[Prognostic model for assessing the human glioma cell malignancy grade based on MDM2, MELK, SOX2, CDK4, DR5 and OCT4 gene expression]. / Prognosticheskaya model' dlya otsenki stepeni zlokachestvennosti kul'tury kletok gliomy cheloveka na osnovanii issledovaniya ekspressii paneli genov MDM2, MELK, SOX2, CDK4, DR5 i OCT4.

Glioma cell cultures are used in basic researches of tumor processes, personalized medicine for selecting treatment regimens depending on individual characteristics of patients and pharmacology for assessing the effectiveness of chemotherapy. Suppression of glioma culture growth without reduction of malignancy grade is common. Drug cancellation may be followed by substitution of precursor cells by more malignant clones. Therefore, analysis of culture cell malignancy grade is important. In the future, intraoperative analysis of glioma cell malignancy grade can be used to select individual therapy. OBJECTIVE: We analyzed the relationship between expression of marker genes TUBB3, CD133, CDK4, CDK6, CIRBP, DR4, DR5, EGFR, FGFR, FSHR, GDNF, GFAP, L1CAM, LEF1, MAP2, MDM2, MELK, NANOG, NOTCH2, OCT4, OLIG2, PDGFRA, PDGFA, PDGFB and SOX2 and glioma cell malignancy grade, as well as created appropriate prognostic model. MATERIAL AND METHODS: We analyzed expression of 25 marker genes in 22 samples of human glioma cultures using quantitative real-time PCR. Statistical analysis was performed using the IBM SPSS Statistics 26.0 software. We used the Kolmogorov-Smirnov and Shapiro-Wilk tests to assess distribution normality. Nonparametric Jonckheere-Terpstra and Spearman tests were applied. RESULTS: We obtained a prognostic model for assessing the grade III and IV glioma cell malignancy based on expression of marker genes MDM2, MELK, SOX2, CDK4, DR5 and OCT4. Predictive accuracy was 83% (Akaike information criterion -55.125).

[Possibilities of differential diagnosis of chronic diseases of the nasal cavity after surgical treatment and topical antibiotic therapy]. / Vozmozhnosti differentsial'noi diagnostiki khronicheskikh zabolevanii polosti nosa posle khirurgicheskogo lecheniya i topicheskoi antibakterial'noi terapii.

OBJECTIVE: The study was performed to assess the endoscopic state of the nasal mucosa after the use of local anti-inflammatory and antibacterial therapy, in particular, Polydexa nasal spray with phenylephrine containing Dexamethasone sodium metasulfobenzoate + Neomycin + Polymyxin B + Phenylephrine, and for the treatment of granulomatosis with polyangiitis. MATERIAL AND METHODS: The study included 940 patients who underwent examination and treatment for chronic rhinosinusitis in the clinic of otorhinolaryngology of I.P. Pavlov SPbSMU surgical treatment of the paranasal sinuses underwent 907 patients. In the postoperative period, the first group (211 patients) underwent toileting of the nasal cavity. The second group (307 patients) received irrigation therapy. The third group (389 patients) received a topical treatment combined of Polydexa with phenylephrine. The dynamics of the condition was assessed on the 1st, 3rd and 7th days of treatment, the evaluation of the effectiveness of the treatment was carried out on the 3rd and 7th days. Differential diagnosis with granulomatosis with polyangiitis was carried out in 33 patients. All patients with granulomatosis with polyangiitis showed signs of chronic rhinosinusitis. Patients were prescribed local anti-inflammatory and antibacterial therapy with Polydexa with phenylephrine for 7 days with endoscopic control of the nasal cavity. CONCLUSION: The use of the combined topical drug Polydexa with phenylephrine in patients with chronic rhinosinusitis and in patients with granulomatosis with polyangiitis has a positive effect, which reduces the clinical manifestations of chronic rhinosinusitis.

A new species of Laelaps Koch, 1836 (Mesostigmata: Laelapidae) parasitic of the sigmodontine rodent Oligoryzomys flavescens Waterhouse, 1837 (Rodentia: Cricetidae): Molecular and morphological characterization.

