[Enhanced Natural Killers with CISH and B2M Gene Knockouts Reveal Increased Cytotoxicity in Glioblastoma Primary Cultures].

In an experimental study using the CRISPR/Cas9 system, "enhanced" NK cell lines with knockout of CISH, the gene for the CIS protein (a negative regulator of NK cytotoxicity), as well as two lines with a knocked-out ß2-microglobulin gene, which provides membrane exposure of MHC class I, were obtained from two parental lines of human natural killers (YT wild type and YT-VAV1^(+) overexpressing the VAV1 cytotoxicity enhancing protein). The knockout efficiency was determined by real-time PCR as well as by flow cytometry with specific antibodies. The resulting CISH^(-/-) or B2M^(-/-) knockout lines were tested for cytotoxicity in primary monolayer cultures of human glioblastoma multiforme. The cytotoxicity of the lines was assessed using a cell analyzer that records the cell index based on cell impedance. YT-CISH^(-/-) has been shown to be significantly more effective than wild-type YT in eliminating primary glioblastoma cells in an in vitro cell monolayer experiment. The cytotoxicity of the YT-VAV1^(+)-CISH^(-/-) and YT-VAV1^(+)B2M^(-/-) lines against glioblastoma cells was the highest, but overall, it did not significantly differ from the initially increased cytotoxicity of the YT-VAV1^(+) line. The lines of NK-like cells obtained may serve as a prototype for the creation of "enhanced" allogeneic and autologous NK- and CAR-NK cells for the immunotherapy of glioblastoma multiforme.

[Stagewise treatment of tandem lesions of the internal carotid artery in patients with ischaemic stroke]. / Étapnoe lechenie tandemnogo porazheniia vnutrennei sonnoi arterii u patsientov s ishemicheskim insul'tom.

AIM: The study was aimed at assessing the results of a stagewise approach to surgical treatment of patients with ischaemic stroke and tandem lesions of coronary arteries. PATIENTS AND METHODS: We analysed the results of treating a total of 19 patients with ischaemic stroke having endured in 2017-2018 surgical interventions for tandem lesions of the internal carotid artery. All 19 patients with tandem lesions underwent as the first stage of surgical treatment recanalization of the intracranial portion of the internal carotid artery. Nine patients were subjected to thromboextraction followed by carotid stenting (Group One), ten underwent thromboextraction and carotid endarterectomy (Group Two). One patient was subjected to simultaneous endovascular intervention at the intra- and extracranial level, in 18 patients reconstruction of the internal carotid artery at the extracranial level (second stage) was performed within a postponed period (3-5 days). In the postoperative period we assessed neurological deficit and the results of 90-day survival. RESULTS: No deterioration of neurological deficit was observed after the first stage of surgical treatment. There were statistically significant differences in the degree of regression of a stagewise treatment at discharge, with predominance in the second group. The 90-day survival rate had no prevalence in either group, despite 1 lethal outcome in a patient with simultaneous thromboextraction and stenting of the extracranial portion of the internal carotid artery. CONCLUSION: Adequate prevention of haemorrhagic transformation of the damaged zone of cerebral tissue in patients with ischaemic stroke caused by tandem lesions of carotid arteries may be ensured by means of selecting stagewise treatment, i. e., postponed elimination of extracranial stenosis of the internal carotid artery. Given a small sample size, the study requires further examination and assessment of the obtained results.

[The effect of music containing binaural beats on daytime fall-asleep dynamics]. / Vliianie muzyki, soderzhashchei binaural'nye bieniia, na dinamiku dnevnogo zasypaniia.

AIM: To test the hypothesis that listening to the music with the effect of binaural beats of theta and delta range during nap decreases sleep latency defined by 2nd slow wave sleep stage appearance, as well as improves its stability. MATERIAL AND METHODS: The experimental set of 20 min duration was established according to the counterbalanced scheme with 21 subjects. Each subject participated in two attempts: one attempt included sound stimulation (music) and another one was sham (silence). RESULTS AND CONCLUSION: The decrease in sleep latency during stimulation is not confirmed reliably. The increase in sleep stability has been confirmed reliably using nonlinear regression model. The findings can be used in the development of non-pharmacologic ways of sleep treatment.

[Radio Frequency Electromagnetic Field Effect on the State of Na+/Ca2+ Exchange in the Isolated Rat Heart].

It has been shown that a single exposure to 171 MHz electromagnetic field with 180 V/m electric field strength and 0.04 mW/kg specific absorption rate significantly alters the Na+/Ca2+ exchange in the isolated rat heart. It is assumed that enhancement of the Na+/Ca2+ exchange towards removing Ca2+ from the cardiomyocytes electromagnetic field exposure is a result of Ca2+ extraction from the sarcoplasmic reticulum and the increase of its intracellular level.

