Background. Photodynamic therapy (PDT) is a promising adjuvant method for the treatment of malignant gliomas (MG), including tumors with continued growth and tumor recurrences. For the clinical application of PDT, it is important to substantiate the effectiveness of the cytodestructive effect of the combined use of laser irradiation (LI) and photosensitizer (PS). Objective. To evaluate the cytodestructive effects of photodynamic exposure with the use of PS chlorine E6 on primary MG cell cultures. Methods. Primary cell cultures were obtained from samples of biopsy material from patients (n = 6) with a verified diagnosis: 3 primary tumors (1 case of diffuse astrocytoma, NOS (G3), 1 – glioblastoma (GB), NOS (G4), 1 – gliosarcoma (G4)) and 3 – with continued tumor growth (1 – diffuse astrocytoma, NOS (G3), 1 – oligodendroglioma, NOS (G3) and 1 – GB, NOS (G4). Groups of cell cultures included: 1) control – cultured in a standard nutrient medium and experimental; 2) cultured with the addition of chlorine E6 (2.0 mg/ml); 3) cultivated without the addition of PS and subjected to LI; 4) cultivated with the addition of chlorine E6 and subsequent exposure to LI. After 24 h, morphological and morphometric studies were carried out. Results. The primary MG cultures were characterized by different growth dynamics; mitotic activity of tumor cells varied from the highest rate in the culture of primary GB to lower values – in cultures of recurrent GB and primary astrocytoma and gliosarcoma, and the lowest – in cultures of continued growth of astrocytoma and oligodendroglioma after combined treatment. Direct exposure to chlorine E6 and LI reduced the total number of cells in the culture and their mitotic activity. The greatest cytodestructive effect was achieved with the combined effect of chlorine E6 and LI: the effective dose in the case of primary astrocytoma cells is 10 J/cm2 in pulse mode; for cells of primary GB and gliosarcoma, recurrent astrocytoma and oligodendroglioma, the effective dose is 25 J/cm2 in pulsed mode. In the case of GB cells, continued growth, a dose of 25 J/cm2 is effective for both continuous and pulsed modes of LI. Conclusions. Primary cell cultures of MG obtained directly from tumor tissue are an adequate model for evaluating the effectiveness of the cytodestructive effect of the combined use of LI and PS for PDT.
We report a lethal case of a female patient who has not undergone surgery for her giant invasive infrasellar/endosellar/suprasellar/parasellar/retrosellar pituitary adenoma. The paper presents results of histological examination of optic nerves and considers the mechanisms of vision loss in prolonged chiasmal and optic nerve compression.
У відділенні психоневрології для дітей із перинатальною патологією та орфанними захворюваннями ДУ «Інститут педіатрії акушерства і гінекології імені академіка О.М. Лук’янової НАМН України» створено мультидисциплінарну команду, яка надає допомогу дітям з орфанними захворюваннями з усієї України. Завдяки сучасним методам клінічної, інструментальної та генетичної діагностики вдається встановити остаточний діагноз. Мета - навести огляд літератури та клінічний кейс дитини з патогенними варіантами генів родини ARID, які, зокрема, призводять до розвитку синдрому Коффіна-Сіріса (КСС), розладу аутистичного спектра та інтелектуальної недостатності, а також до розвитку пухлин нервової системи в дітей. Наведено клінічний випадок дитини з рідкісним захворюванням - КСС, що входить до спектра мутації гена ARID. Застосовано загальноклінічний і неврологічний огляд, електроенцефалографічний моніторинг сну, магнітно-резонансну томографію, повноекзомне секвенування, патогістологічне дослідження. Складність діагностики полягала в тому, що в дитини зі шкірними проявами факоматозу діагностовано за допомогою магнітно-резонансної томографії субепендимальну гігантоклітинну астроцитому (СЕГА), проте не виявлено мутацій, характерних для туберозного склерозу (TSC1, TSC2). Патоморфологічно підтверджено СЕГА I ст. за класифікацією Всесвітньої організації охорони здоров’я. Методом повноекзомного секвенування виявлено дві патогенні мутації гена ARID1A у компаунд гетерозиготному стані. Після об’єднання сукупних клінічних та інструментальних ознак встановлено діагноз - «КСС із СЕГА І ст.». Висновки. Розлади, спричинені мутаціями ARID1A та ARID1B, належать до спектра, що включає синдромальні та несиндромальні випадки інтелектуальної недостатності, розладу аутистичного спектра і КСС із різним поєднанням соматичних і неврологічних симптомів або без них, та об’єднуються під загальним терміном «розлади, пов’язані з ARID». Тільки ґрунтовне вивчення анамнезу, клінічний огляд та сучасні методи інструментальної та генетичної діагностики дають змогу встановити діагноз. Дослідження виконано відповідно до принципів Гельсінської декларації. На проведення досліджень отримано інформовану згоду пацієнта та його батьків. Автори заявляють про відсутність конфлікту інтересів.
