[Surgical treatment of clival and axis bone cysts].

OBJECTIVE: The study objective is to present the tactics of surgical treatment of simple (solitary) bone cysts of the clival region and CII body. MATERIAL AND METHODS: Two patients were operated on for simple clival and axis cysts. RESULTS: The first patient with a simple clival cyst underwent transoral clivectomy and bone cyst evacuation. Postoperative control SCT scans revealed that the cyst cavity was lined with a hemostatic material and biological glue. Restoration of bone structures of the clivus occurred after 8 months. The second patient with a simple cyst of the second cervical vertebra (axis) underwent a two-stage surgical treatment: occipitospondylodesis was carried out first, and transoral removal of the pathological process of the CII body was performed at the second stage. Control scans a year after the surgery revealed the formation of bone tissue in the axis body region, an increased cystic cavity in the odontoid process of the axis, and posterior migration of the odontoid process, which caused compression of the brainstem structures. In connection with this, we performed transoral re-intervention with removal of the cystic separated odontoid process. Postoperative control scans revealed complete removal of the cystic odontoid process and decompression of the dural sac. The article provides a detailed analysis of the modern literature that has not reported any cases of a simple bone cyst located in the clivus or upper cervical vertebrae region and, thus, has not described the tactics for surgical treatment of these complex diseases. The article presents illustrative pre-, intra-, and postoperative images and histological specimens. CONCLUSION: The described cases present successful recurrence-free surgical treatment of simple (solitary) bone cysts located both in the clivus region (1 case) and in the body and odontoid process of the axis (1 case). A feature of the second case was that the patient had an extremely rare combination of a bone cyst of the C2 body and the cystic separated odontoid process of the axis (cystic os odontoideum).

A multi-frequency pulse EPR and ENDOR approach to study strongly coupled nuclei in frozen solutions of high-spin ferric heme proteins.

In spite of the tremendous progress in the field of pulse electron paramagnetic resonance (EPR) in recent years, these techniques have been scarcely used to investigate high-spin (HS) ferric heme proteins. Several technical and spin-system-specific reasons can be identified for this. Additional problems arise when no single crystals of the heme protein are available. In this work, we use the example of a frozen solution of aquometmyoglobin (metMb) to show how a multi-frequency pulse EPR approach can overcome these problems. In particular, the performance of the following pulse EPR techniques are tested: Davies electron nuclear double resonance (ENDOR), hyperfine correlated ENDOR (HYEND), electron-electron double resonance (ELDOR)-detected NMR, and several variants of hyperfine sublevel correlation (HYSCORE) spectroscopy including matched and SMART HYSCORE. The pulse EPR experiments are performed at X-, Q- and W-band microwave frequencies. The advantages and drawbacks of the different methods are discussed in relation to the nuclear interaction that they intend to reveal. The analysis of the spectra is supported by several simulation procedures, which are discussed. This work focuses on the analysis of the hyperfine and nuclear-quadrupole tensors of the strongly coupled nuclei of the first coordination sphere, namely, the directly coordinating heme and histidine nitrogens and the 17O nucleus of the distal water ligand. For the latter, 17O-isotope labeling was used. The accuracy of our results and the spectral resolution are compared in detail to an earlier single-crystal continuous-wave ENDOR study on metMb, and it will be shown how additional information can be obtained from the multi-frequency approach. The current work is therefore prone to become a template for future EPR/ENDOR investigations of HS ferric heme proteins for which no single crystals are available.

Pulsed EPR/ENDOR characterization of perturbations of the Cu(A) center ground state by axial methionine ligand mutations.