Laelaps schatzi sp. nov. (Mesostigmata: Laelapidae) is described on the bases of mites collected from the sigmodontine Oligoryzomys flavescens Waterhouse, 1837 (Rodentia: Cricetidae) in Pereyra, Buenos Aires, Argentina, based on morphological, morphometric and molecular data. Two DNA fragments, mtDNA cytochrome oxidase subunit I (COI) and nuclear 18.S-ITS1-5.8S-ITS2 partial gene sequence were deposited in the public Genbank datebase. The DNA extraction was conducted nondestructively using Chelex®100 modified for this study, to prevent vouchers for further microscopically examination. Laelaps schatzi sp. nov. is similar in general appearance to Laelaps paulistanensis Fonseca, 1936 and Laelaps fonsecai Gettinger, 1992, but the new species is unique in presenting a reticulated surface pattern on the genital shield from level of Zv1 up to posterior margin, and in the presence of one or two additional impair seta/e between J4 setae of dorsal shield. Also, the phylogenetic analyses separate the new species from all other species available in the GenBank of Laelaps.

Effect of Silk Fibroin on Neuroregeneration After Traumatic Brain Injury.

Traumatic brain injury is one of the leading causes of disability among the working-age population worldwide. Despite attempts to develop neuroprotective therapeutic approaches, including pharmacological or cellular technologies, significant advances in brain regeneration have not yet been achieved. Development of silk fibroin-based biomaterials represents a new frontier in neuroregenerative therapies after brain injury. In this study, we estimated the short and long-term effects of silk fibroin scaffold transplantation on traumatic brain injury and biocompatibility of this biomaterial within rat neuro-vascular cells. Silk fibroin microparticles were injected into a brain damage area 1 day after the injury. Silk fibroin affords neuroprotection as judged by diminished brain damage and recovery of long-term neurological functions. We did not detect considerable toxicity to neuro-vascular cells cultured on fibroin/fibroin-gelatin microparticles in vitro. Cultivation of primary cell cultures of neurons and astrocytes on silk fibroin matrices demonstrated their higher viability under oxygen-glucose deprivation compared to 2D conditions on plastic plates. Thus, we conclude that scaffolds based on silk fibroin can become the basis for the creation of constructs aimed to treat brain regeneration after injury.

Protection of Neurovascular Unit Cells with Lithium Chloride and Sodium Valproate Prevents Brain Damage in Neonatal Ischemia/Hypoxia.

Here we studied the cytoprotective effect of lithium chloride and sodium valproate in the in vivo model of neonatal cerebral ischemia/hypoxia and analyzed the influence of these substances on the death of the major neurovascular unit components in experimental ischemia in vitro. Lithium chloride and sodium valproate effectively prevented death of neurons, astrocytes, and endothelial cells in the oxygen-glucose deprivation. This treatment protected the brain of newborn rats from ischemia/hypoxia injury. The results suggest that lithium and sodium valproate can be used for the treatment of neurodegenerative pathologies associated with hypoxia and ischemia in newborns.

CHANGES IN THE NUMBER OF NEURONS, ASTROCYTES AND MICROGLIA IN THE BRAIN AFTER ISCHEMIC STROKE ASSESSED BY IMMUNOHISTOCHEMISTRY AND IMMUNOBLOTTING.

It is known that the mechanisms of damage in the brain after stroke are regulated by combination of several types of cells, primarily of neurons, astrocytes, endothelium and microglia. Ischemic exposure disrupts the balance in the cellular composition of the brain; in the lesion, cells die by necrosis while in tissue surrounding ischemic zone the delayed induction of apoptosis occurs, and namely the ratio of death of different cells determines the clinical outcome of the disease. Thus, the assessment of death of various cell types of the neurovascular unit is an important part of fundamental studies of the mechanisms of brain damage and pre-clinical studies of potential neuroprotective drugs. In this line, we have conducted a comparative study of the two most often used methods: immunohistochemical staining of brain sections, allowing to determine the number and localization of specific cells in the tissue among other types of cells, and immunoblotting that detects specific proteins in the tissue homogenate. We have found that, depending on the type of cells, changes in their number and composition after stroke can be diffuse or localized, which imposes restrictions on the use of any method of estimation of the number of cells in brain tissue. In general, the most preferable is the use of immunohistochemistry, however, with certain limitations, immunoblotting can be used in estimating amounts of astroglia and microglia.

