Chemical Analysis of an Isotopically Labeled Molecule Using Two-Dimensional NMR Spectroscopy at 34 µT.

Low-field nuclear magnetic resonance (NMR) spectroscopy, conducted at or below a few millitesla, provides only limited spectral information due to its inability to resolve chemical shifts. Thus, chemical analysis based on this technique remains challenging. One potential solution to overcome this limitation is the use of isotopically labeled molecules. However, such compounds, particularly their use in two-dimensional (2D) NMR techniques, have rarely been studied. This study presents the results of both experimental and simulated correlation spectroscopy (COSY) on 1-13C-ethanol at 34.38 µT. The strong heteronuclear coupling in this molecule breaks the magnetic equivalence, causing all J-couplings, including homonuclear coupling, to split the 1H spectrum. The obtained COSY spectrum clearly shows the spectral details. Furthermore, we observed that homonuclear coupling between 1H spins generated cross-peaks only when the associated 1H spins were coupled to identical 13C spin states. Our findings demonstrate that a low-field 2D spectrum, even with a moderate spectral line width, can reveal the J-coupling networks of isotopically labeled molecules.

Accidental esophageal intubation via a large type C congenital tracheoesophageal fistula: A case report.

BACKGROUND: Tracheoesophageal fistula (TEF) is a congenital anomaly characterized by interruptions in esophageal continuity with or without fistulous communication to the trachea. Anesthetic management during TEF repair is challenging because of the difficulty of perioperative airway management. It is important to determine the appropriate position of the endotracheal tube (ETT) for proper ventilation and to prevent excessive gastric dilatation. Therefore, the tip of the ETT should be placed immediately below the fistula and above the carina. CASE SUMMARY: A full-term, one-day-old, 2.4 kg, 50 cm male neonate was diagnosed with TEF type C. During induction, an ETT was inserted using video laryngoscope and advanced deeply to ensure that the tip passed over the fistula, according to known strategies. The passage of the ETT through the vocal cords was confirmed via video laryngoscope. However, after inflating the ETT cuff, breath sounds were not heard on bilateral lung auscultation. Instead, gastric sounds were heard. Considering that a large fistula (approximately 6.60 mm × 4.54 mm) located 10.2 mm above the carina was confirmed on preoperative tracheal computed tomography, the possibility of unintentional esophageal intubation was highly suspected. Therefore, we decided to uncuff and withdraw the ETT carefully for repositioning, while monitoring auscultation and end-tidal CO2 simultaneously. At a certain point (9.5 cm from the lip), clear breath sounds and proper end-tidal CO2 readings were suddenly achieved, and adequate ventilation was possible. CONCLUSION: Preanesthetic anatomical evaluation with imaging studies in TEF is necessary to minimize complications related to airway management.

Management of symptomatic cervical facet cyst with cervical interlaminar epidural block: A case report.

BACKGROUND: Symptomatic cervical facet cysts are relatively rare compared to those in the lumbar region. These cysts are usually located in the 7th cervical and 1st thoracic vertebral (C7/T1) area, and surgical excision is performed in most cases. However, facet cysts are associated with degenerative conditions, and elderly patients are often ineligible for surgical procedures. Cervical interlaminar epidural block has been used in patients with cervical radiating symptoms and achieved good results. Therefore, cervical interlaminar epidural block may be the first-choice treatment for symptomatic cervical facet cysts. CASE SUMMARY: A 70-year-old man complained of a tingling sensation in the left hand, focused on the 4th and 5th fingers, for 1 year, and posterior neck pain for over 5 mo. The patient's numeric rating scale (NRS) score was 5/10. The patient was diagnosed with symptomatic cervical facet cyst at the left C7/T1 facet joint. Fluoroscopy-guided cervical interlaminar epidural block at the C7/T1 level with 20 mg triamcinolone and 5 mL of 0.5% lidocaine was administered. The patient's symptoms improved immediately after the block, with an NRS score of 3 points. After 3 mo, his left posterior neck pain and tingling along the left 8th cervical dermatome were relieved, with an NRS score of 2. CONCLUSION: A cervical interlaminar epidural block is a good alternative for managing symptomatic cervical facet cysts.

