Intercellular Conduction Optimizes Arterial Network Function and Conserves Blood Flow Homeostasis During Cerebrovascular Challenges.

OBJECTIVE: Cerebral arterial networks match blood flow delivery with neural activity. Neurovascular response begins with a stimulus and a focal change in vessel diameter, which by themselves is inconsequential to blood flow magnitude, until they spread and alter the contractile status of neighboring arterial segments. We sought to define the mechanisms underlying integrated vascular behavior and considered the role of intercellular electrical signaling in this phenomenon. Approach and Results: Electron microscopic and histochemical analysis revealed the structural coupling of cerebrovascular cells and the expression of gap junctional subunits at the cell interfaces, enabling intercellular signaling among vascular cells. Indeed, robust vasomotor conduction was detected in human and mice cerebral arteries after focal vessel stimulation: a response attributed to endothelial gap junctional communication, as its genetic alteration attenuated this behavior. Conducted responses were observed to ascend from the penetrating arterioles, influencing the contractile status of cortical surface vessels, in a simulated model of cerebral arterial network. Ascending responses recognized in vivo after whisker stimulation were significantly attenuated in mice with altered endothelial gap junctional signaling confirming that gap junctional communication drives integrated vessel responses. The diminishment in vascular communication also impaired the critical ability of the cerebral vasculature to maintain blood flow homeostasis and hence tissue viability after stroke. CONCLUSIONS: Our findings highlight the integral role of intercellular electrical signaling in transcribing focal stimuli into coordinated changes in cerebrovascular contractile activity and expose, a hitherto unknown mechanism for flow regulation after stroke.

Neural systemic impairment from whole-body vibration.

Insidious brain microinjury from motor vehicle-induced whole-body vibration (WBV) has not yet been investigated. For a long time we have believed that WBV would cause cumulative brain microinjury and impair cerebral function, which suggests an important risk factor for motor vehicle accidents and secondary cerebral vascular diseases. Fifty-six Sprague-Dawley rats were divided into seven groups (n = 8): 1) 2-week normal control group, 2) 2-week sham control group (restrained in the tube without vibration), 3) 2-week vibration group (exposed to whole-body vibration at 30 Hz and 0.5g acceleration for 4 hr/day, 5 days/week, for 2 weeks), 4) 4-week sham control group, 5) 4-week vibration group, 6) 8-week sham control group, and 7) 8-week vibration group. At the end point, all rats were evaluated in behavior, physiological, and brain histopathological studies. The cerebral injury from WBV is a cumulative process starting with vasospasm squeezing of the endothelial cells, followed by constriction of the cerebral arteries. After the 4-week vibration, brain neuron apoptosis started. After the 8-week vibration, vacuoles increased further in the brain arteries. Brain capillary walls thickened, mean neuron size was obviously reduced, neuron necrosis became prominent, and wide-ranging chronic cerebral edema was seen. These pathological findings are strongly correlated with neural functional impairments.

[New method to predict cerebral hyperperfusion syndrome after carotid endarterectomy by transcranial Doppler].

OBJECTIVE: To determine the diagnostic value for predicting cerebral hyperperfusion syndrome (CHS) by adding a transcranial Doppler (TCD) measurement at the end of the carotid endarterectomy (CEA) at the operating room. METHODS: Patients who underwent CEA between August 2009 and December 2011 of the prospective clinical trial in whom both intra- and post-operative TCD monitoring were performed were included. The middle cerebral artery velocities pre-clamping, post-declamping and post-operatively were measured by TCD. The intra-operative velocity increase ratio (VR1) was compared to the postoperative velocity increase ratio(VR2) in relation to CHS by calculating the sensitivity,specificity, positive predictive value, negative predictive value. The receiver operating characteristic curve (ROC) were also performed. The area under the curve (AUC) of ROC of VR1 and VR2 were compared.All the data were analyzed using SPSS 20.0 software. RESULTS: VR1 > 100% was identified in 6 patients, while VR2 > 100% was identified in 18 patients, respectively. Ten patients were diagnosed with CHS. The AUC of VR2 (0.728) was higher than AUC of VR1 (0.636). The best fit cutoff point of VR2 was 100%. The sensitivity, specificity, positive predictive value, negative predictive value were 70%, 83%, 39%, 95%, respectively, which demonstrates a better predictive power than VR1. CONCLUSION: Besides the commonly used intra-operative TCD monitoring, additional TCD measurement at the end of the carotid endarterectomy at the operating room is more useful to more accurately predict CHS.

Microanatomical study of the recurrent artery of Heubner.

