Edward P. Quigley

Proton conduction in gramicidin A and in its dioxolane-linked dimer in different lipid bilayers.

Gramicidin A (gA) molecules were covalently linked with a dioxolane ring. Dioxolane-linked gA dimers formed ion channels, selective for monovalent cations, in planar lipid bilayers. The main goal of this study was to compare the functional single ion channel properties of natural gA and its covalently linked dimer in two different lipid bilayers and HCl concentrations (10-8000 mM). Two ion channels with different gating and conductance properties were identified in bilayers from the product of dimerization reaction. The most commonly observed and most stable gramicidin A dimer is the main object of this study. This gramicidin dimer remained in the open state most of the time, with brief closing flickers (tau(closed) approximately 30 micros). The frequency of closing flickers increased with transmembrane potential, making the mean open time moderately voltage dependent (tau(open) changed approximately 1.43-fold/100 mV). Such gating behavior is markedly different from what is seen in natural gA channels. In PEPC (phosphatidylethanolamine-phosphatidylcholine) bilayers, single-channel current-voltage relationships had an ohmic behavior at low voltages, and a marked sublinearity at relatively higher voltages. This behavior contrasts with what was previously described in GMO (glycerylmonooleate) bilayers. In PEPC bilayers, the linear conductance of single-channel proton currents at different proton concentrations was essentially the same for both natural and gA dimers. g(max) and K(D), obtained from fitting experimental points to a Langmuir adsorption isotherm, were approximately 1500 pS and 300 mM, respectively, for both the natural gA and its dimer. In GMO bilayers, however, proton affinities of gA and the dioxolane-dimer were significantly lower (K(D) of approximately 1 and 1.5 M, respectively), and the g(max) higher (approximately 1750 and 2150 pS, respectively) than in PEPC bilayers. Furthermore, the relationship between single-channel conductance and proton concentration was linear at low bulk concentrations of H+ (0.01-2 M) and saturated at concentrations of more than 3 M. It is concluded that 1) The mobility of protons in gramicidin A channels in different lipid bilayers is remarkably similar to proton mobilities in aqueous solutions. In particular, at high concentrations of HCl, proton mobilities in gramicidin A channel and in solution differ by only 25%. 2) Differences between proton conductances in gramicidin A channels in GMO and PEPC cannot be explained by surface charge effects on PEPC membranes. It is proposed that protonated phospholipids adjacent to the mouth of the pore act as an additional source of protons for conduction through gA channels in relation to GMO bilayers. 3) Some experimental results cannot be reconciled with simple alterations in access resistance to proton flow in gA channels. Said differences could be explained if the structure and/or dynamics of water molecules inside gramicidin A channels is modulated by the lipid environment and by modifications in the structure of gA channels. 4) The dioxolane ring is probably responsible for the closing flickers seen in the dimer channel. However, other factors can also influence closing flickers.

Carotid Body Detection on CT Angiography

BACKGROUND AND PURPOSE: Advances in multidetector CT provide exquisite detail with improved delineation of the normal anatomic structures in the head and neck. The carotid body is 1 structure that is now routinely depicted with this new imaging technique. An understanding of the size range of the normal carotid body will allow the radiologist to distinguish patients with prominent normal carotid bodies from those who have a small carotid body paraganglioma. MATERIALS AND METHODS: We performed a retrospective analysis of 180 CTAs to assess the imaging appearance of the normal carotid body in its expected anatomic location. RESULTS: The carotid body was detected in >80% of carotid bifurcations. The normal size range measured from 1.1 to 3.9 mm ± 2 SDs, which is consistent with the reported values from anatomic dissections. CONCLUSIONS: An ovoid avidly enhancing structure at the inferomedial aspect of the carotid bifurcation within the above range should be considered a normal carotid body. When the carotid body measures >6 mm, a small carotid body paraganglioma should be suspected and further evaluated.