Efficacy of ganglion impar block on vulvodynia: Case series and results of mid- and long-term follow-up.

RATIONALE: Vulvodynia is a common chronic gynecological disease that affects approximately 16% of women, although it is rarely diagnosed. However, no known effective treatment exists. The etiology of vulvodynia is unknown and may be heterogeneous and multifactorial, so it is difficult-if not impossible-to improve this condition using 1 treatment method. Reports have shown that vulvodynia has an element of neuropathic pain. Although the role of the sympathetic nervous system in neuropathic pain is controversial, sympathetic nerve blocks have long been used to treat patients with chronic pain giving good results. A ganglion impar block (GIB), a sympathetic nerve block technique, may effectively manage pain and discomfort in patients with vulvodynia. PATIENT CONCERNS: Four patients suffering from chronic vulvar pain for 6 months-10 years were referred by gynecologists. The gynecologists could not identify the cause of the chronic vulvar pain, and symptoms were not improving by conservative therapy with medication. Patients complained of various chronic vulvar pain or discomfort. The initial visual analog scale (VAS) scores were 8 or 9 out of 10, and Leeds assessment of neuropathic symptoms and signs pain scale score was more than 12 out of 24. The review of gynecological medical records confirmed whether they showed allodynia during the cotton swab test and hyperalgesia to pin-prick test. DIAGNOSES: All patients were diagnosed with vulvodynia. INTERVENTIONS: All patients were treated with a GIB, once in 2 patients, 3 times in 1 patient, and 4 times (1 alcoholic neurolysis) in the other patient, under fluoroscopic guidance. OUTCOMES: After the procedures, the VAS score and the leeds assessment of neuropathic symptoms and signs (LANSS) pain scale score were decreased to less than 2 and 5, respectively, in all patients. Follow-up observations for 6 months-2 years revealed that 2 patients' symptoms entirely or nearly entirely improved and did not require further treatment. The pain of the remaining patients were well controlled with medications only. LESSONS: GIB is a good treatment option for patients suffering from chronic pain and discomfort caused by vulvodynia.

Inferior mesenteric plexus block under computed tomography guidance: A case report.

RATIONALE: Inferior mesenteric plexus block is indicated for left-sided lower abdominal pain. However, in patients with terminal cancer, severe abdominal pain can prevent the patient from maintaining the necessary posture during the procedure, and considerable anatomic deformation owing to extensive growth, invasion, and metastasis of the tumor in the abdominal cavity can make the procedure difficult. In these cases, performing the procedures under computed tomography (CT) guidance can ensure greater safety and accuracy. PATIENT CONCERNS: A 63-year-old man was referred for severe left-sided lower abdominal pain. He was unable to lie prone owing to severe lower abdominal pain and right hip surgery performed 15 years ago. His visual analog scale score was 9 out of 10. DIAGNOSES: The patient had terminal pancreatic tail cancer. Compared with abdominal CT findings obtained 50 days ago, hepatic metastasis and peritoneal seeding were still present, infiltration to the tissues around the pancreas and retrogastric area was increased, and most of the abdominal aorta was encased. In addition, metastatic lymph nodes were identified in several areas on the left including the left para-aortic area. However, the lesion causing the pain could not be identified. Therefore, an inferior mesenteric plexus block was performed according to the patient's complaint. INTERVENTIONS: Epidural patient-controlled analgesia was performed first. The patient's pain consequently reduced to a certain level, and the prone position became possible to some extent, so a CT-guided inferior mesenteric plexus block was performed 2 days later. OUTCOMES: After the CT-guided inferior mesenteric plexus block, it became possible to control the patient's pain with a fentanyl patch 75 mcg/hour only, and his visual analog scale score was reduced to 4. After 4 weeks, the patient died without complaints of severe pain as before. LESSONS: CT-guided inferior mesenteric plexus block can be performed in patients with left-sided lower abdominal pain, enabling a safer and more accurate procedure especially in patients with terminal cancer who are unable to lie prone owing to severe lower abdominal pain or with considerable anatomic deformation due to extensive growth, invasion, and metastasis of the tumor in the abdominal cavity.

