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PubMed1998

Evaluation of the echographic contribution to the parasurgical treatment of melanomas of small dimensions.

In this work we underlined the contribution of echography in the parasurgical treatment (photocoagulative) of melanomas of small dimensions. Fourteen patients were examined; some of them were treated with argon laser, others with krypton laser. The echographic examination was performed with the A-B scan technique, naturally accompanied by fluorangiography before and after the operation. An accurate echographic and fluorangiographic follow-up makes it posible to control the evolution of the lesions efficaciously. From this work it is clear how echography gives highly reliable results which allow us, therefore, both to make an adequate therapeutic decision and to evaluate the efficaciousness of the treatment itself.

Adult↗
PubMed1998

Automatic static threshold perimetry is useful for estimating the effects of laser photocoagulation on diabetic maculopathy.

Focal photocoagulation was performed on 18 eyes in 18 diabetes patients; a visual acuity test, fluorescein angiography and automated static threshold perimetry by Octopus automated static perimetry (program 31) were conducted before and 1, 3 and 6 months after krypton laser photocoagulation. Central field sensitivity and total loss were measured by automated static threshold perimetry. Group A (13 eyes) had a total loss of less than 150 dB. In these subjects, mild fluorescein leakage was detected within the area of the vascular arcade. In group B (5 eyes) with a total loss of greater than 150 dB, diffuse fluorescein leakage was also detected outside the vascular arcade. The central field sensitivity was reduced in 2 eyes of group B. The total loss worsened in 8 eyes of group A (61.5%) and 5 eyes of group B (100%). Inspection of photographs gave the impression that the degree of fluorescein leakage and the area of the avascular zone on fluorescein angiography accorded with total loss, thus suggesting that total loss reflects visual functions better than visual acuity or central field sensitivity. Therefore, automated static threshold perimetry appears to be a useful method for estimating visual functions after photocoagulation in diabetic maculopathy patients.

Adult↗
PubMed1992

Vascular remodeling after laser photocoagulation: concurrent regeneration and atrophy.

Laser photocoagulation is associated with paradoxical results: it causes obliteration of vessels, but leads also to the formation of new ones. In an attempt to better understand this dual vascular response we conducted an ultrastructure study of the choroidal vascular repair following krypton laser injury in rats. Three processes were observed: recanalization, neovascularization, and atrophy of both recanalized and newly formed capillaries. Post-lasering repair of the choroidal vasculature can therefore be described as a remodeling process, characterized by both regeneration and involution. The latter appears to be a secondary process of atrophy, contributing to permanent vascular obliteration. These mechanisms might explain why, in spite of initial vascular regeneration, laser photocoagulation treatment has a beneficial effect on choroidal subretinal neovascularization.

Animals↗
PubMed1989

Effects of tumor necrosis factor-alpha on tumor blood flow and hyperthermic treatment.

The impact of recombinant human tumor necrosis factor-alpha (rhTNF-alpha), given alone or in combination with local hyperthermia, on perfusion and growth of a moderately rhTNF-alpha-sensitive rat tumor (DS-carcinosarcoma) was investigated. DS-carcinosarcomas were implanted into the hind foot dorsum of Sprague-Dawley rats. Tumor blood flow (TBF) was measured with the krypton-85 clearance technique. Treatment with either tumor necrosis factor-alpha (0.1-1.0 mg/kg) or hyperthermia (43.3 and 44.3 degrees C, 40 min) can decrease the perfusion of malignant tumors. The TBF reduction was fully established 2 h after rhTNF-alpha injection and lasted for at least 4 h. The application of local hyperthermia (T greater than 42 degrees C) 3 h after rhTNF-alpha administration further diminished tumor blood flow. Volume growth of the tumors was monitored during repeated treatment. rhTNF-alpha (0.2 and 1.0 mg/kg i.v.) combined with hyperthermia (43.3 and 44.3 degrees C, 40 min, starting 3 h after rhTNF-alpha injection) were given every third day from the fifth to the 20th day after tumor implantation. Monotherapies retarded tumor growth in a dose-dependent manner. Combined treatment was superior to either monotherapy leading to local tumor control in 40-50% of the animals treated. It is concluded that local hyperthermia can enhance the efficacy of rhTNF-alpha treatment by further reducing tumor perfusion.

