Blood Flow Stasis Research Articles

Induction of tumour blood flow stasis and necrosis: a new function for epinephrine similar to that of combretastatin A-4 derivative AVE8062 (AC7700).

AVE8062, a derivative of combretastatin A-4, has a strong stanching effect on tumour blood flow (TBF), which leads to complete blockage of nutrient supply to solid tumours and their necrosis. Previously, we reported that TBF stasis is due to increased arteriolar resistance caused by AVE8062 and a lasting decrease in perfusion pressure in tumour-feeding vessels. Here, we measured changes in TBF in rat solid tumour LY80 during continuous administration of AVE8062-like epinephrine or four catecholamines that are unlike AVE8062 (norepinephrine, dopamine, methoxamine, and metaraminol) to the region of increased vascular resistance. Venous administration of 0.3 mg ml−1 epinephrine caused TBF to fall immediately to near zero, where it remained throughout the administration period. With a 30-min drug administration, TBF began to recover immediately when drug administration halted. With a 60-min epinephrine administration, TBF recovered somewhat, but not to the previous level. With drug administration of 120 min, TBF did not recover during the subsequent 8 h. Likewise, 0.1 mg ml−1 epinephrine produced irreversible occlusion after 120 min of administration. In contrast, 120 min of administration of the four other catecholamines resulted in no occlusion. Only the group given 0.3 mg ml−1 epinephrine (not that given methoxamine) showed significantly greater necrosis than the control. We conclude that, for epinephrine to cause irreversible occlusion of these vessels, a marked decrease in perfusion pressure in tumour-feeding blood vessels is necessary and should be maintained for 2 h. This conclusion is consistent with the previously demonstrated mechanism of irreversible arteriole occlusion caused by AVE8062. AVE8062 and epinephrine appear to have the same mechanism of action regarding induction of tumour blood flow stasis.

Mucosal Villus Microcirculatory Disturbances Associated with Rat Intestinal Ischaemia-Reperfusion Injury Are Not Prevented by Tacrolimus

Background/Aims: Microcirculatory disturbances following small intestinal ischaemia-reperfusion (I/R) injury lead to tissue damage that may affect short- and long-term outcome after transplantation. The immunosuppressive drug Tacrolimus (FK506) attenuates I/R injury in a number of organs, raising the possibility that it might be able to control both I/R injury and rejection after small bowel transplantation. However, its effects on intestinal I/R injury have not been evaluated. Methods: PVG rats were subjected to 30 min intestinal ischaemia. Animals received Tacrolimus (1 mg/kg i.p.) 4 and 1 h prior to ischaemia. The mucosa was visualised in an exteriorised ileal segment using in vivo microscopy. FITC-BSA or Acridine orange was used to quantitate macromolecular leak (MML) and leucocyte adhesion respectively every 15 min for 2 h during reperfusion. Heart rate and mean blood pressure (mBP) were monitored throughout the experiment. Results: Ten of 12 untreated animals subjected to intestinal I/R injury failed to survive the 2-hour reperfusion period. MML and leucocyte adhesion were increased in untreated animals (p < 0.001) and blood flow stasis eventually ensued. Similar results were obtained for Tacrolimus pre-treated I/R animals, with 10 of 12 animals again failing to survive reperfusion. Conclusions: Despite previous evidence that Tacrolimus reduces I/R injury in other organs, it did not improve survival rates or prevent villus microcirculatory disturbances following intestinal I/R injury. The severity of microcirculatory damage suffered by the small intestine highlights the importance of alternative therapies to combat I/R in this organ.

