The Future of Neurocritical Care Research: Proceedings and Recommendations from the Fifth Neurocritical Care Research Network Conference.

The Fifth Neurocritical Care Research Network (NCRN) Conference held in Boca Raton, Florida, in September of 2018 was devoted to challenging the current status quo and examining the role of the Neurocritical Care Society (NCS) in driving the science and research of neurocritical care. The aim of this in-person meeting was to set the agenda for the NCS's Neurocritical Care Research Central, which is the overall research arm of the society. Prior to the meeting, all 103 participants received educational content (book and seminar) on the 'Blue Ocean Strategy®,' a concept from the business world which aims to identify undiscovered and uncontested market space, and to brainstorm innovative ideas and methods with which to address current challenges in neurocritical care research. Three five-member working groups met at least four times by teleconference prior to the in-person meeting to prepare answers to a set of questions using the Blue Ocean Strategy concept as a platform. At the Fifth NCRN Conference, these groups presented to a five-member jury and all attendees for open discussion. The jury then developed a set of recommendations for NCS to consider in order to move neurocritical care research forward. We have summarized the topics discussed at the conference and put forward recommendations for the future direction of the NCRN and neurocritical care research in general.

Calponin-Like Protein from Mussel Smooth Muscle Is a Competitive Inhibitor of Actomyosin ATPase.

The goal of this work was to elucidate the mechanism of inhibition of the actin-activated ATPase of myosin subfragment-1 (S1) by the calponin-like protein from mussel bivalve muscle. The calponin-like protein (Cap) is a 40-kDa actin-binding protein from the bivalve muscle of the mussel Crenomytilus grayanus. Kinetic parameters Vmax and KATPase of actomyosin ATPase in the absence and the presence of Cap were determined to investigate the mechanism of inhibition. It was found that Cap mainly causes increase in KATPase value and to a lesser extent the decrease in Vmax, which indicates that it is most likely a competitive inhibitor of actomyosin ATPase. Analysis of Vmax and KATPase parameters in the presence of tropomyosin revealed that the latter is a noncompetitive inhibitor of the actomyosin ATPase.

[Ocular surface viral diseases in children]. / Virusnye zabolevaniya glaznoi poverkhnosti u detei.

UNLABELLED: Over the past 15 years the number of children with inflammatory eye diseases has increased by five-six times. Data analysis of Moscow children's health clinics in 2014 showed that for 40,000 outpatients a viral infection was observed in 49,000 cases, whereas some children suffered from the viral infection twice or thrice. 344 children (0.7 percent) had the viral infection accompanied by keratoconjunctivitis. According to 2015 data, viral infection was observed in 37,957 children, including 325 outpatients (0.8 percent) with keratoconjunctivitis. AIM: To analyze clinical features and treatment options of ocular surface viral diseases in children. MATERIAL AND METHODS: We observed 140 children aged 2 to 13 years with ocular surface viral diseases. RESULTS: Despite the presence of corneal disorders, in 95 percent of children changes were reversible - in 1.5 months corneal opacity was not observed. Yet five percent of children, despite the intensive treatment, had bacterial complications, causing decrease in visual acuity. CONCLUSION: In case of viral infections, ophthalmologists, pediatricians and general practitioners should all be aware of ocular manifestations of these diseases. Even if adequate therapy for ocular surface viral disorders is appointed, in five percent of cases complications are possible, causing decline in visual function. Changes in vision can be a result of general disease manifestation, and only timely and proper treatment will help to relieve the symptoms of inflammation and prevent complications.The results of our observations revealed that the addition of Ophtalmoferon medication to the complex therapy of ocular surface diseases in children showed a high therapeutic efficacy and a good safety profile. This medication, in contrast to other antiviral agents, is available in the form of ready-to-use eye drops, significantly enhancing medication compliance in outpatients.

[Modulation of S-1 conformation and inhibition of the skeletal muscle S-1-ATPase by calponin of the mussel].

