Alexander Potapov

Hiding in the Shadows: CPOX Expression and 5-ALA Induced Fluorescence in Human Glioma Cells

Protoporphyrin IX (PpIX) is widely used in photodynamic diagnosis. To date, the details of molecular mechanisms underlying PpIX accumulation in malignant cells after 5-ALA administration remain unclear. The fluorescence of PpIX was studied in human glioma cells. Several cell cultures were established from glioma tumor tissue to study the differences between fluorescence-positive and fluorescence-negative human glioma tumors. The cell cultures demonstrated fluorescence profiles similar to those of source tumor tissues, which allows us to use these cultures in experimental research. Dynamics of the rates of synthesis and degradation of fluorescent protoporphyrin IX was studied in the cultures obtained. In addition, the expression of CPOX, an enzyme involved in PpIX synthesis, was evaluated. mRNA levels of heme biosynthesis enzymes were analyzed, and PpIX fluorescence proved to correlate with the CPOX protein level, whereas no such correlation was observed at the mRNA level. Fluorescence intensity decreased at low levels of the enzyme, which indicates its critical role in PpIX fluorescence. Finally, the fluorescence intensity proved to correlate with the proliferative activity.

High-resolution mass spectra processing for the identification of different pathological tissue types of brain tumors

The purpose of the work is to demonstrate the possibilities of identifying the different types of pathological tissue identification directly through tissue mass spectrometry. Glioblastoma parts dissected during neurosurgical operation were investigated. Tumor fragments were investigated by the immunohistochemistry method and were identified as necrotic tissue with necrotized vessels, necrotic tissue with tumor stain, tumor with necrosis (tumor tissue as major), tumor, necrotized tumor (necrotic tissues as major), parts of tumor cells, boundary brain tissue, and brain tissue hyperplasia. The technique of classification of tumor tissues based on mass spectrometric profile data processing is suggested in this paper. Classifiers dividing the researched sample to the corresponding tissue type were created as a result of the processing. Classifiers of necrotic and tumor tissues are shown to yield a combined result when the tissue is heterogeneous and consists of both tumor cells and necrotic tissue.

Navigation systems in neurosurgery

When preparing the review, we analyzed publications available at the Medline database; a total of 1,083 publications related to the reviews subject were analyzed. After more careful analysis, we selected 117 publications devoted to the development of neuronavigation in craniocerebral surgery, historical prerequisites of neuronavigation emergence, current trends, and future perspectives of the technique.

Evolution of Surgical Treatment of Chronic Subdural Hematomas

We analyzed results of the surgical treatment of chronic subdural hematomas (CSH) in 148 patients. It’s estimated that removal of hematoma through the burr hole with further closed external drainage is the most adequate method for CSH treatment, especially in the elderly patients. Craniotomy with membranectomy is recommended only in cases of vast dense clots in the hematoma cavity, rarely- in cases with its calcification and also in patients with the real recurrence and insufficiency of the drainage operations.

Great Hospitals of the Russian Federation: National Medical Research Center for Neurosurgery Named After N. N. Burdenko: History and Contemporaneity

In 1929 the surgeon N. N. Burdenko and the neurologist V. V. Kramer founded the first neurosurgical clinic in Moscow, which gained the status of Neurosurgery Institute in 1932. It went through a difficult path of military and peaceful years, overcoming all kinds of obstacles. It was constantly developed, built, and upgraded as it evolved to the National Medical Research Center for Neurosurgery. The history of the center is reviewed in this article, highlighting the notable personalities that have influenced the development of neurologic surgery and its scientific background in the Russian Federation.