[The specific features of the anatomical structure of the artery of labyrinth (an anatomical and topographical study)].

The objective of the present work was to study syntopy of the artery of labyrinth using block-preparations of the posterior cranial fossa, variants of its branching-off from the vertebro-basiliar basin (VBB), and peculiar features of its anatomical structure. A total of 12 block-preparations of the posterior cranial fossa were available for the investigation. They were preliminarily stained with red latex and fixed in a three-point system. These procedures were followed by retrosigmoid craniotomy, opening of dura mater in the supero-lateral part of the cerebellomedulllary cistern, traction of the cerebellum, and blunt separation of the basiliar artery (BA). Variants of branching of the antero-inferior cerebellar artery (AICA) and branching of the artery of labyrinth from AICA were studied. It was shown that the artery of labyrinth branches off from the antero-inferior cerebellar artery in 100% of the cases. The latter artery formed a loop in 14% of the cases (3 ears). The average diameter of the labyrinth artery was 0.32 mm and its mean area 0.06 sq.cm. The artery of labyrinth branched off from the posterior para-stem segment of the antero-inferior cerebellar artery in 42.6% of the cases (9 ears), and from the anterior para-stem segment of AICA in 14.2% of the cases (3 ears). Within the conventional «rhombus¼, the artery of labyrinth was straight in 76.2% of the cases (16 ears) and arc-shaped in 23.8% (4 ears).

[Orbitozygomatic approaches to skull base tumors spreading into the orbit, paranasal sinuses, nasal cavity, and pterygopalatine and infratemporal fossae].

The paper analyzes application of orbitozygomatic approaches at the Department of Skull Base and Craniofacial Surgery of the Burdenko Neurosurgical Institute for a 14-year period. During this time, 723 patients were operated on using the orbitozygomatic approach, which has become the workhorse of surgery for skull base tumors spreading into the orbit, paranasal sinuses, and pterygopalatine and infratemporal fossae. The authors describe seven major modifications of the orbitozygomatic approach that they have used in their practice.

[Orbitozygomatic approaches to the skull base].

The paper is written in the lecture format and dedicated to one of the main basal approaches, the orbitozygomatic approach, that has been widely used by neurosurgeons for several decades. The authors describe the historical background of the approach development and the surgical technique features and also analyze the published data about application of the orbitozygomatic approach in surgery for skull base tumors and cerebral aneurysms.

[Long-term results of surgical treatment of the extracranial arteries aneurysm].

Aneurysms of extracranial arteries are sufficiently rare pathology in the extracranial arteries surgery. The authors aimed to research the frequency of occurrence of the extracranial arteries aneurysm, reasons of the onset, the localization, clinical manifestations and surgical treatment strategy. A retrospective study presents 10 cases of the extracranial arteries aneurysm for the last 5 years. A presence of aneurysmatic dilatation of extra-cranial arteries was detected according to the data of ultrasonic duplex scanning (UDS) and patients were directed to the hospital. The UDS was carried out as a routine procedure because of the presence of earlier acute stroke or the transitory ischemic attack. All the patients were carefully examined. A computed tomography and the recurring UDS were performed and the neurologic status of patients was estimated. An indication to surgical treatment was an increase of the internal carotid artery diameter and the common carotid artery more than 50% or equal to it. The presence of the local saccular aneurysm was one of the reasons. Surgical treatment strategy was determined by the anatomic position and possibilities for the reconstruction. The results of given operations showed, that the surgical treatment strategy should be recommended and could be fully justified in this pathology.

[The influence of application of a low-frequency magnetic field on the serum corticosterone level (an experimental study)].

The results of analysis of the literature publications suggest the necessity of experimental studies aimed at investigation of modulating effect of low-frequency magnetic fields on endocrine organs. The present study was carried out using 200 outbred white male rats (body weight 200-220 g). Corticosterone was measured in blood sera following the application of a low-frequency magnetic field (20 and 53 Hz with induction from 0.4 to 6 mT) generated by a Mini-Expert-T apparatus for induction magnetic therapy during 30 minutes. It was shown that the application of the alternating magnetic field to the adrenal region of the rats in the selected frequency and induction ranges caused a significant increase in the serum corticosterone levels. The results of the present study on the hormonal activity of rat adrenals give reason to consider the influence of the alternating magnetic fields as being modulatory. Analysis of the data thus obtained has demonstrated the non-linear dependence of glucocorticoid activity of the rat adrenal glands on the induction strength of the alternating magnetic field.

