Time course of capillary structure changes in rat skeletal muscle following strenuous eccentric exercise.

AIM: We examined the time course of capillary structure changes in rat skeletal muscle at 1, 3 and 7 days after strenuous eccentric exercise. METHODS: The right gastrocnemius muscles of anaesthetized male Wistar rats were subjected to 300 controlled eccentric contractions using electrical stimulation. The contralateral gastrocnemius muscle was used as control. All morphometric parameters were determined in in situ perfused gastrocnemius muscles in red (Gr, predominantly slow-twitch fibre) and white (Gw, predominantly fast-twitch fibre) portions. RESULTS: Muscle fibre damage was evident on days 1, 3 and 7 in Gr (29.3-53.9% damaged fibres) and Gw (58.9-86.8% damaged fibres) of exercised legs. Electron micrographs of transverse sections did not display collapsed or obstructed capillaries in exercised legs, and capillary endothelial cells retained their normal structures. However, capillary luminal shapes and area were altered in exercised legs on days 1 and 3. The ratio between minimal and maximal capillary diameter in a transverse section (i.e. luminal ellipticity) significantly differed when comparing control (Gr, 0.75 +/- 0.02; Gw, 0.79 +/- 0.03) and exercised legs (Gr, 0.65 +/- 0.03; Gw, 0.66 +/- 0.04) at 1 day after exercise. The mean capillary luminal area was significantly increased in exercised legs after 1 day (Gw, +24.3%) and 3 days (Gr, +31.9%; Gw, +62.2%) compared with control. CONCLUSION: We conclude that (1) capillary endothelial cell structure was maintained in damaged muscles, (2) changes in capillary lumen shapes and distensibility occur in the degenerated muscle up to 3 days after the eccentric contraction period.

Stroke outcome in double-mutant antioxidant transgenic mice.

BACKGROUND AND PURPOSE: Both NO and superoxide cytotoxicity are important in experimental stroke; however, it is unclear whether these molecules act within parallel pathological pathways or as coreagents in a common reaction. We examined these alternatives by comparing outcomes after middle cerebral artery occlusion in male and female neuronal NO synthase (nNOS)-deficient (nNOS-/-) or human CuZn superoxide dismutase-overexpressing (hSOD1+/-) mice and a novel strain with both mutations. METHODS: Permanent middle cerebral artery occlusion was performed by use of the intraluminal filament technique (18 hours). Neurological status was scored, and tissue infarction volume was determined by 2,3,5-triphenyltetrazolium staining and image analysis. RESULTS: Hemispheric infarction volume was reduced in each transgenic strain relative to the genetically matched, wild-type, control cohorts (WT mice): nNOS-/- (80+/-6 mm(3)) and double-mutant (49+/-6 mm(3)) mice versus WT mice (114+/-7 mm(3)) and hSOD1+/- mice (52+/-7 mm(3)) versus WT mice (95+/-5 mm(3)). Human CuZn superoxide dismutase had a larger effect on mean infarction volume (30% of contralateral hemisphere) than did nNOS deficiency (46%). Although infarction volume was less in double-mutant mice compared with nNOS-/- mice, injury was not improved relative to hSOD1+/- mice. There was no difference in histological damage by sex within each strain; however, female nNOS-/- mice were not protected from ischemic injury, unlike male mutants. CONCLUSIONS: Superoxide generation contributes to severe ischemic brain injury in vivo to a greater extent than does neuronally derived NO. In vivo, significant superoxide scavenging by CuZn superoxide dismutase occurs within cellular compartments or through biochemical pathways that are not restricted to, and may be distinct from, neuronal NO/superoxide reaction and peroxynitrite synthesis.

Heme oxygenase-2 acts to prevent neuronal death in brain cultures and following transient cerebral ischemia.

Heme oxygenase (HO) cleaves the heme ring to form biliverdin, which is rapidly reduced to bilirubin, carbon monoxide, and iron. HO1, the first form of the enzyme discovered, is an inducible protein, concentrated in tissues that are exposed to degrading red blood cells and stimulated by hemolysis and numerous other toxic perturbations to eliminate potentially toxic heme. By contrast, HO2 is constitutive and most highly concentrated in neural tissues. Carbon monoxide, formed from HO2, is a putative neurotransmitter in the brain and peripheral autonomic nervous system. HO1 regulates the efflux of potentially toxic iron from cells, as iron efflux is deficient in mice with targeted deletion of HO1 (HO1(-/-)), and transfection of HO1 facilitates iron efflux. Bilirubin appears to be a physiologic neuroprotectant. Activation of HO2 by phorbol esters, that stimulate protein kinase C to phosphorylate HO2, augments production of bilirubin which protects brain cultures from oxidative stress. Bilirubin itself in nanomolar concentrations is neuroprotective, while HO2 deletion (HO2(-/-)) leads to increased neurotoxicity in brain cultures and increased neural damage following transient cerebral ischemia in intact mice. Mechanisms whereby HO2 provides neuroprotection have not been clarified including whether protection is primarily associated with apoptotic or necrotic cell death. Moreover, the generality of neurotoxic stimuli influenced by HO2 has been unclear. We now demonstrate increased neuronal death in cerebellar granule cultures of HO2(-/-) mice with a selective augmentation of apoptotic death. We also demonstrate that HO2 transfection rescues apoptotic death. In intact mice, we show an increased incidence of apoptotic morphology in the penumbra area surrounding the infarct core in HO2(-/-) mice undergoing transient focal ischemia.