The Department of Psychoneurology for Children with Perinatal Pathology and Orphan Diseases at the SI “Institute of Pediatrics, Obstetrics and Gynecology named after academician O.M. Lukyanova of the NAMS of Ukraine” has a multidisciplinary team which provides care to children with orphan diseases from all over Ukraine. Thanks to modern methods of clinical, instrumental and genetic diagnostics, it is possible to establish a definitive diagnosis. Aim - to provide a literature review and a clinical case of a child with pathogenic variants of the ARID family genes, which, in particular, lead to the development of Coffin-Circe syndrome (CCS), autism spectrum disorder and intellectual disability, as well as the development of nervous system tumors in children. The article presents a clinical case of a child with a rare disease - CCS, which is part of the ARID gene mutation spectrum. The general clinical and neurological examination, electroencephalographic sleep monitoring, magnetic resonance imaging, whole-exome sequencing, and pathological examination were performed. The difficulty of diagnosis was that a child with skin manifestations of phacomatosis was diagnosed with subependymal giant cell astrocytoma (SGСA) by magnetic resonance imaging, but there were no mutations characteristic of tuberous sclerosis (TSC1, TSC2). Pathologically, the patient was confirmed to have grade I SGСA according to the World Health Organization classification. Two pathogenic mutations of the ARID1A gene in the compound heterozygous state were detected by whole-exome sequencing. After combining the combined clinical and instrumental signs, the diagnosis of CCS with grade I SGСA was established. Conclusions. The disorders caused by ARID1A and ARID1B mutations belong to a spectrum that includes syndromic and non-syndromic cases of intellectual disability, autism spectrum disorder, and CCS with or without a variety of somatic and neurological symptoms, and are grouped under the general term “ARID-related disorders”. Only a thorough medical history, clinical examination, and modern methods of instrumental and genetic diagnostics can make a diagnosis. The research was carried out in accordance with the principles of the Declaration of Helsinki. Informed consent of the women was obtained for the research. No conflict of interests was declared by the authors.
Chordomas are rare tumors of the axial skeleton that arise from the notochord; they are observed usually at the base of the skull and rarer along the spine. Chordomas are resistant to conventional chemotherapy, that is why it is important, to find the new diagnostic and/or prognostic markers. We have shown recently, that genes of the mitochondrial ribosomal protein S18 (MRPS18) family are differentially expressed in gliomas. The MRPS18-2 oncoprotein is expressed at higher levels in cancerous cells compared to their normal counterpart. Aim. We wanted to investigate the peculiarities of the expression pattern of the MRPS18 family genes in chordoma to better understand their role in cancerogenesis, using an immunohistochemical analysis on chordoma clinical samples. Methods. 15 specimens of chordoma of the base of the skull and 5 samples of the spine localization were studied retrospectively. The specific antibodies against MRPS18-1, MRPS18-2, MRPS18-3, and RB was used for an immunohistochemical analysis. All cases were stained in parallel with appropriate negative control. Results. We found that MRPS18 family proteins are differentially expressed in chordoma tissues. MRPS18-1 showed the strongest signal in all tumor samples. The lowest level of the signal intensity was recorded for the MRPS18-3 protein. In several samples the RB signal was detected in cytoplasm of tumor cells. Importantly, in these samples the strong staining for MRPS18-2 was observed as well. The fine mechanism of sequestering of RB in cytoplasm of cancer cells should be further elucidated. Conclusion, These preliminary results should lead to a larger work on a role of RB-MRPS18 family proteins interaction for chordoma development.
Peripheral nerve injury is an extremely important medical and socio-economic problem. It is far from a solution, despite on rapid development of technologies. To study the effect of long-term electrical stimulation of peripheral nerves, we used a domestically produced electrical stimulation system, which is approved for clinical use. The study was performed on 28 rabbits. ?ontrol of regeneration was carried out after 3 month with morphologic techniques. The use of long-term electrostimulation technology leads to an improvement in the results of the recovery of the nerve trunk after an injury, both directly at the site of damage, when stimulation begins in the early period, and indirectly, after the nerve fibers reach the effector muscle.
