Gaussianity revisited: exploring the Kibble-Zurek mechanism with superconducting rings.

In this paper we use spontaneous flux production in annular superconductors to shed light on the Kibble-Zurek (KZ) scenario. In particular, we examine the effects of finite size and external fields, neither of which is directly amenable to the KZ analysis. Supported by 1D and 3D simulations, the properties of a superconducting ring are seen to be well represented by analytic Gaussian approximations which encode the KZ scales indirectly. Experimental results for annuli in the presence of external fields corroborate these findings.

Zurek-Kibble mechanism for the spontaneous vortex formation in Nb-Al/Al(ox)/Nb Josephson tunnel junctions: new theory and experiment.

New scaling behavior has been both predicted and observed in the spontaneous production of fluxons in quenched Nb-Al/Al(ox)/Nb annular Josephson tunnel junctions (JTJs) as a function of the quench time, tau(Q). The probability f(1) to trap a single defect during the normal-metal-superconductor phase transition clearly follows an allometric dependence on tau(Q) with a scaling exponent sigma = 0.5, as predicted from the Zurek-Kibble mechanism for realistic JTJs formed by strongly coupled superconductors. This definitive experiment replaces one reported by us earlier, in which an idealized model was used that predicted sigma = 0.25, commensurate with the then much poorer data. Our experiment remains the only condensed matter experiment to date to have measured a scaling exponent with any reliability.

Zurek-Kibble domain structures: the dynamics of spontaneous vortex formation in annular Josephson tunnel junctions.

Phase transitions create a domain structure with defects, which has been argued by Zurek and Kibble (ZK) to depend in a characteristic way on the quench rate. We present an experiment to measure the ZK scaling exponent sigma. Using long symmetric Josephson tunnel junctions, for which the predicted index is sigma=0.25, we find sigma=0.27+/-0.05. Further, we agree with the ZK prediction for the overall normalization.

Activation of barium-sensitive inward rectifier potassium channels mediates remote dilation of coronary arterioles.

BACKGROUND: Conducted vasodilation seems to be critical for the functional distribution of blood flow in the skeletal muscle microcirculation. However, this vasoregulatory phenomenon has not been documented in the coronary microcirculation, and its underlying mechanism remains elusive. Because potassium ions are potent metabolic vasodilators in the heart, by activating vascular inward rectifier K(+) (K(ir)) channels, we tested the hypothesis that coronary arterioles exhibit remote vasodilation through activation of this type of channel. METHODS AND RESULTS: Porcine coronary arterioles were isolated, cannulated, and pressurized for in vitro study. Vessels dilated concentration-dependently to extraluminal KCl (5 to 20 mmol/L), bradykinin, adenosine, pinacidil, and sodium nitroprusside. A K(ir) channel blocker, BaCl(2) (30 micromol/L), inhibited vasodilatory responses to KCl and bradykinin but not to adenosine, pinacidil, or nitroprusside. In a flow chamber, localized administration of bradykinin, adenosine, and KCl to the downstream end of the arterioles caused approximately 80% dilation at the site of drug application (local site) and also produced 30% to 60% dilation at the upstream end of arterioles (remote site). Nitroprusside produced a similar dilation at the local site but failed to initiate remote vasodilation. In the presence of Ba(2+), adenosine still dilated the local site, but the local dilations to bradykinin and KCl and the remote dilations to adenosine, bradykinin, and KCl were inhibited. CONCLUSIONS: We demonstrated that some modes of local vasodilation can be conducted to remote sites in coronary arterioles and that local and remote dilations can occur through different vasodilatory mechanisms. Activation of K(ir) channels seems critical for some agonist-induced local vasodilations and also for the initiation and/or transmission of signals causing remote vasodilation.

Measurement of membrane potential and intracellular Ca(2+) of arteriolar endothelium and smooth muscle in vivo.