Animals↗
PubMed1995

Fluorescence cystoscopy following intravesical instillation of 5-aminolevulinic acid: a new procedure with high sensitivity for detection of hardly visible urothelial neoplasias.

Methods have been sought for the in vivo marking of tiny papillary tumors of the bladder and flat urothelial lesions such as dysplasia or carcinoma in situ, which can easily be missed during conventional endoscopy under white light. A new procedure is reported for the fluorescence detection of urothelial dysplasia and early bladder cancer. The method is based on intravesical application of 5-aminolevulinic acid (ALA). ALA if applied exogenously induces accumulation of protoporphyrin IX (PPIX) in the urothelium of the bladder. PPIX is an intensively red fluorescing agent. The mean ratio of fluorescence intensity between urothelial cancer and normal epithelium was found to be 17:1. Fluorescence excitation was achieved by violet light from a krypton ion laser (lambda = 406.7 nm) or from a xenon arc lamp with a bandpass filter system (lambda = 375-440 nm). Both light sources proved to be of equal suitability for fluorescence excitation. Fluorescence microscopy revealed that the PPIX fluorescence is strictly limited to the urothelium. It could not be detected from the submucosa or muscle of the bladder. Bladder wall biopsies were taken from 90 patients with suspicion of bladder cancer under fluorescence view. The fluorescence detection proved to be of high sensitivity (98%). No serious side effects which would preclude further clinical testing, especially no cutaneous photoreaction, were observed. Tumor-associated fluorescence induced by topical ALA application offers new perspectives in the diagnosis and treatment of bladder cancer. In case of suspicious or positive urine cytologic findings, ALA fluorescence cystoscopy may be useful for detecting the precise site of the malignancy. The procedure might be helpful in complete resection or coagulation of urothelial neoplasms. Due to this, diminishing recurrence rates are expected. However, this hypothesis has to be studied in prospective clinical trials.

Administration, Intravesical↗
PubMed1986

[Vision aids for patients with senile exudative macular degeneration. Comparison of spontaneous fibrovascular scars and scotomas induced by photocoagulation].

Aging exudative macular degeneration frequently results in a fibrovascular scar which entails the loss of central vision. This scar implies an often large and deep scotoma which makes it very difficult for the patient to be fitted with low-vision aids. A Goldmann static perimetry study of retinal sensitivity in the treated areas showed that krypton laser at liminar doses does not induce deep scotomas.

Aged↗
PubMed1987

Laser treatment of macular subretinal neovascularizations in angioid streaks.

A serous hemorrhagic maculopathy with a subretinal neovascular membrane appears in about 70% of cases of angioid streaks. In this study a series of 60 eyes (31 patients) was reviewed. In 13 eyes macular subretinal neovascularizations were photocoagulated with blue-green argon laser, green argon laser, dye laser (rhodamine 6G) at 590 nm and red krypton laser. The authors present and discuss the results obtained and suggest that photocoagulation of subretinal new vessels not involving the fovea can be worth performing.

Adolescent↗
PubMed1988

Excimer laser-induced simultaneous ablation and spectral identification of normal and atherosclerotic arterial tissue layers.

A krypton-fluorine excimer laser at a 248-nm wavelength was used to irradiate normal and severely atherosclerotic segments of human postmortem femoral arteries. Single pulses and multiple pulses required for penetration or perforation of the arterial wall were applied with 16 nsec pulse width and 5 J/cm2/pulse energy fluence. The total fluorescence of irradiated and ablated tissue was analyzed in real-time mode by means of spectroscopy. Each laser pulse produced one spectrum that was characteristic of the composition of the tissue layer, which was ablated. Fluorescence spectroscopy indicated a broad-continuum emission between 300 and 700 nm with peak fluorescence of equal intensity at wavelengths of 370 and 460 nm (ratio, 1.004 +/- 0.087) for normal media layers. Atheromas without calcification (lipid, fibrous, and mixed) were found with spectral maxima at the same wavelengths but with significantly reduced intensity at 460 nm (ratio, 1.765 +/- 0.263; p less than 0.001). In contrast to this broad-continuum fluorescence, calcified plaques displayed multiple-line emission with the most prominent peaks at wavelengths of 397, 442, 450, 461, 528, and 558 nm. These fluorescence criteria identified the histologically classified target tissue precisely. Histological examination of the corresponding arterial layers indicated sharply delineated and circumscribed tissue ablation. These results indicate that simultaneous tissue identification (diagnosis) and ablation (treatment) by excimer laser irradiation is feasible under strict laboratory conditions. We conclude that this principle demonstrates the potential for laser beam control by means of target-specific ablation.

