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Biomedical subjects

C A Wilson

Publications and source records attributed to C A Wilson.

At least 19 recordsLinked to original sources

Blood-retinal barrier breakdown caused by diode vs argon laser endophotocoagulation.

We compared the effects of argon and diode laser endophotocoagulation on blood-retinal barrier breakdown using real-time magnetic resonance imaging following intravenous gadolinium-diethylenetriaminepentaacetic acid (Gd-DTPA) injection. Endophotocoagulation was performed on eyes of pigmented rabbits with either the argon or the diode laser to produce ophthalmoscopically similar lesions. Magnetic resonance imaging studies were performed either 2 or 7 days after laser treatment, and coronal T1-weighted proton images were obtained in the first 20 minutes following Gd-DTPA injection. The mean signal intensity over a region of interest in the vitreous cavity was analyzed, and an initial rate analysis was performed on each time-course curve. Two days after treatment, argon laser-treated eyes showed significantly greater leakage of Gd-DTPA than diode laser-treated eyes. The leakage in both groups was substantially reduced by posttreatment day 7. Histopathologic examination performed 2 days following photocoagulation showed less damage of the retinal pigment epithelium and more severe occlusion of the choriocapillaris and deep choroidal vessels in diode laser-treated eyes. These changes may serve to explain the observed differences in Gd-DTPA leakage.

Animals

Measurement of preretinal oxygen tension in the vitrectomized human eye using fluorine-19 magnetic resonance spectroscopy.

We obtained oxygen measurements from a human eye that contained small preretinal droplets of perfluorotributylamine (FTBA) by using fluorine-19 magnetic resonance spectroscopy. These droplets were the remainder of a larger volume of FTBA that was used as an intraoperative retinal tamponade during retinal detachment repair. The spin-lattice relaxation rate ([T1]-1) of the FTBA fluorine nuclide was obtained that could then be related, by direct proportionality, to droplet PO2. With the patient in a supine position, the droplets could be positioned over the macula in the preretinal vitreous space, whereon the FTBA PO2 could be influenced by the preretinal oxygen concentration. Preretinal PO2 values, derived from magnetic resonance spectroscopy, were in the range of 6 to 9 mm Hg, although multiple components were identified that were suggestive of a heterogeneous distribution of PO2 values in the population of droplets. To our knowledge, this investigation is the first to demonstrate the feasibility of this approach to performing long-term, noninvasive preretinal oxygen measurements in the vitrectomized human eye by using small droplets of a liquid perfluorochemical.

Adult

Treatment with intravitreal steroid reduces blood-retinal barrier breakdown due to retinal photocoagulation.

The effect of corticosteroid treatment on blood-retinal barrier breakdown caused by argon-laser panretinal photocoagulation was evaluated in the rabbit eye. One day before photocoagulation, eyes were given either a sub-Tenon (20-mg) or intravitreal (2-mg) injection of triamcinolone acetonide. The severity of blood-retinal barrier breakdown was measured after photocoagulation using rapid sequential magnetic resonance imaging following intravenous administration of gadolinium diethylenetriaminepentaacetic acid. Leakage of gadolinium diethylenetriaminepentaacetic acid into the vitreous space was significantly lower in eyes that received intravitreal triamcinolone acetonide than in control eyes (P = .007); however, sub-Tenon triamcinolone acetonide produced no significant reduction in leakage (P = .65) compared with controls. Fluorescein angiography supported the magnetic resonance imaging findings. We conclude that retinal photocoagulation in the rabbit eye produces blood-retinal barrier breakdown that is partially amenable to corticosteroid treatment.

Animals

Oxygen kinetics in preretinal perfluorotributylamine.