The purpose of this study has been to describe the microanatomy of the recurrent artery of Heubner (RAH) in detail, to deepen anatomical knowledge and aid neurosurgeons in their work. The material was obtained from cadavers (ages 31-75 years) at routine autopsy. A total of 70 human brains (39 male and 31 female) were examined. People who died due to neurological disorders were not included in the study. Right after dissection, the arteries were perfused with acrylic paint emulsion, through the Circle of Willis or electively through the RAH. Brains were fixed in a 10% solution of formaldehyde, sectioned and placed in methyl salicylate for tissue transparency. To obtain corrosion-casts, the vessels were perfused with polyvinyl chloride or Mercox CL-2R resin and corroded using concentrated potassium chloride. The obtained material was analyzed using a stereoscopic light microscope. The RAH was present in 138 hemispheres with a mean of 1.99 RAH per hemisphere (275 RAH in total). The mean RAH length was 25.2 mm and the mean RAH diameter, in its place of origin, was 1 mm. Two to 30 branches (mean=9.4) originated from the stem of the RAH. The number of RAHs showed a negative correlation to the number of arteries from the medial group of lenticulo-striate arteries (LSA) (R=-0.62; p < 0.0001) which branch off the middle cerebral artery (MCA). This study further supports the RAH embryologic theory by Abbie. The RAH, in its extra- and intracerebral course, may join with the middle group of the LSA or directly with the MCA.

[Prognostic value of parameters of cerebral hemodynamics in patients with carotid ischemic stroke].

We studied the dynamics of neurological deficit and parameters of the blood flow through middle cerebral arteries in 97 patients with acute ischemic stroke (IS). Absolute and relative parameters of the blood flow were compared to the severity of neurological deficit in the 1st, 5-7th and 21-23rd days. There was a strong correlation between the asymmetry in blood flow velocity in the first day of II and NIHSS scores in 21-23rd days (r= -0,96; p<0,01). The less pronounced asymmetry in the first day of IS predicted the better recovery of damaged functions in patients with IS.

Ovine middle cerebral artery characterization and quantification of ultrastructure and other features: changes with development.

Regulation of tone, blood pressure, and blood flow in the cerebral vasculature is of vital importance, particularly in the developing infant. We tested the hypothesis that, in addition to accretion of smooth muscle cells (SMCs) in cell layers with vessel thickening, significant changes in smooth muscle structure, as well as phenotype, extracellular matrix, and membrane proteins, in the media of cerebral arteries (CAs) during the course of late fetal development account for associated changes in contractility. Using transmission electron, confocal, wide-field epifluorescence, and light microscopy, we examined the structure and ultrastructure of CAs. Also, we utilized wire myography, Western immunoblotting, and real-time quantitative PCR to examine several other features of these arteries. We compared the main branch ovine middle CAs of 95- and 140-gestational day (GD) fetuses with those of adults (n = 5 for each experimental group). We observed a graded increase in phenylephrine- and KCl-induced contractile responses with development. Structurally, lumen diameter, media thickness, and media cross-sectional area increased dramatically from one age group to the next. With maturation, the cross-sectional profiles of CA SMCs changed from flattened bands in the 95-GD fetus to irregular ovoid-shaped fascicles in the 140-GD fetus and adult. We also observed a change in the type of collagen, specific integrin molecules, and several other parameters of SMC morphology with maturation. Ovine CAs at 95 GD appeared morphologically immature and poorly equipped to respond to major hemodynamic adjustments with maturation.

Interstitial cells from rat middle cerebral artery belong to smooth muscle cell type.

It is now established that non-contractile cells with thin filopodia, also called vascular interstitial cells (VICs), are constitutively present in the media of many, if not all, blood vessels. The aim of this study was to determine the type of cell lineage to which arterial VICs belong using immunocytochemical, and real-time and reverse transcription PCR (RT-PCR). Using RT-PCR, we compared gene expression profiles of single VICs and smooth muscle cells (SMCs) freshly dispersed from rat middle cerebral artery. Both VICs and SMCs expressed the SMC marker, smooth muscle myosin heavy chain (SM-MHC), but did not express fibroblast, pericyte, neuronal, mast cell, endothelial or stem cell markers. Freshly isolated VICs also did not express c-kit, which is the marker for interstitial cells of Cajal in the gastrointestinal tract. Immunocytochemical labelling of contractile proteins showed that VICs and SMCs expressed SM-MHC similarly to the same degree, but VICs in contrast to SMCs had decreased expression of alpha-SM-actin and very low or no expression of calponin. Real-time RT-PCR was consistent with immunocytochemical experiments and showed that VICs had four times lower gene expression of calponin comparing to SMCs, which may explain VICs' inability to contract. VICs had greater expression than SMCs of structural proteins such as non-muscular beta-actin and desmin. The results obtained suggest that VICs represent a subtype of SMCs and may originate from the same precursor as SMCs, but later develop filopodia and a non-contractile cell phenotype.

Determination of wall tension in cerebral artery aneurysms by numerical simulation.