Dysfunction of Kidney Endothelium after Ischemia/Reperfusion and Its Prevention by Mitochondria-Targeted Antioxidant.

One of the most important pathological consequences of renal ischemia/reperfusion (I/R) is kidney malfunctioning. I/R leads to oxidative stress, which affects not only nephron cells but also cells of the vascular wall, especially endothelium, resulting in its damage. Assessment of endothelial damage, its role in pathological changes in organ functioning, and approaches to normalization of endothelial and renal functions are vital problems that need to be resolved. The goal of this study was to examine functional and morphological impairments occurring in the endothelium of renal vessels after I/R and to explore the possibility of alleviation of the severity of these changes using mitochondria-targeted antioxidant 10-(6'-plastoquinonyl)decylrhodamine 19 (SkQR1). Here we demonstrate that 40-min ischemia with 10-min reperfusion results in a profound change in the structure of endothelial cells mitochondria, accompanied by vasoconstriction of renal blood vessels, reduced renal blood flow, and increased number of endothelial cells circulating in the blood. Permeability of the kidney vascular wall increased 48 h after I/R. Injection of SkQR1 improves recovery of renal blood flow and reduces vascular resistance of the kidney in the first minutes of reperfusion; it also reduces the severity of renal insufficiency and normalizes permeability of renal endothelium 48 h after I/R. In in vitro experiments, SkQR1 provided protection of endothelial cells from death provoked by oxygen-glucose deprivation. On the other hand, an inhibitor of NO-synthases, L-nitroarginine, abolished the positive effects of SkQR1 on hemodynamics and protection from renal failure. Thus, dysfunction and death of endothelial cells play an important role in the development of reperfusion injury of renal tissues. Our results indicate that the major pathogenic factors in the endothelial damage are oxidative stress and mitochondrial damage within endothelial cells, while mitochondria-targeted antioxidants could be an effective tool for the protection of tissue from negative effects of ischemia.

Intercellular Transfer of Mitochondria.

Recently described phenomenon of intercellular transfer of mitochondria attracts the attention of researchers in both fundamental science and translational medicine. In particular, the transfer of mitochondria results in the initiation of stem cell differentiation, in reprogramming of differentiated cells, and in the recovery of the lost mitochondrial function in recipient cells. However, the mechanisms of mitochondria transfer between cells and conditions inducing this phenomenon are studied insufficiently. It is still questionable whether this phenomenon exists in vivo. Moreover, it is unclear, how the transfer of mitochondria into somatic cells is affected by the ubiquitination system that, for example, is responsible for the elimination of "alien" mitochondria of the spermatozoon in the oocyte during fertilization. Studies on these processes can provide a powerful incentive for development of strategies for treatment of mitochondria-associated pathologies and give rise a new avenue for therapeutic approaches based on "mitochondrial transplantation".

[Experience of application of ultrasound dissection-aspiration in surgical treatment of extracerebral intracranial tumors].

The results of surgical treatment of 33 patients, suffering intracranial extracerebral tumours, were analyzed. Magnet-resonance tomography of the brain, selective angiography with contrasting of external and internal carotid and vertebral arteries were used for neurovisualization, in several patients-bilaterally. In 19 (57.6%) patients ultrasound dissector Sonoca 300 was applied intraoperatively, permitting to remove the tumour totally in 16 (84.2%) of them. In 3 patients progression of neurological deficiency was noted postoperatively. Application of ultrasound dissection-aspiration in surgical treatment of extracerebral intracranial tumours have had permitted to enhance the surgical interventions radicality, to reduce the operative trauma, postoperative complications rate, cost-effectiveness of the patient management in the early postoperative period, and to raise trustworthy the operated patients quality of life.

[Metabolic syndrome and melatonin].

