Leukotriene B4-Neutrophil Elastase Axis Drives Neutrophil Reverse Transendothelial Cell Migration In Vivo.

Breaching endothelial cells (ECs) is a decisive step in the migration of leukocytes from the vascular lumen to the extravascular tissue, but fundamental aspects of this response remain largely unknown. We have previously shown that neutrophils can exhibit abluminal-to-luminal migration through EC junctions within mouse cremasteric venules and that this response is elicited following reduced expression and/or functionality of the EC junctional adhesion molecule-C (JAM-C). Here we demonstrate that the lipid chemoattractant leukotriene B4 (LTB4) was efficacious at causing loss of venular JAM-C and promoting neutrophil reverse transendothelial cell migration (rTEM) in vivo. Local proteolytic cleavage of EC JAM-C by neutrophil elastase (NE) drove this cascade of events as supported by presentation of NE to JAM-C via the neutrophil adhesion molecule Mac-1. The results identify local LTB4-NE axis as a promoter of neutrophil rTEM and provide evidence that this pathway can propagate a local sterile inflammatory response to become systemic.

Control of low flow regions in the cortical vasculature determines optimal arterio-venous ratios.

The energy demands of neurons are met by a constant supply of glucose and oxygen via the cerebral vasculature. The cerebral cortex is perfused by dense, parallel arterioles and venules, consistently in imbalanced ratios. Whether and how arteriole-venule arrangement and ratio affect the efficiency of energy delivery to the cortex has remained an unanswered question. Here, we show by mathematical modeling and analysis of the mapped mouse sensory cortex that the perfusive efficiency of the network is predicted to be limited by low-flow regions produced between pairs of arterioles or pairs of venules. Increasing either arteriole or venule density decreases the size of these low-flow regions, but increases their number, setting an optimal ratio between arterioles and venules that matches closely that observed across mammalian cortical vasculature. Low-flow regions are reshaped in complex ways by changes in vascular conductance, creating geometric challenges for matching cortical perfusion with neuronal activity.

Pulsatility damping in the microcirculation: Basic pattern and modulating factors.

Blood flow pulsatility is an important determinant of macro- and microvascular physiology. Pulsatility is damped largely in the microcirculation, but the characteristics of this damping and the factors that regulate it have not been fully elucidated yet. Applying computational approaches to real microvascular network geometry, we examined the pattern of pulsatility damping and the role of potential damping factors, including pulse frequency, vascular viscous resistance, vascular compliance, viscoelastic behavior of the vessel wall, and wave propagation and reflection. To this end, three full rat mesenteric vascular networks were reconstructed from intravital microscopic recordings, a one-dimensional (1D) model was used to reproduce pulsatile properties within the network, and potential damping factors were examined by sensitivity analysis. Results demonstrate that blood flow pulsatility is predominantly damped at the arteriolar side and remains at a low level at the venular side. Damping was sensitive to pulse frequency, vascular viscous resistance and vascular compliance, whereas viscoelasticity of the vessel wall or wave propagation and reflection contributed little to pulsatility damping. The present results contribute to our understanding of mechanical forces and their regulation in the microcirculation.

Retinal vasodilatory responses are inversely associated with plasminogen activator inhibitor-1: The African-PREDICT study.

AIMS: Plasminogen activator inhibitor-1 (PAI-1), traditionally associated with fibrinolysis, is increasingly implicated in impaired vascular function. However, studies on its association with microvascular function are limited to the cutaneous and coronary microvascular beds in older and diseased individuals. To better understand its potential involvement in the early stages of disease development, we investigated the associations of retinal vasodilatory responses to flicker light with PAI-1 activity (PAI-1act) in young and healthy individuals. METHODS: We included healthy Black and White women and men (n = 518; aged 20-30 years), and measured plasma PAI-1act and retinal vasodilatory responses to flicker light provocation. We also collected demographic and lifestyle data, measured blood pressure, anthropometry, blood lipids, inflammatory and other biomarkers. RESULTS: In multivariate regression analyses, maximal retinal venular dilation associated independently and inversely with PAI-1act (adj. R2 = 0.11; ß = -0.15; p = 0.001) in the total group. In exploratory subgroup analyses, this association remained in White women (adj. R2 = 0.07; ß = -0.23; p = 0.005), and was more robust with younger age and lower blood pressure and in non-smokers, but also with greater central adiposity, higher low-density lipoprotein cholesterol and inflammation (all p < 0.05). CONCLUSIONS: Our data suggest that in young individuals, PAI-1 may already be associated with subclinical microvascular dysfunction.