The Conduction of Protons in Different Stereoisomers of Dioxolane-Linked Gramicidin A Channels

Two different stereoisomers of the dioxolane-linked gramicidin A (gA) channels were individually synthesized (the SS and RR dimers;. Science. 244:813-817). The structural differences between these dimers arise from different chiralities within the dioxolane linker. The SS dimer mimics the helicity and the inter- and intramolecular hydrogen bonding of the monomer-monomer association of gAs. In contrast, there is a significant disruption of the helicity and hydrogen bonding pattern of the ion channel in the RR dimer. Single ion channels formed by the SS and RR dimers in planar lipid bilayers have different proton transport properties. The lipid environment in which the different dimers are reconstituted also has significant effects on single-channel proton conductance (g(H)). g(H) in the SS dimer is about 2-4 times as large as in the RR. In phospholipid bilayers with 1 M [H(+)](bulk), the current-voltage (I-V) relationship of the SS dimer is sublinear. Under identical experimental conditions, the I-V plot of the RR dimer is supralinear (S-shaped). In glycerylmonooleate bilayers with 1 M [H(+)](bulk), both the SS and RR dimers have a supralinear I-V plot. Consistent with results previously published (. Biophys. J. 73:2489-2502), the SS dimer is stable in lipid bilayers and has fast closures. In contrast, the open state of the RR channel has closed states that can last a few seconds, and the channel eventually inactivates into a closed state in either phospholipid or glycerylmonooleate bilayers. It is concluded that the water dynamics inside the pore as related to proton wire transfer is significantly different in the RR and SS dimers. Different physical mechanisms that could account for this hypothesis are discussed. The gating of the synthetic gA dimers seems to depend on the conformation of the dioxolane link between gAs. The experimental results provide an important framework for a detailed investigation at the atomic level of proton conduction in different and relatively simple ion channel structures.

Covalently linked gramicidin channels: effects of linker hydrophobicity and alkaline metals on different stereoisomers.

The direct role of the dioxolane group on the gating and single-channel conductance of different stereoisomers of the dioxolane-linked gramicidin A (gA) channels reconstituted in planar lipid bilayers was investigated. Four different covalently linked gA dimers were synthesized. In two of them, the linker was the conventional dioxolane described previously (SS and RR channels). Two gAs were covalently linked with a novel modified dioxolane group containing a retinal attachment (ret-SS and ret-RR gA dimers). These proteins also formed ion channels in lipid bilayers and were selective for monovalent cations. The presence of the bulky and hydrophobic retinal group immobilizes the dioxolane linker in the bilayer core preventing its rotation into the hydrophilic lumen of the pore. In 1 M HCl the gating kinetics of the SS or RR dimers were indistinguishable from their retinal counterparts; the dwell-time distributions of the open and closed states in the SS and ret-SS were basically the same. In particular, the inactivation of the RR was not prevented by the presence of the retinal group. It is concluded that neither the fast closing events in the SS or RR dimers nor the inactivation of the RR are likely to be a functional consequence of the flipping of the dioxolane inside the pore of the channel. On the other hand, the inactivation of the RR dimer was entirely eliminated when alkaline metals (Cs(+) or K(+)) were the permeating cations in the channel. In fact, the open state of the RR channel became extremely stable, and the gating characteristics of both the SS and RR channels were different from what was seen before with permeating protons. As in HCl, the presence of a retinal in the dioxolane linker did not affect the gating behavior of the SS and RR in Cs(+)- or K(+)-containing solutions. Alternative hypotheses concerning the gating of linked gA dimers are discussed.

Diagnostic Accuracy of Screening MR Imaging Using Unenhanced Axial CISS and Coronal T2WI for Detection of Small Internal Auditory Canal Lesions

T2-weighted and CISS images were compared with thin-section postcontrast T1 sequences for the detection of small (<10 mm) internal auditory canal schwannomas in 23 patients. The authors concluded that screening noncontrast MR imaging using a combination of axial CISS and coronal T2WI sequences can detect small internal auditory canal lesions with 100% sensitivity and excellent interobserver agreement. BACKGROUND AND PURPOSE: While enhanced T1WI is considered the “gold standard” for detection of internal auditory canal pathology, unenhanced fluid-sensitive sequences have shown high sensitivity for lesion identification. Our purpose was to evaluate the diagnostic accuracy of an unenhanced MR imaging protocol using axial CISS and coronal T2WI for detection of small (10 mm or less) internal auditory canal lesions. MATERIALS AND METHODS: Twenty-three patients with small internal auditory canal lesions and 13 patients without lesions who had undergone MR imaging using the screening protocol and confirmatory gadolinium-enhanced thin section T1WI were identified. Two blinded neuroradiologists retrospectively evaluated all examinations using 1) only axial CISS, 2) only coronal T2WI, and 3) axial and coronal sequences together. Accuracy, specificity, sensitivity, and interobserver agreement were assessed. RESULTS: Median maximum lesion dimension was 4 mm (range, 2–10 mm). Accuracy, specificity, and sensitivity for axial CISS alone were 0.94, 0.96, and 0.91 for observer 1 and 0.94, 0.92, and 1.00 for observer 2. The data for the coronal T2WI sequence only were 0.94, 0.96, and 0.91 for observer 1, and 0.99, 1.00, and 0.96 for observer 2. Using axial and coronal sequences, the data were 0.97, 0.96, and 1.00 for observer 1, and 0.99, 0.98, and 1.00 for observer 2. κ coefficients were 0.84 for the axial sequence only, 0.90 for coronal only, and 0.91 for axial and coronal both. CONCLUSIONS: Screening noncontrast MR imaging using a combination of axial CISS and coronal T2WI sequences can detect small internal auditory canal lesions with 100% sensitivity and excellent interobserver agreement.

