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On the adaptive detection of blood vessels in retinal images.

This paper proposes an automated blood vessel detection scheme based on adaptive contrast enhancement, feature extraction, and tracing. Feature extraction of small blood vessels is performed by using the standard deviation of Gabor filter responses. Tracing of vessels is done via forward detection, bifurcation identification, and backward verification. Tests over twenty images show that for normal images, the true positive rate (TPR) ranges from 80% to 91%, and their corresponding false positive rates (FPR) range from 2.8% to 5.5%. For abnormal images, the TPR ranges from 73.8% to 86.5% and the FPR ranges from 2.1% to 5.3%, respectively. In comparison with two published solution schemes that were also based on the STARE database, our scheme has lower FPR for the reported TPR measure.

Algorithms↗

[Photocoagulation of the retinal feeder vessels of a chorioretinal anastomosis in age-related macular degeneration].

The presence of a chorioretinal anastomosis in the setting of age-related macular degeneration is known as a sign of poor prognosis. No treatment has proven to be effective. We describe a 71-year-old female patient presenting with exudative age-related macular degeneration, a chorioretinal anastomosis with two retinal feeder vessels (arteriole and veinule), a serous retinal pigment epithelium detachment, and a suspected early subretinal neovascular membrane. She was treated with ICG-guided laser photocoagulation directed to the hot spot, with treatment-zone enlargement directed to the retinal feeder vessels and followed up for 6 months. Although a second laser treatment for the reperfused subretinal neovascular membrane was needed, the clinical and angiographic end result was beneficial. Visual acuity improved by two lines, the chorioretinal anastomosis was occluded, and the pigment epithelium reattached. Taking into account the low therapeutic success rate described in the literature, we suggest that the specific treatment of the retinal feeder vessels, which to our knowledge has not been described before, may be a valuable treatment option.

Aged↗

[Expression of basic fibroblast growth factor and fibroblast growth factor receptor 1 in the experimental retinal vein occlusion model].

Retinal ischemia promotes retinal neovascularization. Vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) are important growth factors for neovascularization. We did experimental retinal vein occlusion (RVO) and examined the expression of basic FGF and FGF receptor 1(one of the basic FGF receptors) by in situ hybridization. We used adult pigmented rats (Brown-Norway strain). Dye laser photocoagulation (577 nm) was applied to the retinal arteries and veins within two disc diameters of the optic nerve head to injure the retinal vessels. After one week, laser photocoagulation was applied to only the retinal veins to occlude them (RVO model). As a control, laser photocoagulation was applied to the posterior retina avoiding the retinal vessels. After treatment, the eyes were removed and 10 microns thick cryostat-cut chorioretinal section were used for in situ hybridization with probes as mentioned above. In the RVO model, expression of messenger RNA of basic FGF (b-FGF) and FGF receptor 1 increased in the inner nuclear layer and the inner segment of the photoreceptors, and appeared in the retinal vessel wall in the early stage. This shows that b-FGF and FGF receptor 1 increased in the ischemic retina, and were produced on the retinal vessel wall. This suggests that b-FGF may be involved in protection, regeneration, and proliferation of the retinal vascular endothelial cells in retinal circulatory disturbance.

Animals↗

Hyperglycemia affects flicker-induced vasodilation in the retina of healthy subjects.

Flickering light stimulation of retinal photoreceptors induces retinal vessel dilation in humans. In the present study the effect of high blood glucose levels on this neuro-vascular mechanism was investigated in 12 healthy young male subjects. Blood glucose levels were consecutively increased during 30 min to 100, 200 and 300 mg/dl and kept at the respective level for the following 30 min using hyperglycemic insulin clamps. Eight Hertz flickering light was applied to the fundus at the end of each glucose plateau during continuous retinal vessel diameter measurements with the Zeiss retinal vessel analyser (RVA). During normoglycemia (100 mg/dl) flickering light induced a significant vasodilation of retinal arteries (+2.8+/-0.4%, p<0.0001) and veins (+2.6+/-0.4%, p<0.0001). At 300 mg/dl blood glucose the flicker response in retinal veins was significantly decreased by 55% (p=0.015 versus 100 mg/dl). The modified RVA employed in the present study provides high sensitivity and is capable of studying flicker-induced retinal vasodilation. Using this technique the present study confirms that flickering light stimulation of the human retina induces vasodilation in retinal vessels in healthy subjects. In addition, our data indicate that the retinal vessel response to flickering light stimulation is significantly reduced during hyperglycemia in humans. The relevance of this finding for diabetes-related eye disease remains to be shown.