Blood flow velocity comparison in the eye capillaries and postcapillary venules between normal pregnant and non-pregnant women.

BACKGROUND: There is no consensus on how much and at what diameters the blood flow velocity changes in the female microcirculation during normal pregnancy. METHODS: A non-contact, digital slit-lamp biomicroscopy system was used to measure axial blood velocity (Vax) and diameter (D) in the conjunctival microcirculation of 28 normal non-pregnant women (Control Group), 17 women in the first semester of their normal pregnancy (Group 1) and 16 women in the third trimester of their normal pregnancy (Group 2). Blood volume flow (Q) was estimated from Vax and D. Microvessels were classified as "capillaries" (CAP) with D < 9 µm, "postcapillary venules of size 1" (PC1) with 9 ≤ D < 14 µm and "postcapillary venules of size 2" (PC2) with 14 ≤ D ≤ 24 µm. RESULTS: The women groups did not differ significantly in age, diastolic and systolic pressure and diameter of each size. Taking as baseline the capillary Vax of 0.51 mm/s of the Control Group, there was a statistically significant (p < 0.001) increase to 0.74 mm/s (45%) in Group 1 and to 0.95 mm/s (86%) in Group 2. This significant Vax increase in capillaries (CAP) was a consistent finding irrespective of the exact vessel size cut-off value for discriminating CAP from PC1. There was no statistical difference in Vax among groups at postcapillary venules of size 2 (PC2). Statistical conclusions for blood volume flows were similar to velocities. CONCLUSIONS: Normal pregnancy increases significantly axial blood velocity (Vax) in capillaries (CAP) with diameter <9 µm.

Arterial Supercharging Is More Beneficial to Flap Survival Due to Quadruple Dilation of Venules.

BACKGROUND: "Choke vessels" are communicating conduits between adjacent perforasomes in the skin. Most researches focus mainly on the arterial aspect of the "choke vessels" and neglect the venous aspect, an imbalance needed to be addressed. MATERIALS AND METHODS: The study was divided into parts I, II, and III. Part I was for observation of the vascular morphological evolution in the choke zone after flap harvest in rats. Part II was for determination of the importance of the dilation of the arterial and venous components of "choke vessels" by preserving the iliolumbar artery (ILA group) or vein (ILV group). A laser Doppler flowmeter and a speckle flow imaging system were adopted for monitoring the hemodynamic impact caused by the different manipulation. Part III was for corroboration of part II by manipulation of other vessels. RESULTS: In part I, the arteries and veins between the iliolumbar and intercostal perforasomes underwent modest dilation, whereas the venules between the veins nearly quadrupled in diameter. In part II, flaps in the ILA group were much more intensive in blood perfusion than flaps in the ILV group. The flap necrosis rate was 0.31 ± 0.07 in the ILV group, being significantly larger than 0.10 ± 0.03 in the ILA group. Part III confirmed that venous superdrainage is less efficacious in reducing flap necrosis than arterial supercharging, in which the position of the additional artery was far more important than the diameter. CONCLUSIONS: The extensive dilation of the venous component of choke vessels makes a more potent compensatory role for venous drainage after flap harvest, indicating arterial supercharging is better in augmenting flap viability than venous superdrainage.

Excitability and contractility in arterioles and venules from the urinary bladder.

The urinary bladder performs two key physiological functions: (1) to store urine, and (2) void urine at an appropriate time. While these two functions seem simple, both processes exert prolonged stretch and compressive forces on the urinary bladder vasculature that are greater than those seen by vessels in any other hollow organ. To compensate for these forces, the urinary bladder vasculature has adapted several key features that maintain blood flow during bladder filling and prevent damaging pressure fluctuations during emptying. This chapter first describes key anatomical features of the urinary bladder vasculature and how these features aid in maintaining blood flow in the milieu of the functioning bladder. Next, we investigate the mechanisms regulating excitability of urinary bladder arterioles with emphasis on the development and regulation of myogenic tone. We then discuss the physiological significance and excitability of urinary bladder capillaries and venules, and their important roles in maintaining tissue perfusion. Finally, the functionality of the urinary bladder vasculature will be explored in terms of bladder dysfunction, to understand if lower urinary tract symptoms associated with disease can be considered vascular in nature. Also included are perspectives on the urinary bladder itself as a model for understanding ischemia/reperfusion injury and the possibility that the urinary bladder holds a key to mitigating deleterious effects that result when blood flow is occluded and rapidly restored to other organs.