Preoperative rigid laryngoscopic examination and modified jaw thrust manoeuver during fibreoptic-assisted tracheal intubation for general anaesthesia.

Preoperative laryngoscopic examination of the airway informs general anaesthesia management and planning. However, the same glottic opening view cannot always be obtained during direct laryngoscopy of anaesthetised patients. In this case report, a patient underwent preoperative rigid laryngoscopy due to medical history, and no problems were anticipated in performing tracheal intubation; however, the direct laryngoscopic view was a Grade 4 on the Cormack-Lehane Scale after anaesthesia induction. A jaw thrust manoeuvre to facilitate fibreoptic-assisted nasotracheal intubation was not feasible. In order to compensate, a modified method of jaw thrust was implemented, where both thumbs were placed on the floor of the patient's mouth, leading to a successful result. Safe airway management should be implemented with proper planning based on a careful preoperative evaluation.

Dynamic nuclear polarisation of liquids at one microtesla using circularly polarised RF with application to millimetre resolution MRI.

Magnetic resonance imaging in ultra-low fields is often limited by mediocre signal-to-noise ratio hindering a higher resolution. Overhauser dynamic nuclear polarisation (O-DNP) using nitroxide radicals has been an efficient solution for enhancing the thermal nuclear polarisation. However, the concurrence of positive and negative polarisation enhancements arises in ultra-low fields resulting in a significantly reduced net enhancement, making O-DNP far less attractive. Here, we address this issue by applying circularly polarised RF. O-DNP with circularly polarised RF renders a considerably improved enhancement factor of around 150,000 at 1.2 µT. A birdcage coil was adopted into an ultra-low field MRI system to generate the circularly polarised RF field homogeneously over a large volume. We acquired an MR image of a nitroxide radical solution with an average in-plane resolution of 1 mm. De-noising through compressive sensing further improved the image quality.

Ultralow-field and spin-locking relaxation dispersion in postmortem pig brain.

PURPOSE: To investigate tissue-specific differences, a quantitative comparison was made between relaxation dispersion in postmortem pig brain measured at ultralow fields (ULF) and spin locking at 7 tesla (T). The goal was to determine whether ULF-MRI has potential advantages for in vivo human brain imaging. METHODS: Separate specimens of gray matter and white matter were investigated using an ULF-MRI system with superconducting quantum interference device (SQUID) signal detection to measure T1ULF at fields from 58.7 to 235.0 µT and using a commercial MRI scanner to measure T1ρ7T at spin-locking fields from 5.0 to 235.0 µT. RESULTS: At matched field strengths, T1ρ7T is 50 to 100% longer than T1ULF. Furthermore, dispersion in T1ULF is close to linear between 58.7 and 235 µT, whereas dispersion in T1ρ7T is highly nonlinear over the same range. A subtle elbow in the T1ULF dispersion at approximately 140 µT is tentatively attributed to the local dipolar field of macromolecules. It is suggested that different relaxation mechanisms dominate each method and that ULF-MRI has a fundamentally different sensitivity to the macromolecular structure of neural tissue. CONCLUSIONS: Ultralow-field MRI may offer distinct, quantitative advantages for human brain imaging, while simultaneously avoiding the severe heating limitation imposed on high-field spin locking. Magn Reson Med 78:2342-2351, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Dynamic nuclear polarization in the hyperfine-field-dominant region.

