Effects of High-LET Radiation Exposure and Hindlimb Unloading on Skeletal Muscle Resistance Artery Vasomotor Properties and Cancellous Bone Microarchitecture in Mice.

Weightlessness during spaceflight leads to functional changes in resistance arteries and loss of cancellous bone, which may be potentiated by radiation exposure. The purpose of this study was to assess the effects of hindlimb unloading (HU) and total-body irradiation (TBI) on the vasomotor responses of skeletal muscle arteries. Male C57BL/6 mice were assigned to control, HU (13-16 days), TBI (1 Gy (56)Fe, 600 MeV, 10 cGy/min) and HU-TBI groups. Gastrocnemius muscle feed arteries were isolated for in vitro study. Endothelium-dependent (acetylcholine) and -independent (Dea-NONOate) vasodilator and vasoconstrictor (KCl, phenylephrine and myogenic) responses were evaluated. Arterial endothelial nitric oxide synthase (eNOS), superoxide dismutase-1 (SOD-1) and xanthine oxidase (XO) protein content and tibial cancellous bone microarchitecture were quantified. Endothelium-dependent and -independent vasodilator responses were impaired in all groups relative to control, and acetylcholine-induced vasodilation was lower in the HU-TBI group relative to that in the HU and TBI groups. Reductions in endothelium-dependent vasodilation correlated with a lower cancellous bone volume fraction. Nitric oxide synthase inhibition abolished all group differences in endothelium-dependent vasodilation. HU and HU-TBI resulted in decreases in eNOS protein levels, while TBI and HU-TBI produced lower SOD-1 and higher XO protein content. Vasoconstrictor responses were not altered. Reductions in NO bioavailability (eNOS), lower anti-oxidant capacity (SOD-1) and higher pro-oxidant capacity (XO) may contribute to the deficits in NOS signaling in skeletal muscle resistance arteries. These findings suggest that the combination of insults experienced in spaceflight leads to impairment of vasodilator function in resistance arteries that is mediated through deficits in NOS signaling.

Neurovascular coupling is not mediated by potassium siphoning from glial cells.

Neuronal activity evokes localized changes in blood flow, a response termed neurovascular coupling. One widely recognized hypothesis of neurovascular coupling holds that glial cell depolarization evoked by neuronal activity leads to the release of K+ onto blood vessels (K+ siphoning) and to vessel relaxation. We now present two direct tests of this glial cell-K+ siphoning hypothesis of neurovascular coupling. Potassium efflux was evoked from glial cells in the rat retina by applying depolarizing current pulses to individual cells. Glial depolarizations as large as 100 mV produced no change in the diameter of adjacent arterioles. We also monitored light-evoked vascular responses in Kir4.1 knock-out mice, where functional Kir K+ channels are absent from retinal glial cells. The magnitude of light-evoked vasodilations was identical in Kir4.1 knock-out and wild-type animals. Contrary to the hypothesis, the results demonstrate that glial K+ siphoning in the retina does not contribute significantly to neurovascular coupling.

Adjunct brachytherapy: a new concept to prevent intimal hyperplasia after surgical endarterectomy?

OBJECTIVE: Endarterectomy represents a therapeutical option for patients with advanced coronary artery disease. The mid-term results are compromised by restenosis due to neointima formation. The aim of this study was to evaluate a new treatment concept - endarterectomy with consecutive gamma-irradiation - in a rat model. METHODS: Male Sprague-Dawley rats underwent left carotid endarterectomy with removal of intima: control (n=10) or were irradiated with 15 Gray (Gy) (n=13) or 20 Gy (n=10) postoperatively and compared with sham-operated rats (n=10). After 3 weeks, carotid arteries were perfusion-fixed and vessel compartment areas were measured. Transmission electron microscopy and immunohistochemical staining were used to confirm neointima formation. RESULTS: Three weeks after endarterectomy, neointimal hyperplasia was found in the control group (0.07+/-0.04 mm(2)). After irradiation, a dose-dependent reduction of neointima was observed (0.003 mm(2) at 15 Gy and 0.0007 mm(2) at 20 Gy, P<0.0001). However, immunohistochemical staining revealed that thin re-endothelialization after irradiation was not inhibited. CONCLUSIONS: Gamma-irradiation significantly suppressed neointimal hyperplasia in a rat model of surgical endarterectomy. Despite inhibition of intimal hyperplasia, re-endothelialization after adjuvant brachytherapy was present. Adjuvant brachytherapy may be therefore a new concept to prevent restenosis after endarterectomy in patients.