Role of Pericytes in the Initiation and Propagation of Spontaneous Activity in the Microvasculature.

The microvasculature is composed of arterioles, capillaries and venules. Spontaneous arteriolar constrictions reduce effective vascular resistance to enhance tissue perfusion, while spontaneous venular constrictions facilitate the drainage of tissue metabolites by pumping blood. In the venules of visceral organs, mural cells, i.e. smooth muscle cells (SMCs) or pericytes, periodically generate spontaneous phasic constrictions, Ca2+ transients and transient depolarisations. These events arise from spontaneous Ca2+ release from the sarco-endoplasmic reticulum (SR/ER) and the subsequent opening of Ca2+-activated chloride channels (CaCCs). CaCC-dependent depolarisation further activates L-type voltage-dependent Ca2+ channels (LVDCCs) that play a critical role in maintaining the synchrony amongst mural cells. Mural cells in arterioles or capillaries are also capable of developing spontaneous activity. Non-contractile capillary pericytes generate spontaneous Ca2+ transients primarily relying on SR/ER Ca2+ release. Synchrony amongst capillary pericytes depends on gap junction-mediated spread of depolarisations resulting from the opening of either CaCCs or T-type VDCCs (TVDCCs) in a microvascular bed-dependent manner. The propagation of capillary Ca2+ transients into arterioles requires the opening of either L- or TVDCCs again depending on the microvascular bed. Since the blockade of gap junctions or CaCCs prevents spontaneous Ca2+ transients in arterioles and venules but not capillaries, capillary pericytes appear to play a primary role in generating spontaneous activity of the microvasculature unit. Pericytes in capillaries where the interchange of substances between tissues and the circulation takes place may provide the fundamental drive for upstream arterioles and downstream venules so that the microvasculature network functions as an integrated unit.

Association between blood pressure and retinal arteriolar and venular diameters in Chinese early adolescent children, and whether the association has gender difference: a cross-sectional study.

BACKGROUND: To establish the independent association between blood pressure (BP) and retinal vascular caliber, especially the retinal venular caliber, in a population of 12-year-old Chinese children. METHODS: We have examined 1501 students in the 7th grade with mean age of 12.7 years. A non-mydriatic fundus camera (Canon CR-2, Tokyo, Japan) was used to capture 450 fundus images of the right eyes. Retinal vascular caliber was measured using a computer-based program (IVAN). BP was measured using an automated sphygmomanometer (HEM-907, Omron, Kyoto, Japan). RESULTS: The mean retinal arteriolar caliber was 145.3 µm (95% confidence interval [CI], 110.6-189.6 µm) and the mean venular caliber was 212.7 µm (95% CI, 170.6-271.3 µm). After controlling for age, sex, axial length, BMI, waist, spherical equivalent, birth weight, gestational age and fellow retinal vessel caliber, children in the highest quartile of BP had significantly narrower retinal arteriolar caliber than those with lower quartiles (P for trend< 0.05). Each 10-mmHg increase in BP was associated with narrowing of the retinal arterioles by 3.00 µm (multivariable-adjusted P < 0.001), and the results were consist in three BP measurements. The association between BP measures and retinal venular caliber did not persist after adjusting for fellow arteriolar caliber. And there was no significant interaction between BP and sex, age, BMI, and birth status. CONCLUSIONS: In a large population of adolescent Chinese children, higher BP was found to be associated with narrower retinal arterioles, but not with retinal venules. Sex and other confounding factors had no effect on the relationship of BP and retinal vessel diameter.

Altered Bulbar Conjunctival Microcirculation in Response to Contact Lens Wear.