Dynamic nuclear polarization (DNP) allows measuring enhanced nuclear magnetic resonance (NMR) signals. Though the efficiency of DNP has been known to increase at low fields, the usefulness of DNP has not been throughly investigated yet. Here, using a superconducting quantum interference device-based NMR system, we performed a series of DNP experiments with a nitroxide radical and measured DNP spectra at several magnetic fields down to sub-microtesla. In the DNP spectra, the large overlap of two peaks having opposite signs results in net enhancement factors, which are significantly lower than theoretical expectations and nearly invariant with respect to magnetic fields below the Earth's field. The numerical analysis based on the radical's Hamiltonian provides qualitative explanations of such features. The net enhancement factor reached 325 at maximum experimentally, but our analysis reveals that the local enhancement factor at the center of the rf coil is 575, which is unaffected by detection schemes. We conclude that DNP in the hyperfine-field-dominant region yields sufficiently enhanced NMR signals at magnetic fields above 1 µT.

T 1 relaxation measurement of ex-vivo breast cancer tissues at ultralow magnetic fields.

We investigated T 1 relaxations of ex-vivo cancer tissues at low magnetic fields in order to check the possibility of achieving a T 1 contrast higher than those obtained at high fields. The T 1 relaxations of fifteen pairs (normal and cancerous) of breast tissue samples were measured at three magnetic fields, 37, 62, and 122 µT, using our superconducting quantum interference device-based ultralow field nuclear magnetic resonance setup, optimally developed for ex-vivo tissue studies. A signal reconstruction based on Bayesian statistics for noise reduction was exploited to overcome the low signal-to-noise ratio. The ductal and lobular-type tissues did not exhibit meaningful T 1 contrast values between normal and cancerous tissues at the three different fields. On the other hand, an enhanced T 1 contrast was obtained for the mucinous cancer tissue.

Two-dimensional NMR spectroscopy of (13)C methanol at less than 5 µT.

Two-dimensional (2D) spectroscopy is one of the most significant applications of nuclear magnetic resonance (NMR). Here, we demonstrate that the 2D NMR can be performed even at a low magnetic field of less than 5µT, which is ten times less than the Earth's magnetic field. The pulses used in the experiment were composed of circularly polarized fields for coherent as well as wideband excitations. Since the excitation band covers the entire spectral range, the simplest two-pulse sequence delivered the full 2D spectrum. At 5µT, methanol with (13)C enriched up to 99% belongs to a strongly coupled regime, and thus its 2D spectrum exhibits complicated spectral correlations, which can be exploited as a fingerprint in chemical analysis. In addition, we show that, with compressive sensing, the acquisition of the 2D spectrum can be accelerated to take only 45% of the overall duration.

Toward a brain functional connectivity mapping modality by simultaneous imaging of coherent brainwaves.

Matching the proton-magnetic-resonance frequency to the frequency of a periodic neural oscillation (e.g., alpha or gamma band waves) by magnetic resonance imaging techniques, enables direct visualization of brain functional connectivity. Functional connectivity has been studied by analyzing the correlation between coherent neural oscillations in different areas of the brain. In electro- or magneto-encephalography, coherent source reconstruction in a source-space is very tricky due to power leaking from the correlation among the sources. For this reason, most studies have been limited to sensor-space analyses, which give doubtful results because of volume current mixing. The direct visualization of coherent brain oscillations can circumvent this problem. The feasibility of this idea was demonstrated by conducting phantom experiments with a SQUID-based, micro-Tesla NMR/MRI system. We introduce an experimental trick, an effective step-up of the measurement B-field in a pulse sequence, to decouple the magnetic resonance signal from the strong magneto-encephalographic signal at the same frequency.

Strong pulsed excitations using circularly polarized fields for ultra-low field NMR.

A pulse, which is produced by a single coil and thereby has a linear polarization, cannot coherently drive nuclear spins if the pulse is stronger than the static field B0. The inaccuracy of the pulse, which arises from the failure of the rotating wave approximation, has been an obstacle in adopting multiple pulse techniques in ultra-low field NMR where B0 is less than a few µT. Here, we show that such a limitation can be overcome by applying pulses of circular polarization using two orthogonal coils. The sinusoidal nutation of the nuclear spins was experimentally obtained, which indicates that coherent and precise controls of the nuclear spins can be achieved with circularly polarized pulses. Additional demonstration of the Carl-Purcell-Meiboom-Gill sequence verifies the feasibility of adopting multiple pulse sequences to ultra-low field NMR studies.