Malignant gliomas of the brain are a global medical and social problem with a trend toward a steady increase in morbidity and mortality rates. A method that enables the visual identification of tumor tissue and simultaneously selectively destroys it is photodynamic therapy, which involves the introduction of a photosensitizer (PS) followed by its activation at a certain wavelength of light. The selectivity of the accumulation of PS in the tumor tissue of the malignant gliomas is one of the key issues in the problem of increasing the effectiveness of photodynamic therapy. Objective: to compare the effects of photodynamic exposure using PS chlorin E6 on human glioblastoma (GB) cells of the U251 line and non-malignant human embryonic kidney cells of the HEK293 line. Material and methods. Groups of cell cultures were formed depending on the conditions of cultivation and exogenous influence: 1) control - cultivated in a standard nutrient medium (Modified Eagle's Medium (MEM)) with L-glutamine, 1 mmol of sodium pyruvate, 10% fetal bovine serum) and experimental: 2) cultivated under the conditions of adding chlorin E6 (concentrations 1.0 and 2.0 μg/ml); 3) cultivated on a nutrient medium without the addition of PS and exposed to laser irradiation (LI) (λ=660 nm, power in the range 0.4-0.6 W, dose in the range 10-75 J/cm2, continuous or pulse mode); 4) cultured under conditions of chlorin E6 addition and subsequent exposure to LI (power in the range 0.4-0.6 W, dose in the range 10-75 J/cm2, continuous or pulse mode). After exposure to the specified experimental factors, dynamic observation with microphotographic registration was performed for 24 h, followed by microscopic and micrometric studies (number of viable cells, total number of cells, mitotic index (MI,%)). Results. PS chlorin E6 is incorporated into the cytoplasm of cells of U251 and HEK293 cell lines, the intensity of fluorescence is comparable. Upon exposure to chlorin E6 (1.0 and 2.0 μg/ml), cytodestructive and antimitotic effects are increased in a dose-dependent manner in the culture of human GB cells of the U251 line. The cytodestructive effect of chlorin E6 on cell cultures of the HEK293 line is less pronounced, but the antimitotic effect is comparable in both types of cell cultures. Under the influence of LI, cytodestructive and antimitotic effects increase in a dose-dependent manner in the culture of human GB cells of the U251 line. The level of cytodestructive and antimitotic effects is significantly lower in the cultures of non-neoplastic HEK293 cells. The most significant drop in the mitotic activity of GB U251 cells (~100%) was recorded at the lowest LI dose of 25 J/cm2, power of 0.6 W in pulse mode. For HEK293 cells, the most significant decrease in mitotic activity (~80%) was recorded at LI with a power of 0.6 W and dose of 75 J/cm2 in continuous mode. Under the combined effect of chlorin E6 (1 and 2 μg/ml, pre-incubation of 4 h) and LI in different modes, the viability of tumor cells in U251 culture decreases in a dose-dependent manner; the smallest dose of LI to achieve the maximum cytotoxic effect is 25 J/cm2, with a power of 0.6 W in pulse mode when using chlorin E6 at a concentration of 2 μg/ml. The specified characteristics of photodynamic exposure do not cause irreversible effects in HEK293 cultures (reference cells). Conclusions. An effective mode of photodynamic exposure to achieve a cytodestructive and antimitotic effect in the culture of human GB cells of the U251 line, which is relatively safe for non-malignant cells, has been established: the combined application of a laser irradiation dose of 25 J/cm2, with a power of 0.6 W in pulse mode during the preliminary incubation of the cell culture with chlorin E6 at a concentration of 2 μg/ml for 4 h.
Peripheral nerve injury is an extremely important problem during the war in Ukraine. In the overall pattern of injury, 5% of patients have peripheral nerve injuries and 1% have brachial plexus injuries. Under conditions of hostilities, this indicator increases to 70% or more. The victims are mainly young people of working age, which indicates the great medical and socio-economic significance of the problem. Materials and methods. The study was conducted on 28 rabbits Group 1 (n=7): suture of the sciatic nerve and implantation of the non-working antenna of the electrical stimulation device. Group 2 (n=7): sciatic nerve suture + implantation of an electric stimulator antenna in the same route as in group 1 and the beginning of stimulation on the 2nd day. Group 3 (n=7): sciatic nerve suture + implantation of an electric stimulator antenna in the same route as in group 1 and the beginning of stimulation 3 weeks after operation, when the first signs of regeneration occurred. Group 4 (n=7): autograft of the sciatic nerve + implantation of an electric stimulator antenna and the beginning of stimulation at a time point that will coincide with the beginning of signs of reinnervation of the effector muscle. Сontrol of regeneration was carried out in 12 weeks. The axial cylinder (AC) diameter, myelin thickness (MS) and MS/AC ratio were analyzed using electron mycroscopy. Results. It was showed a statistically significant increase of AC and MS indicators in the study group 2 relatively to comparison group 1, respectively, by 1.8 and 1.75 times. The increase of AC and MS in group 2, relatively to the comparison group and a visual decrease at the ultrastructural level of the number of destructively changed myelin sheaths (strengthening of reparative and regenerative processes) were detected. Conclusion. Therefore, long-term invasive electrostimulation of the damaged peripheral nerve has a positive effect on the regeneration of the neuromuscular complex.
