Protein-Ligand Interaction Analyses with Nuclear Magnetic Resonance Spectroscopy Enhanced by Dissolution Triplet Dynamic Nuclear Polarization.

Solution-state nuclear magnetic resonance spectroscopy (NMR) is a powerful method for the analysis of intermolecular interactions within a biomolecular system. However, low sensitivity is one of the major obstacles of NMR. We improved the sensitivity of solution-state 13C NMR for the observation of intermolecular interactions between protein and ligand using hyperpolarized solution samples at room temperature. Eutectic crystals composed of 13C-salicylic acid and benzoic acid doped with pentacene were hyperpolarized by dynamic nuclear polarization using photoexcited triplet electrons, and a 13C nuclear polarization of 0.72 ± 0.07% was achieved after dissolution. The binding of human serum albumin and 13C-salicylate was observed with several hundred times sensitivity enhancement under mild conditions. The established 13C NMR was applied for pharmaceutical NMR experiments by observation of the partial return of the 13C chemical shift of salicylate by competitive binding with other non-isotope-labeled drugs.

Prediction of 1H Singlet Relaxation via Intermolecular Dipolar Couplings Using the Molecular Dynamics Method.

Dissolution dynamic nuclear polarization has been applied in various fields, including chemistry, biology, and medical science. To expand the scope of these applications, the nuclear singlet state, which is decoherence-free against dipolar relaxation between spin pairs, has been studied experimentally, theoretically, and numerically. The singlet state composed of proton spins is used in several applications, such as enhanced polarization preservation, molecular tagging to probe slow dynamic processes, and detection of ligand-protein complexes. In this study, we predict the lifetimes of the nuclear spin states composed of proton spin pairs using the molecular dynamics method and quantum chemistry simulations. We consider intramolecular dipolar, intermolecular dipolar between solvent and solute, chemical shift anisotropy, and spin-rotation interactions. In particular, the relaxation rate of intermolecular dipolar interactions is calculated using the molecular dynamics method for various solvents. The calculated values and the experimental values are of the same order of magnitude. Our program would provide insight into the molecular design of several NMR applications and would be helpful in predicting the nuclear spin relaxation time of synthetic molecules in advance.

In vitro measurement of tissue integrity during saccular aneurysm embolizations for simulator-based training.

In the domain of endovascular neurosurgery, the measurement of tissue integrity is needed for simulator-based training and for the development of new intravascular instruments and treatment techniques. In vitro evaluation of tissue manipulation can be achieved using photoelastic stress analysis and vasculature modeling with photoelastic materials. In this research we constructed two types of vasculature models of saccular aneurysms for differentiation of embolization techniques according to the respect for tissue integrity measurements based on the stress within the blood vessel model wall. In an aneurysm model with 5 mm dome diameter, embolization using MicroPlex 10 (Complex 1D, with 4 mm diameter loops), a maximum area of 3.97 mm² with stress above 1 kPa was measured. This area increased to 5.50 mm² when the dome was touched deliberately with the release mechanism of the coil, and to 4.87 mm² for an embolization using Micrusphere, (Spherical 18 Platinum Coil). In a similar way trans-cell stent-assisted coil embolization was also compared to human blood pressure simulation using a model of a wide-necked saccular aneurysm with 7 mm diameter. The area with stress above 1kPa was below 1 mm² for the pressure simulation and maximized at 3.79 mm² during the trans-cell insertion of the micro-catheter and at 8.92 mm² during the embolization. The presented results show that this measurement system is useful for identifying techniques compromising tissue integrity, comparing and studying coils and embolization techniques for a specific vasculature morphology and comparing their natural stress variations such as that produced by blood pressure.

Subcutaneous hematoma associated with manual cervical massage during carotid artery stenting. A case report.