Adult↗

Angiopoietin-2 plays an important role in retinal angiogenesis.

Angiopoietin 2 (Ang2) expression in the retina is increased during physiologic and pathologic neovascularization suggesting that it may be involved. In this study, we used Ang2-deficient mice to test that hypothesis. Mice deficient in Ang2 showed delayed and incomplete development of the superficial vascular bed of the retina, which develops primarily by vasculogenesis, and complete absence of the intermediate and deep vascular beds which develop by angiogenesis. In addition to incomplete retinal vascular development, Ang2-deficient mice showed lack of regression of the hyaloid vasculature, resulting in a phenotype that mimics infants with persistent fetal vasculature (PFV), a relatively common congenital abnormality. Exposure to high levels of oxygen resulted in partial regression of the retinal vessels, indicating that oxygen-induced regression of retinal vessels does not require Ang2. When these oxygen-exposed mice with few retinal vessels were moved to room air, there was no ischemia-induced retinal neovascularization. These data support the hypothesis that Ang2 plays a critical role in physiologic and pathologic angiogenesis, and physiologic, but not oxygen-induced vascular regression. The data also suggest that infants with PFV should be examined for genetic modifications that would be expected to cause perturbations in Tie2 signaling.

Angiopoietin-2↗

IGF-I is critical for normal vascularization of the human retina.

Experimental and clinical studies suggest that GH and IGF-I may be involved in neovascularization of the retina in diabetes and retinopathy of prematurity. However, the role of GH and IGF-I has not been well established in normal retinal vessel development in humans. Therefore, we examined retinal vessel morphology by digital image analysis of ocular fundus photographs in 13 patients with genetic defects of the GH/IGF-I axis and low levels of IGF-I during and after normal retinal vessel growth. Eleven patients (four females and seven males aged 10-49 yr) had defects of the GH receptor (Laron syndrome). One male (20 yr) had a partial deletion of the IGF-I gene, and one female (14 yr) had a single allele deletion of the IGF-I receptor gene. Patients with defects in the GH/IGF-I axis had significantly less retinal vascularization as evidenced by lower number of vascular branching points (median 23, range 16-25), compared with the reference group of 100 normal controls (median 28, range 19-40, P < 0.001). All 13 individuals had vascular branching points below the median of the reference group. This is the first study to provide genetic evidence for a role of the GH and IGF-I system in retinal vascularization in humans.

Adolescent↗

Diameter variations of retinal blood vessels during and after treatment with hyperbaric oxygen.

AIMS: To quantify retinal vascular change during and after hyperbaric oxygenation (HO) for 6x5 weekly 90 minute treatments. METHODS: Fundus photographs were taken before, during, and after HO at 2.5 atmospheres absolute pressure (ATA) on days 1, 2, 3, 10, 20, 29, and 30 of treatment on three patients using a specially developed hand held ophthalmoscope with a digital colour camera. Blood vessel diameter was estimated on red free retinal images. The mean of three measurements of arterioles and venoles close to the optic disc was calculated. Consistency and repeatability of the method was verified by estimating the diameter of the vessels by three measurements in each of seven images taken within 70 seconds on the same person. Analysis of variance with Bonferroni correction for multiple comparisons was conducted to ascertain whether significant intergroup differences existed. RESULTS: Breathing 100% oxygen at 2.5 ATA constricts retinal arterioles by 9.6% (standard deviation 0.3%) and venoles by 20.6% (SD 0.3%) of their size in air at ambient pressure. Constriction escalates during treatment. Ten minutes after the HO, arterioles dilate to 94.5% (SD 0.3%) and venoles to 89.0% (SD 0.3%) of their primary size. This pattern is the same for each day of measurement. Heart frequency falls continually during HO. Systolic, diastolic, and mean arterial pressures stay constant. CONCLUSION: Exposure to hyperbaric oxygen causes constriction of the retinal vessels. It is found that this constriction is constant through the series of treatments. This suggests that oxygen or products thereof are responsible for the vascular changes during and after hyperbaric oxygenation probably through autoregulation of the retinal vessels.