Angioplasty, Balloon↗
PubMed1976

Progressive perfusion impairment during prolonged low flow myocardial ischemia in dogs.

Recent studies have shown that after total coronary artery occlusion, there is impaired "reflow" of blood accompanied by myocardial and capillary endothelial cell swelling. To investigate the effect of prolonged low flow myocardial ischemia on coronary vascular resistance, regional hypoperfusion of the distal left anterior descending coronary artery was studied in 31 autonomically blocked dogs on right heart bypass. Heart rate, aortic pressure, and, during ischemia, left ventricular end-diastolic pressure were held constant. The distal left anterior descending coronary artery was perfused at a substantially reduced perfusion pressure which resulted in an antegrade coronary blood flow that usually was between 3% and 7% (0.5-1 ml/min) of control. When relative hypothermia (33-34 degrees C) was induced in nine dogs, left anterior descending coronary artery vascular resistance did not change during 2.5-3 hours of low flow ischemia. Under euthermic conditions (37-40 degrees C) in 17 dogs there was a consistent progressive increase in distal left anterior descending coronary artery vascular resistance starting at 90 minutes (median) after onset of ischemia. By 110-140 minutes ischemic antegrade flow decreased by 35 +/- 4% (SEM) (P less than 0.01). Directionally similar flow changes were observed in six euthermic experiments using the krypton-85 washout technique. Light microscopy did not reveal hemorrhage as a cause of the increased vascular resistance. The perfusion impairment did not occur in two euthermic, nonischemic hearts. In five dogs elevation of serum osmolality by 23 +/- 11 mOsmol/liter with mannitol attenuated the progressive decrease in flow. Thus, a progressive perfusion defect exists in the ischemic low flow state in the heart which presumably contributes to the extent of eventual necrosis.

Animals↗
PubMedDate not supplied

The effects of calcium antagonism on the epicerebral circulation in early vasospasm.

We have studied the effects of the calcium antagonist verapamil on the epicerebral arteriovenous transit time and regional epicerebral circulation of dogs by direct measurement of arterial diameters, fluorescein angiography, and krypton-85 regional epicerebral blood flow analysis. A large craniectomy was performed and vasoconstriction was induced by the subarachnoid injection of human platelet-rich plasma (PRP) pretreated with 25 mu M of ADP to cause maximum aggregation. Once vasoconstriction was established, verapamil (0.1 mg/kg) was topically applied to the perforated arachnoid. The PRP-ADP produced a mean decrease in the arterial diameters of 38.2 +/- 1.6% (p less than 0.01) at 10 minutes after its injection and verapamil produced a mean dilatation of 19.5 +/- 2.5% (p less than 0.01), compared to control values. Regional epicerebral blood flow was 54.9 +/- 3.4 ml/100 g/min in the control state, 34.8 +/- 3.2 ml/100 g/min (p less than 0.01) during vasospasm, and 78.2 +/- 4.5 ml/100 g/min (p less than 0.01) after verapamil. Fluorescein angiography, after verapamil, demonstrated a mean acceleration of the arteriovenous circulation time of 4.5 +/- 0.8 seconds (p less than 0.01) compared to the spasm value. We concluded that the topical application of verapamil can dilate previously constricted cortical arteries and that this dilatation is associated with acceleration of the epicerebral transit time and increased cerebral blood flow.

Administration, Topical↗
PubMedDate not supplied

Influence of cerebral vasoconstricting and vasodilating agents on blood flow in regions of focal ischemia.