Morphometric and histological changes in the vascular wall after external radiation for the prevention of intimal hyperplasia.

BACKGROUND: Recent reports describe spontaneous dissections and aneurysms after coronary and peripheral artery irradiation for the prevention of intimal hyperplasia. We investigated histological changes and the vasomotor reaction in the vascular wall after external radiation for the prevention of intimal hyperplasia in rabbits. MATERIALS AND METHODS: The aorta was experimentally injured in 34 rabbits who were then assigned to one of three groups: irradiation with 20 Gy; with 25 Gy; and a control group with no irradiation. Before the arterial injury and 45 days later, vasomotor function was assessed with an intravascular ultrasound catheter. The aorta was resected for morphometric and histological studies. RESULTS: After injury and irradiation, vasomotor responses were significantly lower in the two irradiated groups (P < 0.05). Intimal thickness and the intima/media ratio were significantly lower in irradiated groups. In the irradiated group histological examination showed reduced intimal proliferation with an intact endothelium. In the 20-Gy irradiated group the vascular media contained necrotic areas, and in the 25-Gy irradiated group, severe fibrosis. CONCLUSION: After arterial injury, external irradiation at 20 and 25 Gy effectively reduces aortic intimal and medial thickening. Histological changes include recasting with necrosis and fibrosis causing a decreased vasomotor response. Further investigations are needed to confirm medial necrosis and replacement with fibrosis. Because the irradiation doses in this study match those currently used and also recommended for experimental and clinical use, if confirmed in humans parietal recasting might possibly explain the reported spontaneous dissections and aneurysm formation after irradiation.

Endovascular irradiation impairs vascular functional responses in noninjured pig coronary arteries.

PURPOSE: To assess the effects of endovascular irradiation on vascular structure and function in pig coronary arteries in the absence of vascular injury. METHODS AND MATERIALS: Vasomotor responses to contractions of KCl and prostaglandin F2alpha (PGF2alpha), relaxations to endothelium-dependent (substance P, A23187) and -independent (sodium nitroprusside, SNP) agents; endothelial morphology and superoxide anion (02*-) production were investigated in control (naive), sham and irradiated (20 Gy, 32P) arteries 1 month after irradiation. RESULTS: Contractions to KCl and PGF2alpha in the presence of L-NAME were significantly decreased, relaxations to substance P and A23187 were abolished and SNP-induced relaxation was potentiated in irradiated arteries compared to naive and sham-treated vessels. Scanning electron microscopy (SEM) revealed enlarged endothelial cells (ECs) exhibiting surface microvilli. O2*- production was significantly increased in irradiated vessels (437.0 +/- 37.3 vs. 126.0 +/- 11.6 RLU/s/mg tissue, P < .01). CONCLUSIONS: One month after brachytherapy, normal pig coronary arteries showed abnormal vascular reactivity, altered endothelial morphology and increased production of O2*-. Lack of relaxation to substance P and A23187 reflects ionizing radiation-mediated damage to ECs, whereas potentiation of relaxation to SNP suggests additional deleterious effects on medial smooth muscle cells (SMCs). Increased O2*- production might have contributed to endothelial dysfunction by scavenging nitric oxide (NO).

Vitamin C prevents radiation-induced endothelium-dependent vasomotor dysfunction and de-endothelialization by inhibiting oxidative damage in the rat.

1. The present study was undertaken to determine whether endothelial function or morphology was altered in aortic rings of rats after irradiation, to investigate the mechanism of radiation effects on the endothelium and to examine the effect of vitamin C treatment against radiation-induced damage of the endothelium. 2. Female Sprague-Dawley rats were randomized into four groups (control, radiation, radiation + vitamin C, radiation + vitamin C + NG-nitro-L-arginine methyl ester (L-NAME); n = 10 for each group and n = 7 for the control group) and were irradiated with 10 Gy of 137Cs as a radiation source. Segments of the thoracic aorta were obtained and isometric tension, levels of 8-hydroxydeoxyguanosine (OH-dG) and immunohistochemical staining were measured. 3. Irradiation significantly impaired the acetylcholine-induced vasodilation of aortic segments, an effect that could be prevented by pretreatment with vitamin C (500 mg/kg per day). This beneficial effect of vitamin C was abolished by the addition of L-NAME (100 microg/kg per day), an inhibitor of nitric oxide (NO) synthesis. Irradiation significantly increased the level of OH-dG in the aorta (1.02 +/- 0.27 vs 2.61 +/- 0.78 OH-dG/105 deoxyguanosine (dG) for control and irradiated tissues, respectively; P < 0.01), an increase that was prevented by vitamin C treatment (1.59 +/- 0.23 OH-dG/105 dG; P < 0.01). Irradiation caused significant de-endothelialization (von Willebrand factor (vWF) staining was 93 +/- 7 vs 100% in irradiated and control tissues, respectively; P < 0.05) and this was prevented by vitamin C treatment (vWF staining 98 +/- 3%; P < 0.05). 4. Radiation caused endothelial damage and impaired NO production through oxidative injury, resulting in a selective impairment of endothelial-dependent vasodilation that could be prevented by vitamin C, partly through anti-oxidant mechanisms.