Aged↗

Nitric oxide regulates retinal vascular tone in humans.

The purpose of the present study was to investigate the contribution of basal nitric oxide (NO) on retinal vascular tone in humans. In addition, we set out to elucidate the role of NO in flicker-induced retinal vasodilation in humans. Twelve healthy young subjects were studied in a three-way crossover design. Subjects received an intravenous infusion of either placebo or NG-monomethyl-L-arginine (L-NMMA; 3 or 6 mg/kg over 5 min), an inhibitor of NO synthase. Thereafter, diffuse luminance flicker was consecutively performed for 16, 32, and 64 s at a frequency of 8 Hz. The effect of L-NMMA on retinal arterial and venous diameter was assessed under resting conditions and during the hyperemic flicker response. Retinal vessel diameter was measured with a Zeiss retinal vessel analyzer. L-NMMA significantly reduced arterial diameter (3 mg/kg: -2%; 6 mg/kg: -4%, P < 0.001) and venous diameter (3 mg/kg: -5%; 6 mg/kg: -8%, P < 0.001). After placebo infusion, flicker induced a significant increase in retinal vessel diameter (P < 0.001). At a flicker duration of 64 s, arterial diameter increased by 4% and venous diameter increased by 3%. L-NMMA did not abolish these hyperemic responses but blunted venous vasodilation (P = 0.017) and arterial vasodilation (P = 0.02) in response to flicker stimulation. Our data indicate that NO contributes to basal retinal vascular tone in humans. In addition, NO appears to play a role in flicker-induced vasodilation of the human retinal vasculature.

Adult↗

Retinal blood flow during hyperoxia in humans revisited: concerted results using different measurement techniques.

Retinal vasculature shows pronounced vasoconstriction in response to hyperoxia, which appears to be related to the constant oxygen demand of the retina. However, the exact amount of blood flow reduction and the exact time course of this phenomenon are still a matter of debate. We set out to investigate the retinal response to hyperoxia using innovative techniques for the assessment of retinal hemodynamics. In a total of 48 healthy volunteers we studied the effect of 100% O(2) breathing on retinal blood flow using two methods. Red blood cell movement in larger retinal veins was quantified with combined laser Doppler velocimetry and retinal vessel size measurement. Retinal white blood cell movement was quantified with the blue field entoptic technique. The time course of retinal vasoconstriction in response to hyperoxia was assessed by continuous vessel size determination using the Zeiss retinal vessel analyzer. The response to hyperoxia as measured with combined laser Doppler velocimetry and vessel size measurement was almost twice as high as that observed with the blue field technique. Vasoconstriction in response to 100% O(2) breathing occurred within the first 5 min and no counterregulatory or adaptive mechanisms were observed. Based on these results we hypothesize that hyperoxia-induced vasoconstriction differentially affects red and white blood cell movement in the human retina. This hypothesis is based on the complex interactions between red and white blood cells in microcirculation, which have been described in detail for other vascular beds.

Adult↗

Upregulation of chemokine expression in the retinal vasculature in ischemia-reperfusion injury.