We describe a patient with subcutaneous hematoma associated with manual cervical massage during carotid artery stenting.A 73-year-old man with left cervical carotid artery stenosis presented with left amaurosis fugax. We performed carotid artery stenting using distal embolic protection with balloon occlusion. Dual antiplatelet therapy was maintained in the periprocedural period and an anticoagulant agent was administered during the procedure. Because the aspiration catheter became entrapped by the stent, it did not reach the distal side of the stenotic lesion, and manual compression of the cervical region was therefore performed. Immediately afterwards, a subcutaneous hemorrhage occurred in the cervical region. There was no postoperative dyspnea due to enlargement of the hematoma, which was absorbed spontaneously.Cervical subcutaneous hematoma can occur in the cervical region due to cervical massage in patients who are receiving adjuvant antiplatelet therapy and anticoagulation therapy.

Uneven distribution of the luxS gene within the genus Campylobacter.

Polymerase chain reaction (PCR) amplification was performed on 20 isolates of five Campylobacter species using a degenerate primer pair designed in silico to generate a product of the luxS gene or its homologue from Campylobacter organisms. Although the primer pair successfully amplified products of approximately 500 base pairs (bp) with the eight isolates of C. jejuni and C. coli and some of C. upsaliensis and C. fetus, it failed to amplify fragments with all four isolates of C. lari (two urease-negative C. lari; two urease-positive thermophilic campylobacters). When Southern blot hybridisation analysis was carried using the mixed luxS gene fragments prepared from the C. jejuni, C. coli, C. upsaliensis and C. fetus strains as a probe, all C. jejuni, C. coli, C. upsaliensis and C. fetus isolates gave positive signals, but no positive signal was detected with any C. lari isolate. These results clearly indicate that C. jejuni, C. coli, C. upsaliensis and C. fetus carry the luxS gene or its homologue. However, no luxS gene or its homologue was identified to occur in the C. lari genome. Although autoinducer-2 assays were positive in C. jejuni, C. coli, C. upsaliensis and C. fetus isolates, it was negative with all the C. lari isolates examined. In addition, a biofilm formation assay demonstrated that biofilm formation in the C. lari species does not appear to correlate with the occurrence of the luxS gene because biofilm formation occurred among some isolates of C. lari.

Photoelastic stress analysis in blood vessel phantoms: three-dimensional visualization and saccular aneurysm with bleb.

BACKGROUND: The photoelastic effect is used for stress measurement during endovascular surgery simulation for quantitative evaluation of catheter trajectory in in vitro environments. By extending the capabilities of this sensing technology, its potential for intravascular tools evaluation will increase. METHODS: In this research the error introduced by stress direction on magnitude measurements was studied, then stress measurements were made in the phantom modelling of a saccular aneurysm with bleb. To visualize three-dimensionally the stress field changes produced by a guide wire in a phantom wall, a scanner and an algorithm relying on maximum likelihood-expectation maximization are proposed. Three-dimensional fields at different pressure level were compared with the stress field surrounding the guide wire. RESULTS: The maximum error in stress magnitude measurements due to stress direction was 2.52%. Stress local maximum was detected in the bleb phantom before rupture. Three-dimensional visualization was obtained in vasculature phantom with average errors of 10.73%, 4.55%, 3.18% for inner pressures of 80, 120, 160 mmHg, respectively. Stress measurement in the neighbourhood of the guide wire is equivalent to applying an inner pressure of 120 mmHg. CONCLUSIONS: For the presented polariscope, the weak influence of stress direction in magnitude measurements was confirmed. In vasculature phantoms, the three-dimensional visualization of stress eliminated birefringence visualization distortion and enabled more comprehensive comparison of stress produced by intravascular tools with stress produced by normal blood pressure.

(2)H-decoupling-accelerated (1)H spin diffusion in dynamic nuclear polarization with photoexcited triplet electrons.

In dynamic nuclear polarization (DNP) experiments applied to organic solids for creating nonequilibrium, high (1)H spin polarization, an efficient buildup of (1)H polarization is attained by partially deuterating the material of interest with an appropriate (1)H concentration. In such a dilute (1)H spin system, it is shown that the (1)H spin diffusion rate and thereby the buildup efficiency of (1)H polarization can further be enhanced by continually applying radiofrequency irradiation for deuterium decoupling during the DNP process. As experimentally confirmed in this work, the electron spin polarization of the photoexcited triplet state is mainly transferred only to those (1)H spins, which are in the vicinity of the electron spins, and (1)H spin diffusion transports the localized (1)H polarization over the whole sample volume. The (1)H spin diffusion coefficients are estimated from DNP repetition interval dependence of the initial buildup rate of (1)H polarization, and the result indicates that the spin diffusion coefficient is enhanced by a factor of 2 compared to that without (2)H decoupling.