The effect of ionizing irradiation on vasomotor reactivity in the rat thoracic aorta in vitro.

PURPOSE: Ionizing irradiation inhibits restenosis in animal models and human. Vasomotor tone preservation during and after radiation therapy is of clinical importance. We therefore investigated vascular reactivity following radiation therapy. METHODS AND MATERIALS: Wistar Sabra rats were treated with a single dose of 1000 cGy external X-ray irradiation. Vascular reactivity of 192 segments of rat thoracic aorta was studied in vitro in four groups (12 rats in each group, four segments from each aorta). Immediately after in vivo irradiation, immediately after ex vivo irradiation, 1 month after irradiation, and no irradiation (control). RESULTS: Vasoconstriction to phenylephrine (PE) 10(-9)-10(-5) M or KCl 118.0 mM in all the irradiated groups was similar to controls. Endothelium-dependent vasorelaxation to acetylcholine (ACh) 10(-9)-10(-5) M in segments studied immediately after in vivo irradiation was increased compared to controls at all concentrations (109.7+/-35. and 90.0+/-40.0%, respectively, at 10(-5) M, P=.006). Endothelium-independent relaxation to nitroglycerin 10(-9)-10(-5) M in all irradiated groups was similar to controls. CONCLUSIONS: External-ionizing irradiation with 1000 cGy in the rat aortic model induces acute and transient increase in endothelium-dependent relaxation to ACh, and does not alter vasoconstriction and endothelium-independent relaxation.

[The temperature reaction of rabbits to local microwave exposures]. / Temperaturnaia reaktsiia krolikov na lokal'nye vozdeistviia mikrovolnami.

A rapid increase of skin temperature after discontinuation of local microwave (MW) exposure of the heart and thyroid indicates a vasomotor response in the exposed and adjacent zones. The infrared radiation (IR) of the particular parts of skin surface before and after local MW exposures (460 MHz, current density 10, 120 and 230 mW/cm2) of the rabbit heart and thyroid regions was registered by IR thermograph. The investigations were performed on un-narcotized, narcotized (nembutal 40 mg/kg) animals, on rabbits with anaesthetized skin receptors (lidocaine 100 mg) and on carcasses. The changes observed depended on the exposure site, functional status of the rabbits and MW current density. The energy-depending reactions are attributed to direct myocardial reactions and extracardiac vegetative responses in the cardiac and thyroid exposures, respectively.

Effects of hypnotic suggestions on ultraviolet B radiation-induced erythema and skin blood flow.

Results from both animal and human studies have indicated that inflammatory skin reactions such as the flare response to histamine prick test involve a neurogenic regulatory component. It is still unknown to which degree inflammation induced by ultraviolet (UV) radiation may be mediated by the central nervous system. To investigate this, the effect of hypnotic suggestions to increase and decrease the response to UVB radiation on erythema and cutaneous blood flow was investigated in 10 highly hypnotizable subjects. The results showed a significant effect of hypnotic suggestions on cutaneous blood flow compared with the responses of a control group. For erythema no conclusive evidence was found. The results indicate the possibility of separate regulatory mechanisms behind central nervous system influence on UVB-induced erythema and skin blood flow respectively, and further investigations are needed.

Effects of high-dose intracoronary irradiation on vasomotor function and smooth muscle histopathology.

A significant component of restenosis after coronary angioplasty is due to medial proliferation. Targeted ablation of the proliferating cells by ionizing radiation may prevent restenosis. We delivered high-dose intracoronary gamma-irradiation in porcine coronary arteries and assessed vasomotor function acutely and at 32 days, with pathological analysis at 32 days. Changes in luminal area were assessed by intravascular ultrasound. Irradiated segments acutely displayed vasoconstriction to acetylcholine, with loss of smooth muscle response to nitroglycerin. Restudy revealed restoration of normal vasodilatory response to acetylcholine but persistent loss of response to nitroglycerin. Histopathology at 32 days revealed minor neointima formation without luminal compromise and diffuse fibrosis of the smooth muscle layer. The surrounding myocardium was normal. Focal medial fibrosis without significant endothelial or myocardial damage can be achieved via this technique; intracoronary irradiation, therefore, may be an effective way of impairing the restenosis process.