Regional cerebral blood flow (rCBF) measurements with krypton-85 (100 separate determinations) were compared in squirrel monkeys anesthetized with sodium pentobarbital (a cerebral vasoconstrictor) and halothane (a cerebral vasodilator) before, during, and after middle cerebral artery (MCA) occlusion. Prior to MCA occlusion, a normal physiological response to alterations in arterial carbon dioxide tensions (Paco2) was demonstrated in both groups of monkeys; the cerebral vascular resistance was significantly lower in those anesthetized with halothane. During ischemia, there was loss of autoregulation and a failure to respond to alterations in Paco2 in both groups. Flow in the ischemic region remained uniform in the barbiturate group but decreased progressively in the halothane group, suggesting a "paradoxical response" to the dilating agent. Reactive hyperemia (luxury perfusion) was demonstrated in both groups after restoration of flow. The use of a beta-emitting isotope ensured that measurements in regions of ischemia accurately reflected rCBF and were free of the artifacts ("look through" and Compton scatter) related to use of a gamma-emitting indicator.

Animals↗
PubMedDate not supplied

Use of hydrogen for measurement of regional cerebral blood flow: problem of intercompartmental diffusion.

The extreme diffusibility of hydrogen, compared with xenon or krypton, may create serious artifacts when it is used to measure local blood flow with a tissue electrode. The errors are greatest when hydrogen is given by intra-arterial slug injection, and when the electrode is within 2mm of another tissue compartment, CSF, or air. These all appear to be a consequence of intercompartmental diffusion which can occur at rates of the same order of magnitude as clearance from the tissue by blood flow. No matter how small the electrode, the ultimate spatial resolution of the method appears to be about 2mm unless quantitative account is taken of diffusion. An important precaution in use of the method is to obtain homogeneous tissue saturation by prolonged inhalation administration.

Animals↗
PubMed1990

Left lower lobe ventilation is reduced in patients with cardiomegaly in the supine but not the prone position.

To determine the effect of the heart on regional ventilation, Krypton-81m (81mKr) tomographic (SPECT) ventilation scans were recorded in seven patients with cardiomegaly and four normal subjects in the supine and prone positions. All patients had a cardiothoracic ratio of greater than 0.50 and clear lung fields radiographically. Using standard gamma camera tomographic reconstruction techniques, images of transaxial slices were obtained during a 360 degree rotation around the thorax of the subject breathing the radioactive gas 81mKr. The transaxial images, acquired over 10 min were aligned in each posture at the level of the cardiac apex, mid-heart, and aortic arch and were matched in relation to a radioactive marker on the chest wall and to anatomic landmarks. A horizontal line (gravity independent and parallel to the couch) was drawn on the transaxial section through the dorsal regions of the right and left lung. Counts per resolution element (12 to 15 mm) were plotted along this line and the ratios of the peak values in right and left lung compared. These ratios represent differences in regional ventilation per unit lung volume. In controls the mean left-to-right (L/R) peak count ratio varied from 0.91 to 1.00 at the three levels (range: 0.76 to 1.04); there were no significant differences between supine and prone. In patients with cardiomegaly the mean (+/- SEM) L/R peak count ratio at cardiac apex, mid-heart, and aortic arch was 0.46 (+/- 0.08), 0.55 (+/- 0.07), and 0.89 (+/- 0.08) when supine and 1.04 (+/- 0.07), 1.05 (+/- 0.05), and 1.08 (+/- 0.07) when prone, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1991

Enrichment of subpopulations of respiratory epithelial cells using flow cytometry.