We have developed an intensity analysis technique for fluorescence microscopy that allows us to measure, in real time, the diameter and the membrane potential or intracellular calcium ([Ca(2+)]i) of in vivo arteriolar endothelium or smooth muscle. Cheek pouch arterioles of anesthetized hamsters were luminally or abluminally labeled with Di-8-ANEPPS, a voltage-sensitive dye, or Fura PE3, a calcium indicator. The peak fluorescence intensities of the images were used to locate the endothelium or smooth muscle. The changes in membrane potential or [Ca(2+)]i were determined based on the ratiometric analysis of fluorescence intensity of the endothelium or smooth muscle. Membrane depolarization of the smooth muscle using KCl caused a decrease in the ratio of emission, 620 nm/560 nm ( approximately 6 mV/% ratio). The ratio of excitation, 340 nm/380 nm, increased with increasing free Ca(2+). Methacholine, a muscarinic receptor agonist, caused arteriolar dilation (12.2 +/- 0.9 µm). It produced hyperpolarization of the endothelium and smooth muscle (2.8 +/- 0.6% and 2.3 +/- 0.3% in ratio). Methacholine also induced an increase in [Ca(2+)]i (11.0 +/- 1.1% in ratio) of the endothelium. In contrast, methacholine caused a biphasic change in [Ca(2+)]i of the smooth muscle, a rapid reduction (-3.4 +/- 0.2% in ratio) followed by a prolonged increase (2.4 +/- 0.2% in ratio). These results demonstrate that the peak intensity analysis can be used to determine in real time the changes in membrane potential or [Ca(2+)]i of in vivo endothelium or smooth muscle.

Nonvasomotor influence of sodium nitroprusside on arteriolar remote response to methacholine.

Vascular communication functions to facilitate blood distribution within tissues and can be demonstrated as conducted vasomotor responses. This study was designed to determine if local application of sodium nitroprusside (SNP) would affect arteriolar function at remote sites. In the cheek pouch of anesthetized hamsters, local application of SNP and nifedipine caused arteriolar dilation only at the site of application (7.1 +/- 0.5 and 7.4 +/- 0.6 microm), but not at remote sites. The application of SNP enhanced subsequent remote, nitric oxide (NO)-independent dilation in response to methacholine, which was applied at a site upstream from the SNP application site (6.7 +/- 0.7 versus 4.5 +/- 0.7 microm for methacholine alone). This potentiating effect was also observed following application of 3-morpholinosydnonimine, but not following nifedipine. This nonvasomotor influence of SNP was not affected by N(omega)-nitro-L-arginine (L-NA), tetrodotoxin (TTX), Gap 27 peptide or halothane. Attenuated local dilation in response to methacholine by L-NA could be partially recovered following downstream application of SNP, suggesting that SNP-induced potentiation was associated with enhanced vasodilatory signals at the methacholine application site. Thus, our results suggest that SNP induces nonvasomotor signals in arterioles to affect the network distribution of blood flow. Intrinsic NO, TTX-sensitive Na(+) channels and gap junctional communication do not seem to play a major role in the conduction of the nonvasomotor signals.

Technique for using video microscopy and indicator dilution for repeated measurements of cardiac output in small animals.

BACKGROUND: The authors developed an indicator dilution technique for small animals to repeatedly determine cardiac output and blood volume without cardiac instrumentation or blood sampling. METHODS: Observations were made in the hamster (N = 32, 70 mg/kg pentobarbital) cremaster using in vivo fluorescence videomicroscopy. Fluorescein isothiocyanate-conjugated bovine serum albumin (10 mg/ml) was injected as a bolus dose (right jugular) while video recording the light intensity in a 20-microm arteriole (intensified charge-coupled device [CCD] camera at fixed gain). The intensity signal was analyzed over time (background subtracted) and calibrated to the dye concentration. The ex vivo calibration was performed using a constant optical path length (20 microm) and a range of dye and hematocrit concentrations. In vivo tube hematocrit was determined using standard methods with fluorescently labeled erythrocytes. Thus, quenching of the fluorescence signal by hemoglobin was corrected for the calibration, and the plasma space in the arteriole was determined. The steady state dye concentration measured by the light intensity at 2 min was not different from the dye concentration found by direct spectrophotometric analysis of the plasma. RESULTS: Cardiac index was calculated as milliliters of blood per minute per kilogram body weight. The calculated cardiac index was 359 +/- 18 ml.min(-1).kg(-1), which is not different from the reported values for hamsters. Cardiac output was increased twofold when enough intravenous nitroprusside or nitroglycerine was injected to decrease mean arterial pressure from 90 to 70 mmHg. Cardiac output was elevated during dobutamine infusion (16 microg.kg(-1).min(-1)) and decreased during esmolol infusion (50, 75.kg(-1).min(-1)). Blood volume determined from the steady state dye concentrations was 6.2 +/- 0.5 ml/100 g body weight, within the normal range for hamsters. CONCLUSIONS: Fluorescent dye dilution and video microscopy can be used to repeatedly determine cardiac output or blood volume in small animals.