Total compensation of pulse transients inside a resonator.

The profile of rf pulses that nuclear spins experience inside a resonator deviates from that of rf voltage signals generated by a NMR spectrometer according to users' pulse programming, when change of the profile in time is comparable to or shorter than the time constant of the resonator. In our previous work [Takeda et al., J. Magn. Reson. 197 (2009) 242-244], we proposed active compensation of rf pulse transients, in which the amplitude transient of the rf pulse can be suppressed without sacrificing the Q factor of the probe. Here we extend the idea of active compensation toward total compensation of the amplitude as well as phase transients. By measuring the transient response of the probe to a given excitation using a pickup coil, the response function determining the transient behavior of the probe is numerically obtained. Then, by numerically solving the convolution equation with the help of Laplace transformation, one can obtain the amplitude and phase profiles of the pulse that should be programmed in the spectrometer in order to apply the rf pulses to the nuclear spins as intended. Accurate rf pulsing based on this idea is experimentally demonstrated, and prospect and requirements for coping with the receiver dead-time problem are discussed.

Autonomous catheter insertion system using magnetic motion capture sensor for endovascular surgery.

BACKGROUND: In order to reduce fluoroscope usage in endovascular surgery, there is a need to develop autonomous catheter insertion systems. METHODS: We propose a system for tracking the position and speed of a catheter using a magnetic motion capture sensor to provide feedback to a catheter-driving mechanism, to perform autonomous catheter insertion in major vasculature. Catheter insertion speed control and path reconstruction experiments were performed with the system inside a silicone model of major vasculature to simulate surgery. RESULTS: The system controlled the catheter for speeds of 6.14 mm/s and reproduced a two-dimensional path inside the silicone blood vessel phantom with less than 7 mm of error. CONCLUSIONS: We found that error in speed control rises as a result of friction between the catheter and the model wall. Path reconstruction error depends on the model's cross-sectional diameter, the properties of the catheter insertion mechanism, the magnetic sensor and the system guidance technique.

Numerical evaluation method for catheter prototypes using photo-elastic stress analysis on patient-specific vascular model.

BACKGROUND: To date, no quantitative analysis has been developed to evaluate catheter performance inside the vascular lumen. METHODS: An evaluation system for endovascular tools was built with a polyurethane elastomer vascular model inside a polariscope and a catheter driving system. This robotic system reproduced a catheter insertion trajectory inside the vascular model, using a surgical catheter and three catheter prototypes used for motion capture on endovascular surgery simulation. Birefringence is produced by photo-elastic characteristics of the polyurethane elastomer when the material is submitted external stress. The birefringence produced by the catheter on the vascular model wall was recorded and represented numerically by the correlation between consecutive frames of the registered video. RESULTS: Correlation values between frames showed that the performance of the prototypes was lower than that of the medical use catheter. The performance of prototypes was reduced by microcoils on their tips. CONCLUSIONS: This methodology opens new options to evaluate medical catheters and physicians skills. opyright

Extracranial vertebral artery aneurysm associated with neurofibromatosis type 1. A case report.

SUMMARY: Extracranial vertebral artery aneurysm associated with neurofibromatosis (NF1) is very rare. The aneurysm was successfully treated by endovascular trapping of the aneurysm and proximal vertebral artery with coils.

New simple method for purification of class I alcohol dehydrogenase.