Attenuation of Proton Currents by Methanol in a Dioxolane-Linked Gramicidin A Channel in Different Lipid Bilayers

The mobility of protons in a dioxolane-linked gramicidin A channel (D1) is comparable to the mobility of protons in aqueous solutions (Cukierman, S., E. P. Quigley, and D. S. Crumrine. 1997. Biophys. J. 73:2489-2502). Aliphatic alcohols decrease the mobility of H+ in aqueous solutions. In this study, the effects of methanol on proton conduction through D1 channels were investigated in different lipid bilayers and at different HCl concentrations. Methanol attenuated H+ currents in a voltage-independent manner. Attenuation of proton currents was also independent of H+ concentrations in solution. In phospholipid bilayers, methanol decreased the single channel conductance to protons without affecting the binding affinity of protons to bilayers. In glycerylmonooleate membranes, the attenuation of single channel proton conductances qualitatively resembled the decrease of conductivities of HCl solutions by methanol. However, in both types of lipid bilayers, single channel proton conductances through D1 channels were considerably more attenuated than the conductivities of different HCl solutions. This suggests that methanol modulates single proton currents through D1 channels. It is proposed that, on average, one methanol molecule binds to a D1 channel, and attenuates H+ conductance. The Gibbs free energy of this process (DeltaG0) is approximately 1.2 kcal/mol, which is comparable to the free energy of decrease of HCl conductivity in methanol solutions (1.6 kcal/mol). Apolar substances like urea and glucose that do not transport protons in HCl solutions and do not permeate D1 channels decreased solution conductivity and single channel conductance by a considerably larger proportion than methanol. Cs+ currents through D1 channels were considerably less (fivefold) attenuated by methanol than proton currents. It is proposed that methanol partitions inside the pore of gramicidin channels and delays the transfer of protons between water and methanol molecules, causing a significant attenuation of the single channel proton conductance. Gramicidin channels offer an interesting experimental model to study proton hopping along a single chain of water molecules interrupted by a single methanol molecule.

Gating and permeation in ion channels formed by gramicidin A and its dioxolane-linked dimer in Na(+) and Cs(+) solutions.

Abstract. The association of two gramicidin A (gA) peptides via H-bonds in lipid bilayers causes the formation of an ion channel that is selective for monovalent cations only. In this study, two gAs were covalently linked with a dioxolane group (SS dimer). Some functional properties of natural gA channels were compared to that synthetic dimer in Na+- or Cs+-containing solutions. The SS dimer remained in the open configuration most of the time, while natural gA channels had a relatively brief mean open time. Single channel conductances to Na+ (gNa) or Cs+ (gCs) in the SS dimer were smaller than in natural gA. However, gNa was considerably more attenuated than gCs. This probably results from a tight solvation of Na+ by the dioxolane linker in the SS channel. In Cs+ solutions, the SS had frequent closures. By contrast, in Na+ solutions the synthetic dimer remained essentially in the open state. The mean open times of SS channels in different solutions (Topen,Na > Topen,Cs > Topen,H) were inversely proportional to the single channel conductances (gH > gCs > gNa). This suggests that ion occupancy inside the pore stabilizes the open configuration of the gA dimer. The mean closed time of the SS dimer was longer in Cs+ than in H+ solutions. Possible mechanisms for these effects are discussed.