Localized adenovirus-mediated gene transfer into vascular smooth muscle in the hamster cheek pouch.

OBJECTIVE: Our purpose was to develop a method for adenovirus delivery to the hamster cheek pouch to experimentally target gene transfer in tissue used for microvascular studies. METHODS: Separate constructs were tested with transgenes for lacZ or green fluorescent protein (GFP) driven by three promoters: RSV, CMV, and SM22. With university approval, adenovirus was delivered in anesthetized (pentobarbital, 70 mg/kg) hamsters (n = 28) by using either a vascular systemic injection or tissue infiltration (interstitial space behind the pouch). During 3 days, animals receiving infiltration gained the expected weight, whereas those receiving vascular injection lost weight; no other behavior changes were noted. RESULTS: On day 3 postadenoviral delivery (infiltration), expression of lacZ (histology, beta-galactosidase) or GFP (fluorescence microscopy) was confirmed across the tissue (CMV and RSV promoters) and exclusively in vascular smooth muscle cells (specific SM22 promoter), without evidence of tissue inflammation. In vitro microvascular experiments verified normal responses in the cheek pouch of day 3 postadenoviral delivery animals. We tested local dilation to methacholine, adenosine, remote dilation to methacholine, adenosine, nitroprusside, and LM609 (alpha(v)beta3 integrin agonist), flow-dependent dilation, and flow recruitment. CONCLUSIONS: Thus, this method enables targeted, cell-specific gene transfer to one tissue important for microvascular studies, without significant systemic exposure and without adverse inflammation.

Microcirculatory basis for nonuniform flow delivery with intravenous nitroprusside.

BACKGROUND: The purpose of this study was to determine the effects of systemic infusions of nitroglycerin and sodium nitroprusside on flow distribution and wall shear stress in the microcirculation. METHODS: With university approval, the cremaster muscle of 28 anesthetized (70 mg/kg pentobarbital given intraperitoneally) hamsters (Harlan Sprague Dawley: Syrian; weight, 121+/-11 g [mean +/- SDD) was observed using in vivo fluorescence microscopy. Arteriolar diameter, erythrocyte flux, and velocity were measured for a feed arteriole and its sequential branches. Observations were made during control (mean arterial pressure, 88+/-4 mm Hg) and after 30 min of intravenous delivery of sodium nitroprusside or nitroglycerin, titrated to decrease mean arterial pressure by 20 mm Hg. RESULTS: Sodium nitroprusside significantly dilated select upstream portions of the network (23+/-2.6 to 29+/-2.6 microm); no arterioles were dilated with nitroglycerin. Erythrocyte flux into the feed (i.e., inflow into the arteriolar network) and into the sequential branches (i.e., distribution within the network) were evaluated. With nitroglycerin, inflow decreased significantly from 1,560+/-335 to 855+/-171 cells/s, and flux into the branches decreased evenly. With sodium nitroprusside, inflow increased significantly to 2,600+/-918 cells/s, yet cells were "stolen" from upstream branches (a decrease from 425+/-67 to 309+/-87 cells/s in the first branch). Excess flow passed into a downstream "thorough-fare channel," significantly increasing flux from 347+/-111 to 761+/-246 cells/s. Wall shear stress decreased uniformly with nitroglycerin infusion, with a decrease in the feed from 8.8+/-2.5 to 6+/-1.7 dyn/cm2. With sodium nitroprusside, variable changes occurred that were location specific within the network. For instance, at the inflow point to the network, wall shear stress changed from 8.3+/-2.5 to 4.2+/-3.3 dyn/cm2. CONCLUSIONS: Nitroglycerin infusion promoted homogeneity of flow. Sodium nitroprusside significantly increased the heterogeneity of flow within this arteriolar network; an anatomic location for steal induced by sodium nitroprusside is identified.

Network vascular communication initiated by increases in tissue adenosine.

