure (i.o.p) that can develop during the first 24 hours after surgery. DESIGN: Randomized controlled trial. ANIMALS: 32 adult dogs undergoing elective unilateral or bilateral phacoemulsification. PROCEDURE: Dogs were randomly assigned to 1 of 4 groups with 8 dogs/group: phacoemulsification and intracameral administration of 0.5 ml of 0.01% carbachol at the end of surgery; phacoemulsification, intraocular lens implantation, and intracameral administration of 0.5 ml of 0.01% carbachol; phacoemulsification and intracameral administration of 0.5 ml of balanced salt solution; and phacoemulsification, intraocular lens implantation, and intracameral administration of 0.5 ml of balanced salt solution. Intraocular pressure was measured at 3 and 6 hours and the morning after surgery. Aqueous flare was also measured 6 hours and the morning after surgery. RESULTS: None of the dogs treated with carbachol developed postoperative ocular hypertension (i.e., IOP > 27 mm of Hg), whereas 12 of 16 control dogs had ocular hypertension 3 hours after surgery. Intraocular pressure 3 hours after surgery was not significantly associated with phacoemulsification time or phacoemulsification power or with whether the dog received an intraocular lens implant. Severity of aqueous flare was similar for treated and control dogs. CLINICAL IMPLICATIONS: Results suggested that intracameral administration of 0.01% carbachol at the end of surgery was a safe and efficacious method of preventing the postoperative increase in IOP associated with phacoemulsification in dogs.

Animals↗

PubMed1998

The effect of continuous light on refractive error and the ocular components of the rat.

Phototoxic induced degeneration of the rat retina is a well-documented phenomenon resulting in losses of photoreceptors and their cell bodies, and an overall retinal thinning. This process may serve as a test of the hypothesis that the retinoscopic reflex originates from the inner limiting membrane of the retina. Retinal thinning should produce myopia in the absence of any other ocular component changes in a stable, mature eye if the inner limiting membrane model is correct. Phototoxic retinal degeneration was induced in 10 albino rats by exposure to 19 days of continuous light (1,800 cd m-2). Another 10 albino rats exposed to 12-hr on/12-hr off cycled light served as controls. Before and after the exposure to constant light, measures were made of refractive state by cycloplegic retinoscopy, corneal curvature and lens curvature by Purkinje image photography, and axial length of the globe by A-scan ultrasonography. Comparing pre- to post-exposure values, phototoxic degeneration resulted in a mean (+/- S.D.) myopic shift of -5.10 +/- 2.12 D (P < 0.002). The corneal curvature also steepened significantly (0.17 +/- 0.11 mm, equivalent to -6.0 D; P < 0.004), while the posterior curvature of the crystalline lens flattened by 0.21 +/- 0.22 equivalent mm (P < 0.027), and the axial length shortened by 0.11 +/- 0.11 equivalent mm (all tests Wilcoxon signed-rank; P < 0.025). Phototoxic rats underwent a mean retinal thinning of 49.6 mu compared to controls (Kruskal-Wallis test; P < 0.0008). No refractive or ocular parameters changed significantly in the controls. Phototoxic degeneration in the rat has optical consequences beyond simple retinal thinning. The size of the eye and the curvature of refractive surfaces can be altered in a mature eye well after the completion of development. The multiple changes which occur prevent phototoxic retinal thinning from serving as a test of the inner limiting membrane model for retinoscopic reflections.

Animals↗

PubMed1998

Effect of substance P, insulin-like growth factor-1 and vasoactive intestinal polypeptide on corneal re-epithelialization in galactosemic rats.

PURPOSE: The purpose of the study was to examine the effect of topical application of substance P (SP), insulin-like growth factor-1 (IGF-1) and vasoactive intestinal polypeptide (VIP) on corneal re-epithelialization in galactosemic rats. METHODS: Experimental galactosemia was induced by feeding a diet containing 30% galactose for 4-6 months. The corneal epithelium was debrided bi-laterally by scraping with a blunted scalpel blade. One eye (control) received only a saline solution whilst the other eye received a solution of SP and/or IGF-1 or VIP. A single drop of control or test solution was administered 4 times daily until wound closure. Corneas were stained with fluorescein and videotaped under ultraviolet illumination at regular time intervals after debridement. After digitizing the video image, the wound area was calculated using an image analysis program (NIH Image). RESULTS: Corneal re-epithelialization was significantly delayed in galactosemic compared to normal animals. Rates of healing were 1.3 +/- 0.07 mm2/hour and 1.02 +/- 0.02 mm2/hour for normal and galactosemic animals, respectively. Topical application of SP in concentrations ranging from 25 pg/ml up to 250 microg/ml had no significant effect on the rate of corneal re-epithelialization. Similarly, IGF-1 (1 microg/ml) or VIP (1 microg/ml) when applied alone did not affect re-epithelialization. Furthermore, resurfacing of the debrided area was not affected by co-application of SP (250 microg/ml) and IGF-1 or VIP. CONCLUSIONS: Independent or combined topical application of SP, VIP or IGF-1 at the concentrations tested, does not modulate corneal epithelial wound healing in rats with galactosemia induced by 30% galactose.

Animals↗

PubMed1998

The effect of elevated extracellular glucose on migration, adhesion and proliferation of SV40 transformed human corneal epithelial cells.

PURPOSE: To examine the effect of elevated extracellular glucose, thus simulating diabetes, on migration, adhesion and proliferation of SV40 transformed human corneal epithelial (HCE) cells. METHODS: HCE cells were maintained in serum supplemented media containing 5 mM, 17.5 mM or 38 mM D-glucose. Cell migration was determined using Blind well chambers fitted with fibronectin/collagen I coated filters. In adhesion experiments, cells were allowed to adhere to extracellular matrix protein-coated wells for 90 min at 37 degrees C. Non-adherent cells were removed by washing, then the fluorochrome calcein-AM was added to quantitate the number of attached cells. Proliferation was determined by plating the cells at low density, then quantitating viable cells with calcein-AM 5 to 7 days later. RESULTS: Raising extracellular glucose from 5 mM to 17.5 mM significantly increased cell migration by 42%. When glucose was further raised to 38 mM, migration was not significantly different from that in 5 mM glucose. Adhesion to fibronectin and collagen I (but not IV) was significantly increased (62% and 32% respectively) when cells were cultured in 17.5 mM glucose. Similarly, proliferation was increased by 44%. Adhesion and proliferation tended to be decreased at 38 mM compared to 17.5 mM glucose, but not significantly so. In the presence of 5 mM glucose and mannitol (12.5 mM or 33 mM), neither migration, adhesion nor proliferation were significantly different from that in 5 mM glucose alone. CONCLUSION: Elevated extracellular glucose modulates migration, adhesion and proliferation of HCE cells. The effects are dependent on the concentration of glucose and are not due to changes in osmolality since mannitol failed to produce similar results. Our in vitro findings suggest that high-glucose effects may directly contribute to the etiology of impaired corneal wound healing in diabetes.

Cell Adhesion↗