Simultaneous optical mapping of intracellular free calcium and action potentials from Langendorff perfused hearts.

The cardiac action potential (AP) controls the rise and fall of intracellular free Ca2+ (Ca(i)), and thus the amplitude and kinetics of force generation. Besides excitation-contraction coupling, the reverse process where Ca(i) influences the AP through Ca(i)-dependent ionic currents has been implicated as the mechanism underlying QT alternans and cardiac arrhythmias in heart failure, ischemia/reperfusion, cardiac myopathy, myocardial infarction, congenital and drug-induced long QT syndrome, and ventricular fibrillation. The development of dual optical mapping at high spatial and temporal resolution provides a powerful tool to investigate the role of Ca(i) anomalies in eliciting cardiac arrhythmias. This unit describes experimental protocols to map APs and Ca(i) transients from perfused hearts by labeling the heart with two fluorescent dyes, one to measure transmembrane potential (Vm), the other Ca(i) transients. High spatial and temporal resolution is achieved by selecting Vm and Ca(i) probes with the same excitation but different emission wavelengths, to avoid cross-talk and mechanical components.

Engraftment of connexin 43-expressing cells prevents post-infarct arrhythmia.

Ventricular tachyarrhythmias are the main cause of sudden death in patients after myocardial infarction. Here we show that transplantation of embryonic cardiomyocytes (eCMs) in myocardial infarcts protects against the induction of ventricular tachycardia (VT) in mice. Engraftment of eCMs, but not skeletal myoblasts (SMs), bone marrow cells or cardiac myofibroblasts, markedly decreased the incidence of VT induced by in vivo pacing. eCM engraftment results in improved electrical coupling between the surrounding myocardium and the infarct region, and Ca2+ signals from engrafted eCMs expressing a genetically encoded Ca2+ indicator could be entrained during sinoatrial cardiac activation in vivo. eCM grafts also increased conduction velocity and decreased the incidence of conduction block within the infarct. VT protection is critically dependent on expression of the gap-junction protein connexin 43 (Cx43; also known as Gja1): SMs genetically engineered to express Cx43 conferred a similar protection to that of eCMs against induced VT. Thus, engraftment of Cx43-expressing myocytes has the potential to reduce life-threatening post-infarct arrhythmias through the augmentation of intercellular coupling, suggesting autologous strategies for cardiac cell-based therapy.

Propagated endothelial Ca2+ waves and arteriolar dilation in vivo: measurements in Cx40BAC GCaMP2 transgenic mice.

To study endothelial cell (EC)- specific Ca(2+) signaling in vivo we engineered transgenic mice in which the Ca(2+) sensor GCaMP2 is placed under control of endogenous connexin40 (Cx40) transcription regulatory elements within a bacterial artificial chromosome (BAC), resulting in high sensor expression in arterial ECs, atrial myocytes, and cardiac Purkinje fibers. High signal/noise Ca(2+) signals were obtained in Cx40(BAC)-GCaMP2 mice within the ventricular Purkinje cell network in vitro and in ECs of cremaster muscle arterioles in vivo. Microiontophoresis of acetylcholine (ACh) onto arterioles triggered a transient increase in EC Ca(2+) fluorescence that propagated along the arteriole with an initial velocity of approximately 116 microm/s (n=28) and decayed over distances up to 974 microm. The local rise in EC Ca(2+) was followed (delay, 830+/-60 ms; n=8) by vasodilation that conducted rapidly (mm/s), bidirectionally, and into branches for distances exceeding 1 mm. At intermediate distances (300 to 600 microm), rapidly-conducted vasodilation occurred without changing EC Ca(2+), and additional dilation occurred after arrival of a Ca(2+) wave. In contrast, focal delivery of sodium nitroprusside evoked similar local dilations without Ca(2+) signaling or conduction. We conclude that in vivo responses to ACh in arterioles consists of 2 phases: (1) a rapidly-conducted vasodilation initiated by a local rise in EC Ca(2+) but independent of EC Ca(2+) signaling at remote sites; and (2) a slower complementary dilation associated with a Ca(2+) wave that propagates along the endothelium.