The application of flow cytometry to enrich airway epithelial cell subpopulations is described. A complementary epithelial cell preparative technique is also outlined. The ability of the airway epithelium to protect the lung from environmental insults results from a complex interaction among the different cells that form its matrix. The separation of the different epithelial cell types is an essential step in the studies of mechanisms of the controlling factors of cell repair, cell differentiation, and neoplastic transformation. Epithelial cells of the New Zealand white rabbit trachea are prepared using enzymatic digestion and microdissection. Small sections of tracheal wall are dissected into pieces approximately 10 mm2. The mucosa is dissected and placed in 0.15% hyaluronidase for 40 min at 22 degrees C. Mucus is removed, and the mucosa is then placed in 0.1% pronase at 37 degrees C for 30 min. With careful dissection, the epithelium can be dissected from the mucosa in 10-mm2 sheets. Sheets of epithelial cells are placed in 6 ml of an enzymatic solution containing collagenase, 0.2% bovine serum albumin, 0.04% soya bean trypsin inhibitor, 0.06 ml of 1 M Hepes buffer for 3 h at 37 degrees C. The cells are gently pipetted during the 3-h period, yielding a suspension of viable cells. Subpopulations of these different cell types are enriched using an Orthocytofluorograph 50111. A krypton ion laser was used for excitation of cells at 488 nm. Forward-angle and 90 degrees scatter were gated on the histogram. The purification of the ciliated, basal, and secretory cells was 90%, 97%, and 94%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2003

Reduced choroidal neovascular membrane formation in matrix metalloproteinase-2-deficient mice.

PURPOSE: Findings in studies have suggested a role for matrix metalloproteinase (MMP)-2 in angiogenesis, including choroidal neovascularization (CNV). To investigate further, the current study was conducted to observe the formation of experimental CNV in MMP-2-deficient mice. METHODS: CNV was induced in wild-type and MMP-2-deficient mice by krypton laser photocoagulation of the fundus. The time-course of expression of MMP-2 mRNA after laser treatment was determined by in situ hybridization with anti-sense and sense cRNA probes. MMP-2 protein distribution was determined by immunohistochemistry. Ten days after treatment, the extent of CNV was evaluated on hematoxylin-eosin stained serial sections. The maximum height of the CNV lesions was calculated by image analysis of digitized histologic images. RESULTS: Expression of MMP-2 mRNA was detected in the CNV lesions at day 3 after laser treatment and peaked at day 5, after which it slowly declined. MMP-2 mRNA expression appeared to be highest at the margins of the membrane. Immunostaining for MMP-2 confirmed the presence of MMP-2 protein in the CNV lesions. The CNV lesions of MMP-2-deficient mice showed that relative thickness was reduced by 31% compared with wild-type mice (P = 0.006). CONCLUSIONS: The present study demonstrated that MMP-2 mRNA and protein are upregulated during experimental CNV in the mouse. The marked difference in thickness of the CNV membrane between wild-type and MMP-2-deficient mice shows that MMP-2 is involved in the formation of experimental CNV in the mouse. These results suggest that pharmacologic targeting of MMPs, including MMP-2, may reduce formation of CNV in conditions such as age-related macular degeneration.

Animals↗
PubMed2004

Subthreshold transpupillary thermotherapy reduces experimental choroidal neovascularization in the mouse without collateral damage to the neural retina.

PURPOSE: Transpupillary thermotherapy (TTT) is currently being evaluated for treatment of choroidal neovascularization (CNV) in age-related macular degeneration. To optimize TTT for CNV, the effect was analyzed of invisible (subthreshold) or visible (threshold) doses of TTT on the normal mouse retina and on experimental CNV. METHODS: TTT was delivered to the normal retina of 42 mice with a diode laser at increasing power settings (50, 60, 70, or 80 mW), to obtain thermal lesions ranging from invisible (subthreshold) to visible (threshold) burns. CNV was induced in 53 mice by krypton laser photocoagulation of the fundus, after which the CNV lesions were treated with TTT (50, 60, or 80 mW). Eyes were enucleated 7 days after TTT and prepared for histology, and the CNV complex was evaluated on hematoxylin-eosin stained serial sections by measuring the maximum height of the CNV lesions. Ultrastructural changes were examined by transmission electron microscopy. RESULTS: Increasing the TTT laser power yielded gradually more visible effects. At 50 mW, which induced subthreshold burns, no damage was seen in the neural retina, retinal pigment epithelium (RPE), or choroid at any time point. By contrast, eyes treated with higher power exhibited progressively more damage to the neural retina, including a complete disruption of the outer nuclear layer. When TTT was applied to the laser-induced CNV lesions, the height of lesions was significantly reduced (P < 0.001) in response to all three power settings at 7 days after treatment. The mean relative thickness of the CNV lesion was 3.29 +/- 0.89 in untreated mice, whereas in TTT-treated mice it was 1.69 +/- 0.35, 1.69 +/- 0.41 and 1.70 +/- 0.17 at power settings of 50, 60, and 80 mW, respectively. The overlying neural retina showed no apparent damage with the 50- or 60-mW settings, whereas outer nuclear layer disruption occurred with a power of 80 mW. Electron microscopy confirmed the presence of vascular occlusion at 1 day and a fibrotic scar at 7 days after TTT. CONCLUSIONS: Subthreshold TTT can effectively occlude newly formed vessels and cause regression of the experimental CNV complex without damaging the neural retina. The results demonstrate the importance of using subthreshold laser power in experimental and clinical evaluation of TTT.