[Anatomic rationale for a transmaxillary access to the cavernous sinus].

UNLABELLED: W. Couldwell et al. were the first to propose a transmaxillary access to the cavernous sinus in 1997. The authors showed that this approach was low-invasive and cosmetic and it ensured visualization of different nervous formations of the cavernous sinus and the intracavernous segment of the internal carotid artery. This study was undertaken to study microsurgical anatomy, to simulate a transmaxillary access, to demonstrate its expediency, and to assess the use of endoscopic techniques when this access was applied. The study was conducted in 3 steps: 1) a craniometric study on 33 skulls and 25 craniograms to examine the craniological and geometric parameters of the anatomy of the osseous structures included into the transmaxillary access; 2) simulation of the access on the osseous structures of the skull (2 sides); by including anterior and posterior maxillotomy and bone drilling-out around the round foramen; 3) microsurgical preparation--dissection was performed on 3 head samples (5 sides) at the Laboratory of Microneurosurgical Anatomy, Acad. N. N. Burdenko Research Institute of Neurosurgery, Russian Academy of Medical Sciences. Endoscopy was tested when the transmaxillary access was applied. The results were as follows: 1. The depth of the access failed to correlate with the shape of the skull. The operative observation angle averaged 18-23 degrees. 2. Simulation of the transmaxillary access on the dried skull made it possible to visualize the medial portion of the infratemporal fossa, by enlarging the pterygpid-maxillary fissure. The bone drilling-out boundaries for the skull base were defined. 3. Microsurgical dissection after removal of the posterior maxillary sinus wall and opening the pterygopalatine fossa. The topography of the maxillary artery and nerve was studied. After drilling out the bone of the skull base, the lower wall of the cavernous sinus was crescent. The cavernous sinus was opened as far as possible both above the maxillary nerve and between the second and third branches of the trigeminal nerve. CONCLUSIONS: 1. The access is deep and narrow, yet low-traumatic. 2. It may be the access of choice in removing a small pathological focus in the pterygopalatine fossa, round foramen or lower portions of the cavernous fossa. 3. The access may be used to approach the medial portion of the infratemporal fossa. 4. The described stepwise microsurgical anatomy and internal guiding lines in the retromaxilllary space permit one to perform surgical operations with confidence. 5. With this access, there is no guidance over the great vessel (internal carotid artery). 6. The access passes through the vestibule of the mouth; in this connection its application is undesirable at surgery for intradural abnormalities.

[Urgent surgical correction of ventricular septal defects in children of first months of life].

The article presents the results of urgent surgical correction of ventricular septal defects (VSDs) and a range of associated congenital heart diseases (CHDs) in 123 children of the first 6 months of life, hospitalized between 1988 and 2002 in a severe or critical condition. In 102 patients the progression of clinical symptoms was caused by a big single VSD and in 21 patients--by multiple VSDs. 70 patients had significant associated CHDs. Significant extracardiac anomalies were revealed in 75 patients. The condition of 55.3% of children had deteriorated due to pneumonia. Correction of defects with cardiopulmonary bypass was performed after urgent examination and intensive therapy. Hemodynamically significant VSD were closed in 117 (95.2%) patients. Two patients were reoperatied due to the presence of residual VSD. Partial or full simultaneous correction of associated CHDs was performed in 68 patients. 93.5% of patients with isolated VSDs survived the operation. The survival rate of patients with associated CHDs and extracardiac anomalies was 81.5%. Postoperative mortality risk factors were found to be critical condition at hospitalization and preoperative activity of infectious process (P < 0.01 and < 0.05, respectively). Accumulation of experience and introduction of a special approach allowed an 8-time decrease of the hospital mortality, which has not exceeded 3.4% within last 3 years. Good and excellent follow-up results were obtained in 98% of the survived patients within 10 to 67 months after operation. Precise diagnostics of associated CHDs and extracardiac anomalies, intensive preoperative treatment and weighed estimation of indications and risk factors allow significant improvement in the results of primary correction of VSD and accompanying cardiovascular anomalies in children of the first months of life in a heavy and critical condition upon admission.

[Some mechanisms of adenosine protective effect in the "calcium paradox"]. / Nekotorye mekhanizmy zashchitnogo deistviia adenozina pri kal'tsievom paradokse.