Hepatic outflow obstruction: enhancement patterns of the liver on MR angiography

Our purpose was to present the enhancement patterns of the liver on MR angiography in patients with hepatic outflow obstruction. Twenty-three patients with Budd-Chiari syndrome (4 in acute stage and 19 in chronic stage of the disease) were examined with 3D contrast-enhanced MR angiography. During early and late portal venous phase of MR angiography the pattern of parenchymal enhancement was assessed on source images. The enhancement patterns were evaluated under 4 groups as following: (a) central (b) peripheral (c) patchy and (d) homogeneous enhancement. The morphologic changes in the liver (lobar hypertrophy or atrophy, hepatic surface irregularities) were also recorded. In the acute stage global liver enlargement (75%) with caudate hypertrophy (100%) and central enhancement of the liver (75%) were suggestive findings of the hepatic outflow obstruction. The left lobe hypertrophy (53%) associated with the caudate lobe hypertrophy (72%) and irregular surface (26%) were predominant in the chronic stage of the disease. The enhancement patterns seen in chronic disease were variable and reflected the persistent stasis of the portal blood flow (patchy enhancement in 32% of the patients) or the altered hemodynamics of the liver due to the development of subcapsular collaterals (peripheral enhancement in 21% of the patients). Homogeneous enhancement of the liver in Budd-Chiari syndrome may indicate the chronicity of the outflow obstruction (37%) and shows a more stable hepatic perfusion that occurs after the formation of intra and extrahepatic collateral veins. The morphological and perfusional features on multiphase contrast-enhanced MR angiography are valuable in understanding the effects of the hepatic outflow obstruction on the liver parenchyma.

Effects of FK409 on intestinal ischemia-reperfusion injury and ischemia-induced changes in the rat mucosal villus microcirculation.

The small intestine is extremely sensitive to ischemia-reperfusion (I/R) injury and a range of microcirculatory disturbances contribute to tissue damage. Nitric oxide (NO) seems to be involved in tissue protection after I/R injury. This study therefore assessed the effects of the NO donor, FK409, on intestinal I/R injury and changes induced in intestinal microcirculation. PVG rats were subjected to 30-min intestinal ischemia with a subgroup of animals receiving FK409 (10 mg/kg i.v.) 30 min before ischemia and 30 min postreperfusion. Controls underwent sham surgery. The mucosal surface was visualized via an incision made in an exteriorized ileal segment and FITC-BSA or acridine orange was used to quantitate macromolecular leak (MML) and leukocyte adhesion, respectively. MML from, and numbers of adherent leukocytes within, individual villi were determined every 15 min for 2 hr after removal of the vessel clamp. Heart rate and mean blood pressure (mBP) were monitored throughout the experiment. Eleven of 12 untreated animals subjected to intestinal I/R injury failed to survive the 2 hr reperfusion period, whereas all 12 FK409-treated animals survived. MML and leukocyte adhesion were increased in untreated animals (P<0.001), and blood flow stasis eventually ensued. Although FK409 decreased mBP (P<0.001), MML and leukocyte adhesion were significantly (P<0.001) reduced, and villus blood flow was maintained throughout the observation period. FK409 prevented mortality after intestinal I/R, significantly reduced leukocyte adhesion, and maintained blood flow after intestinal ischemia and may therefore be of value in reducing tissue damage and improving outcome after small bowel transplantation.

Current trends in the management of thromboembolic events.

Thrombus formation on a disrupted atherosclerotic plaque is the main pathogenetic mechanism for the acute coronary syndromes of myocardial infarction and unstable angina. Myocardial infarction results from an acute total occlusion of the artery, while unstable angina is secondary in most cases to mural thrombus formation. Thrombus formation has also been implicated in chronic atherosclerotic disease progression, and in restenosis following coronary angioplasty. Therapeutic measures to treat thrombus rely on the ability of drugs either to prevent thrombus extension, to dissolve its fibrin component, or to prevent further platelet aggregation. Following arterial vessel wall injury, platelets adhere to the subendothelial collagen. Glycoprotein Ib on the platelet membrane is the primary receptor for this single layer of platelets, and it binds von Willebrand factor as its extracellular ligand. Platelet aggregation involves activation of several different pathways leading to exposure of the glycoprotein IIb/IIIa receptor on the platelet. These receptors bind either fibrinogen or von Willebrand factor, leading to platelet‐ platelet interaction and the formation of a white platelet thrombus. The luminal component of thrombus is a mixture of platelets and fibrin. The occlusive tail of the thrombus is composed of red blood cells and fibrin (red thrombus). Both plaque disruption and erosion of a plaque involve the presence of an inflammatory process at the site of thrombus formation. This inflammatory process consists primarily of activated macrophages and T lymphocytes. Thrombus formation depends on at least three factors: (i) the depth of vessel wall injury and the thrombogenicity of the exposed plaque; (ii) the prothrombotic state; and (iii) haemodynamic factors that promote vasoconstriction and stasis of blood flow. Traditionally, plaque rupture has been considered to result mainly from the physical pressure exerted upon the plaque surface. The plaque's vulnerability to rupture depends upon the size and consistency of its atheromatous core. Davies …