Metabolic syndrome (MS) is characterized by the following symptoms: obesity, AH, dyslipidemia, insulin resistance. Pathophysiologically, MS is underlain by disorders of many biochemical and physiological processes, such as elevated levels of low density lipoproteins, hyperstimulation of pancreatic b-cells, increased insulin secretion, substitution of lipid metabolism for carbohydrate one, overgrowth of adipose tissue, excess production of adiponectin, leptin and other signal molecules and a rise in their local intravascular concentration, weight gain. Endogenous and exogenous melatonin inhibits these pathophysiological mechanisms, normalizes metabolism, equilibrates insulin secretion, prevents pancreatic hyperfunction, phosphorylates insulin receptors, inactivates active oxygen and nitrogen species including those produced in LDLP metabolism. Melatonin has specific MT1 and MT2 receptors localized in all body cells. Due to this, it exerts combined preventive action in patients with MS. Recently, melatonin has been reported to have therapeutic effect in MS; it may be recommended to treat this condition.

Neural progenitor and hemopoietic stem cells inhibit the growth of low-differentiated glioma.

The effects of neural progenitor and hemopoietic stem cells on C6 glioma cells were studied in in vivo and in vitro experiments. Considerable inhibition of proliferation during co-culturing of glioma cells with neural progenitor cells was revealed by quantitative MTT test and bromodeoxyuridine incorporation test. Labeled neural progenitor and hemopoietic stem cells implanted into the focus of experimental cerebral glioma C6 survive in the brain of experimental animals for at least 7 days, migrate with glioma cells, and accumulate in the peritumoral space. Under these conditions, neural progenitor cells differentiate with the formation of long processes. Morphometric analysis of glioma cells showed that implantation of neural progenitor and hemopoietic stem cells is accompanied by considerable inhibition of the growth of experimental glioma C6 in comparison with the control. The mechanisms of tumor-suppressive effects of neural and hemopoietic stem cells require further investigation.

Characterisation of aflatoxin and deoxynivalenol exposure among pregnant Egyptian women.

Mycotoxins such as the aflatoxins and deoxynivalenol (DON) are frequent contaminants of food. Aflatoxin B1 (AFB1) and DON affect the immune system and restrict growth; additionally AFB1 is carcinogenic. To date there are limited descriptive biomarker data concerning maternal exposures during pregnancy, and none on co-exposures to these mycotoxins. This survey was a cross-sectional assessment providing descriptive data on the concentrations of serum aflatoxin-albumin (AF-alb), urinary aflatoxin M1 (AFM1), and urinary DON for 98 pregnant women from Egypt, in relation to diet and socioeconomic status, during the third trimester. AF-alb was detected in 34 of 98 (35%) samples, geometric mean (GM) of positives = 4.9 pg AF-lys mg(-1) albumin (95% confidence interval (CI) = 4.1-5.8 pg mg(-1)), and AFM1 in 44 of 93 (48%) samples, GM of positives = 19.7 pg mg(-1) creatinine (95%CI = 14.8-26.3 pg mg(-1)). AF-alb and AFM1 levels were positively correlated (R = 0.276, p = 0.007). DON was detected in 63 of 93 (68%), GM of positives = 2.8 ng mg(-1) (95%CI = 2.1-3.6 ng mg(-1)). Aflatoxin and DON biomarkers were observed in 41% of the subjects concurrently. The frequency and level of these biomarkers in Egyptian women were modest compared with known high-risk countries. However, this study represents the first biomarker survey to report on the occurrence of DON biomarkers in an African population, in addition to the co-occurrence of these two potent mycotoxins. This combined exposure may be of particular concern during pregnancy given the potential of toxin transfer to the foetus.

[Results of operator's work during space flight (experiment "pilot") under different work-and-rest cycles].