New opportunities of magnetic-resonance imaging: an algorithm of CSD-HARDI tractography in reconstruction of the brainstem reticular formation fibers

The study purpose was to develop a technique for intravital visualization of the brainstem reticular formation fibers in healthy volunteers using magnetic resonance imaging (MRI). MATERIAL AND METHODS The study included 21 subjects (13 males and 8 females) aged 21 to 62 years. The study was performed on a magnetic resonance imaging scanner with a magnetic field strength of 3 T in T1, T2, T2-FLAIR, DWI, and SWI modes. A CSD-HARDI algorithm was used to identify thin intersecting fibers of the reticular formatio. RESULTS We developed a technique for reconstructing the reticular formation pathways, tested it in healthy volunteers, and obtained standard quantitative indicators (fractional anisotropy (FA), apparent diffusion coefficient (ACD), fiber length and density, and axial and radial diffusion). We performed a comparative analysis of these indicators in males and females. There was no difference between these groups and between indicators for the right and left brainstem. Our findings will enable comparative analysis of examination results in patients with brain pathology accompanied by brainstem injury, which may help predict the outcome. This work was supported by a grant of the Russian Foundation for Basic Research (#16-04-01472).

The role of lipid metabolism disorders, atypical isoforms of protein kinase C, and mutational status of cytosolic and mitochondrial forms of isocitrate dehydrogenase in carcinogenesis of glial tumors

The relationship between molecular genetic and metabolic disorders is one of the challenges of modern oncology. In this review, we consider lipid metabolism and its changes as one of the factors of oncogenesis of glial tumors. Also, we demonstrate that the genome and the metabolome are interconnected by a large number of links, and the metabolic pathways, during their reorganization, are able to drastically affect the genetic structure of the cell and, in particular, cause its tumor transformation. Our own observations and analysis of the literature data allow us to conclude that mass spectrometry is a highly accurate current method for assessing metabolic disorders at the cellular level. The use of mass spectrometry during surgery allows the neurosurgeon to obtain real-time data on the level of specific molecular markers in the resected tissue, thereby bringing intraoperative navigation techniques to the molecular level. The generation of molecular fingerprints for each tumor significantly complements the available neuroimaging, molecular genetic, and immunohistochemical data.

In vitro terahertz spectroscopy of gelatin-embedded human brain tumors: a pilot study

We have performed the in vitro terahertz (THz) spectroscopy of human brain tumors. In order to fix tissues for the THz measurements, we have applied the gelatin embedding. It allows for preserving tissues from hydration/dehydration and sustaining their THz response similar to that of the freshly-excised tissues for a long time after resection. We have assembled an experimental setup for the reflection-mode measurements of human brain tissues based on the THz pulsed spectrometer. We have used this setup to study in vitro the refractive index and the amplitude absorption coefficient of 2 samples of malignant glioma (grade IV), 1 sample of meningioma (grade I), and samples of intact tissues. We have observed significant differences between the THz responses of normal and pathological tissues of the brain. The results of this paper highlight the potential of the THz technology in the intraoperative neurodiagnosis of tumors relying on the endogenous labels of tumorous tissues.

Long association tracts of the human white matter: an analysis of 18 hemisphere dissections and in vivo HARDI-CSD tractography