Нейрошкірний меланоз - рідкісне захворювання, що належить до групи факоматозів, характеризується наявністю гігантських меланоцитарних невусів на шкірі та меланоцитарною інфільтрацією паренхіми і оболонок мозку, а також високим ризиком неврологічних ускладнень і малігнізації. Мета - проаналізувати відомості сучасної фахової літератури щодо етіології, патогенезу, підходів до діагностики та лікування рідкісного орфанного захворювання - нейрошкірного меланозу (хвороби Рокітанського); описати клінічні випадки цього захворювання. Наведено огляд наукової літератури та власний випадок встановлення діагнозу і лікування двох пацієнтів. У першому випадку описано клінічну історію хлопчика віком 1 рік 4 місяці з нейрошкірним меланозом та аномалією Денді-Вокера, ускладненими частими фармакорезистентними епілептичними нападами. У другому випадку - історію хвороби хлопчика віком 14 років з уперше діагностованим нейрошкірним меланозом, ускладненим прогресуючою обструктивною гідроцефалією та генералізованими тоніко-клонічними нападами. Особливої уваги заслуговують МРТ-ознаки, зокрема, специфічні зміни в Т2-зваженому режимі, що виявляє типові ділянки гіперінтенсивного сигналу в паренхімі мозку та від твердої оболонки (лептомеланоз). Саме дані нейровізуалізації з високою імовірністю дають змогу припустити правильний діагноз, скоротивши час і витрати на додаткові обстеження. Патогенетичне лікування цього захворювання відсутнє. Для симптоматичного лікування обструктивної гідроцефалії застосовують оперативне втручання, а за наявності епілептичних нападів - антиконвульсивну терапію. Висновки. Наведене клінічне спостереження підтверджує складність діагностичного пошуку при орфанних захворюваннях нервової системи. У діагностиці нейрошкірного меланозу ключове місце належить даним об’єктивного огляду та методам нейровізуалізації (КТ і МРТ головного та спинного мозку). Дослідження виконано відповідно до принципів Гельсінської декларації. На проведення досліджень отримано інформовану згоду батьків дітей. Автори заявляють про відсутність конфлікту інтересів.
Objective: to study the effect of photodynamic exposure with the use of chlorine E6 in cell cultures of the standardized human glioblastoma (GB) cell line U251 under different modes of laser irradiation (LI) in vitro. Materials and methods. Groups of cell cultures of the U251 line were formed, depending on conditions of cultivation and exogenous influence: 1) control – cultivated in a standard nutrient medium (MEM with L-glutamine, 1 mml sodium pyruvate, 10% fetal calf serum) and experimental: 2) cultivated under conditions of adding a photosensitizer chlorine E6 (1.0, 2.0 and 3.0 μg/ml); 3) cultured in a nutrient medium without adding chlorine E6 and subjected to LI (intensity in the range 0.4–0.6 W, dose in the range 25–90 J/cm2, continuous or pulse mode); 4) cultivated under the conditions of adding chlorine E6 and subsequent exposure to LI in the specified modes. Intravital dynamic observation with photo-registration (fluorescence and light microscopy, survey staining methods, intravital staining with a vital dye (0.2% trypan blue solution), morphometric studies (mitotic index, numerical density of viable cells) were carried out. Results. Cell cultures of the human GB U251 line are characterized by the formation of peculiar intercellular connections (reticular histoarchitectonics) of tumor cells with high polymorphism and proliferation activity. Chlorine E6 is incorporated into the cytoplasm of U251 cells with preservation of fluorescence intensity for 72 hours (observation period). The fluorescence intensity of chlorine E6, incorporated by non-tumorally transformed cells of the rat fetal brain (E14-16), is much weaker. Under the influence of chlorine E6 (1.0, 2.0 and 3.0 μg/ml), cytodestructive processes in U251 cell culture increase in a dose-dependent manner with a progressive loss of viability and a decrease of mitotic index. After exposure to LI in the studied regimes the viability of U251 cells decreases in a dose-dependent manner already 1 h after exposure, with a further decrease after 24 h (the most significant (~30%) – at doses of LI 75–90 J/cm2 in the pulse mode). Under the combined exposure of chlorine E6 (2.0 μg/ml) and LI, the viability of U251 cells decreases in a dose-dependent manner already 1 hour after exposure (by 4.5–10.0 times), the most significant (~80%) – at doses of LI 75–90 J/cm2 in pulse mode. After 24 h of observation under all modes of combined exposure of chlorine E6 and LI, viable cells in U251 cultures were not detected. Conclusions. Sufficient effectiveness of the cytodestructive effect of chlorine E6 (2.0 μg/ml, preincubation for 6–24 h) and the lowest studied dose of LI (25 J/cm2) in the pulse mode in the cell culture of human GB U251 line was established. The use of vital dye provides an opportunity to record cytotoxic effects in the culture of U251 tumor cells at an early stage (within 1 h after exposure to chlorine E6 and LI).