PURPOSE: This study was conducted to determine blood flow velocities and corresponding vessel diameters to characterize the response of the bulbar conjunctival microvasculature to contact lens wear. METHODS: A functional slit-lamp biomicroscope (FSLB), an adapted traditional slitlamp, was used to image the temporal bulbar conjunctiva of 22 healthy subjects before and after 6 hr of contact lens wear. All of the measurable venules on the conjunctiva were processed to yield vessel diameters and blood flow velocities. RESULTS: The average blood flow velocity increased from 0.51±0.20 to 0.65±0.22 mm/sec (P<0.001) after 6 hr of lens wear. The blood flow velocity distribution showed a velocity increase that correlated with the vessel diameter increase from the baseline (r=0.826, P<0.05). This pattern maintained a similar trend after 6 hr of lens wear (r=0.925, P<0.05), and increased velocities were found across all of the vessel diameter ranges (P<0.001). CONCLUSIONS: Blood flow velocity increases across all of the vessel diameter ranges in response to contact lens wear. Functional slitlamp biomicroscope is capable of characterizing the bulbar microvascular response to contact lens wear.

Mechanisms underlying spontaneous constrictions of postcapillary venules in the rat stomach.

Postcapillary venules (PCVs) play a critical role in regulating capillary hydrostatic pressure, but their contractile mechanisms are not well understood. We examined the properties of spontaneous vasomotion and corresponding Ca(2+) transients in gastric PCV. In the rat gastric submucosa, changes in PCV diameter and intracellular Ca(2+) dynamics were visualised by video tracking system and fluorescent Ca(2+) imaging, respectively, while PCV morphology was examined by immunohistochemistry. Stellate-shaped PCV mural cells expressing α-smooth muscle actin exhibited synchronised spontaneous Ca(2+) transients to develop vasomotion which was abolished by nifedipine (1 µM), cyclopiazonic acid (10 µM), or Ca(2+)-activated Cl(-) channel inhibitors (100 µM niflumic acid, 1 µM T16Ainh-A01). A gap junction blocker (3 µM carbenoxolone) disrupted the synchrony of spontaneous Ca(2+) transients amongst PCV mural cells and attenuated spontaneous vasomotion. Low chloride solution ([Cl(-)]0 = 12.4 mM) also disrupted the synchrony of spontaneous Ca(2+) transients and abolished vasomotion. Na(+)-K(+)-Cl(-) co-transporter inhibitors (10 µM bumetanide, 30 µM furosemide) suppressed spontaneous Ca(2+) transients and vasoconstrictions. A phosphodiesterase type 5 (PDE5) inhibitor (1 µM tadalafil) disrupted the spontaneous Ca(2+) transient synchrony and abolished vasomotion in a nitric oxide (NO)-dependent manner. Thus, gastric PCVs exhibit spontaneous vasomotion, resulting from synchronised spontaneous Ca(2+) transients within a network of stellate-shaped PCV mural cells. An active Cl(-) accumulation partly via Na(+)-K(+)-Cl(-) co-transport appears to be fundamental in maintaining depolarisation upon the opening of Ca(2+)-activated Cl(-) channels that triggers Ca(2+) influx via voltage-dependent L-type Ca(2+) channels. Basal PDE5 activity may continuously counteract vaso-relaxing effects of endothelial NO to maintain spontaneous vasomotion.

A statistical model of the penetrating arterioles and venules in the human cerebral cortex.

OBJECTIVE: Models of the cerebral microvasculature are required at many different scales in order to understand the effects of microvascular topology on CBF. There are, however, no data-driven models at the arteriolar/venular scale. In this paper, we develop a data-driven algorithm based on available data to generate statistically accurate penetrating arterioles and venules. METHODS: A novel order-based density-filling algorithm is developed based on the statistical data including bifurcating angles, LDRs, and area ratios. Three thousand simulations are presented, and the results validated against morphological data. These are combined with a previous capillary network in order to calculate full vascular network parameters. RESULTS: Statistically accurate penetrating trees were successfully generated. All properties provided a good fit to experimental data. The k exponent had a median of 2.5 and an interquartile range of 1.75-3.7. CBF showed a standard deviation ranging from ±18% to ±34% of the mean, depending on the penetrating vessel diameter. CONCLUSIONS: Small CBF variations indicate that the topology of the penetrating vessels plays only a small part in the large regional variations of CBF seen in the brain. These results open up the possibility of efficient oxygen and blood flow simulations at MRI voxel scales which can be directly validated against MRI data.