Previous studies have relied on various electrodes or probes to monitor preretinal oxygen tension in an effort to gain insight into retinal oxygenation. In order to corroborate and extend the results of such studies, we developed a relatively non-invasive method of determining preretinal oxygen tension using 19F nuclear magnetic resonance (NMR) spectroscopy. Small liquid perfluorocarbon (LPFC) droplets were injected into the preretinal vitreous space of the rabbit eye. The T1 value obtained from the fluorine nuclide could then be used to determine preretinal oxygen tension (PO2) with a high degree of sensitivity, since the fluorine spin-lattice relaxation rate (T1)-1 in LPFCs is directly proportional to PO2 under conditions of no flow and known temperature. In the present study, we investigated the oxygen uptake and clearance rates from small preretinal droplets of the LPFC perfluorotributylamine (FTBA) in response to step changes in arterial PO2. At all FTBA volumes examined (2, 10 and 100 microliters), the oxygen uptake and clearance curves were well approximated by a simple exponential equation with mean time constants 9.8/15.3, 21.4/19.4 and 77.7/45.3 min (uptake/clearance), respectively. Following return to normoxemic (baseline) conditions, FTBA droplets provided a preretinal PO2 of 39.4 +/- 9.2 mmHg (mean +/- S.D., n = 12). The 19F NMR method provides a measure of steady-state preretinal PO2 that independently verifies and complements information obtained using oxygen-sensitive microelectrodes or probes. However, the long time constants for oxygen uptake and clearance, particularly in FTBA volumes on the order of 10 microliters and greater, may represent a practical limitation of this method for determining rapid oxygen flux in the preretinal vitreous space.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Behavioural effects in adulthood of neonatal manipulation of brain serotonin levels in normal and androgenized females.

5HT concentrations in the hypothalamus are higher in females than males over the second week of life and this differentiation is testosterone-dependent. We have investigated the possible influence of 5HT over this period on the development of systems that control adult behaviour, in particular those influenced by neonatal testosterone. Neonatal androgenization (250 micrograms/pup testosterone propionate; TP; on day 1 postpartum) induced a masculine pattern of behaviour in females ovariectomised in adulthood and bearing a TP implant. The neonatal treatment reduced exploration, motor activity and female sexual behaviour and increased anxiety, orientation toward the incentive female and male sexual behaviour. Depletion of 5HT by pCPA (100 mg/kg days 8-16 postpartum) enhanced the TP-induced increment in locomotion and female sexual behaviour and increased sexual orientation toward the incentive female, while 5HTP (20 mg/kg days 8-16 postpartum) antagonised the reduction in exploration by TP. Thus 5HT may normally exert an inhibitory control on the action of neonatal testosterone on exploration, motor activity and sexual behaviour. Neonatal PCPA treatment also had a marked anxiolytic effect which was independent of the presence of T as it was noted in normal and androgenized females and previously has been observed in intact males. This might indicate a primary control by a serotonergic system on the development of the systems controlling anxiety.

5-Hydroxytryptophan

The requirements for viral entry differ from those for virally induced syncytium formation in NIH 3T3/DTras cells exposed to Moloney murine leukemia virus.

Moloney murine leukemia virus (Mo-MuLV) has the unique ability to infect different cells via either a low-pH-dependent or a pH-independent entry pathway. Only the pH-independent mechanism of Mo-MuLV entry has been associated with Mo-MuLV-induced syncytium formation. We have now identified a transformed cell line (NIH 3T3/DTras) which efficiently forms syncytia when exposed to Mo-MuLV, yet is low pH dependent for Mo-MuLV entry. Treatment of NIH 3T3/DTras cells with chloroquine, an agent which raises endosomal pH, blocks Mo-MuLV entry, but not Mo-MuLV-induced syncytium formation. This demonstrates that fusion which accompanies viral entry and fusion which is responsible for syncytium formation occur as independent processes in these cells. In addition, we determined that neither inherent differences in the Mo-MuLV receptor nor reduced affinity for Mo-MuLV gp70 can account for resistance of NIH 3T3 cells to Mo-MuLV-induced syncytium formation.

3T3 Cells

In vivo effects of interleukin-1 beta on blood leukocytes in BB rats prone or resistant to diabetes.

Previous studies have determined that daily low dose injections of the potent cytokine interleukin-1 beta (IL-1 beta) decreased the frequency of insulin-dependent diabetes mellitus (IDDM) in diabetes-prone (DP) BB rats. In contrast, high dose injections induced an earlier than normal onset. In this study we tested whether the effects of daily human recombinant IL-1 beta injections on leukocyte subsets were associated with its modulation of IDDM onset in BB rats. Prior to the onset of IDDM in DP BB rats, high dose IL-1 beta induced leukocytosis (P less than 0.05), neutrophilia (P less than 0.01), and monocytosis (P less than 0.001). At the onset of IDDM, lymphocyte (P less than 0.01) and neutrophil (P less than 0.001) numbers were increased in high dose treated DP rats but not in rats given saline or low dose IL-1 beta. In 60-day-old diabetes-resistant (DR) BB rats, neurophilia was induced by both low (P less than 0.05) and high (P less than 0.001) dose IL-1 beta without the development of IDDM. At 130 days of age, when the rats were killed, it was discovered that 14/22 (64%) IL-1 beta injected DR rats developed neutralizing IL-1 beta antibodies. Significantly lower neutrophil numbers were observed in high dose DR rats which developed IL-1 beta antibodies compared with those which did not (P = 0.032). Thus, neutrophilia was dissociated from high IL-1 beta acceleration of IDDM onset.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Quantitative determination of the partial oxygen pressure in the vitrectomized rabbit eye in vivo using 19F NMR.