Neurocutaneous melanosis is a rare disease belonging to the group of phakomatoses, characterized by the presence of giant melanocytic nevi on the skin and melanocytic infiltration of the parenchyma and meninges, as well as a high risk of neurological complications and malignancy. Purpose - to analyze the information of modern specialized literature on the etiology, pathogenesis, approaches to diagnosis and treatment of a rare orphan disease - neurocutaneous melanosis (Rokitansky's disease) and to describe clinical cases of this disease. A review of the scientific literature and a personal case of diagnosis and treatment of two patients are given. In the first case, the clinical history of a boy aged 1 year and 4 months is presented with neurocutaneous melanosis and Dandy-Walker anomaly, complicated by frequent drug-resistant epileptic seizures. In the second case, a description of the medical history of a 14-year-old boy with newly diagnosed neurocutaneous melanosis complicated by progressive obstructive hydrocephalus and generalized tonic-clonic seizures is presented. MRI features deserve special attention, namely specific changes in the T2-weighted mode, which reveals typical areas of hyperintense signal in the brain parenchyma and from the dura mater (leptomelanosis). It is the neuroimaging data with a high probability that gives reason to assume the correct diagnosis, reducing time and costs for additional examinations. There is no pathogenetic treatment for this disease. For the symptomatic treatment of obstructive hydrocephalus, surgical intervention is used, and in the presence of epileptic seizures, anticonvulsant therapy is used. Conclusions. The given clinical observation confirms the complexity of the diagnostic search for orphan diseases of the nervous system. Objective examination data and neuroimaging methods (CT and MRI of the brain and spinal cord) play a key role in the diagnosis of neurocutaneous melanosis. The research was carried out in accordance with the principles of the Helsinki Declaration. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.
Pediatric brain tumors currently show the highest incidence among solid childhood malignancies and, together with leukemia, are the leading cause of death from cancer in childhood. Embryonal brain tumors are the most common and frequent type of childhood brain cancer and are usually characterized by an extremely aggressive course of the disease with the worst outcomes in most cases. There is an urgent need for specific refined molecular diagnostics, which would help to develop personalized treatment. In the present review paper, the latest molecular characteristics of various classified forms of embryonal brain tumors were analyzed in detail. Overexpression of the MYC and MYCN genes is characteristic of many embryonal brain tumors, leading to enhanced cell proliferation and disturbances in the cell cycle. The functioning of the SWI2/SNF2 chromatin remodeling complex are distorted in such malignancies as well. Noteworthy, LIN28 and MYC discussed here are involved in the induction of pluripotency. We have to mention that molecular mechanisms underlying the development of embryonal brain tumors of the central nervous system (CNS) are still not well understood. Thus, it is important to uncover such mechanisms with the aim to provide a better prognosis of the course of disease and to create personalized therapy.
Aim. The objective of this study is to analyze the clinical course of lumbosacral spinal nerve root anomalies / variants Type I according to the A. Neidre and I. MacNab’s classification, along with a retrospective MRI data analysis before surgery, focusing on the unique branching and topography of the spinal nerve roots. Additionally, we aim to address the relevance of correlating preoperative visualized features with the presence of anatomical anomalies attributed to these spinal nerve root variations, and their implications on the clinical presentation. Our study aims to compare our findings with the global diagnostic experience, classification, and specific aspects of performing discectomies based on individual anatomical characteristics of the lumbosacral region. Results. We present data from our own observation, when during surgery, a probable verification of a Type I lumbosacral spinal nerve root anomaly / variation according to the A. Neidre and I. MacNab’s classification was discovered in a patient. The anomaly appeared as a dense mass resembling a nerve root in the structural characteristics but with a significantly larger diameter. It emerged from the dural space at the level of the upper third of the L5 arch and then divided into two parts. A thinner bundle, comparable in diameter to a normal nerve root, exited through the L5-S1 intervertebral foramen, while a considerably thicker bundle followed a path parallel to the axis of the dural sac and exited through the S1-S2 intervertebral foramen. Comparison with global clinical observations revealed that despite the significant clinical and surgical implications, several aspects of this anomaly remain poorly studied and require further exploration and systematization. Conclusions. The lumbosacral nerve root anomalies (LSNRA) demonstrate a variety of structural variations and branching patterns, which are reflected in different classifications. Among the professional community, the classification of LSNRA, developed by A. Neidre and I. MacNab in 1983 and supplemented by S. M. Burke et al. in 2013, is recognized as the most relevant and frequently applied. While changes in vertebral bone structures can be easily diagnosed using modern imaging methods, preoperative diagnosis of LSNRA proves to be challenging, and the informativeness of standard MRI is limited, as confirmed by the clinical case presented. The dissonance between prominent neurological symptoms and relatively minor disc protrusion allows for the potential presence of a root variation/anomaly when planning the lumbar spine discectomy. The most informative diagnostic method for LSNRA is MR neurography. Among the radiological signs of Type I lumbоsacral nerve root anomalies, the following are considered valid: “corner sign”, “fat crescent sign”, and “parallel sign”. The verification of LSNRA is predominantly intraoperatively based on a series of characteristic topographicanatomical features, including atypical locations of the root’s origin from the dural sac, “unusual” nerve root exit angle, even up to a right angle, and technical challenges in root mobilization during routine lumbar discectomy procedures.