BACKGROUND Anatomy of the conduction tracts of the cerebral cortex has been studied for a long time. Invention of diffusion tensor tractography renewed interest in this subject. The objectives of this work were to develop and improve protocols for dissection of the long association tracts of the human brain with studying the features of their segmentation, topography, and variability, compare the obtained data with the MR tractography data, and prepare for further clinical and anatomical studies. MATERIAL AND METHODS We used 18 cerebral hemispheres (from 10 males and 8 females; 9 left and 9 right hemispheres). The mean age of cadavers was 68 years. Specimen were fixated in accordance with the Klingler technique. Immediately after collection, specimens were placed in a 10% formalin solution for at least 4 weeks. After that, the pia was removed; specimens were frozen at -20 °C for a week and then unfrozen in a 96% ethanol solution for a day. We performed 10 lateral dissections, 2 lateral dissections with isolation of the frontal aslant tract, 2 basal dissections, 1 combined basolateral dissection, 2 frontal dissections, and 1 medial dissection. At the time of dissection and after it, specimens were stored in a 96% ethanol solution. Modified, disposable, therapeutic wooden spatulas were used for manipulations. A microscope (magnification of 6-40x) was used in 2 lateral and 2 basal dissections. MR tractography (HARDI-CSD) was carried out in 5 healthy volunteers using a GE Signa HDxt MRI scanner a field strength of 3.0 T. RESULTS We clearly identified the following fascicles: the arcuate fascicle (AF) and superior longitudinal fascicle (SLF) in 6/6 hemispheres on the right and in 5/6 hemispheres on the left, the inferior longitudinal fascicle (ILF) in 3/6 hemispheres on the left and in 4/6 hemispheres on the right, the uncinate fascicle (UF) in 4/4 hemispheres on the left and in 4/4 hemispheres on the right, and the inferior fronto-occipital fascicle (IFOF) in 4/4 hemispheres on the left and in 3/4 hemispheres on the right. Identification was less successful in the case of the frontal aslant tract (FAT) in 1/2 hemispheres on the left and in 0/2 hemispheres on the right. The used technique failed to identify the vertical occipital fascicle (VOF) of Wernicke, a segment of the superior longitudinal fascicle SLF I, and the middle longitudinal fascicle (MdLF). The MR tractography HARDI-CSD data were compared with the dissection data. We described in detail segmentation of the superior longitudinal, arcuate, and inferior fronto-occipital fascicles. Contradictory data were obtained for the superior longitudinal fascicle: a two-segment structure (SLFh and SLFv) was found in most (10/12) specimens, while a three-segment structure was revealed in the other (2/12) specimens (identified SLF II and SLF III). In the arcuate fascicle, the ventral and dorsal segments were successfully identified in 2/12 cases (1 left and 1 right), whereas identification failed in the other cases. During dissection of the inferior fronto-occipital fascicle, we could identify its surface layer in 1 of 8 cases (left) and its deep layer in one more case (left). CONCLUSION Examination of the long association tracts using the Klingler technique has significant limitations in the fiber intersection areas (sagittal striatum). The frontal aslant tract was least studied; we proposed a special anterior dissection technique for its isolation. The superior longitudinal fascicle can have both the two-segment (10/12) and three-segment (2/12) structure. Investigation of the segmental anatomy of the long association tracts will be continued in further dissections. When planning neurosurgical interventions in the projection areas of the long association tracts, both preoperative HARDI-tractography and anatomical dissections ex vivo, based on the proposed protocols, can be recommended for the operating surgeon to master a three-dimensional picture of the tract topography.

Хирургическая анатомия периинсулярных ассоциативных проводящих путей. Часть I. Система верхнего продольного пучка

AIM To study the peri-insular association tract anatomy and define the permissible anatomical boundaries for resection of glial insular tumors with allowance for the surgical anatomy of the peri-insular association tracts. MATERIAL AND METHODS In an anatomic study of the superior longitudinal fascicle system (SLF I, SLF II, SLF III, arcuate fascicle), we used 12 anatomical specimens (6 left and 6 right hemispheres) prepared according to the Klinglers fiber dissection technique. To confirm the dissection data, we used MR tractography (HARDI-CSD-tractography) of the conduction tracts, which was performed in two healthy volunteers. RESULTS Except the SLF I (identified in 7 hemispheres by fiber dissection), all fascicles of the SLF system were found in all investigated hemispheres by both fiber dissection and MR tractography. The transcortical approach to the insula through the frontal and (or) parietal operculum is associated with a significant risk of transverse transection of the SLF III fibers passing in the frontal and parietal opercula. The most optimal area for the transcortical approach to the insula is the anterior third of the superior temporal gyrus that lacks important association tracts and, consequently, a risk of their injury. The superior peri-insular sulcus is an intraoperative landmark for the transsylvian approach, which enables identification of the SLF II and arcuate fascicle in the surgical wound. CONCLUSION Detailed knowledge of the peri-insular association tract anatomy is the prerequisite for neurosurgery in the insular region. Our findings facilitate correct identification of both the site for cerebral operculum dissection upon the transcortical approach and the intraoperative landmarks for locating the association tracts in the surgical wound upon the transsylvian approach to the insula.