Características clínicas y prevalencia de los estados de hipercoagulabilidad en pacientes con enfermedad tromboembólica venosa

Blood flow stasis, an alteration in the coagulation process or endothelium injury the most common mechanisms involved in venous thromboembolic diseases (TED) and constitute the so-called hypercoagulable states (HS). The HS have been classified into two groups: primary and secondary and several of them can be measured. Our purpose was to investigate the prevalence in Spain of measurable HS in patients with venous TED and to know the clinical picture of pulmonary embolism in these patients. Sixty of 175 consecutive patients diagnosed with deep venous thrombosis and pulmonary embolism fulfilled criteria to study a HS. The study was performed 1 month after anticoagulant therapy was finished. Seventeen (28%) of the 60 patients had one HS that was a primary disorder in 14 and a secondary one in 3. The increase of PAI-1 level was the most common; recurrence of thrombotic events and familial history were frequent in these selected patients. The prevalence of hypercoagulable states in venous thromboembolic disease was high in this series. The clinical picture was similar to those to be expected and PAI-1 was the most frequent marker of hypercoagulable states.

Comparison of thermal damage calculated using magnetic resonancethermometry, with magnetic resonance imaging post-treatment and histology,after interstitial microwave thermal therapy of rabbit brain

Clinical application of high-temperature thermal therapy as a treatment forsolid tumours requires an accurate and close to real-time method forassessing tissue damage. Imaging methods that detect structural changesduring heating may underestimate the extent of thermal damage. This is dueto the occurrence of delayed damage manifested at tissue locations exposedto temperatures lower than those required to cause immediate structuralchanges. An alternative approach is to measure temperature and thencalculate the expected damage based on the temperature history at eachtissue location. Magnetic resonance (MR) imaging methods now allowtemperature maps of the target and surrounding tissues to be generated inalmost real-time. The aim of this work was to evaluate whether thermaldamage zones calculated on the basis of MR thermometry maps measured duringheating correspond to actual tissue damage as measured after treatment byhistological methods and MR imaging.Four male rabbits were treated with high-temperature thermal therapydelivered in the brain by a single microwave antenna operating at 915 MHz.MR scanning was performed before, during and after treatment in a 1.5 Twhole-body scanner. Temperature maps were produced using the protonresonance frequency (PRF) shift method of MR thermometry. In addition,conventional T1-weighted and T2-weighted spin-echo images wereacquired after treatment. Thermal damage zones corresponding to cell death,microvascular blood flow stasis and protein coagulation were calculatedusing an Arrhenius analysis of the MR temperature/time course data. Thecalculated zones were compared with the lesions seen on histopathologicalexamination of the brains which were removed within 6-8 h of treatment.The results showed that calculated damage zones based on MR thermometryagreed well with areas of damage as assessed using histology after heatingwas completed. The data suggest that real-time calculations of finalexpected thermal damage based on an Arrhenius analysis of MR temperaturedata may provide a useful method of real-time monitoring of thermal therapywhen combined with conventional T2-weighted images taken aftertreatment.

Structural Remodeling of the Left Atrial Appendage in Patients with Chronic Non-Valvular Atrial Fibrillation: Implications for Thrombus Formation, Systemic Embolism, and Assessment by Transesophageal Echocardiography