BACKGROUND AND PURPOSE: Cerebral artery aneurysms rupture when wall tension exceeds the strength of the wall tissue. At present, risk-assessment of unruptured aneurysms does not include evaluation of the lesions shape, yet clinical experience suggests that this is of importance. We aimed to develop a computational model for simulation of fluid-structure interaction in cerebral aneurysms based on patient specific lesion geometry, with special emphasis on wall tension. METHODS: An advanced isogeometric fluid-structure analysis model incorporating flexible aneurysm wall based on patient specific computed tomography angiogram images was developed. Variables used in the simulation model were retrieved from a literature review. RESULTS: The simulation results exposed areas of high wall tension and wall displacement located where aneurysms usually rupture. CONCLUSIONS: We suggest that analyzing wall tension and wall displacement in cerebral aneurysms by numeric simulation could be developed into a novel method for individualized prediction of rupture risk.

On the presence of neurotrophin p75 receptor on rat sympathetic cerebrovascular nerves.

Although the presence of neurotrophin p75 receptor on sympathetic nerves is a well-recognised feature, there is still a scarcity of details of the distribution of the receptor on cerebrovascular nerves. This study examined the distribution of p75 receptor on perivascular sympathetic nerves of the middle cerebral artery and the basilar artery of healthy young rats using immunohistochemical methods at the laser confocal microscope and transmission electron microscope levels. Immunofluorescence methods of detection of tyrosine hydroxylase (TH) in sympathetic nerves, p75 receptor associated with the nerves, and also S-100 protein in Schwann cells were applied in conjunction with confocal microscopy, while the pre-embedding single and double immunolabelling methods (ExtrAvidin and immuno-gold-silver) were applied for the electron microscopic examination. Immunofluorescence studies revealed "punctuate" distribution of the p75 receptor on sympathetic nerves including accompanying Schwann cells. Image analysis of the nerves showed that the level of co-localization of p75 receptor and TH was low. Immunolabelling applied at the electron microscope level also showed scarce co-localization of TH (which was intra-axonal) and p75. Immunoreactivity for p75 receptor was present on the cell membrane of perivascular axons and to a greater extent on the processes of accompanying Schwann cells. Some Schwann cell processes were adjacent to each other displaying strong immunoreactivity for p75 receptor; immunoreactivity was located on the extracellular sites of the adjacent cell membranes suggesting that the receptor was involved in cross talk between these. It is likely that variability of locations of p75 receptor detected in the study reflects diverse interactions of p75 receptor with axons and Schwann cells. It might also imply a diverse role for the receptor and/or the plasticity of sympathetic cerebrovascular nerves to neurotrophin signalling.

Lipid-soluble smoke particles damage endothelial cells and reduce endothelium-dependent dilatation in rat and man.

BACKGROUND: Cigarette smoking is a strong risk factor for vascular disease and known to cause dysfunction of the endothelium. However, the molecular mechanisms involved are still not fully understood. METHODS: In order to reveal the direct effects of lipid-soluble smoke particles on the endothelium, ring segments isolated from rat mesenteric arteries and human middle cerebral arteries (MCA) obtained at autopsy were incubated for 6 to 48 hrs in the presence of dimethylsulphoxide (DMSO)-soluble particles from cigarette smoke (DSP), i.e. lipid-soluble smoke particles. The endothelial microstructure was examined by transmission electron microscopy. The endothelial function was evaluated by acetylcholine (ACh)-induced endothelium-dependent vasodilatation, using a sensitive myograph. RESULTS: After DSP treatment, the arterial endothelium was swollen and loosing its attachment. In functional tests, the total ACh-induced dilatation, the nitric oxide (NO)-mediated and the endothelium-derived hyperpolarization factor (EDHF)-mediated dilatations were significantly decreased by DSP in a time- and concentration-dependent manner (p < 0.05). Nicotine, an important compound in cigarette smoke had, in an equivalent concentration as in DSP, no such effects (p > 0.05). Similar results were obtained in the human MCA. CONCLUSION: Thus, we demonstrate that the lipid-soluble smoke particles, but not nicotine, caused damage to arterial endothelium and reduced the endothelium-dependent dilatation in man and rat.

Mechanisms involved in the early increase of serotonin contraction evoked by endotoxin in rat middle cerebral arteries.