The effect of axial ligand mutation on the Cu(A) site in the recombinant water soluble fragment of subunit II of Thermus thermophilus cytochrome c oxidase ba(3) has been investigated. The weak methionine ligand was replaced by glutamate and glutamine which are stronger ligands. Two constructs, M160T0 and M160T9, that differ in the length of the peptide were prepared. M160T0 is the original soluble fragment construct of cytochrome ba(3) that encodes 135 amino acids of subunit II, omitting the transmembrane helix that anchors the domain in the membrane. In M160T9 nine C-terminal amino acids are missing, including one histidine. The latter has been used to reduce the amount of a secondary T2 copper which is most probably coordinated to a surface histidine in M160T0. The changes in the spin density in the Cu(A) site, as manifested by the hyperfine couplings of the weakly and strongly coupled nitrogens, and of the cysteine beta-protons, were followed using a combination of advanced EPR techniques. X-band ( approximately 9 GHz) electron-spin-echo envelope modulation (ESEEM) and two-dimensional (2D) hyperfine sublevel correlation (HYSCORE) spectroscopy were employed to measure the weakly coupled (14)N nuclei, and X- and W-band (95 GHz) pulsed electron-nuclear double resonance (ENDOR) spectroscopy for probing the strongly coupled (14)N nuclei and the beta-protons. The high field measurements were extremely useful as they allowed us to resolve the T2 and Cu(A) signals in the g( perpendicular) region and gave (1)H ENDOR spectra free of overlapping (14)N signals. The effects of the M160Q and M160E mutations were: (i) increase in A( parallel)((63,65)Cu), (ii) larger hyperfine coupling of the weakly coupled backbone nitrogen of C153, (iii) reduction in the isotropic hyperfine interaction, a(iso), of some of the beta-protons making them more similar, (iv) the a(iso) value of one of the remote nitrogens of the histidine residues is decreased, thus distinguishing the two histidines, and finally, (v) the symmetry of the g-tensor remained axial. These effects were associated with an increase in the Cu-Cu distance and subtle changes in the geometry of the Cu(2)S(2) core which are consistent with the electronic structural model of Gamelin et al. (Gamelin, D. R.; Randall, D. W.; Hay, M. T.; Houser, R. P.; Mulder, T. C.; Canters, G. W.; de Vries, S.; Tolman, W. B.; Lu, Y.; Solomon, E. I. J. Am. Chem. Soc. 1998, 120, 5246-5263).

A Q-band pulse EPR/ENDOR spectrometer and the implementation of advanced one- and two-dimensional pulse EPR methodology.

A versatile high-power pulse Q-band EPR spectrometer operating at 34.5--35.5 GHz and in a temperature range of 4--300 K is described. The spectrometer allows one to perform one- and two-dimensional multifrequency pulse EPR and pulse ENDOR experiments, as well as continuous wave experiments. It is equipped with two microwave sources and four microwave channels to generate pulse sequences with different amplitudes, phases, and carrier frequencies. A microwave pulse power of up to 100 W is available. Two channels form radiofrequency pulses with adjustable phases for ENDOR experiments. The spectrometer performance is demonstrated by single crystal pulse ENDOR experiments on a copper complex. A HYSCORE experiment demonstrates that the advantages of high-field EPR and correlation spectroscopy can be combined and exploited at Q-band. Furthermore, we illustrate how this combination can be used in cases where the HYSCORE experiment is no longer effective at 35 GHz because of the shallow modulation depth. Even in cases where the echo modulation is virtually absent in the HYSCORE experiment at Q-band, matched microwave pulses allow one to get HYSCORE spectra with a signal-to-noise ratio as good as at X-band. Finally, it is shown that the high microwave power, the short pulses, and the broad resonator bandwidth make the spectrometer well suited to Fourier transform EPR experiments.

Multiphoton resonances in pulse EPR

Two- and three-photon electron spin echoes of a two-level system are observed using, in addition to the microwave excitation, a linearly polarized radio-frequency field oriented along the static magnetic field B(0). Such multiphoton echoes are detected when the sum of the energies of one microwave and one or two radio-frequency photons are equal to the difference between energies of two spin states. The multiphoton character of the echoes is confirmed by measuring the spin nutation frequency as a function of the angle between the radio-frequency field and B(0), and monitoring the echo amplitude as a function of the radiation field strengths. Floquet theory, usually applied for the description of multiphoton resonances with an odd number of photons, is extended to the case where an even number of photons participate in the transition. Copyright 2000 Academic Press.