Stroke in estrogen receptor-alpha-deficient mice.

BACKGROUND AND PURPOSE: Recent evidence suggests that endogenous estrogens or hormone replacement therapy can ameliorate brain damage from experimental stroke. Protective mechanisms involve enhanced cerebral vasodilation during ischemic stress as well as direct preservation of neuronal viability. We hypothesized that if the intracellular estrogen receptor subtype-alpha (ERalpha) is important to estrogen's signaling in the ischemic brain, then ERalpha-deficient (knockout) (ERalphaKO) female mice would sustain exaggerated cerebral infarction damage after middle cerebral artery occlusion. METHODS: The histopathology of cresyl violet-stained tissues was evaluated after reversible middle cerebral artery occlusion (2 hours, followed by 22 hours of reperfusion) in ERalphaKO transgenic and wild-type (WT) mice (C57BL/6J background strain). End-ischemic cerebral blood flow mapping was obtained from additional female murine cohorts by using [(14)C]iodoantipyrine autoradiography. RESULTS: Total hemispheric tissue damage was not altered by ERalpha deficiency in female mice: 51.9+/-10.6 mm(3) in ERalphaKO versus 60.5+/-5.0 mm(3) in WT. Striatal infarction was equivalent, 12.2+/-1.7 mm(3) in ERalphaKO and 13.4+/-1.0 mm(3) in WT mice, but cortical infarction was paradoxically smaller relative to that of the WT (20.7+/-4.5 mm(3) in ERalphaKO versus 30.6+/-4.1 mm(3) in WT). Intraocclusion blood flow to the parietal cortex was higher in ERalphaKO than in WT mice, likely accounting for the reduced infarction in this anatomic area. There were no differences in stroke outcomes by region or genotype in male animals. CONCLUSIONS: Loss of ERalpha does not enhance tissue damage in the female animal, suggesting that estrogen inhibits brain injury by mechanisms that do not depend on activation of this receptor subtype.

Heme oxygenase-2 is neuroprotective in cerebral ischemia.

Heme oxygenase (HO) is believed to be a potent antioxidant enzyme in the nervous system; it degrades heme from heme-containing proteins, giving rise to carbon monoxide, iron, and biliverdin, which is rapidly reduced to bilirubin. The first identified isoform of the enzyme, HO1, is an inducible heat-shock protein expressed in high levels in peripheral organs and barely detectable under normal conditions in the brain, whereas HO2 is constitutive and most highly concentrated in the brain. Interestingly, although HO2 is constitutively expressed, its activity can be modulated by phosphorylation. We demonstrated that bilirubin, formed from HO2, is neuroprotectant, as neurotoxicity is augmented in neuronal cultures from mice with targeted deletion of HO2 (HO2(-/-)) and reversed by low concentrations of bilirubin. We now show that neural damage following middle cerebral artery occlusion (MCAO) and reperfusion, a model of focal ischemia of vascular stroke, is substantially worsened in HO2(-/-) animals. By contrast, stroke damage is not significantly altered in HO1(-/-) mice, despite their greater debility. Neural damage following intracranial injections of N-methyl-d-aspartate (NMDA) is also accentuated in HO2(-/-) animals. These findings establish HO2 as an endogenous neuroprotective system in the brain whose pharmacologic manipulation may have therapeutic relevance.

Restenosis after transluminal angioplasty for atherosclerotic vertebral and subclavian artery stenosis.

A total of 34 stenotic lesions of the vertebral or subclavian artery were treated in 29 patients using percutaneous transluminal angioplasty (PTA). Seventeen patients were followed angiographically for up to 2 years after PTA. Of 17 patients, five (29.4%) had restenosis which developed between 3 and 6 months after the procedure. Of these patients, two were treated by repeat angioplasty uneventfully, two were treated by surgical reconstruction, and the fifth is being observed without treatment while the patient remains asymptomatic. Neointimal fibromuscular hyperplasia as seen in the surgically resected specimens was thought to be the major cause of restenosis after PTA. Indications of PTA for vertebrobasilar and subclavian artery restenosis are discussed.

Poly(ADP-ribose) polymerase gene disruption renders mice resistant to cerebral ischemia.