Animals↗
PubMed2004

Stimulation of neovascularization by the anti-angiogenic factor PEDF.

PURPOSE: Examine the effect of (pigment epithelium-derived growth factor; PEDF) on laser-induced choroidal neovascularization (CNV). METHODS: Adult C57Bl/6 mice were anesthetized and four laser spots were placed in each quadrant of the fundus with a krypton red laser (614 nm, 50 microm, 0.05 second, 200 mW). Animals were treated with various doses of PEDF administered with miniosmotic pumps implanted subcutaneously. Seven days after laser treatment, mice were perfused with 3% FITC high-molecular-weight dextran, the eyes enucleated, and neovascularization analyzed by confocal microscopy. Data were recorded as the volume of the neovascular complex. The effect of PEDF on endothelial cell migration, vascular tube formation in synthetic basement membrane, and VEGF production was also determined. RESULTS: Mice receiving a lower dose of PEDF (90 microg/mL) had significantly decreased areas of CNV. A high dose of PEDF (360 microg/mL) significantly increased CNV, whereas an intermediate dose (180 microg/mL) of PEDF had no effect. PEDF inhibited endothelial cell migration and vascular tube formation at lower doses (0.5-5 microg/mL). High doses of PEDF (25-50 microg/mL) stimulated endothelial cell migration, enhanced vascular tube formation in vitro, and stimulated VEGF production from endothelial cells. Neutralizing anti-VEGF antibody completely reversed the stimulatory effects of high doses of PEDF on CNV in vivo. CONCLUSIONS: PEDF demonstrates opposing effects on CNV and endothelial cell function. Whereas low doses are inhibitory, high doses can augment the development of the neovasculature. These results suggest that the effects of PEDF on neovascularization are more complex than originally believed and that caution should be exercised when PEDF therapies are considered.

Animals↗
PubMed1976

Diaphragmatic blood flow and energy expenditure in the dog. Effects of inspiratory airflow resistance and hypercapnia.

To investigate the mechanisms which enable the diaphragm to preserve ventilation when the work of breathing is elevated, we measured diaphragmatic blood flow (Q di) and oxygen consumption (VO2 di) in lightly anesthetized dogs. The animals were studied when they breathed quietly, when they inhaled 5% CO2 in 21% or 14% O2, or when they inhaled these gas mixtures through moderate to severe inspiratory resistances. Q di was determined from the integrated diaphragmatic arteriovenous difference of krypton-85, by the Kety-Schmidt technique. VO2 di was calculated as the product of Q di and the diaphragmatic arteriovenous oxygen difference ([A-V]O2 di). Alteration in these parameters consequent to augmentation of ventilatory effort were compared with concomitant alterations in diaphragmatic electrical activity (EMG di) and an inspiratory pleural pressure-time index (PPTI). Addition of inspiratory resistances combined with inhalation of CO2 usually increased Q di and consistently increased VO2 di, EMG di, and PPTI, the maximum increases being approximately 400-1,600% above control levels. In individual animals, as inspiratory resistance was increased, VO2 di, EMG di, and PPTI rose in direct proportion to each other. In the group as a whole, during resistance breathing the oxygen requirements of the diaphragm were met by a combination of increased [A-V]O2 di and Q di. Unlike other skeletal muscles, oxygen extraction tended to plateau at peak loads, whereas blood flow continued to rise as PPTI and VO2 di increased. We conclude that augmentation of perfusion permits the diaphragm to sustain high levels of contractile effort when the work of breathing is increased.

Airway Resistance↗