Cardiac beat-to-beat alternations driven by unusual spiral waves.

Alternans, a beat-to-beat temporal alternation in the sequence of heartbeats, is a known precursor of the development of cardiac fibrillation, leading to sudden cardiac death. The equally important precursor of cardiac arrhythmias is the rotating spiral wave of electro-mechanical activity, or reentry, on the heart tissue. Here, we show that these two seemingly different phenomena can have a remarkable relationship. In well controlled in vitro tissue cultures, isotropic populations of rat ventricular myocytes sustaining a temporal rhythm of alternans can support period-2 oscillatory reentries and vice versa. These reentries bear "line defects" across which the phase of local excitation slips rather abruptly by 2pi, when a full period-2 cycle of alternans completes in 4pi. In other words, the cells belonging to the line defects are period-1 oscillatory, whereas all of the others in the bulk medium are period-2 oscillatory. We also find that a slowly rotating line defect results in a quasi-periodic like oscillation in the bulk medium. Some key features of these phenomena can be well reproduced in computer simulations of a nonlinear reaction-diffusion model.

Monte Carlo simulation of 3D mapping of cardiac electrical activity with spinning slit confocal optics.

Optical techniques used to map transmembrane potential and intracellular Ca2+ activities of intact hearts are restricted to the surface and cannot resolve activity in deeper layers due to the lack of depth resolution. The recent development of spinning slit confocal optics offers advantages of depth resolution as well as high-speed confocal imaging which are necessary for millisecond-scale, depth-resolved mapping of membrane potential and/or intracellular Ca2+ concentration. Here, we show simulated confocal optics derived from confocal slits on a high-speed spinning disk using Monte Carlo method with a numerical heart tissue model and find that depth-resolved optical mapping is feasible down to around 800 microm below the surface using 670-nm excitation light. The numerical model shows that (1) a minimum slit separation, which is found to be a function of depth of the focal plane and the numerical aperture of the objective lens, for minimum background noise exists and (2) narrower slit widths result in slightly greater depth resolution but has a negative impact in significantly lower overall fluorescence intensity. An experimental test of this optics has been performed by imaging two overlapping layers of fluorescent beads and the result confirms the expected depth resolution in non-scattering medium. These results will be able to serve as a benchmark on how a 3D-imaging system can be expected to perform and what kind of theoretical depth-resolution can be expected from it.

Complex-periodic spiral waves in confluent cardiac cell cultures induced by localized inhomogeneities.

Spatiotemporal wave activities in excitable heart tissues have long been the subject of numerous studies because they underlie different forms of cardiac arrhythmias. In particular, understanding the dynamics and the instabilities of spiral waves have become very important because they can cause reentrant tachycardia and their subsequent transitions to fibrillation. Although many aspects of cardiac spiral waves have been investigated through experiments and model simulations, their complex properties are far from well understood. Here, we show that intriguing complex-periodic (such as period-2, period-3, period-4, or aperiodic) spiral wave states can arise in monolayer tissues of cardiac cell culture in vitro, and demonstrate that these different dynamic states can coexist with abrupt and spontaneous transitions among them without any change in system parameters; in other words, the medium supports multistability. Based on extensive image data analysis, we have confirmed that these spiral waves are driven by their tips tracing complex orbits whose unusual, meandering shapes are formed by delicate interplay between localized conduction blocks and nonlinear properties of the culture medium.

Regular and alternant spiral waves of contractile motion on rat ventricle cell cultures.

We demonstrate that meandering as well as regular spiral waves can form in a well-controlled culture layer of rat ventricle cells and that the meandering spiral wave, in particular, can generate an alternant rhythm. These observations are made possible by a newly developed, noninvasive phase contrast macro-optics that is simple but highly effective in visualizing the contractile motion of the populations of cardiac cells.