Insular gliomas account for 25% of all low-grade and 10% of high-grade gliomas. Neurosurgical treatment of insular gliomas involves achieving the maximum possible volume of tumor removal while ensuring high quality of life. The anatomical proximity of functionally important brain structures and the involvement of important insular arteries and veins limits the possibility of radical removal of tumors. The key to the effectiveness of surgical intervention in insular gliomas is the selection and implementation of adequate surgical access surgical access. The most commonly used approach to insular gliomas is transsylvian-transinsular. The implementation of this approach is largely determined by the individual characteristics of the venous system of the sylvian fissure, since it is characterized by extreme anatomical variability in particular, the type of outflow direction dominance, the number of veins, their size, type of branching, drainage, collateral connections. The review presents data on the informativeness of modern methods of instrumental research in the assessment of the venous system of the sylvian fissure and insula with the aim of planning surgery for insular gliomas. Methods of preserving venous collectors of the sylvian fissure and possible complications associated with the exclusion of draining veins from the circulation are described.
o account morphological characteristicsMaterials and methods. Retrospective analysis of the results of diagnosis and treatment of patients with CSDH treated in the neurotrauma department of the Romodanov Institute of Neurosurgery NAMS of Ukraine and the City Hospital for Emergency and Emergency Medicine of Zaporizhzhya in 2010‒2019. Among the patients were 80 elderly people, 32 senile and 6 long-lived people. Multispiral computed tomography (MSCT) was performed in 86.7 % of patients, magnetic resonance imaging (MRI) in 13.3 %. In 118 cases, surgical intervention was performed (various types and volume), in 67 % ‒ a morphological study of the fragments of the capsule of the CSDH was performed.Results. The indication for surgery in elderly and senile persons with CSDH is the presence of hematomas with a volume of ≥100 cm3, regardless of the severity of the dislocation syndrome. In 95 % of observations, accompanying somatic diseases were detected. Control of treatment effectiveness was carried out with the help of MRI (in 73.5 % of cases) and CT (in 26.5 %). According to the results of histological studies, three variants of the structure of the capsule of CSDH have been established (according to the density of the location of vessels (the level of expression of the vascular endothelial growth factor (VEGF)) and the ratio of mesenchymal cells), which, together with clinical data, makes it possible to determine the genesis of CSDH. Also, several variants of the structure of the capsule of CSDH were identified, depending on the timing of the diagnosis and morphological examination. Analysis of the relative number and ratio of different cell pools (lymphocytes, fibroblasts, fibrocytes), quantification of immunopositive cells with a reaction to VEGF antibodies, as well as determination of the number of newly formed vessels per unit area makes it possible to predict the course and risk of recurrence of CSDH. Conclusions. The clinical and diagnostic characteristic of CSDH, characteristic of the elderly and senile, is the presence of hematomas with a volume of ≥100 cm3 and the discrepancy between the severity of the condition, clinical and neurological manifestations, and the degree of expressiveness of dislocation-compression changes according to MSCT/MRI data. It is effective to remove CSDH through 1 – 2 milling holes. Microcraniotomy is considered appropriate for a hematoma with complex neuroimaging architecture (multiple septa, layers, trabeculae), in some cases of hematoma recurrence. If the inner surface of the capsule is turned to the blood clot and is represented by a layer of spindle-shaped cells, this indicates a non-traumatic origin of the hematoma. A staged complex clinical and diagnostic examination for CSDH has both clinical and purely practical significance (especially expert), justifying the costs of treatment and insurance payments, ensures the social adaptation of the patient and brings the solution of these issues closer to international standards. Indications, criteria for choosing tactics and the sequence of performing procedures taking into account risks, prognostic clinical-radiological and structural signs are based mainly on recommendations and consensuses and require further detailed study.