Left atrial appendage (LAA) is frequently the site of thrombus formation in patients with chronic atrial fibrillation (AF). Transesophageal echocardiography and hematologic studies have identified blood flow stasis (spontaneous echogenic contrast) and abnormal coagulation (increased serum fibrinogen) as important predisposing factors to formation of LAA thrombi. However, the third component of the Virchow's triad, i.e., endothelial abnormalities, has not been adequately studied. Accordingly, we studied, at necropsy, the LAA morphology in 46 hearts of patients with ( n = 22) and without ( n = 24) chronic AF. Compared to patients without AF, those with AF had significantly larger LAA volumes (1.7% 1.1 vs. 5.4% 3.7 mL, p = 0.0002), and larger luminal surface area of the bisected LAA (4.4% 1.8 vs. 7.1% 4.5 cm 2, p = 0.01). However, both the absolute and relative surface area of the transected pectinate muscles were reduced in patients with AF (2.6% 1.1 vs. 1.8% 1.0 cm 2, p = 0.02 and 38% 15 vs. 21% 14%, p = 0.0003). In addition, in most patients (73%) with chronic AF, the LAA showed significant endocardial thickening with fibrous and elastic tissue (endocardial fibroelastosis) compared to those without AF (13%, p < 0.0001). Endocardial fibroelastosis resulted in a smooth LAA luminal surface and encased the pectinate muscles. These findings suggest that LAA remodeling (dilation, stretching, and reduction in pectinate muscle volume, as well as endocardial fibroelastosis) occurs frequently in chronic AF and may contribute to the increased risk of thrombus formation and systemic embolism. Additionally, the information may have relevance in interpreting transesophageal echocardiographic images of the LAA in patients with chronic AF.

Mast cell stabilization prevents ethanol-induced rat gastric mucosal injury: mechanisms of protection.

We previously demonstrated that 60% ethanol increased macromolecular leakage and induced lesion formation in areas of permanent flow stasis within gastric mucosal vessels. Mast cells and their mediators have been implicated in acute mucosal injury. Fluorescent in vivo microscopy was used to assess the effects of ketotifen, a mast cell stabilizer, and pyrilamine, a histamine (H1)-receptor antagonist, on ethanol-induced rat gastric mucosal injury. Experiments were carried out on anaesthetized rats pretreated orally with ketotifen (1 mg/kg) or pyrilamine (30 mg/kg). Fluorescein isothiocyanate-bovine serum albumin (FITC-BSA; 0.2 mL/100 g), a marker for quantitating macromolecular leakage was administered intra-arterially. Ethanol (60%) or distilled water was applied topically to the gastric mucosa. Macromolecular leakage of FITC-BSA, vessel diameters and leucocyte activity were quantified using image analysis. Pretreatment with ketotifen or pyrilamine, followed by ethanol, caused no change in macromolecular leakage compared with controls. Both compounds prevented blood flow stasis in all areas and no lesion formation was observed. However, increased leucocyte activity and increases in vessel diameter were observed following pretreatment with ketotifen and pyrilamine, respectively. The data suggest that vasoactive substances released from mast cells may be involved in the aetiology of ethanol-induced gastric mucosal damage. The prevention of these normal physiological responses to injury may lead to the employment of other microcirculatory mechanisms of defence.

Analysis of acute vascular damage after photodynamic therapy using benzoporphyrin derivative (BPD).

Benzoporphyrin derivative monoacid ring A (BPD-MA, verteporfin) is currently under investigation as a photosensitizer for photodynamic therapy (PDT). Since BPD exhibits rapid pharmacokinetics in plasma and tissues, we assessed damage to tumour and muscle microvasculature when light treatment for PDT was given at short times after injection of photosensitizer. Groups of rats with chondrosarcoma were given 2 mg kg−1 of BPD intravenously 5 min to 180 min before light treatment of 150 J cm−2 690 nm. Vascular response was monitored using intravital microscopy and tumour cure was monitored by following regrowth over 42 days. For treatment at 5 or 30 min after BPD injection, blood flow stasis was limited to tumour microvasculature with lesser response in the surrounding normal microvasculature, indicating selective targeting for damage. No acute changes were observed in vessels when light was given 180 min after BPD injection. Tumour regression after light treatment occurred in all animals given PDT with BPD. Long-term tumour regression was greater in animals treated 5 min after BPD injection and least in animals given treatment 180 min after drug injection. The correlation between the timing for vascular damage and cure implies that blood flow stasis plays a significant role in PDT-induced tumour destruction. © 1999 Cancer Research Campaign

Epi-illumination fluorescent light microscopy for the in vivo study of rat hepatic microvascular response to cryothermia.