AIMS: The purpose of this study was to develop a new simple method for purification of rat class I alcohol dehydrogenase (ADH, EC 1.1.1.1). METHODS AND RESULTS: Immobilized p-hydroxyacetophenone was used as a ligand for affinity chromatography for the initial purification step after ammonium sulfate precipitation of the cytosolic fraction of rat liver. Then the eluant was separated by using ion-exchange chromatography, and homogenous class I ADH, as judged by the results of SDS-PAGE and confirmed by the results of the amino-acid sequence of peptides degraded from a 39 kDa protein, was obtained with a high yield (57%). The purified ADH showed kinetic constants of 1.3 mmol/l for Km and 62.4 per min for Kcat with ethanol as a substrate. ADH was also successfully purified from yeast by a similar method using p-hydroxyacetophenone affinity chromatography. CONCLUSIONS: This simple method involving only two chromatographic procedures may be very useful for purification of ADH.

Local hyperthermia enhances thrombosis in aneurysms containing platinum coils.

SUMMARY: Despite recent technical advances in embolization of cerebral aneurysms with platinum coils, some aneurysms eventually resulted in incomplete packing with remnant neck or dome filling. Such a situation with a remaining inflow zone may pose a risk of rupture and subsequent regrowth. Metals characteristically generate heat under high-frequency alternating magnetic fields (AMF). We used this property to induce local hyperthermia and promote thrombogenesis in incompletely packed aneurysms. Glass model aneurysms packed with coils were subjected to AMF to investigate the correlation between weight of platinum and temperature elevation and the correlation between flow rates of water through the model and temperature elevation. Next, activated coagulation time (ACT) of blood obtained from dogs was studied at various temperatures. Finally, side-wall aneurysms created in the canine carotid artery using a venous patch were packed with platinum coils. Change in temperature and angiographic changes were investigated after AMF application. In the glass model, the weight of platinum was correlated with elevation of temperature, and a negative logarithmic correlation was evident between flow rate and elevation of temperature. Elevation of blood sample temperature tended to shorten ACT. In canine carotid aneurysms, elevation of intra-aneurysmal temperature was confirmed and sufficient elevation of temperature was found to promote angiographically evident thrombogenesis of the remnant space after AMF application. Local hyperthermia may be useful in completing luminal obliteration of aneurysms after coil embolization. It may particularly useful for ruptured aneurysms to prevent the early rerupture.

Assessing Prognosis of Areas of Acute Cerebral Ischemia Using Perfusion CT.

SUMMARY: Vascular recanalization by the vascular reconstruction method can dramatically improve ischemic symptoms in patients with acute cerebral ischemia. However, this treatment method is frequently associated with haemorrhagic complications. The indications for this therapeutic approach have been described in a number of studies in the literature. The present paper discusses the possibility of assessing the prognosis of ischemic areas using perfusion CT (PCT) by comparing the results obtained before and after thrombolytic therapy. Twenty-six patients underwent vascular reconstruction at our hospital between July 2002 and March 2004. Of these patients, six who underwent PCT before treatment and showed adequate recanalization following vascular reconstruction were included in the present study. PCT images were obtained using the first-pass bolus-tracking method with a 16-row multislice helical CT scanner. Areas of cerebral ischemia were evaluated by CT before and after vascular reconstruction. A region of interest was placed in the area showing low density in CT images before vascular reconstruction. The mean average CBF (mL/min/100 g), CBV (mL/100 g), and MTT (s) values were calculated in areas with and without cerebral infarction after vascular reconstruction. The %CBF, % CBV, and %MTT values relative to the normal side were evaluated with reference to the time until recanalization. Transarterial vascular reconstruction resulted in full recanalization in four patients and partial recanalization in two. The mean time from onset to recanalization was 284.7 +/- 63.27 minutes and was not longer than six hours in any patient. The patient prognosis results in terms of GOS were GR in two patients, MD in three patients, and SD in one patient. Based on comparison of the time after examination to recanalization, the %CBF showed a significant positive correlation in the salvaged area (Y = 47.321 + 2.491 x %CBF:R(2) = 0.792, p < 0.05). A significant correlation was not observed in %CBV, but %MTT showed a significant negative correlation (Y = 269.45 - 0.356 x %MTT:R(2) = 0.794, p < 0.05). The %CBF and %MTT results obtained by PCT performed before transarterial vascular reconstruction suggest that it may be possible to estimate the time before vascular reconstruction and the relationship with prognosis. These findings are expected to help ensure the appropriate application of vascular reconstruction and to provide useful information for developing optimal therapeutic protocols, thus reducing complications. In addition, because the results are based on the time after examination, the appropriate therapeutic approach can be determined even when the time of onset of ischemia is uncertain.