A novel 40 kDa protein has been detected in native thin filaments from catch muscles of the mussel Crenomytilus grayanus. In this study, using skeletal muscle actin and S-1, we investigated the effects of the mussel 40-kDa actin-binding protein on the acto · S-1 ATPase activity. On increasing the 40-kDa actin-binding protein (CaP-40) concentration, the actin-activated ATPase activity decreased, and was inhibited 80% at a CaP-40 to actin ratio of 0.5. Polarized fluorimetry technique and glycerinated muscle fibers were used to study effects of CaP-40 on the orientation and mobility of fluorescent label 1.5-IAEDANS specifically bound to CyS-707 of myosin subfragment-1 in the absence of nucleotide, and in the presence of MgADP or MgATP. We have concluded that CaP-40 binding to actin affects the strong binding of myosin to actin but has no effect on the weak binding. Thus, the influence of the CaP-40 on the formation of strong actomyosin binding forms A · M and A · M · ADP manifests itself by a decrease in the relative content of myosin cross-bridges strongly bound with actin, which probably results in a decrease in the relative content of "switch on" actin monomers in thin filaments. This suggests that, as calponin CaP-40 selects its target the phase of strong actomyosin binding binding which preceded by a phase generating power stroke.

Effect of ions of potassium and lithium on NO synthase expression in the human adrenal cortex.

The expression of endothelial and inducible NO synthase in the human adrenal glands was studied under a change in the concentration of K(+), which plays a regulatory role in aldosterone secretion. K(+) ions stimulated the expression of both isoforms of NO synthase in the human adrenal cortex. A stimulatory effect of K(+) on NO synthase is probably related to activation of the calmodulin system and potassium-induced translocation of protein kinase C. Lithium produced n inhibitory effect on both isoforms of NO synthase, which suggests that protein kinase C serves a major regulator of expression in the human adrenal glands.

[Meiotic segregation of chromosomes 13 and 14 of heterozygous Robertsonian translocation der(13;14) (q10; q10) carriers].

Meiotic segregation of chromosomes 13 and 14 was assessed by fluorescence in situ hybridization on sperm of five heterozygous carriers of the most frequent Robertsonian translocation der(13;14). Alternate segregation mode was predominant (mean 78.2 +/- 5.7%). The prevalence of balanced sperm varied from 69.4 to 86.5%. Adjacent segregation mode was detected in 18.64 +/- 4.90% of sperm; 3:0 mode was detected in 2.48 +/- 1.20% of sperm. These results are informative for reproductive counseling of Robertsonian translocation der(13;14) carriers, providing information for assessment of probability of receiving normal/balanced embryos in assisted reproduction cycles.

Nuclear export and expression of human T-cell leukemia virus type 1 tax/rex mRNA are RxRE/Rex dependent.

Human T-cell leukemia virus type 1 (HTLV-1) is a complex retrovirus associated with the lymphoproliferative disease adult T-cell leukemia/lymphoma (ATL) and the neurodegenerative disorder tropical spastic paraparesis/HTLV-1-associated myelopathy (TSP/HAM). Replication of HTLV-1 is under the control of two major trans-acting proteins, Tax and Rex. Previous studies suggested that Tax activates transcription from the viral long terminal repeat (LTR) through recruitment of cellular CREB and transcriptional coactivators. Other studies reported that Rex acts posttranscriptionally and allows the cytoplasmic export of unspliced or incompletely spliced viral mRNAs carrying gag/pol and env only. As opposed to HIV's Rev-responsive element (RRE), the Rex-responsive element (RxRE) is present in all viral mRNAs in HTLV-1. However, based on indirect observations, it is believed that nuclear export and expression of the doubly spliced tax/rex RNA are Rex independent. In this study, we demonstrate that Rex does stimulate Tax expression, through nuclear-cytoplasmic export of the tax/rex RNA, even though a Rex-independent basal export mechanism exists. This effect was dependent upon the RxRE element and the RNA-binding activity of Rex. In addition, Rex-mediated export of tax/rex RNA was CRM1 dependent and inhibited by leptomycin B treatment. RNA immunoprecipitation (RNA-IP) experiments confirmed Rex binding to the tax/rex RNA in both transfected cells with HTLV-1 molecular clones and HTLV-1-infected T cells. Since both Rex and p30 interact with the tax/rex RNA and with one another, this may offer a temporal and dynamic regulation of HTLV-1 replication. Our results shed light on HTLV-1 replication and reveal a more complex regulatory network than previously anticipated.