[Regulation of vasomotor tone of small skeletal muscle veins by intrinsic mechanisms]. / Vázizomkisvénák vazomotortónusának intrinszik szabályozómechanizmusai.

In many developed countries the prevalence of venous disorders and its consequences are higher than that of arterial diseases. Thus it is very important to understand the exact physiological and pathophysiological function of small veins and their control mechanisms. Small veins and venules have an important role in the regulation of capillary fluid exchange, as well as return of the venous blood into the heart. However, there is only limited knowledge available regarding the role of local mechanisms controlling the vasomotor tone and diameter of small veins. In the last decade the authors focused on the elucidation of these mechanisms in isolated skeletal muscle venules of rats. Their results suggest that the tone of small veins is controlled by the integration of several mechanisms, activated by the intraluminal pressure and flow/wall shear stress, in addition to numerous local mediators synthesized and released from the smooth muscle and endothelium. These mechanisms are involved - in a complex manner - in the control of postcapillary resistance, thus regulation of tissue blood supply, venous return and consequently in the modulation of the cardiac output, as well.

Functional properties of submucosal venules in the rat stomach.

Venules in the stomach may have intrinsic properties for maintaining active microcirculation drainage even during gastric filling. Properties of spontaneous and nerve-mediated activity of submucosal venules in the rat stomach were investigated. Changes in vasodiameter and intracellular Ca(2+) in venular smooth muscle cells (SMCs) were monitored by video tracking and Fluo-8 Ca(2+) imaging, respectively. Venular SMCs developed synchronous spontaneous Ca(2+) transients and corresponding rhythmic constrictions of the venules. Nominally Ca(2+)-free solution or an L-type Ca(2+) channel blocker (1 µM nifedipine) disrupted the Ca(2+) transient synchrony and abolished spontaneous constrictions. Spontaneous constrictions were also prevented by inhibitors of sarcoplasmic reticulum Ca(2+)-ATPase (10 µM cyclopiazonic acid (CPA)), IP3 receptors (100 µM 2-APB) or Ca(2+)-activated Cl(-) channels (100 µM niflumic acid). Transmural nerve stimulation (TNS) induced a long-lasting venular constriction that was abolished by α-adrenoceptor antagonist (1 µM phentolamine), while TNS evoked a sympathetic transient constriction of arterioles that was abolished by a combination of phentolamine and a P2 purinoceptor antagonist (10 µM pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS)). Consistently, P2X1 purinoceptor immunoreactivity was detected in arteriolar but not venular SMCs. Primary afferent nerve stimulation (300 nM capsaicin) caused a venular dilatation by releasing calcitonin gene-related peptide. Thus, Ca(2+) release from the sarcoplasmic reticulum may play a fundamental role in the generation of spontaneous Ca(2+) transients, while electrical coupling amongst venular SMCs via L-type Ca(2+) channel activation appears to be critical for Ca(2+) transient synchrony as well as spontaneous contractions. Sympathetic venular constrictions appear to be exclusively mediated by noradrenaline due to the lack of P2X1 receptor in venular SMCs.

Elevated CXCL1 expression in gp130-deficient endothelial cells impairs neutrophil migration in mice.

Neutrophils emigrate from venules to sites of infection or injury in response to chemotactic gradients. How these gradients form is not well understood. Some IL-6 family cytokines stimulate endothelial cells to express adhesion molecules and chemokines that recruit leukocytes. Receptors for these cytokines share the signaling subunit gp130. We studied knockout mice lacking gp130 in endothelial cells. Unexpectedly, gp130-deficient endothelial cells constitutively expressed more CXCL1 in vivo and in vitro, and even more upon stimulation with tumor necrosis factor-α. Mobilization of this increased CXCL1 from intracellular stores to the venular surface triggered ß2 integrin-dependent arrest of neutrophils rolling on selectins but impaired intraluminal crawling and transendothelial migration. Superfusing CXCL1 over venules promoted neutrophil migration only after intravenously injecting mAb to CXCL1 to diminish its intravascular function or heparinase to release CXCL1 from endothelial proteoglycans. Remarkably, mice lacking gp130 in endothelial cells had impaired histamine-induced venular permeability, which was restored by injecting anti-P-selectin mAb to prevent neutrophil rolling and arrest. Thus, excessive CXCL1 expression in gp130-deficient endothelial cells augments neutrophil adhesion but hinders migration, most likely by disrupting chemotactic gradients. Our data define a role for endothelial cell gp130 in regulating integrin-dependent adhesion and de-adhesion of neutrophils during inflammation.