A possibility of preventing the "calcium paradox" with the aid of adenosine was studied as well as some mechanisms of adenosine effect upon the heart in case of the "calcium paradox". Adenosine was found to suppress release of amino acids from the heart in perfusion with calcium-free medium, to efficiently prevent disorders in the energy-dependent functions of mitochondrion and myoglobin release from the heart in reperfusion with Ca2+ -containing solution. Adenosine was also found to increase 2-10-fold lactate release from the heart. Adenosine seems to be able to activate glycolysis. Iodine acetate was shown to completely suppress the adenosine ability to decrease amino acid release from the heart perfused with calcium-free medium. Under conditions of iodine acetate blocking of glycolysis was found to possess no protective properties against cytolysis in the "calcium paradox". The heart mitochondria isolated in the end of the experiment revealed low values of free or phosphorylating respiration and complete dissociation of oxidation. Also a protective effect of adenosine in inhibition of Na+, K+ -ATPhase with Strophantinum, was studied.

[Dependence of myocardial contracture on energy resources during the calcium paradox]. / Zavisimost' kontraktury miokarda pri "Kal'tsievom paradokse" ot énergeticheskikh resursov.

Perfusion of the rat isolated hearts with calcium-free and calcium containing solution revealed a complex and deep myocardial damage called the calcium paradox. The reperfusion of the rat heart with calcium rich media resulted in myoglobin loss from the heart, significant decreasing of ATP and phosphocreatine level, complete uncoupling of respiration and phosphorylation in mitochondria, occurrence of myocardial contracture. Decreasing of sodium level to 30 mM--80 mM in calcium free media exacerbates the heart damage due to the calcium paradox with absence of contracture. Addition of phosphocreatine (1 mM, 5 mM, 10 mM) evoked some restoration of ATP contents in the tissue with appearance of significant contracture. Phosphocreatine exacerbated the loss of myoglobin from the heart subjected to the calcium paradox. A discrepancy between myocardial contracture and degree of cellular damage has been observed during the calcium paradox.

[Skull base meningiomas spreading into the infratemporal fossa: clinical picture, diagnosis, and treatment policy]. / Meningiomy osnovaniia cherepa, rasprostraniaiushchiecia v podvisochnuiu iamku: klinika, diagnostika, taktika lecheniia.

From 1997 to 2004, the Academician N. N. Burdenko Research Institute of Neurosurgery has operated on 54 patients with intracranial meningiomas spreading into the infratemporal fossa. Fifteen patients were operated on for the first time. Thirty-nine patients had undergone surgical interventions on the average 3 times (from 2 to 8). All the patients were operated on via different orbitozygomatic approaches depending on the extent of the process. Opening the upper and lower palpebral fissures and the round foramen with resection, if required, the pterygoid processes suffice to remove tumors from the areas of the upper and lower palpebral fissures, which spread into the sphenoid and maxillary sinuses. If there are tumors at the site of the base of the anterior surface of the pyramid, and the articular bursa, it is expedient to open the oval and spinous foramens, to resect the external portions of the fundus of the middle cranial fossa and, if required, the articular process of the lower jaw. By taking into account the X-ray and histological patterns, it may be stated that invasion of meningiomas is not always accompanied by the development of hyperostosis. According to our findings, extracranial growth of meningiomas points to the invasion of osseous structures of the middle cranial fossa. Furthermore, if meningiomas grow into the infratemporal fossa, they frequently involve the muscles, nerves, and mucosa. After removing the tumors spreading to the infratemporal fossa, the optimum plastic repairs of defects of the base of the skull are as follows: hermetic closure of basal defect of the dura mater with a free fat flap, by fixing it with sutures and fibrin-thrombin glue with additional plastic repair of skull base defect with local displaced tissues on a pedicle (with a temporal muscular fascioperiosteal flap, a Bisch fat flap). Further policy of management of these patients is a complicated problem. It depends on the radicalism of an operation and the invasiveness of the process. The histobiological features of infiltrative meningiomas should be studied and this will determine management policy. Conceivably, the use of postoperative radiation therapy will be substantiated in a definite group of patients.

[A broad subfrontal access to the tumors in the base of the skull]. / Rasshirennyi subfrontal'nyi dostup k opukholiam osnovaniia cherepa.