The present study investigated the mechanisms involved in the increased 5-hydroxytryptamine (5-HT) vasoconstriction observed in rat middle cerebral arteries exposed in vitro to lipopolysaccharide (LPS, 10 microg x ml-1) for 1-5 h. Functional, immunohistochemical and Western blot analysis and superoxide anion measurements by ethidium fluorescence were performed. LPS exposure increased 5-HT (10 microm) vasoconstriction only during the first 4 h. In contrast to control tissue, indomethacin (10 microm), the COX-2 inhibitor NS 398 (10 microm), the TXA2/PGH2 receptor antagonist SQ 29548 (1 microm) and the TXA2 synthase inhibitor furegrelate (1 microm) reduced 5-HT contraction of LPS-treated arteries from hour one. The iNOS inhibitor aminoguanidine (0.1 mm) increased 5-HT contraction from hour three of LPS incubation. The superoxide anion scavenger superoxide dismutase (SOD, 100 U ml-1) and the H2O2 scavenger catalase (1000 U ml-1), as well as the respective inhibitors of NAD(P)H oxidase and xanthine oxidase, apocynin (0.3 mm) and allopurinol (0.3 mm), reduced 5-HT contraction after LPS incubation. LPS induced an increase in superoxide anion levels that was abolished by PEG-SOD. Subthreshold concentrations of the TXA2 analogue U 46619, xanthine/xanthine oxidase and H2O2 potentiated, whereas those of sodium nitroprusside inhibited, the 5-HT contraction. COX-2 expression was increased at 1 and 5 h of LPS incubation, while that of iNOS, Cu/Zn-SOD and Mn-SOD was only increased after 5 h. All the three vascular layers expressed COX-2 and Cu/Zn-SOD. iNOS expression was detected in the endothelium and adventitia after LPS. In conclusion, increased production of TXA2 from COX-2, superoxide anion and H2O2 enhanced vasoconstriction to 5-HT during the first few hours of LPS exposure; iNOS and SOD expression counteracted that increase at 5 h. These changes can contribute to the disturbance of cerebral blood flow in endotoxic shock.

Progression of middle cerebral artery occlusive disease and its relationship with further vascular events after stroke.

BACKGROUND: Serial changes of flow velocities of transcranial Doppler ultrasound (TCD) in symptomatic middle cerebral artery (MCA) occlusive disease may be related to the occurrence of further vascular events, but prospective data are lacking. METHODS: We conducted a prospective study on patients with cerebral ischemia who were hospitalized with symptomatic MCA stenosis or occlusion. We repeated TCD examinations 6 months after the initial examinations and recorded any stroke or coronary events during this period. The changes of MCA flow velocities were categorized as normalized artery, stable artery, and progressed artery, which were determined according to the changes of MCA velocities at 6 months. RESULTS: We studied 143 consecutive patients who had relevant MCA occlusive diseases (107 with stenosis and 36 with occlusion). At 6 months, the velocities in the MCA returned to normal in 42 patients (29%), they were stable in 80 patients (62%), and they progressed in 13 patients (9%). The number of clinical events varied significantly among the 3 groups: there were 2 patients (4.8%) with clinical events in the normal group, 11 patients (12.5%) with clinical events in the stable group, and 5 patients (38.5%) with clinical events in the progressed group (P=0.004). The 18 recurrent events included 10 recurrent strokes, 5 transient ischemic attacks, and 3 acute coronary syndromes. CONCLUSIONS: Progression of MCA occlusive diseases is associated with an increased risk of vascular events. Further studies are required to establish the value of serial TCD examinations in predicting future clinical events.

[Morphometrical analyze of the middle cerebral artery system at the 13-15 weeks fetuses]. / Analiza morfometrica a sistemului arterei cerebrale mijlocii la fatul de 13-15 saptamâni.

Tele-encephalization process is accompanied by the appearance and progressive complication of the middle cerebral artery system. The aim of our study is to analyze the morphometrical parameters of the middle cerebral artery branches in the beginning of the edification of its system. We used 162 cerebral hemispheres from 88 fetuses aged of 13-15 weeks. Middle cerebral artery system was injected with a gelatin-China ink mixture and images recorded by means of a Zeiss surgical microscope. Parameters evaluation (length, proximal and distal diameters, external surface, volume, angles of bifurcation) was realized with KS-300 program. At this early age middle cerebral artery system has only 4-5 generations of branches usually resulting from acute angle bifurcations.

Endothelin in the middle cerebral artery: a case of multiple system atrophy.

In this study, we show the changes in the wall of the middle cerebral artery of a subject who suffered multiple system atrophy with autonomic failure. An electron-immunocytochemical approach was employed to reveal the presence of endothelin-1. Our results demonstrate the presence of immunoreactive endothelin-1 in the endothelial cells of the intima, vascular smooth muscle cells and macrophages of the media and neointima, and perivascular nerves/axons varicosities at the adventitial-medial border of the artery. It is concluded that endothelin-1 may, therefore, play a number of roles within diseased cerebral artery. The finding of endothelin-1-positive varicosities of autonomic innervation to this artery suggests an influence of neural endothelin on vascular smooth muscle in multiple system atrophy with autonomic failure. However, the presence of features such as neointima formation, wall irregularities and foam cells suggest the coexistence of atherosclerosis.