Vascular communication of vasomotor signals appears to coordinate the distribution of tissue blood flow. This study was performed to determine whether elevated tissue concentrations of adenosine or nitric oxide could induce vascular communicating signals. To test this, remote arteriolar responses were tested when drugs were applied either directly to an arteriole ( approximately 20 microm diameter), or into the tissue in a region (with no vessels over 10 microm in diameter) that was 500 microm away from the arteriole and that bore no defined relationship to the flow path of the remote arteriole. In anesthetized hamster cheek pouch (n = 25), or cremaster muscle (n = 10), remote arteriolar responses were measured in response to nitric oxide (NO) donors (10(-5) to 10(-3) M), adenosine (10(-5) to 10(-3) M), or papaverine (10(-5) to 10(-2) M) applied for 40-120 s. Papaverine caused no remote response when applied directly while adenosine and NO donors caused similar, late-onset (10-20 s), dose-dependent, remote responses in both preparations. Remarkably however, only adenosine initiated a consistent remote arteriolar dilation when applied to the tissue site. Thus, increases in tissue adenosine may be critical for vascular communication of metabolic demands without regard to the specific blood flow path.

Pharmacologic study of muscarinic receptor subtypes and arteriolar dilations: a comparison of conducted and local responses.

Arteriolar relaxation caused by the application of muscarinic agonists is mediated by multiple factors. One factor causes dilation only at the point of drug microapplication (local response), and a second factor causes responses remote (500 microm away) from the site of application (conducted response). This study was performed to determine if different muscarinic subtypes mediate the two responses. Arterioles of anesthetized hamster cheek pouch were studied with videomicroscopy. Muscarinic antagonists methscopolamine, scopolamine, pirenzepine, 4-DAMP (4-diphenylacetoxy-N-methylpiperidine methiodide), and AFDX-116 [(11-2[[2-[(diethylamino)methyl]-1-piperidinyl]acetyl]-5, 11-dihydro-6H-pyrido [2,3-b][1,4]benzodiazepin-6-one)] were cumulatively applied, and the K(B) for each antagonist was determined for the local and conducted responses caused by methacholine microapplication (10(-4) M, 5 s). The pK(B) (local, conducted) were not significantly different for the two responses when using scopolamine (10.5, 10.4). When the antagonist AFDX-116 (5.6, 6.3), selective for muscarinic receptor (m2) subtype was applied, the K(B) was greater for the conducted response. The pK(B) was greater, however, for the local response when the m1 subtype-selective pirenzepine (7.7, 6.9) or m3 subtype-selective 4-DAMP (10.1, 9.8) was applied. Thus the antagonist pK(B) ratio for on the local and conducted responses depends on the subtype selectivity of the antagonist. These data strongly suggest that different receptors are involved in the two responses.

Volatile anesthetic agents and vascular communication in the microcirculation of the hamster cheek pouch.

BACKGROUND: There is communication between tissue and the vascular network involved in regulating distribution of blood flow. Signals generated by the tissue are communicated upstream to create a coordinated network response in unison with other controllers of blood flow, such as myogenic and flow-dependent responses. METHODS: This vascular communication was modeled with the microapplication of methacholine (10(-4) M) or potassium chloride solution (KCl; 100 mM) to arterioles (40-60 microm in diameter) of the cheek pouch of anesthetized hamsters and viewed with videomicroscopy. Local and conducted (500 microm upstream) responses were measured. Halothane or isoflurane (1%, 2%, and 3%) was equilibrated with the superfusion solution and applied to the entire tissue. Responses to KCl and methacholine were then repeated in the presence of an anesthetic agent. RESULTS: Halothane and isoflurane increased the resting diameter of the arterioles. They also decreased the methacholine-initiated dilations. To test for the effects of increased resting diameter on the dilations, 0%, 5% and 10% oxygen alone was applied to the pouch to alter the tone, and the methacholine responses were repeated. The dilations decreased with oxygen-induced increases in resting diameter, but the conducted dilation decreased to a lesser extent than was seen with the volatile anesthetic agents. Neither halothane nor isoflurane decreased constrictions caused by KCl. CONCLUSIONS: Decreased methacholine-initiated conducted dilations caused by halothane and isoflurane were not due to decreases in cell-cell communication because KCl conducted responses persisted. Therefore, cell-cell vascular communication appears intact in the presence of clinical concentrations of halothane and isoflurane.