[Effects of continuous laser on arterial vasomotoricity in vitro]. / Effets du laser continu sur la vasomotricité artérielle in vitro.

The effects of a continuous-wave argon laser irradiation on arterial vasomotricity were studied isometrically on 19 aortic segments obtained from rabbits. The segments had a resting tension of 1 g and were suspended in a perfusion chamber filled with a Krebs-bicarbonate buffer solution at 37 degrees C gassed with 95 p. 100 O2 and 5 p. 100 CO2. Irradiation was performed with an argon laser emitting principally at 488-514 nm, via a flexible optical fibre 200 microns in diameter. The tension of each specimen, measured with a strain gauge, was recorded at the same time as the arterial wall temperature, measured by a thermistor probe. The integrity of the endothelium was checked by scanning electron microscopy and acetylcholine test. The endothelium of 5 specimens was deliberately abrased, and this was checked by the same methods. Laser irradiations of more than 1 watt regularly (n = 34) induced a contraction the kinetics of which were parallel to those of the arterial wall heating and usually reversible after the end of irradiation. With irradiations of less than 0.1 watt, the parietal temperature never rose by more than 5 degrees C, and the vasomotor response always (n = 41) was a relaxation which was reversible after irradiation. At intermediate levels of power, the vasomotor response was biphasic and of low amplitude (n = 100). At all levels, similar responses were given by specimens whose endothelium had been removed. Thus, at the powers used for angioplasty, irradiation of the arterial wall by continuous-wave argon laser induces and intense vasoconstriction which is independent of the endothelium.(ABSTRACT TRUNCATED AT 250 WORDS)

Metabolic and vasomotor responses of rhesus monkeys exposed to 225-MHz radiofrequency energy.

A previous study showed a substantial increase in the colonic temperature of rhesus monkeys (Macaca mulatta) exposed to radiofrequency (RF) fields at a frequency near whole-body resonance and specific absorption rates (SAR) of 2-3 W/kg. The present experiments were conducted to determine the metabolic and vasomotor responses during exposures to similar RF fields. We exposed five adult male rhesus monkeys to 225 MHz radiation (E orientation) in an anechoic chamber. Oxygen consumption and carbon dioxide production were measured before, during, and after RF exposure. Colonic, tail and leg skin temperatures were continuously monitored with RF-nonperturbing probes. The monkeys were irradiated at two carefully-controlled ambient temperatures, either cool (20 degrees C) or thermoneutral (26 degrees C). Power densities ranged from 0 (sham) to 10.0 mW/cm2 with an average whole-body SAR of 0.285 (W/kg)/(mW/cm2). We used two experimental protocols, each of which began with a 120-min pre-exposure equilibration period. One protocol involved repetitive 10-min RF exposures at successively higher power densities with a recovery period between exposures. In the second protocol, a 120-min RF exposure permitted the measurement of steady-state thermoregulatory responses. Metabolic and vasomotor adjustments in the rhesus monkey exposed to 225 MHz occurred during brief or sustained exposures at SARs at or above 1.4 W/kg. The SAR required to produce a given response varied with ambient temperature. Metabolic and vasomotor responses were coordinated effectively to produce a stable deep body temperature. The results show that the thermoregulatory response of the rhesus monkey to an RF exposure at a resonant frequency limits storage of heat in the body. However, substantial increases in colonic temperature were not prevented by such responses, even in a cool environment.

[Heterogeneity of neurocytes of different brain regions to repeated superhigh-frequency irradiation]. / Geterogennost' reaktsii neirotsitov razlichnykh otdelov golovnogo mozga na mnogokratnoe sverkhvysokochastotnoe obluchenie.

The study was based on the analysis of morphological changes in the brain of 45 rats exposed for 10 days to ultrahigh frequency field of moderate gravity. Detailed studies of the state of the basophilic substance and the volume of the nuclei in the course of the neurocyte swelling were undertaken. The response of the neurocytes of various brain divisions was found to be heterogenous. The most pronounced were dystrophic changes in the sincipital area of the cortex. A lack of correlation between the character of the nuclear swelling and the degree of chromatolysis in the thalamic neurocytes was noted.