To elucidate the hepatic microvascular response to cryothermia, we studied the liver microcirculation of Sprague-Dawley rats after one and two 4-minute freeze-thaw cycles using intravital fluorescence microscopy. Irrespective of the number of freeze-thaw cycles applied, the nature of hepatic microvascular injury was characterized by complete stasis of sinusoidal blood flow within the central part of the cryolesions and heterogeneous sinusoidal perfusion in a critically perfused border zone located at the periphery of the lesions. Analysis over time (2 hours) revealed a successive shutdown of sinusoidal perfusion within this critically perfused border zone, which was caused by intravascularly lodging cell aggregates, blocking the lumen of individual sinusoids. The aggregates consisted of parenchymal cells and cell fragments, but did not include leukocytes or platelets. Strikingly, microvascular perfusion failure was associated with Ito cell disintegration and marked dilation of sinusoids (15.6 +/- 0.8 microm vs. 8.8 +/- 0.8 microm; P <.05). This excludes sinusoidal constriction as the cause of nutritive perfusion failure, and may indicate dysfunction of Ito cell-regulated vasomotor control by cryothermia. However, because circulating cell aggregates were frequently observed plugging individual microvessels, dilation of sinusoids may just be the result of passive distension caused by outflow blockade. Analysis of hepatic tissue at 8 weeks after cryothermia did not reveal regeneration and microvascular remodeling, but loss of hepatic tissue, which corresponded well with the tissue area presenting with sinusoidal perfusion failure during the initial observation period after cryothermia. The fact that there was no recovery of sinusoidal perfusion over the initial 2-hour observation period, but loss of tissue after 8 weeks, supports the view that cryothermia induces injury not only by direct low-temperature-mediated action, but also through ischemia caused by irreversible deterioration of the microcirculation.

Reversal and protection against indomethacin-induced blood stasis and mucosal damage in the rat jejunum by a beta3-adrenoceptor agonist.

In the rat, indomethacin causes jejunal villous shortening, microvascular distortion, and blood stasis prior to ulceration. The beta3-adrenoceptor agonist CL316,243 (CL) prevents both the early histological changes and ulceration. To test the hypothesis that the beta3-adrenoceptor agonist CL316,243 exerts its protective effect by prevention and/or reversal of blood flow changes in the rat jejunum exposed to indomethacin. In anaesthetized rats, jejunal villous blood flow was measured in surface capillaries using fluorescence microscopy. Stasis of superficial capillary blood flow was induced by combined topical and i.v. indomethacin (100 microg/mL, 2.8 x 10(-4) M). To examine the effect of CL on blood stasis, CL was applied either i.v. (1 mg/kg) or luminally (100 microg/mL, 2.5 x 10(-5)M) at the onset of stasis. Prophylactic protection was assessed by giving i.v. CL simultaneously with indomethacin. Results were compared with controls which received luminal saline applied at blood stasis. The effect of i.v. CL (1 mg/kg) alone, or luminal CL (100 microg/mL) alone on basal villous blood flow was also examined. The small intestines were perfusion-fixed with 10% formol saline, and removed for histology, n = 5 for all groups. Luminal CL given at stasis reversed indomethacin-induced stasis within 10 min, whereas i.v. CL did not. Pretreatment with i.v. CL prevented the onset of stasis. Basal blood flow was raised slightly only by luminal CL. The beta-adrenoceptor agonist CL316,243 can protect against indomethacin-induced blood stasis in rat jejunal villi.

Managing Atrial Fibrillation to Prevent Its Major Complication

Atrial fibrillation is an acute or chronic arrhythmia that occurs postoperatively or during intense emotional stress, exercise, or acute alcohol intoxication. More than 10% of Americans aged 75 and older have atrial fibrillation, which is common in elderly patients with cardiopulmonary disorders. During atrial fibrillation, uncoordinated electrical activity leads to ineffective atrial contraction, reduced atrial filling time, and decreasing cardiac output. Blood flow stasis may cause thrombi to form in the quivering atria. Cardioversion may be indicated to convert an unstable patient into sinus rhythm. However, if cardioversion converts the patient's status to sinus rhythm, thrombi may become dislodged and propelled into the bloodstream as emboli. Occlusion of a cerebral blood vessel often follows, leading to stroke. Because patients with longstanding atrial fibrillation are predisposed to stroke, anticoagulation therapy (usually with heparin, warfarin, or aspirin) should be initiated 3 weeks prior to cardioversion. Proper anticoagulation can usually prevent ischemic stroke.

Acute blood stasis reduces interstitial uptake of albumin from intestinal microcirculatory networks.