We have quantitatively determined the PO2 within the vitreous space of the vitrectomized rabbit eye in vivo using 19F NMR and perfluorotributylamine (FTBA). The results of the present work are in good agreement with previous measurements of PO2 within the nonvitrectimized rabbit eye obtained using oxygen microelectrodes. In this study, good precision and accuracy were achieved by: (1) having minimal flow effects present, (2) optimizing the signal-to-noise through the use of neat FTBA and surface coil transmission and detection, and (3) performing an inversion-recovery pulse sequence, with adiabatic pulses, to optimize the dynamic range of the T1 experiment. Possible deleterious effects on the measured T1 due to the vitreal temperature gradient are discussed. To the best of our knowledge the results of this study demonstrate, for the first time, a quantitative determination of intraocular PO2 in vivo using 19F NMR and FTBA.

Animals

Experimental traction retinal detachment in the cat.

We developed a reproducible model of traction retinal detachment (TRD) in the cat eye by creating a serous retinal detachment and then injecting 2.5 x 10(5) kitten dermal fibroblasts into the vitreous cavity at the site of a retinal wound. Serous detachments were produced by exposing an area of retina to focused light after intravenous injection of rose bengal (a photosensitizing dye). TRD developed rapidly within the first 2 weeks after fibroblast injection, accompanied by the formation of vitreoretinal strands and, to a lesser degree, epiretinal and/or subretinal proliferation. Histopathology demonstrated fibroblasts within the vitreous or along the posterior hyaloid face. Focal deposits of fibroblasts were occasionally found on the inner surface of the retina and/or in the subretinal space. Fibroblast proliferation was confirmed by uptake of radiolabeled thymidine. Deposition of collagen was noted at as early as 3 days after fibroblast injection. Neovascularization was not observed. Control eyes that did not receive fibroblasts showed resolution of serous detachment without retinal traction. In all eyes, retinal degeneration and thinning were seen in the area of previous photodynamic treatment. In this model of TRD, anteroposterior traction (due to vitreous strands) predominates, as is observed in experimental posterior penetrating ocular injury induced by intravitreal blood injection, which also results in vitreous strand formation. Our model, however, enables clinical assessment of TRD in the cat without the media opacification produced by vitreous blood.

Animals

Interleukin-1 beta regulation of islet and thyroid autoimmunity in the BB rat.

Daily injections of high dose human recombinant interleukin-1 beta (IL-1 beta) accelerated the onset of both insulin-dependent diabetes mellitus and lymphocytic thyroiditis in genetically prone BB rats. In diabetes-resistant BB rats, high dose IL-1 beta failed to induce diabetes. Additionally, the presence of neutralizing IL-1 beta antibodies in these rats strongly correlated with inhibition of lymphocytic thyroiditis. Since low dose IL-1 beta protects diabetes-prone rats from IDDM, we conclude that IL-1 beta is a potent modulator of autoimmune diabetes and thyroid disease in genetically susceptible rats.

Adrenal Glands

Viral and cellular factors governing hamster cell infection by murine and gibbon ape leukemia viruses.

Hamster cells are resistant to infection by most retroviruses, including Moloney murine leukemia virus (MoMLV) and gibbon ape leukemia viruses (GaLVs). We have constructed MoMLV-GaLV hybrid virions to identify viral and cellular determinants responsible for the inability of GaLV and MoMLV to infect hamster cells. The substitution of MoMLV core components for GaLV core components circumvents the resistance of hamster cells to infection by GaLV, demonstrating that hamster cells have receptors for GaLV but are not efficiently infected by this primate retrovirus because of a postpenetration block. In contrast, hamster cells are apparently resistant to MoMLV infection because although they bear a receptor for MoMLV, the receptor is nonfunctional. Treatment of CHO K1 or BHK 21 hamster cells with the glycosylation inhibitor tunicamycin allows the cells to be infected by MoMLV. The construction of MoMLV-GaLV hybrid virions that can efficiently infect resistant cells has allowed the identification of viral and cellular factors responsible for restricting infection of hamster cells by MoMLV and GaLV.