The correlation between operator's job quality and crew work-rest cycle (WRC) aboard the International space station was studied. The experiment involved 10 Russian members of ISS missions 17-24 at the age of 35 to 51 yrs. Mission duration varied from 163 to 200 days, averaging 180 days. Each cosmonaut carried out several "pilot" test sessions. The number of sessions per mission ranged from 4 to 6. The procedure consisted of simulating manual operation of transport vehicle Soyuz on the stages of hang-up, berthing and docking with the ISS. Objective job quality parameters were accuracy of the Soyuz and ISS relative motion control and time for completion which actually characterized work rate. WRC intensity was judged by the data of monitoring at the Moscow Mission Control Center. The results lend support to the dependence of operator's efficiency on WRC. In operators aimed at the highest accuracy this dependence manifested itself in work rate parameters; work accuracy but not rate was more WRC-dependent in operators aimed at doing their job fast. In other words, WRC intensity impacted mostly those job qualities that operator considered to be of secondary importance.

Activation of expression of brain-derived neurotrophic factor at the site of implantation of allogenic and xenogenic neural stem (progenitor) cells in rats with ischemic cortical stroke.

Ischemic stroke was modeled in the sensorimotor zone of the brain cortex in adult rats. Rat embryonic nervous tissue, neural stem cells from human olfactory epithelium, and rat fibroblasts (cell control) were implanted into the peri-infarction area of rats of different groups immediately after stroke modeling. Expression of BDNF mRNA was analyzed 7 days after surgery by real-time PCR. BDNF expression in cell preparation before their implantation was minimum. The expression of BDNF mRNA increased by 5-6 times in the areas of implantation of rat fibroblasts and human olfactory epithelium and by 23 times in the area of implantation of rat embryonic nervous tissue compared to periinfarction areas without cell implantation. These findings confirm the possibility of realization of the therapeutic effects of neural stem cells via expression of trophic factors.

Reaction of oligoglia to spinal cord injury in rats and transplantation of human olfactory ensheathing cells.

In experiments on rats with lateral TVIII hemisection of the spinal cord and transplantation of ensheating olfactory cells, we studied structural changes at the lesion site and adjacent rostral and dorsal regions of the spinal cord. The state of oligodendrocytes and myelin fibers and motor function in experimental animal were analyzed. Open field testing (BBB test) showed that motor functions steadily increased (by 13% on average) within the interval from day 21 to day 53 after transplantation. Histological examination showed that groups of transplanted cells carrying human nuclear marker (HNu(+)cells) were still present at the lesion site 30 days after surgery. Some of these cells migrated in the rostral and caudal directions from the injection site to a distance up to 6 mm. At the initial period after hemisection, the number of oligodendrocytes (O4(+)-cells) in the immediate vicinity to the lesion site decreased 2-fold, but no significant changes in the number of neurons were found in the rostral and dorsal fragments of the spinal cord compared to the corresponding parameter in controls. Sixty days after transplantation, the cross-section area in the rostral part of the spinal cord at a distance of 3 mm from damage site increased by 15.3% compared to the control. The number of O4(+)-cells at the lesion site and in adjacent rostral and caudal parts of the spinal cord by 22.8% surpassed that in the control group. The number of remyelinated axons also increased. These findings suggest that the absence of pronounced structural changes in the rostral and caudal parts of the spinal cord compared to lesion site at early stages after damage and cell transplantation. At the same time, pronounced activation of oligodendrocytes in this region suggests their involvement together with Schwann cells into remyelination of regenerating axons, which can serve as a factor of partial restoration of motor functions after spinal cord injury.

Study of the efficiency of transplantation of human neural stem cells to rats with spinal trauma: the use of functional load tests and BBB test.

Human ensheating neural stem cells of the olfactory epithelium were transplanted to adult male rats immediately after contusion trauma of the spinal cord at T9 level rostrally and caudally to the injury. Voluntary movements (by a 21-point BBB scale), rota-rod performance, and walking along a narrowing beam were monitored weekly over 60 days. In rats receiving cell transplantation, the mean BBB score significantly increased by 11% by the end of the experiment. The mean parameters of load tests also regularly surpassed the corresponding parameters in controls. The efficiency of transplantation (percent of animals with motor function recovery parameters surpassing the corresponding mean values in the control groups) was 62% by the state of voluntary motions, 37% by the rota-rod test, and 32% by the narrowing beam test. Morphometry revealed considerable shrinking of the zone of traumatic damage in the spinal cord and activation of posttraumatic remyelination in animals receiving transplantation of human neural stem cells.