Temporary blood flow stoppage occurs in a greater percentage of the capillaries when blood flow to organs is reduced. Previous studies on the small intestine have suggested that acute blood stasis (< or = 10 min) results in expression of negative charge, not present when blood flow is brisk, on the luminal surface of mucosal capillaries. Negative surface charge would tend to reduce transcapillary passage of albumin from blood to interstitium, since albumin is also negatively charged. Here we test the hypothesis that acute blood stasis reduces the interstitial uptake of albumin from mucosal capillary networks in rat small intestine in situ. Animals were subjected to two treatments, which included intestinal blood flow and acute stasis. After each treatment, fluorescent albumins were perfused into the intestinal circulation, and then interstitial fluorescence was recorded using fluorescence microscopy. Images were later quantified by computer analysis. After brisk blood flow, but not after acute blood stasis, fluorescence rapidly appeared in the interstitium and resulted in higher interstitial fluorescence intensity values. These results may have relevance to the mechanisms by which albumin flux in the small intestine is synchronized with digestion and fasting, which are associated with high and low intestinal blood flow, respectively.

The effects of thrombocytopenia on vessel stasis and macromolecular leakage after photodynamic therapy using photofrin.

Several studies have reported thrombus formation and/or the release of specific vasoactive eicosanoids, suggesting that platelet activation or damage after photodynamic therapy (PDT) may contribute to blood flow stasis. The role of circulating platelets on blood flow stasis and vascular leakage of macromolecules during and after PDT was assessed in an intravital animal model. Sprague-Dawley rats bearing chondrosarcoma on the right hind limb were injected intravenously (i.v.) with 25 mg/kg Photofrin 24 h before light treatment of 135 J/cm2 at 630 nm. Thrombocytopenia was induced in animals by administration of 3.75 mg/kg of rabbit anti-rat platelet antibody i.v. 30 min before the initiation of the light treatment. This regimen reduced circulating platelet levels from 300,000/mm3 to 20,000/mm3. Reductions in the luminal diameter of the microvasculature in normal muscle and tumor were observed in control animals given Photofrin and light. Venule leakage of macromolecules was noted shortly after the start of light treatment and continued throughout the period of observation. Animals made thrombocytopenic showed none of these changes after PDT in either normal tissues or tumor. The lack of vessel response correlated with the absence of thromboxane release in blood during PDT. These data suggest that platelets and eicosanoid release are necessary for vessel constriction and blood flow stasis after PDT using Photofrin.

Superoxide dismutase reduces islet microvascular injury induced by streptozotocin in the rat.

Intravenous administration of streptozotocin (STZ) leads to permanent diabetes mellitus in rats. We investigated the possible role of islet microcirculatory changes and free radical formation in this animal model. In vivo fluorescence microscopy was performed for 4 h after administration of STZ. Vascular permeability, capillary blood flow, and endothelial leukocyte adhesion were measured in endocrine and exocrine pancreatic tissue. The earliest microcirculatory event was an increase in vascular permeability in pancreatic islets, with a peak 1 h after STZ administration. The difference between islet and exocrine tissue light intensity was +15.8 +/- 5.6% at t = 60 min. Islet blood flow velocity significantly decreased after 3 h, whereas blood flow in the exocrine pancreas was not affected. Complete stasis of islet blood flow was observed only in rats receiving STZ. Neither increased leukocyte adhesion to islet vascular endothelium nor ischemia-reperfusion phenomena were observed. Prophylactic administration of the radical scavenger superoxide dismutase prevented STZ-induced damage to the islet microcirculation in the initial phase of this model. We conclude that STZ leads to severe microcirculatory disturbances within pancreatic islets in rats. Apparently, these changes are mediated at least in part by free oxygen radicals.

Morphological changes elicited in skeletal muscle by a Nd: YAG laser scalpel and electrocautery during surgical reduction of the human tongue

A Nd:YAG laser scalpel was used for the surgical reduction of a human hyperplastic tongue. This instrument combines a fine cutting precision with haemostatic properties, whereby loss of blood is minimized and the surgeon's field of view unimpeded by flooding from the damaged capillary bed. The coagulative properties of Nd:YAG laser light are, however, insufficient to effect blood flow stasis in larger calibre vessels (arteries > 2 mm; veins > 3-5 mm), such as those located at the base of the tongue. For this purpose, bipolar diathermy (electrocautery) was employed. The ultrastructural changes incurred by skeletal muscle fibres using these two "heat" sources were compared. The damage profile elicited using each modality was similar: coagulation of myofilamentous proteins leads to destruction of fibrillar architecture with concomitant loss of periodic banding; on moving away from the wound margin, characteristic features are gradually restored. As the severity of these heat-induced effects decreases, there is a corresponding increase in superimposed dislocation and tearing phenomena induced by post-treatment swelling.