PURPOSE: To evaluate chemokine expression at various retinal sites after ischemia-reperfusion injury, using reverse transcription-polymerase chain reaction (RT-PCR) analysis of selected tissue obtained by laser capture microdissection. METHODS: Retinal ischemia was produced in Lewis rats by increasing intraocular pressure for 75 minutes. At 3, 6, 12, and 24 hours after reperfusion, RT-PCR was used to measure the levels of monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, interleukin (IL)-8, and interferon-gamma-inducible 10-kDa protein (IP-10) mRNA expression in the ganglion cell layer (GCL), inner nuclear layer (INL), outer nuclear layer (ONL), and retinal vessels, after laser capture microdissection of these retinal layers. These chemokines were further localized by immunohistochemical methods, using antibodies specific to MCP-1 and MIP-1alpha. Leukocyte infiltration into the retina was detected with immunostaining for leukocyte common antigen. RESULTS: Ischemia-reperfusion induced expression of MCP-1, MIP-1alpha, and MIP-1beta mRNA in the retinal vessels 3 hours after reperfusion. Six hours after reperfusion, expression of these chemokines and IL-8 mRNA was seen in the GCL and INL. Twelve hours after reperfusion, IP-10 mRNA expression was seen in the GCL and INL. Immunoreactive MCP-1 and MIP-1alpha were detected in the GCL, INL, and the retinal vessels 24 hours after reperfusion. No chemokine mRNA expression or immunoreactivity was detected in the ONL at any time. Leukocyte infiltration was noted at 12 hours, increasing markedly 24 hours after reperfusion. CONCLUSIONS: Ischemia-reperfusion retinal injury results in generation of highly chemotactic agents, initially in the retinal vasculature, then in the other inner retinal layers. Such differential chemokine expression may play a role in leukocyte recruitment and selective leukocyte infiltration in the inner retina, leading to retinal damage primarily localized to the ganglion cells and other inner neuronal structures.

Animals↗

Plasma cortisol in men--relationship with atherosclerosis of retinal arteries.

The production of cortisol increases in acute stress but the effects of chronic stress on plasma cortisol are still controversial. Stress on the other hand plays a role in coronary artery disease (CAD) and carotid atherosclerosis. Since there is no data about plasma cortisol and atherosclerosis of the retinal arteries, the purpose of this study was to explore the relationship between plasma cortisol in 101 adult males with the degree of their retinal vessels atherosclerosis. The results were compared with those in 47 matched apparently healthy men with no retinal vessels changes. The atherosclerotic changes of retinal vessels were determined by direct ophthalmoscopy and graded (1-4) according to Scheie. Morning plasma cortisol levels were determined by radioimmunoassay using commercial kits. The results were compared by using chi-square test. No association between morning plasma cortisol concentrations and retinal vessels atherosclerosis could be found. The results of this study do not support a role for physiological levels of plasma cortisol in the development of atherosclerosis, at least of the retinal arteries, in men.

Age Factors↗

Retinal branch vessel occlusion in acute intermittent porphyria.

Three patients with acute intermittent porphyria were noted to have retinal branch vessel occlusion. Branch "vein" occlusion, segmental optic atrophy, and soft exudate were the most common ocular manifestation. Two patients had labile elevated hypertension. When patients present with retinal branch vessel occlusion and a constellation of bizarre symptoms that might include hypertension, abdominal pain, acute psychotic behavior and/or cutaneous photosensitivity, the diagnosis of acute intermittent porphyria should be considered.

Abdomen↗

Development of the primate retinal vasculature.

Human and macaque retinae have similar retinal vascular anatomy. The general features of the retinal vascular anatomy of these two primates have much in common with more widely studied animal models such as rat and cat. However, primates are unique amongst mammals in having a region in temporal retina specialized for high visual acuity, which includes the fovea centralis (or 'fovea'). Several features distinguish the fovea from other parts of the retina, including a very high local density of cone photoreceptors, a high density of inner retinal cells during development, and an absence of retinal blood vessels. The retinal vascular complex comprises a number of cell types, in addition to vascular endothelial cells, including pericytes, microglia, astrocytes-none of which is intrinsic to the retina. In addition, amacrine-like cells make bouton-like associations with retinal vessels and may be involved in the autoregulation of blood flow. During development endothelial cells 'invade' the retina, accompanied by a population of microglial cells; glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes are also seen associated with the developing vasculature, and are in advance of the vascular front by a few hundred microns. Recent findings indicate that astrocytes at the vascular front proliferate in response to factors released by endothelial cells, including leukemia inhibitory factor. Better understood is the role of GFAP-immunoreactive astrocytes just in advance of the developing vessels. These astrocytes are sensitive to hypoxia and in response release vascular endothelial growth factor (VEGF) which in turn promotes the migration, differentiation and proliferation of vascular endothelial cells. This hypoxia/VEGF-mediated process of migration, proliferation and differentiation appears common to the retinae of a variety of species, including human. However, in human and macaque retina, different mechanisms appear to govern the development of the retinal vessels growing along the horizontal meridian of the retina towards the central area, which contains the fovea. Despite the relatively advanced state of differentiation and maturation of cells in the central area compared with the periphery, the growth of retinal vessels into the central area has been described as 'retarded', and the incidence of cell proliferation associated with these vessels is lower than in peripheral vessels. Furthermore, neither retinal vessels nor their accompanying astrocytes grow into a circumscribed region which, at a later stage, develops into the foveal depression. These observations suggest that molecular markers define the foveal region and inhibit cell proliferation and vascular growth at the fovea and, perhaps, along the horizontal meridian. The findings also suggest that at the fovea, the retina is adapted morphologically to its blood supply, since in the vicinity of the fovea, the development of retinal vessels is retarded or inhibited. The limitations on vascularization of central retina has implications for its vulnerability to degenerative changes, as seen in age-related macular degeneration.