Noncontrast computed tomographic Hounsfield unit evaluation of cerebral venous thrombosis: a quantitative evaluation

IntroductionOur objective is to determine the utility of noncontrast Hounsfield unit values, Hounsfield unit values corrected for the patient’s hematocrit, and venoarterial Hounsfield unit difference measurements in the identification of intracranial venous thrombosis on noncontrast head computed tomography.MethodsWe retrospectively reviewed noncontrast head computed tomography exams performed in both normal patients and those with cerebral venous thrombosis, acquiring Hounsfield unit values in normal and thrombosed cerebral venous structures. Also, we acquired Hounsfield unit values in the internal carotid artery for comparison to thrombosed and nonthrombosed venous structures and compared the venous Hounsfield unit values to the patient’s hematocrit.ResultsA significant difference is identified between Hounsfield unit values in thrombosed and nonthrombosed venous structures. Applying Hounsfield unit threshold values of greater than 65, a Hounsfield unit to hematocrit ratio of greater than 1.7, and venoarterial difference values greater than 15 alone and in combination, the majority of cases of venous thrombosis are identifiable on noncontrast head computed tomography.ConclusionAbsolute Hounsfield unit values, Hounsfield unit to hematocrit ratios, and venoarterial Hounsfield unit value differences are a useful adjunct in noncontrast head computed tomographic evaluation of cerebral venous thrombosis.

Chiasmal stroke in patient with atrial fibrillation and complete occlusion of right internal carotid artery

F abian et al (1) recently published a case of “Chiasmal stroke following open-heart surgery” in the Journal, which described a 62-year-old man with total loss of vision in his right eye and loss of the temporal half of the vision in the left eye, and an otherwise normal neurologic examination. We report another case of chiasmal stroke. A 59-year-old man with probable myocarditis and a several month history of intermittent shortness of breath was diagnosed with atrial fibrillation with rapid ventricular rate and started on anticoagulation. Eight days later he experienced sudden complete vision loss in the right eye and loss of the temporal half of the visual field in the left eye. He was found to have an atrial thrombus and complete occlusion of his right internal carotid artery. Three days after acute vision loss, visual acuity was of no light perception, right eye, and 20/30, left eye. The right pupil was amaurotic. All other aspects of the examination, including appearance of the optic nerves, were normal. Automated perimetry demonstrated complete temporal field loss in the left eye respecting the vertical median (Fig. 1). His neurologic examination was normal. The day after vision loss, MRI of the brain showed an abnormality in the anterior chiasm on the right (Fig. 2). In addition, there was occlusion of the right internal carotid artery, an acute small left cerebellar infarct, and white matter changes consistent with small vessel ischemia. This patient had normal laboratory values for rapid plasma reagin and anti-neutrophil cytoplasmic antibody. Acute chiasmal infarction is a rare event, presumably because of its redundant and complex blood supply. To date, the only other report of chiasmal infarct was in a patient with acquired immunodeficiency syndrome, mucormyosis, and diabetes mellitus (2). We believe that the cause of our patients infarction was stroke, with embolus from a left atrial thrombus. Julie B. Shelton, MD John A Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City Department of Neurology, University of Utah Clinical Neurosciences Center, Salt Lake City Kellogg Eye Center, Department of Ophthalmology University of Michigan, Ann Arbor Kathleen B. Digre, MD Bradley J. Katz, MD, PhD Judith E. A. Warner, MD John A Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City Department of Neurology, University of Utah Clinical Neurosciences Center, Salt Lake City Edward P. Quigley, MD, PhD Neuroradiology Section, Department of Radiology University of Utah School of Medicine, Salt Lake City julieshe@med.umich.edu

GoPro Hero Cameras for Creation of a Three-Dimensional, Educational, Neurointerventional Video

Neurointerventional education relies on an apprenticeship model, with the trainee observing and participating in procedures with the guidance of a mentor. While educational videos are becoming prevalent in surgical cases, there is a dearth of comparable educational material for trainees in neurointerventional programs. We sought to create a high-quality, three-dimensional video of a routine diagnostic cerebral angiogram for use as an educational tool. A diagnostic cerebral angiogram was recorded using two GoPro HERO 3+ cameras with the Dual HERO System to capture the proceduralist’s hands during the case. This video was edited with recordings from the video monitors to create a real-time three-dimensional video of both the actions of the neurointerventionalist and the resulting wire/catheter movements. The final edited video, in either two or three dimensions, can serve as another instructional tool for the training of residents and/or fellows. Additional videos can be created in a similar fashion of more complicated neurointerventional cases. The GoPro HERO 3+ camera and Dual HERO System can be used to create educational videos of neurointerventional procedures.