Studies on gastric mucosal microcirculation. 1. The nature of regional variations induced by ethanol injury.

The focal nature of gastric ulcers raises the possibility of underlying regional disturbances in gastric mucosal microcirculation. This study employed fluorescent in vivo microscopy with the aim of directly investigating the response of several areas of the gastric mucosa to 60% ethanol. Changes in macromolecular leakage of fluorescein labelled albumin, vessel diameter, and acridine red labelled leucocyte adhesion and rolling were assessed over a period of two hours. A total of 0.5 ml 60% ethanol was topically applied for five minutes to the exteriorised gastric mucosa of anaesthetised rats. RATS: Three distinct patterns of response were found. Areas of lesion formation were small and occurred within five minutes. These areas showed persistent blood flow stasis throughout the course of the experiment, increased leakage (p < 0.02), and no leucocyte adhesion. Peripheral to the lesion, sustained leakage (p < 0.02) was found with adherence of leucocytes (p < 0.01) after lesion formation. Sites more remote to any lesion showed transient leakage and significant numbers of 'rolling' leucocytes (p < 0.01) were observed again after the lesion had formed. Despite widespread exposure of the entire gastric mucosa to 60% ethanol the resultant mucosal injury was limited. Widespread vascular damage was found reflected by macromolecular leakage, the pattern of which showed regional variation.

Vascular effects of photodynamic therapy.

Vascular damage and blood flow stasis are consequences of photodynamic therapy (PDT) of solid tumors using many photosensitizers. Microvascular stasis and resulting hypoxia are effective means to produce cytotoxicity and tumor regression. The observation of blood flow stasis after photodynamic therapy results from a combination of damage to sensitive sites within the microvasculature and the resulting physiological responses to this damage. A generalized hypothesis for the mechanisms leading to vessel stasis begins with perturbation and damage to endothelial cells during light treatment of photosensitized tissues. Endothelial cell damage leads to the establishment of thrombogenic sites within the vessel lumen and this initiates a physiological cascade of responses including platelet aggregation, the release of vasoactive molecules, leukocyte adhesion, increases in vascular permeability, and vessel constriction. These effects from damage combine to produce blood flow stasis.

Photodynamic treatment of human endothelial cells promotes the adherence of neutrophils in vitro.

The effects of photodynamic treatment (PDT) on venules include vascular leakage accompanied by oedema formation, vasoconstriction and blood flow stasis. The goal of this study was to gain insight into the mechanism underlying these vascular events by studying one of the earliest observations after PDT, granulocyte adhesion, in an in vitro model. For this purpose human umbilical vein endothelial cells (HUVECs) preincubated with Photofrin II (PII) were illuminated with red light and incubated with neutrophils. PDT led to a dramatic change in the morphology of the endothelial cells. Clearly, neutrophils adhered to the subendothelial matrix and their adherence coincided with an increase in the percentage of exposed subendothelial matrix by the gradual contraction of endothelial cells. Furthermore, the increase in adherence was dependent on drug dose, illumination time and the time delay after PDT. The neutrophil adherence could be inhibited by anti-beta2-integrin antibodies, which suggests that the alphaL-, alphaM- or alphaX-beta2 receptors of the neutrophil mediated this phenomenon. At 4 degrees C or by preincubation of the neutrophils with staurosporin, their adherence to the subendothelial matrix exposed by PDT of endothelial cells could be prevented. Apparently, activation of the beta2-integrin receptor by interaction with the subendothelial matrix is necessary for the increased binding of neutrophils. Taken together, these in vitro findings suggest that the PDT-induced contraction of the endothelial cells permits neutrophil adherence to the subendothelial matrix. It is conceivable that a similar mechanism contributes to the initial adherence of granulocytes to the vessel wall as observed after PDT in vivo.