Animals↗

[Measurement of the diameter of segments of retinal branch vessels in digital fundus images - An experimental study of the method and reproducibility].

PURPOSE: The aim of this study was the evaluation of new algorithms to measure diameter of segments of retinal branch vessels offline in local dependence and independent by observer. Methods 360 flashed fundus images (camera FF 450 ZEISS Germany, Visualis IMEDOS GmbH Weimar/Germany) of 12 eyes of healthy volunteers (10 independent sessions containing 3 images for every eye) were analysed. Algorithms detect the vessel diameter along the vessel course automatically. Corresponding segments of a retinal artery and vein were examined (mean length of the segment 2.5 mm) in every image of one eye. Results The marked arterial segment was detected automatically in 359 pictures and the venous segment in all pictures. The mean vessel diameter was detected in single pictures with a mean coefficient of variation (CV) for arteries of 3.4 % and for veins of 2.7 %. The differences of arterial and venous diameter between images were not significant. Analyzing sessions the CV of the mean vessel diameter were reduced for arteries to 2.7 % and for veins to 2.5 %. The standard deviation of the mean vessel diameter was independent of the vessel diameter (branch vessels with diameter of 120 to 200 micrometer). The mean CV of the vessel diameter at single locations were 5.7 % for arteries and 3.8 % for veins. Conclusions The new algorithms are useful for retinal vessel analysis, if there are no questions concerning the dynamic of vessel behaviour.

Adult↗

Effect of histamine and cimetidine on retinal and choroidal blood flow in humans.

Intravenous administration of histamine causes an increase in choroidal blood flow and retinal vessel diameter in healthy subjects. The mechanism underlying this effect remains to be elucidated. In the present study, we hypothesized that H2 receptor blockade alters hemodynamic effects of histamine in the choroid and retina. Eighteen healthy male nonsmoking volunteers were included in this randomized, double-masked, placebo-controlled two-way crossover study. Histamine (0.32 microg.kg(-1).min(-1) over 30 min) was infused intravenously in the absence (NaCl as placebo) or presence of the H2 blocker cimetidine (2.3 mg/min over 50 min). Ocular hemodynamic parameters, blood pressure, and intraocular pressure were measured before drug administration, after infusion of cimetidine or placebo, and after coinfusion of histamine. Subfoveal choroidal blood flow and fundus pulsation amplitude were measured with laser-Doppler flowmetry and laser interferometry, respectively. Retinal arterial and venous diameters were measured with a retinal vessel analyzer. Retinal blood velocity was assessed with bidirectional laser-Doppler velocimetry. Histamine increased subfoveal choroidal blood flow (+14 +/- 15%, P < 0.001), fundus pulsation amplitude (+11 +/- 5%, P < 0.001), retinal venous diameter (+3.0 +/- 3.6%, P = 0.002), and retinal arterial diameter (+2.8 +/- 4.2%, P < 0.01) but did not change retinal blood velocity. The H2 antagonist cimetidine had no significant effect on ocular hemodynamic parameters. In addition, cimetidine did not modify effects of histamine on choroidal blood flow, fundus pulsation amplitude, retinal venous diameter, and retinal arterial diameter compared with placebo. The present data confirm that histamine increases choroidal blood flow and retinal vessel diameters in healthy subjects. This ocular vasodilator effect of histamine is, however, not altered by administration of an H2 blocker. Whether the increase in blood flow is mediated via H1 receptors or other hitherto unidentified mechanisms remains to be elucidated.