Azide binding to the trinuclear copper center in laccase and ascorbate oxidase.

Azide binding to the blue copper oxidases laccase and ascorbate oxidase (AO) was investigated by electron paramagnetic resonance (EPR) and pulsed electron-nuclear double resonance (ENDOR) spectroscopies. As the laccase : azide molar ratio decreases from 1:1 to 1:7, the intensity of the type 2 (T2) Cu(II) EPR signal decreases and a signal at g approximately 1.9 appears. Temperature and microwave power dependent EPR measurements showed that this signal has a relatively short relaxation time and is therefore observed only below 40 K. A g approximately 1.97 signal, with similar saturation characteristics was found in the AO : azide (1:7) sample. The g < 2 signals in both proteins are assigned to an S = 1 dipolar coupled Cu(II) pair whereby the azide binding disrupts the anti-ferromagnetic coupling of the type 3 (T3) Cu(II) pair. Analysis of the position of the g < 2 signals suggests that the distance between the dipolar coupled Cu(II) pair is shorter in laccase than in AO. The proximity of T2 Cu(II) to the S = 1 Cu(II) pair enhances its relaxation rate, reducing its signal intensity relative to that of native protein. The disruption of the T3 anti-ferromagnetic coupling occurs only in part of the protein molecules, and in the remaining part a different azide binding mode is observed. The 130 K EPR spectra of AO and laccase with azide (1:7) exhibit, in addition to an unperturbed T2 Cu(II) signal, new features in the g parallel region that are attributed to a perturbed T2 in protein molecules where the anti-ferromagnetic coupling of T3 has not been disrupted. While these features are also apparent in the AO : azide sample at 10 K, they are absent in the EPR spectra of the laccase : azide sample measured in the range of 6-90 K. Moreover, pulsed ENDOR measurements carried out at 4.2 K on the latter exhibited only a reduction in the intensity of the 20 MHz peak of the 14N histidine coordinated to the T2 Cu(II) but did not resolve any significant changes that could indicate azide binding to this ion. The lack of T2 Cu(II) signal perturbation below 90 K in laccase may be due to temperature dependence of the coupling within the trinuclear : azide complex.

A W-band pulsed ENDOR spectrometer: setup and application to transition metal centers.

The design and performance of a 95 GHz pulsed W-band EPR/ENDOR spectrometer is described with emphasis on the ENDOR part. Its unique feature is the easy and fast sample exchange at 4.2 K for frozen solution and single crystal samples. In addition, the microwave bridge power output is relatively high (maximum 267 mW), which allows the application of short microwave pulses. This increases the sensitivity in echo experiments because of the broader excitation bandwidth and the possibility of employing short pulse intervals, as long as the dead time does not increase significantly with the power. The spectrometer features two microwave and radiofrequency (0.1-220 MHz, 3 kW pulse power) channels and a 6 T superconducting magnet in a solenoid configuration. The magnet is equipped with cryogenic sweep coils providing a sweep range of +/-0. 4 and +/-0.2 T for a center field of 0-4 and 4-6 T, respectively. The spectrometer performance is demonstrated on Cu(II) centers in single crystals, a zeolite polycrystalline sample, and a protein frozen solution.

[Autoantibodies against NMDA-receptors in the blood of patients with acute disorders of cerebral circulation]. / Autoantitela k NMDA-retseptoram v krovi bol'nykh s ostrymi narusheniiami mozgovogo krovoobrashcheniia.

Concentration of autoantibodies to human brain NMDA-receptor in blood serum of patients with acute brain ischemic stroke was significantly higher than in healthy persons and patients with other neurological diseases. Average content of cholesterol, lipid peroxides and glutamate in the serum were close in all patient groups investigated. It was suggested that the content of autoantibodies to human brain NMDA-receptor could be used as biochemical criteria for acute brain stroke patient's condition control and their treatment efficiency.