The use of direct electrical stimulation to produce shortening of the denervated circular oculus in experimental facial nerve palsy (FNP) allows for complete closure of the ocular slit. The aim of this study is to study morphological changes in the nerves under the influence of direct electrical stimulation with biphasic pulses over a long period of time. Materials and methods: A model of a single facial nerve injury with consequent paralysis of orbicularis oculi muscle was created in the experiment on 12 adult animals. The animals were divided into 2 groups: main and control. In the main group, the stimulating part of the bioelectric device for eye closure was implanted, after which the ability to work in real-time mode to perform long-duration stimulation of orbicularis oculi muscle was tested. The control group did not receive any effect effects on orbicularis oculi muscle. After 30 days, all the animals underwent orbicularis oculi muscle elimination on the side of the facial nerve palsy with the following morphological examination. Results: An intergroup comparative analysis using the nonparametric U-Mann-Whitney criterion for changes in the Ki-67 proliferation index revealed a statistically significant difference between the comparison group and the experimental group, that was affected by the electrical stimulation with the increase of the parameter for the group with electrical stimulation according to the control group from 28,3 (25,0; 30.8) to 30.9 (28.8; 33.3), or 1.1 times (p=0.04); Analysis of variation in mitochondrial photocurrent density revealed statistically significant differences between the control group and the experimental group, with the increase of this parameter for the group with electrical stimulation compared with the control group from 10.5 (6.3; 14.4) to 15.8 (12.8 17,6), i.e. by 1.5 times (p=0.0004); a statistically significant increase of the parameter for the group with electrical stimulation versus the control group from 48,9 (40,6; 63.0) to 64.4 (60.0; 72.4), i.e. by a factor of 1.3 (p = 0.00005). Conclusions: The assessment of the effects of electrostimulation on the musculoskeletal apparatus after neuroraphy (in the experiment) revealed structural and ultrastructural changes, indicating the increased morphofunctional activity of renewed regeneration of muscle fibers in the study under the influence of direct electrical stimulation. In addition, no specific pathological changes were detected under the influence of direct electrostimulation for 30 days.
Purpose: to evaluate changes in the structure of synaptic contacts in various types of focal brain pathology. Materials and methods. The results of treatment of 40 cases of supratentorial focal lesions of the brain (FLB) were retrospectively evaluated. The cases are divided into groups: 30 gliomas of various degrees of malignancy and 5 consequences of TBI, 5 epilepsy. All patients underwent surgical interventions. The synaptic plasticity of axo-dendritic and axo-spiny asymmetric synapses of neurons of the VI-VII layers of the frontotemporal cortex was studied by electron microscopy. Morphometric analysis was carried out on a computer image analyzer САИ-01АВН using the software "Kappa opto-electronics GmbH" using the STATISTICA 7 program package. The results. It was established that the density of synapses decreased in glioblastomas (GB) and craniocerebral injury (ССІ). Qualitative changes demonstrate the plasticity of architectonics of synapse, in particular due to the increase in the number of perforated synaptic contacts. Maximum thickening and diffuse stratification of the postsynaptic seal indicates a violation of the functional capacity of the postsynaptic component of the contacts. A decrease in the number of synaptic vesicles was revealed in ССІ and GB, with their rearrangement, which is probably a manifestation of synaptic dysfunction. The latter proves the irreversibility of destructive local changes and is unfavorable criterion. The risk of the formation of destructive-degenerative changes in the synaptic apparatus is 7.64 times higher in DA, 3.17 times higher in GB, and 17.31 times higher in ССІ compared to cases of epilepsy, with GB significantly increases by 13.5 times compared to DA. Therefore, the assessment of the structural features of neuroplasticity should take into account the morphogenesis of the BM in comparison with clinical data Conclusions. In the zones of invasive growth of gliomas of various degrees of malignancy and in ССІ and epilepsy, the indicators of synaptic plasticity differ statistically significantly. The density of placement of synapses is lower in GB and ССІ. The probability of non-reversibility of destructive-degenerative changes of synapses according to the number of SVs in FLB correlates with the degree of glioma differentiation with a sensitivity of 81.0% and a specificity of 76.0%. According to the structural changes of synaptic connections in tumors, probable differences between the variants have been proven: GB and DA, the sensitivity of the discriminant model is 85.0%, the specificity is 74.0%, which is an indirect evidence of the growth rate of the tumor mass and its destructive effect on the surrounding brain matter. The obtained results are important in assessing the prognosis of the further course of the disease.