Nitric oxide (NO) and peroxynitrite, formed from NO and superoxide anion, have been implicated as mediators of neuronal damage following focal ischemia, but their molecular targets have not been defined. One candidate pathway is DNA damage leading to activation of the nuclear enzyme, poly(ADP-ribose) polymerase (PARP), which catalyzes attachment of ADP ribose units from NAD to nuclear proteins following DNA damage. Excessive activation of PARP can deplete NAD and ATP, which is consumed in regeneration of NAD, leading to cell death by energy depletion. We show that genetic disruption of PARP provides profound protection against glutamate-NO-mediated ischemic insults in vitro and major decreases in infarct volume after reversible middle cerebral artery occlusion. These results provide compelling evidence for a primary involvement of PARP activation in neuronal damage following focal ischemia and suggest that therapies designed towards inhibiting PARP may provide benefit in the treatment of cerebrovascular disease.

Nontoxic embolic liquids for treatment of arteriovenous malformations.

Interventional radiology is becoming one of the standard treatments of arteriovenous malformation (AVM). Cyanoacrylate derivatives and polymer solutions are widely used to occlude the AVM nidus by their injection through a catheter, but they are far from satisfactory embolic liquids. For instance, cyanoacrylate derivatives sometimes glue the catheter to the artery, resulting in serious complications; in addition, the organic solvents used to dissolve polymers cause damage to the surrounding brain tissue of the AVM. Therefore, we attempted to develop embolic liquids by dissolving poly(2-hydroxyethyl methacrylate-co-methyl methacrylate) in Iopamiron with an addition of a small amount of ethyl alcohol. This new embolic liquid is not cytotoxic and is easily injected into the AVM through a thin, long catheter to effectively occlude the AVM.

Intraarterial infusion of high-concentration papaverine damages cerebral arteries in rats.

PURPOSE: To determine the appropriate concentration of papaverine for therapeutic intraarterial infusion against cerebral vasospasm. METHODS: We investigated histopathologic changes in cerebral arteries and brain tissue of normal Wistar rats that had received infusions of papaverine via the carotid artery. Rats were infused with 0.20 mL papaverine (concentration, 0.4% to 4.0%) via the internal carotid artery. Injury to the vascular wall was evaluated by transmission electron microscopy; pathologic changes in cerebral tissue were studied by light microscopy. RESULTS: Neither brain necrosis nor brain edema was seen under light microscopy at any concentration. At 4.0% papaverine concentration, degeneration of endothelial cells and medial smooth muscle, including vacuole formation, was observed under electron microscopy. At 1.4% concentration, degeneration of endothelial cells was seen. Extravasation of Evans blue dye was noted when drug concentration exceeded 1.4%. At 0.8% concentration, no histopathologic change was noted. CONCLUSION: On the basis of these results, we recommend a papaverine concentration of 0.8% or less for intraarterial infusion.

Interlocking detachable coils for spontaneous vertebral arteriovenous fistula.

We report a case of a spontaneous vertebral arteriovenous fistula treated with interlocking detachable coils (IDC). The patient had presented with paroxysmal dizziness and a cervical bruit. Angiography demonstrated a high flow arteriovenous fistula of the right extracranial vertebral artery. Embolisation was performed via the transarterial approach using 2 IDCs. The advantage of IDCs in the treatment of high flow arteriovenous fistulae is discussed.

Histological changes in brain tissue and vasculature after intracarotid infusion of organic solvents in rats.

Organic solvents, such as ethanol or dimethyl sulphoxide (DMSO), have been used in liquid embolic agents. To investigate the effects of these solvents on the cerebral blood vessels and cerebral tissue, we subjected Wistar rats weighing 250-300 g to internal carotid artery infusion of 0.2 ml diluted ethanol (10%, 40% or 70%) or anhydrous DMSO (100%). Some rats were sacrificed 5 min after the infusion and the remainder at 10 days. Rats injected with ethanol at high concentration or DMSO showed extensive exudation of Evans blue at the site of injection 5 min after infusion, together with full-thickness necrosis of the wall of vessels and swelling of brain cells. In contrast, rats injected with 10% or 40% ethanol solution showed necrosis of only the intimal layer and partial necrosis of the medial layer and no brain swelling was observed. These findings suggest that ethanol at low concentration can be used as a relatively safe solvent for liquid embolic substances.

Autoperfusion balloon catheter for treatment of vertebral artery stenosis.

Percutaneous transluminal angioplasty (PTA) of the vertebral artery was performed with an autoperfusion balloon catheter in five patients. There were no complications in the form of embolic episodes or neurological deficits due to brain ischaemia during inflation. In critical cases with insufficient collateral circulation during temporary occlusion, the use of an autoperfusion balloon catheter may expand the indications for PTA in patients with ischaemic cerebrovascular disease.

Embolization of dural arteriovenous fistulas with interlocking detachable coils.

Our clinical experience with interlocking detachable coils for the embolization of high-flow dural arteriovenous fistulas is reported. Interlocking detachable coils are useful for transarterial and transvenous embolizations of dural arteriovenous fistulas because (a) immediate coil detachment is possible, (b) the coils can be replaced easily, (c) detached coils rarely migrate, and (d) fewer interlocking detachable coils than conventional fiber coils are required for successful embolization.