Adult↗

Intravenous administration of diphenhydramine reduces histamine-induced vasodilator effects in the retina and choroid.

PURPOSE: Intravenous administration of histamine causes an increase in choroidal blood flow (ChBF) and retinal vessel diameters in healthy subjects. The receptor mediating this response has not yet been identified. The present study was undertaken to clarify whether H1 receptor blockade with diphenhydramine affects the hemodynamic response of histamine in the choroid and the retina. METHODS: A randomized, double-masked, placebo-controlled, two-way crossover study was performed in 18 healthy, male, nonsmoking subjects. Histamine (0.32 microg/kg per minute over 30 minutes) was infused intravenously in the absence (NaCl as placebo) or presence of the H1 blocker diphenhydramine (1.0 mg/min over 50 minutes). Ocular hemodynamic parameters, blood pressure, and intraocular pressure were measured before drug administration, after infusion of diphenhydramine or placebo, and after co-infusion of histamine. Subfoveal ChBF and fundus pulsation amplitude (FPA) were measured with laser Doppler flowmetry and laser interferometry, respectively. Retinal arterial and venous diameters were measured with a retinal vessel analyzer. Retinal blood velocity was assessed with bidirectional laser Doppler velocimetry. RESULTS: Administration of histamine caused a decrease in mean arterial pressure by -4% +/- 9% (ANOVA P = 0.01). This effect was blunted by coadministration of diphenhydramine (ANOVA, P = 0.04). Histamine significantly increased FPA and subfoveal ChBF. Coadministration of diphenhydramine significantly reduced this effect (ANOVA; FPA P = 0.001, ChBF P = 0.049). Histamine significantly increased retinal arterial diameter by +3.5% +/- 4.5% and retinal venous diameter by +3.7% +/- 2.8%. Again, coadministration of diphenhydramine significantly reduced the vasodilative effect to +0.3% +/- 5.5% in retinal arteries (ANOVA, P = 0.00006) and to +0.9% +/- 2.5% in retinal veins (ANOVA, P = 0.004). CONCLUSIONS: The present data confirm that histamine increases ChBF and retinal vessel diameters in healthy subjects. Administration of the H1 receptor blocker diphenhydramine significantly reduced histamine-induced changes in ocular perfusion parameters. These results strongly indicate that in the retina and choroid, H1 receptors are involved in the histamine-mediated hemodynamic effects in vivo.

Adult↗

Laser-induced experimental vascular occlusion using liposome-encapsulated ADP.

The therapeutic occlusion of retinal vessels is often helpful in treating various pathological conditions. We compared the combined effects of argon laser photocoagulation and adenosine diphosphate (ADP) released from temperature-sensitive liposomes with argon laser photocoagulation alone on occlusion of retinal vessels in pigmented rats. In Group A, 8 eyes were treated with liposome-encapsulated ADP and laser photocoagulation. In Group B, 8 eyes were treated with laser photocoagulation alone. The laser parameters (power, spot size, exposure time) were maintained at the same levels for both groups. The laser was focused on the retinal vessels at the optic nerve head. The treated retinal vessels were observed at time zero, day 1, day 4, and weekly for a period of 3 months. At time zero, 6 of 8 eyes were totally occluded in Group A, with best results obtained at 80 mW. Only 1 of 8 eyes in Group B achieved total occlusion. After 3 months, 4 of 8 eyes in Group A remained totally occluded; no eyes in Group B were occluded. Complete and permanent occlusion of retinal vessels can be achieved by using ADP and laser photocoagulation of lower power density than traditional laser photocoagulation alone.

Adenosine Diphosphate↗

[The preprocessing of subtraction and the enhancement for biomedical image of retinal blood vessels].

Image segmentation is still a difficult problem since its effect would vary with the subjects processed. An approach of subtracting background from the entire image of retinal blood vessels presented in this paper. The background subtraction is based on the real image itself taken photographically and is not dependent on the prior knowledge of system for recording image, the approach achieves the grayscale enhancement of retinal blood vessels in preprocessing and provides a quality image for the next process of binarization. This experiment in the preprocessing of subtraction shows good enhancement effect.

Algorithms↗