Role of high endothelial venule-expressed heparan sulfate in chemokine presentation and lymphocyte homing.

Lymphocyte homing to peripheral lymph nodes (PLNs) is mediated by multistep interactions between lymphocytes and high endothelial venules (HEVs). Heparan sulfate (HS) has been implicated in the presentation of chemokines on the surface of HEVs during this process. However, it remains unclear whether this cell surface presentation is a prerequisite for lymphocyte homing. In this study, we generated conditional knockout (cKO) mice lacking Ext1, which encodes a glycosyltransferase essential for HS synthesis, by crossing Ext1(flox/flox) mice with GlcNAc6ST-2-Cre transgenic mice expressing Cre recombinase in HEVs. Immunohistochemical studies indicated that HS expression was specifically eliminated in PLN HEVs but retained in other blood vessels in the cKO mice. The accumulation of a major secondary lymphoid tissue chemokine, CCL21, on HEVs was also abrogated without affecting CCL21 mRNA levels, indicating that HS presents CCL21 on HEVs in vivo. Notably, a short-term lymphocyte homing assay indicated that lymphocyte homing to PLNs was diminished in the cKO mice by 30-40%. Consistent with this result, contact hypersensitivity responses were also diminished in the cKO mice. The residual lymphocyte homing to PLNs in the cKO mice was dependent on pertussis toxin-sensitive Gi protein signaling, in which lysophosphatidic acid-mediated signaling was partly involved. These results suggest that chemokine presentation by HS on the surface of HEVs facilitates but is not absolutely required for lymphocyte homing.

Venous oxygen saturation is reduced and variable in central retinal vein occlusion.

PURPOSE: To estimate the presence and variability of retinal hypoxia in patients with central retinal vein occlusion (CRVO). METHOD: Hemoglobin oxygen saturation was measured in retinal vessels of both eyes in 14 patients with unilateral CRVO. The noninvasive spectrophotometric retinal oximeter is based on a fundus camera and simultaneously captures two images at 570 nm and 600 nm wavelengths. Five of the patients were followed with repeated retinal oximetry images over time. RESULTS: The mean oxygen saturation in retinal venules was 31 % ±12 % in CRVO eyes and 52 % ±11 % in unaffected fellow eyes (mean ±SD, n = 14, p < 0.0001). The arteriovenous difference was 63 % ±11 % in eyes with CRVO and 43 % ±7 % in fellow eyes (p < 0.0001). The variability of retinal venous oxygen saturation was substantial within and between eyes affected by CRVO. Venular oxygen saturation improved with treatment and over time in all five patients that were followed. CONCLUSION: CRVO eyes are hypoxic compared to fellow eyes and arteriovenous difference in hemoglobin oxygen saturation is increased. This is consistent with tissue hypoxia resulting from reduced blood flow. Further studies are needed to understand the correlation between hypoxia, severity of disease and prognosis.

The effects of topical and intravenous JM-1232(-) on cerebral pial microvessels of rabbits.