Вступ. При плануванні хірургічного видалення церебральних судинних мальформацій (ЦСМ) функціонально важливих зон головного мозку (ФB3-ГМ) актуальним, на наш погляд, є запитання, чи входить до її складу функціонально спроможна тканина мозку, від чого залежать клінічні результати і прогноз безпеки тотальної резекції ЦСМ. Питання безпеки хірургічного видалення має першочергове значення. Згідно даних літератури, поміж судинами артеріовенозних мальформацій (ABМ) і кавернозних мальформацій (KМ) паренхіми головного мозку немає. Мета. Оцінити значення МР-трактографії при вирішенні тактики хірургічного лікування ЦСМ шляхом співставлення із даними морфологічних досліджень. Методи дослідження. Проведено огляд літератури і сучасних рекомендацій щодо хірургічного лікування КМ і АВМ, локалізованих у ФB3-ГМ та стовбурі головного мозку. Четверо пацієнтів із ЦСМ ФB3-ГМ вибрані нами ретроспективно за наявністю даних МР-трактографії та гістологічних препаратів, отриманих при хірургічному видаленні (2 KМ i 2 ABМ). Методом світлової мікроскопії проведено дослідження мікропрепаратів, забарвлених за класичною (гематоксилін-еозином) та спеціальними методиками (за Масоном та Хартом). Результати. Дані МР-трактографії у наших пацієнтів показали наявність провідних шляхів в складі тканин KМ і ABM та зниження фракційної анізотропії в зонах зацікавлення, що свідчить про їх втягнення у патологічний процес. При світловій мікроскопії препаратів ЦСМ нами виявлено тканину мозку поміж патологічними судинами. Це пояснює, на наш погляд, візуалізацію провідних шляхів в складі ЦСМ при МР-трактографії. Висновки. При плануванні операцій з приводу ЦСМ необхідно звертати увагу на результати візуалізації провідних шляхів. Згідно MP-трактографії та морфологічних даних, в деяких випадках у структурі КМ або АВМ присутня паренхіма мозку, що може потребувати додаткової уваги при прийнятті рішення щодо радикальності резекції ЦСМ ФB3-ГМ.
Medulloblastoma is one of the most prevalent brain tumors in children. Due to alterations in the gene expression patterns, medulloblastomas display diversity in the transcriptional, genetic, and clinical markers. However, these markers are few. Hence, there is an urgent need for other molecular, preferentially, non-invasive markers to propose the personalized treatment. One of the putative markers can be the mitochondrial ribosomal protein MRPS18-2. Purpose: In the present work we aimed to study expression of the MRPS18 family genes at mRNA and protein levels, in serum and tissue of the medulloblastoma. Materials and methods. To do so, a real-time quantitative polymerase chain reaction (qPCR) was used to assess the relative expression of RB and MRPS18 family genes at mRNA levels in patient sera and tissue. Protein signals were detected by immunohistochemistry. The relative expression of MRPS18 genes was lower, when assessed in serum of the tumor patient compared with the control. Results. Thus, MRPS18-1 expression level, detected in serum, is up to 8.5-folds lower than in the control sample, while in tissue it is quite similar in both samples. The MRPS18-2 gene was detected at up to 26-fold lower levels in the serum of the tumor patient. Importantly, MRPS18-2 and MRPS18-3 are elevated by 13- and 7.2-fold, respectively, in tumor tissue, compared to the control. Moreover, the MRPS18-2 protein signal is dramatically elevated in medulloblastoma cells, compared with the conditionally healthy brain tissue. Concluding, the members of the MRPS18 protein family, especially MRPS18-2, are the putative candidates for molecular prognostic markers. More experiments should be done on a study on this family, and on a large cohort.
Treatment of malignant gliomas of the brain remains a serious problem on a global scale, despite intensive research into the causes and mechanisms of their progression. When using traditional surgical approaches and imaging methods tumor cell infiltrates may be overlooked, as a result of which, malignant gliomas relapse often occurs near the marginal region of the surgical cavity. A method that allows visual identification of tumor tissue and at the same time provides an opportunity to selectively destroy it is photodynamic therapy (PDT) ‒ a two-stage treatment that includes the introduction (intravenous, intraperitoneal, local or oral) of a light-sensitive chemical agent (photosensitizer (PS)) followed by its activation at a certain wavelength of light.The principle of PDT is based on the cytotoxic effects caused by PS, which selectively accumulates in malignant tumor cells and is activated by light rays of the appropriate wavelength, generating singlet oxygen and free radicals, which trigger photochemical reactions in tumor cells with subsequent destruction of protein structures. Tumor tissue has a higher affinity for PSs. PSs are divided into 1st, 2nd and 3rd generation molecules. So far, 3rd generation PSs have not yet been approved for clinical use. In vitro and in vivo experimental studies confirmed the effectiveness of PDT of brain tumors using 2nd generation PSs.The simultaneous use of surgery under the control of fluorescence and PDT enables both the visualization of tumor cells and their selective destruction. Regardless of PDT, PSs are used for the purpose of auxiliary delineation of tumor borders for maximum tumor removal during fluorescence-guided surgery.The review examines the development of PDT in a historical aspect, the contribution of domestic scientists, in particular, scientists of the Institute of Neurosurgery named after acad. A. P. Romodanov, National Academy of Medical Sciences of Ukraine to the development of the problem of PDT in neuro-oncology; preclinical studies of PDT and experimental approaches to increase the efficiency of PDT are characterized. Analysis of data from clinical trials confirms that using PDT as an adjunctive treatment of malignant gliomas administered immediately after maximal resection is safe, reduces the risk of recurrence by targeting residual tumor cells in the resection cavity, improves survival and quality of life of patients. The absence of information on the development of resistance to multiple PDT sessions suggests the possibility of repeated treatments of tumor cells not removed during surgery.