Cumulative conducted vasodilation within a single arteriole and the maximum conducted response.

The vascular network functions to distribute blood flow to the tissues that require it, and conducted vasodilation may facilitate this function. Experiments on arterioles in anesthetized hamster cheek pouch modeled the conducted responses that may come from a series of neighboring capillary modules and determined whether cumulative conducted responses could thereby maximally dilate upstream arterioles. Methacholine (10(-5) M) was simultaneously microapplied on an arteriole (resting diameter, approximately 22 microns; maximum diameter, approximately 47 microns) from one to four micropipettes spaced 100 microns apart, and with each added pipette the conducted dilation increased (up to a maximum dilation of approximately 5 microns). Increasing the methacholine 10-fold (10(-4) M) did not further increase the conducted response. The conducted response could also not be increased by lengthening the duration of microapplication. Yet, dilations that were not cumulative along a single arteriole became cumulative when initiated instead on adjacent arterioles. Therefore, these data demonstrate that conducted dilation along a single arteriole is limited and, if this model is correct, suggest that neighboring capillary modules may communicate only a limited conducted response to the network.

Components of methacholine-initiated conducted vasodilation are unaffected by arteriolar pressure.

Conducted vasodilation occurs remotely from a site of microapplication of a drug. Intravascular pressure is required for a conducted response in vivo, yet in vitro studies in unpressurized arterioles show pressure is not essential. To determine how pressure affects conducted vasodilation, intra-arteriolar pressure was controlled within an in situ isolated segment (average length 950 +/- 96 microns, average baseline diameter 28 +/- 2.1 microns) of arterioles in the hamster cheek pouch. Methacholine (10(-4) M, 5 s) was microapplied either onto the isolated segment or remotely, with local and conducted vasodilation measured at both locations. Increasing pressure in the lumen of the segment (0-80 cmH2O) increased the segment local dilation to methacholine, and the segment-conducted dilation plateaued (at 4.1 +/- 0.8 micron) when segment pressure reached 20 cmH2O. Any local (16 +/- 1.5 microns) and conducted (4.4 +/- 1.3 microns) dilations viewed outside the segment were unaffected by segment pressure and persisted in its absence. Thus segment pressure affected only electromechanical transduction of the conducted response. Thus vasomotor signals move throughout the vasculature regardless of tone, but tone is essential to transduce the response.

Remote effects of pressure changes in arterioles.

Blood is distributed to match the demands of the tissue in accordance with the local effectors of pressure, flow, nerves, and metabolites. Influences of these effectors are integrated and communicated to larger vessels to create a coordinated upstream response that allows for the blood required to meet the metabolic demands of the tissue. Each effector's contribution to the communicated response is unknown. In the present study in situ segments of arteriole, within the cheek pouch of the anesthetized hamster, were isolated from the pressure and flow of the surrounding vasculature while maintaining electrotonic continuity. Pressure was transiently increased or decreased (-70 to +120 cmH2O) for 60 s. These pressure changes within the isolated segment caused myogenic responses within the isolated segment as well as changes in diameter at remote arteriolar locations (ranging from -8 to 5 microns) that were outside the isolated segment and insulated from the changes in pressure. The size of the remote response correlated significantly with the change in pressure inside the isolated segment. This demonstrates that the effects of pressure changes in arterioles are communicated to neighboring portions of the vasculature.

Arteriolar endothelial cell barrier separates two populations of muscarinic receptors.

The endothelium of arterioles can function as a barrier to diffusion of hydrophilic molecules when studied in vitro. Thus a substance applied to one side of the arteriole is relatively ineffective in reaching receptors on the opposite side of the vessel wall unless it is lipid soluble. To study the receptor populations on the two sides of the arteriolar endothelium, we used micropipettes to apply methacholine (MCh; 1.0 microM), either luminally or adventitially, for 5 s to the arterioles of the cheek pouch of pentobarbital-anesthetized hamsters. MCh equally dilated the arterioles regardless of the side of application. That different populations of receptors are located on either side of the arteriole was shown by the fact that adventitially applied hydrophilic methscopolamine was ineffective in blocking the effects of the luminally applied MCh but completely blocked the effects of abluminally applied MCh. In contrast, the luminal population of receptors was easily blocked by adventially applied scopolamine, which is lipophilic. Separate and independent populations of receptors in the vessel wall suggests the potential for differential control between humoral and adventitial sources of vasoactive metabolites.