BACKGROUND: JM-1232(-) is a novel anesthetic agent which acts through gamma-aminobutyric acid receptors. Cerebral pial vascular effects of JM-1232(-) are unknown. We thus evaluated topical and intravenous effects of JM-1232(-) on cerebral pial microvessels in rabbits, and the extent to which carbon dioxide (CO2) reactivity is preserved. METHODS: Closed cranial windows were used to visualize cerebral pial circulation in 29 Japanese white rabbits. In the first experiment, the cranial window was superfused with increasing concentrations of JM-1232(-): 10(-11), 10(-9), 10(-7), 10(-5) mol/L, n = 8 per concentration. In the second experiment, we examined the effects of an intravenous bolus of 1 mg/kg bolus of JM-1232(-), followed by the continuous infusion at 0.3 mg/kg/minute on cerebral pial vascular alteration (n = 9). In the third, we examined CO2 reactivity of cerebral pial vessels under JM-1232(-) (n = 6) or sevoflurane anesthesia (n = 6). RESULTS: Topical application of JM-1232(-) did not change pial venular diameter, and constricted arterials only at the highest concentration. Intravenous administration of JM-1232(-) produced cerebral pial constriction which gradually diminished over time. Under intravenous administration of JM-1232(-) and inhaled sevoflurane, diameters of vessels increased in parallel with CO2 partial pressure. Slopes of linear regression and correlation coefficients in arterioles and venules were comparable for JM-1232(-) anesthesia and sevoflurane anesthesia. CONCLUSIONS: Topical application of JM-1232(-) had little effect on cerebral pial vessels. Intravenous administration produced vasoconstriction of cerebral pial arterioles and venules, however those changes were clinically unimportant. In addition, JM-1232(-) did not impair CO2 responsiveness. At least from the perspective of vascular reactivity, JM-1232(-) thus appears safe for neurosurgical patients.

Albumin modulates S1P delivery from red blood cells in perfused microvessels: mechanism of the protein effect.

Removal of plasma proteins from perfusates increases vascular permeability. The common interpretation of the action of albumin is that it forms part of the permeability barrier by electrostatic binding to the endothelial glycocalyx. We tested the alternate hypothesis that removal of perfusate albumin in rat venular microvessels decreased the availability of sphingosine-1-phosphate (S1P), which is normally carried in plasma bound to albumin and lipoproteins and is required to maintain stable baseline endothelial barriers (Am J Physiol Heart Circ Physiol 303: H825-H834, 2012). Red blood cells (RBCs) are a primary source of S1P in the normal circulation. We compared apparent albumin permeability coefficients [solute permeability (Ps)] measured using perfusates containing albumin (10 mg/ml, control) and conditioned by 20-min exposure to rat RBCs with Ps when test perfusates were in RBC-conditioned protein-free Ringer solution. The control perfusate S1P concentration (439 ± 46 nM) was near the normal plasma value at 37 °C and established a stable baseline Ps (0.9 ± 0.4 × 10(-6) cm/s). Ringer solution perfusate contained 52 ± 8 nM S1P and increased Ps more than 10-fold (16.1 ± 3.9 × 10(-6) cm/s). Consistent with albumin-dependent transport of S1P from RBCs, S1P concentrations in RBC-conditioned solutions decreased as albumin concentration, hematocrit, and temperature decreased. Protein-free Ringer solution perfusates that used liposomes instead of RBCs as flow markers failed to maintain normal permeability, reproducing the "albumin effect" in these mammalian microvessels. We conclude that the albumin effect depends on the action of albumin to facilitate the release and transport of S1P from RBCs that normally provide a significant amount of S1P to the endothelium.

Effect of uneven red cell influx on formation of cell-free layer in small venules.

This study examined how the uneven influx of red blood cells (RBCs) from feeding vessels influences formation of cell-free layer (CFL) in the downstream vessel of a venular bifurcation. Spatio-temporal variations of the CFL width along the downstream vessel (19-41-µm inner diameter, D) were determined at 0.5D intervals from 0.5D to 3.0D away from the bifurcation. Upstream flow conditions were quantified by the ratio of volume flow rates (Q*=Q(High)/Q(Low)) between high flow (Q(High)) and low flow feeding (Q(Low)) vessels. The RBC aggregation level in the rats was adjusted to be at healthy human levels by infusing Dextran 500. Our results suggested that the CFL formation process could be seen only from 2.0D away from the bifurcating point. The mean CFL width at the wall adjacent to the feeding vessel with a higher flow rate was consistently greater than that at the opposite wall, leading to an asymmetric CFL formation in the vessel. A positive relation (P<0.05) between the asymmetry of the CFL width and the volume flow rate ratio (Q*) was found. Our numerical prediction showed that flow resistance in the venular network could be significantly increased by the asymmetric formation of CFL downstream and this effect might become more pronounced under pathological flow conditions such as hyper-aggregating and/or low shear conditions.