[Influence of the neural stem cell transplantation on the restoration of CNS functions in rats with cortical stroke].

Ischemic stroke was modeled in white pedigreeless rats by the superficial blood vessel devascularization in the sensorimotor cortex. The preparations of neural progenitors--rat embryonic neural stem cells (rENSC) and human olfactory epithelium-derived neural stem cells (hOENSC) and differentiated fibroblasts ("cell control") were transplanted at the perimeter of the devascularized region. These cells marked with vital tracer stayed alive in the brain parenchyma for at least 16 days. The monitoring of contralateral forepaw motor deficit during 8 weeks demonstrated that only rats with rENSC transplantation had the stable and significant improvement of performance in cylinder test and swimming test (forepaw inhibition test) in comparison to "cell controls" and rats without cell transplantation. The maximal difference in the relative values (the efficacy) was 25% to the end of the experiment. There was no difference in the indicators of vibrissae-elicited forelimb placing test between experimental groups. The methodological approach used makes it possible to broaden the study of mechanisms of neural stem cells' therapeutic effect in stroke.

[General condition evaluations by members of ISS missions 1 to 15].

Traditionally crew questioning about general wellbeing is a part of their psychic monitoring. Information provided during compasses at the Earth's request is included in the neuropsychic section of the weekly crew health report prepared by the medical support group at the Moscow Mission Control Center. Purpose of the work was to analyze the structure content of comments on well-being made by the members of ISS-1-15 crews as well as emotionality of these comments excited by heavy work-rest schedule and length of space duties. Most often well-being evaluations were positive varying within the range from non-committal "all right" and reserved "It's OK" to exaggerated "terrific" "great" "fine"and "perfect"which were more common to astronauts than cosmonauts. Cosmonauts' comments had the seal of personality It was recognized that the demanding work/rest schedule had little impact on the comments and that they did not change with time during 24-wk missions.

[Peculiarities of hemostasis, metabolic activity of platelets, and rate of aspirin resistance in patients with chronic heart failure after aorto-coronary bypass surgery].

Aim of the study was to investigate in dynamics peculiarities of hemostasis including platelet aggregation and activity of NADP-dependent dehydrogenases in platelets, as well as prevalence of resistance to aspirin in patients with functional class II-III chronic heart failure (CHF). We determined parameters of vascular thrombocytic and coagulative hemostasis, the state of intracellular metabolism of platelets as assessed by activity of NADP-dependent dehydrogenases in 46 men (age 45 - 72 years) with NYHA class II (n=16) and III (n=30) CHF before and in 12 - 14 days after coronary artery bypass grafting (CABG). After CABG all patients received aspirin (75 - 150 mg/day). Patients with ischemic CHF had moderate activation of intravascular coagulation, endotheliosis, elevation of fibrinogen and plasminogen levels. These processes were significantly augmented after CABG and were most pronounced in class II CHF. At the background of therapy with aspirin in 36 patients (78.3%) ADP and adrenaline induced platelet aggregation was reduced 2 - 3 times. In 10 patients (21.7%) aggregation remained at initial level or even increased what evidenced for resistance to aspirin. Among patients with functional classes II and III CHF portion of resistant to aspirin was 12.5 and 26.6%, respectively. In these patients most striking changes in intracellular metabolism of platelets were revealed. These changes manifested as derangements of energetic and plastic processes in the cell. Thus aspirin resistant patients with CHF comprise a group with risk of development of atherothrombosis of coronary arteries, arteriovenous grafts and arterial conduits.

Immunoliposomal containers as systems of directed transport of minor interfering RNA into Schwann cells.

Immunoliposomal container system for targeted delivery of minor interfering RNA into Schwann cells was developed. Monoclonal antibodies to myelin basic protein served as the vector. Analysis by sandwich ELISA of myelin basic protein showed specific suppression of the target protein gene expression in Schwann cells after incubation with PEG-conjugated immunoliposomes loaded with minor interfering RNA blocking the synthesis of myelin basic protein. Prospects of effective use of this system for targeting of minor interfering RNA for the treatment of diseases of genetic etiology are discussed.