Microcirculatory responses to exogenous endothelial cell-derived relaxing factor.

Endothelium-derived relaxing factor (EDRF) plays an important role in the vasodilatory responses of large blood vessels. However, such a role has yet to be conclusively shown for the microvasculature. In this study we tested the sensitivity of arterioles in the cheek pouch of pentobarbital-anesthetized hamsters to the EDRF-dependent agonists bradykinin and A23187, as well as to exogenous EDRF from cultured bovine aortic endothelial cells. The pouch superfusion fluid was arranged to first pass through a column containing endothelial cells and then on to the tissue. Bradykinin (10-30 nM) or A23187 (0.3 microM) was introduced either upstream or downstream to the endothelial cells, and the resultant responses were measured with video microscopy. Bradykinin and A23187 both caused a dose-dependent release of a microvessel dilator from cultured endothelial cells. We take this dilator to be EDRF based on the characteristics of the responses to the stimuli. Indomethacin (7.7 microM) was present in the superfusate to eliminate the production of cyclooxygenase products from the endothelial cells, and the magnitude of the response was diminished if the superfusate was first passed through a 3-min delay coil before arrival at the pouch. The arterioles dilated to the direct application of bradykinin in a dose-dependent fashion. They did not respond however to the direct application of A23187. These studies demonstrate that arteriolar smooth muscle is able to respond to exogenous EDRF and support the premise that EDRF may play an active role in the regulation of blood flow in the microcirculation.

Arteriolar smooth muscle responses are modulated by an intramural diffusion barrier.

Arterioles (40-80 micron diameter) were isolated from the hamster cheek pouch, cannulated at both ends, and perfused with 3-(N-morpholino)propanesulfonic acid (MOPS)-buffered physiological salt solution (PSS). The vessels were observed with an inverted microscope and video system, and arteriolar diameter was measured. Arterioles were found to be 100 times more responsive to the alpha 1-adrenoceptor agonist phenylephrine when applied to the adventitial surface than when applied to the luminal surface. In contrast, SKF 89748-A, also an alpha 1-adrenoceptor selective agonist, but with a much greater lipid solubility than phenylephrine, was equipotent from either surface of the arteriole. We hypothesized that the difference between the two drugs was due to the ability of SKF 89748-A to permeate a diffusion barrier in the arteriolar wall because of its lipid-solubility. To test this hypothesis, a spectrum of antagonists with different sites of action and lipid solubilities was tested. The alpha-adrenoceptor antagonists phentolamine and benextramine and the muscarinic receptor antagonists atropine, scopolamine, and methscopolamine were all found to be more potent at blocking the action of appropriate agonists when applied to the same surface of the arteriole as the agonist than when applied to the opposite surface. Octanol-water partition coefficients were measured for each of the compounds, and these were found to be highly correlated with the ratio of luminal potency to adventitial potency for each of the drugs tested. These data support the hypothesis that the endothelial cell layer in these arterioles forms a barrier to the diffusion of small, water-soluble molecules from the lumen to the smooth muscle cell layer. Such a barrier may have a significant effect on arteriolar reactivity.

Hypodipsia and hypernatremia in congenital hydrocephalus.

The authors describe a case of hypodipsia and severe hypernatremia most probably secondary to hydrocephalus in a 22-year-old man in the absence of abnormalities of ADH secretion or metabolism. The patient became hypernatremic only in situations when the decreased spontaneous fluid intake was insufficient to replace that lost caused by sweating or vomiting. Adequate hydration returned the sodium value to normal.

Giant cell arteritis in a 20-year-old black man: a cause of intermittent calf claudication.

Giant cell arteritis in a young black man is an extremely unusual occurrence. A 20-year-old black man came for treatment of bilateral leg claudication that had been present for a 2-month period. His medical and angiographic evaluation led to an arterial biopsy that demonstrated giant cell arteritis. The patient was treated with corticosteroids and his condition has subsequently improved. Unusual variants of giant cell arteritis are discussed.