Computational fluid dynamics for brain circulation and aneurysm with therapeutic devices.

SUMMARY: We analysed fluid dynamics at brain arteries having multiple inflow and out flow like Willis ring based on clinical imaging modalities. In addition, we analysed fluid dynamics with therapeutic devices like coils and stents to simulate their influences to blood flow. 3D CTA and MRA obtained three-dimensional structures of the brain vessels. The centreline was obtained from the three dimensional structure. Diameter of the blood vessels was measured by 3D CTA/MRA then smooth surfaced blood vessel models were created. For the fluid analysis, we developed a home brew software which can display parameters such as streamline, etc. In addition, our CFD (computational fluid dynamics) software can work in collaboration with a CAD (computer aided design) software which we also developed (VCAD: Volume CAD). So, therapeutic devices such as coils, balloons and stents could be placed in the models and CFD analysis could be performed placing devices in the models. The flow pattern in the complicated vascular structure could be calculated such as Willis ring which has multiple inputs like ICA, VA and multiple outlets like MCA and PCA with communicating arteries. CFD with therapeutic devices could also analysed with our system. CFD including communicating arteries will assist the simulation of parent artery occlusion. CFD with therapeutic devices is helpful not only for simulation for embolization, but will help us to design therapeutic devices under computer simulation.

Fine Structure of Detached Coils Observed by Micro-CT.

SUMMARY: Detached GDC was observed by high resolutional industrial microCT, whose resolution was higher than 0.1 mm. Unexpected destruction of the coils (kinking) was detected and unraveling also clearly visualized. Much higher resolution can improve safe and effectivity of GDC procedure.

Treatment Strategy for Cerebral Aneurysms Based on the Evidence of the Efficacy of GDC Embolization.

SUMMARY: The authors reviewed 531 patients with cerebral aneurysms treated with Guglielmi detachable coils (GDCs) over 5 years to clarify both the advantages and disadvantages of embolization based on the evidence of complications by aneurysm profile. There were 52 technical complications, 25 of which resulted in unfavorable patient outcomes. Intraoperative rupture, the most serious complication exacerbating the patient's condition, occurred in 19 patients, 4 of whom expired. All of these aneurysms were very small and were mostly located in the AcomA and PICA portions. Thirteen patients encountered thromboembolic complications, 6 of whom were elderly with acute ruptured aneurysms at MCA and the tip of BA. For large or giant aneurysms manifesting the mass effect, particularly those in the ICA-C2 portion compressing the optic nerve, the saccular packing did little to ameliorate the symptoms, and subsequent surgical or endovascular trapping was needed. Therefore, saccular embolization of endovascularly difficult, very small AcomA aneurysms and large C2 aneurysm with visual symptoms should be used sparingly based on a risk-benefit assessment.

Treatment strategy for aneurysms of the posterior cerebral artery.

SUMMARY: The authors carried out a retrospective review of the records of 12 patients with aneurysms of the posterior cerebral artery (PCA). Four were asymptomatic, 1 presented with a mass effect, and 7 with a subarachnoid haemorrhage (SAH). Of the 7 ruptured aneurysms, 3 were embolized and 2 were clipped. However, 2 patients died from rebleeding before any treatment. Of the 5 unruptured aneurysms, 1 was embolized with coils but the remaining 4 have been conservatively observed. No aneurysms have ruptured during the follow-up period, and 3 have thrombosed spontaneously. According to our results, the PCA aneurysms should be treated aggressively in the early phase. Although the preservation of the anatomical integrity of the PCA should naturally be one of the prime objectives, PCA occlusion may sometimes be inevitable when treating large or fusiform aneurysms. On the other hand, conservative therapy is one of the options for the treatment of incidentally encountered unruptured ones, because these have the possibility of spontaneous thrombosis.