3T3 Cells

Interaction of estradiol, alpha-melanocyte-stimulating hormone, and dopamine in the regulation of sexual receptivity in the female rat.

Ovariectomized and adrenalectomized rats kept in a reversed lighting system received either 5 micrograms (once) or 1 microgram estradiol benzoate (EB) daily for 2, 3, or 4 days. These treatments induced sexual receptivity in a proportion of the rats, and alpha-melanocyte-stimulating hormone (alpha-MSH; 20 micrograms/rat s.c.) enhanced this proportion. In rats made receptive by 5 micrograms EB alone, the dopamine (DA) activity in preoptic area and arcuate nucleus was significantly reduced, but the alpha-MSH concentrations were not affected 54-56 h after EB treatment. Addition of alpha-MSH significantly increased the DA activity in ventromedial nucleus (VMN) and zona incerta, but this action is unlikely to account for its stimulation of sexual receptivity, since this effect was not blocked by 0.1 mg/kg haloperidol. Treatment with 100 mg/kg Dopa, which induces a presynaptic DA action, stimulated the receptivity in the EB-primed rats and selectively increased the concentration of alpha-MSH in the VMN, while 50 mg/kg Dopa plus 50 mg/kg benserazide, which induces a postsynaptic DA activity, inhibited the receptivity and reduced the alpha-MSH levels in the VMN. It is suggested that estradiol stimulates the female sexual receptivity by reducing the activity of a dopaminergic system in arcuate nucleus and preoptic area. alpha-MSH does not appear to affect this action, although it enhances the effect of steroids on the sexual behaviour, probably at the level of the VMN. The secretion of alpha-MSH may be under a dopaminergic inhibitory control, and the peptide may autoregulate its own secretion via this dopaminergic system which is sited in zona incerta and VMN.

3,4-Dihydroxyphenylacetic Acid

Treatment of experimental preretinal neovascularization using photodynamic thrombosis.

Retinal or preretinal neovascularization (NV) is the result of many ischemic conditions of the retina and is an important factor leading to severe visual loss in diabetic retinopathy. Panretinal photocoagulation does not always control its growth or bleeding sequelae. A potential new treatment modality, photodynamic therapy (PDT), was evaluated for limiting the progression of experimental NV in the rabbit eye. The NV was produced by injecting cultured dermal fibroblasts into the preretinal vitreous space after combined enzymatic and mechanical vitreolysis. This method results in traction retinal detachment with a rapid and consistent growth of NV. After administration of the photosensitizing dye rose bengal (20 mg/kg intravenously), PDT was done using a slit-lamp light source focused through a fundus contact lens (45 J/cm2). The NV was treated on two separate occasions during the active phase of growth (on days 13 and 21 after fibroblast injection). Control animals were exposed to light before injection of rose bengal. Eight randomly assigned animals in each group were followed between treatments and for 28 days after the second treatment. The appearance of NV was documented by frequent photography and fluorescein angiography. The PDT resulted in thrombosis of NV for at least 3 days. Reperfusion, however, was consistently noted at 7 days. Thrombosis was associated with a delay in the growth and maturation of NV fronds, which resumed after reperfusion. Twenty-eight days after the second treatment, NV in both experimental and control eyes had undergone atrophy. At that time (the conclusion of follow-up), however, the size of treated NV fronds (estimated from computerized image analysis of fluorescein angiograms) was significantly less than that of controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Neonatal serotonin reduction alters the adult feminine sexual behaviour of golden hamsters.

HPLC analysis of hamster hypothalamic 5HT indicated higher levels in females than in males on day 12 after birth. Levels of 5HT and 5HIAA could be reduced in both sexes by pCPA administration. Male and female hamster pups were treated on days 1-7 or 7-14 after birth with either pCPA, 5HTP or buffer, and tested for feminine and masculine sexual behaviour in adulthood. 5HTP had no effect on behaviour in either sex. pCPA had no effect on masculine sexual behaviour nor did it affect feminine sexual behaviour when given between days 1-7. When administered on days 7-14, pCPA significantly decreased the time that females spent displaying feminine sexual behaviour, while significantly increasing it in males. We, therefore, suggest that serotonin may be modulating a neural substrate already differentiated by androgens.

Aging