A total of 30 patients, aged 16 to 67, with neoplasms in the brain base and in paranasal sinuses were operated on, during 1997-2002, at Burdenko's Research Institute for Neurosurgery of the Russian Academy of Medical Sciences (RAMS). A majority of patients (21) had meningiomas. Other observations included: fibrous dysplasia of the anterior cranial fossa in 3 patients, chondroma in 2 patients, angiofibroma in 2 patients, pituitary adenoma in 1 patient and adamantinoma in 1 patient. The application of diluted subfrontal approach and of its modifications is indicated for extradural tumors located in the anterior cranial fossa and extending into the latticed main sinus of the nasopharynx. No increasing neurological symptomatology was noticed in any patients. Nasal liquor was not noted either. A seizure of an essential reduction of exophthalmos was registered in cases with crania-orbital neoplasm. It restored postoperatively in all patients with malfunctions of nasal breathing. Two patients died. The death reasons are not directly related with the approach technique. The diluted subfrontal approach is effective in extensive tumors of the anterior cranial fossa involving the latticed and main sinuses. Our experience of using the diluted subfrontal approach showed its efficiency not only in extensive extradural neoplasms but also in large meningiomas of the anterior cranial fossa extending into the latticed sinus, main bone platform, tubercle of sella turcica and into both optic canals. The discussed approach ensures essentially reduced brain traction as compared with the traditional subfrontal approach. The hermetic closure of defects formed in the brain basis is an important surgery stage involving the diluted subfrontal approach. This approach is, under modern conditions, a valuable addition to the methods, which are generally accepted in neurosurgery, and it has undoubtedly an indisputable perspective.

[Dependence of myocardium injury on extracellular K+ concentration during calcium paradox]. / Zavisimost' povrezhdenii serdtsa ot vnekletochnoi kontsentratsii K+ pri "kal'tsievom paradokse".

It is well-known that the first stage of the calcium paradox involves decreasing of Na+ gradient. The decreased sodium gradient is a cause of activation of the Na(+)-Ca+ exchange and formation of cardiac injury during the calcium repletion. Potassium ions are natural extracellular activators of Na(+)-pump. It has been shown that heart perfusion by Ca(2+)-free medium evoked extrusion from cells of hydrophilic amino acids whose transport-depends on sodium gradient. The heart reperdusion with Ca(2+)-containing agent leads to myofibrillar contracture and extensive myoglobin release. The simultaneous events are: elevation in tissue water contents, decreasing of intracellular concentration of adeninnucleotides, uncoupling of oxidation and phosphorylation in mitochondria. The decreasing of K+ level to 0.5 mM exacerbates myocardial damage during the calcium paradox, despite absence of myocardial contracture. The elevation of K+ (to 10 mM or 20 mM) attenuated the calcium paradox development in the heart. The elevated K+ concentration protected isolated heart from extensive myoglobin release, development of myocardial contracture. The high K+ concentrations alleviate mitochondrial damage and elevate contents of adeninnucleotide in the tissue. The positive effect of the elevated K+ concentration can be completely blocked by strophanthine, the selective Na+, K(+)-pumb blocker.

Dobutamine prevents experimental postintoxication liver cirrhosis in mice.

Various chronic inflammatory and necrotic processes in the liver parenchyma are accompanied by pathological morphofunctional changes, which are associated with hepatocyte death and hyperplasia of the connective tissue. Regeneration of the liver parenchyma should include not only prevention of fibrosis, but also stimulation of hepatocyte proliferation. The adrenoceptor agonist dobutamine stimulated proliferative activity of cultured hepatocytes and prevented the development of postintoxication liver cirrhosis in mice produced by chronic poisoning with CCl(4).

[Access to the anterior cranial fossa through the frontal sinus]. / Dostup k opukholiam perednei cherepnoi iamki cherez lobnuiu pazukhu.

In 1998 to 2001, the Academician N. N. Burdenko Research Institute of Neurosurgery operated on 15 patients aged 20 to 65 years who had bulky processes in the base of the skull and paranasal sinuses by applying an access through the frontal sinus. Most patients (n = 7) had meningiomas. The others had osteoma of the base of anterior cranial fossa (n = 2), chondroma (n = 2), angiofibroma (n = 1), fibroma (n = 1), esthesioneuroepithelioma (n = 1), and neurinoma (n = 1). The main criteria for choosing this access were the site and extent of a process, the sizes of the frontal sinus, and no signs of acute or chronic rhinosinusitis. The length of frontal sinuses in the most measurement was 3.5 to 5.0 cm, their width was 7 to 10 cm. All patients showed no progression of neurological symptoms. Three patients had a decrease in visual acuity from 1.0 to 0.8; with vascular therapy, visual disorders regressed during 2 months. Olfaction became worse in 4 cases, there was anosmia in 2 cases. Nasal liquorrhea was not observed. When there were tumors in the anterior cranial fossa, an access through the anterior wall of the frontal sinus permits radical removal of both intra- and extracranial parts of a tumor with closure of the defect with a periostal flap.