40-kDa protein from thin filaments of the mussel Crenomytilus grayanus changes the conformation of F-actin during the ATPase cycle.

Polarized fluorimetry was used to study in ghost muscle fibers the influence of a 40-kDa protein from the thin filaments of the mussel Crenomytilus grayanus on conformational changes of F-actin modified by the fluorescent probes 1,5-IAEDANS and FITC-phalloidin during myosin subfragment (S1) binding in the absence of nucleotides and in the presence of MgADP or MgATP. The fluorescence probes were rigidly bound with actin, which made the absorption and emission dipoles of the probes sensitive to changes in the orientation and mobility of both actin monomer and its subdomain-1 in thin filaments of the muscle fiber. On modeling different intermediate states of actomyosin, the orientation and mobility of oscillators of the dyes were changed discretely, which suggests multistep changes in the actin conformation during the cycle of ATP hydrolysis. The 40-kDa protein influenced the orientation and mobility of the fluorescent probes markedly, suppressing changes in their orientation and mobility in the absence of nucleotides and in the presence of MgADP, but enhancing these changes in the presence of MgATP. The calponin-like 40-kDa protein is supposed to prevent formation of the strong binding state of actomyosin in the absence of nucleotides and in the presence of MgADP but to activate formation of this state in the presence of MgATP.

[Common and individual in EEG spectral differences while performing verbal and spatial tasks].

It is shown previously that types of carried-out spatial-figurative and verbal-logic tasks can be distinguished by means of the trained automatic classifier, using individual for each examinee characteristics of EEG power spectra. Research with application of a subset of the same tasks, but aimed to identify the EEG spectral power group differences has been carried out with 31 participants. Contrast distribution of statistically significant differences on frequency bands qualitatively coincides with ranging of the bands by estimates of efficiency of recognition with the trained classifier, except for a range theta. Alpha bands are most involved. Results of comparison of the general (group) and individual indicators correspond to ideas of more significant involvement of mechanisms of semantic memory in the solution of verbal tasks, and also point to possible differences in balance of internal and external attention at realization of verbal-logic and spatial-figurative activity.

[Obesity and diabetes mellitus].

New literature data and the results of own researches concerning the role of excessive body weight and the development of type 2 diabetes mellitus in humans are presented in the analytical review. Inaccordance with current insights, obesity and type 2 diabetes are considered diseases of inflammatory nature, characterized by systemic chronic low-grade inflammation, where different kinds of cytokines are cardinally involved. Unfavourable life style, i.e. excessive, high-energy, and irrational nutrition--an excessive consumption of animal fats and foods containing the high amount of glucose and starch with an insufficient use of high fiber vegetables, fish and vitamin D, and also sedentary, inactive life style leads to adipocyte hypertrophy and migration of M1 macrophages into the adipose tissue (AT). As a result, there is a low-grade inflammation accompanied by an increased production of proinflammatory cytokines (IL-1, IL-6, TNF-α, etc.), adipokines (leptin, resistin, visfatin etc.) and chemokines (CCL2, CCL5, CCL26 and CX3C). Under the influence of these cytokines, on the one hand, IR "is emerged", and on the other--there is apoptosis of the ß-cells, that should be followed by the occurrence of clinically diagnosed type 2 diabetes. However, there is also the opposite system in humans, protecting the organism from the development of type 2 diabetes, and including an increase in the formation of M2 macrophages and the increased formation of secretion of antidiabetic cytokines (IL-4, IL-10, IL-13, etc.) and adiponectin.

[Diagnostics of the primary multiple forms of the colorectal cancer].

The issue analyses the diagnostics of the repeated malignant lesions of the colorectal region. The study covers the two decades period (1992-2011 yy). Of the observed patients with primary colorectal tumors, 238 showed the repeated lesions of the region. The article focuses on the on-time diagnostics and differential approach to the repeated malignancies of the colorectal region.

40-kDa actin-binding protein of thin filaments of the mussel Crenomytilus grayanus inhibits the strong bond formation between actin and myosin head during the ATPase cycle.

Mobility and spatial orientation of a novel 40-kDa actin-binding protein from the smooth muscle of the mussel Crenomytilus grayanus was studied by polarized fluorometry. The influence of this protein on orientation and mobility of the myosin heads was investigated during modeling the different stages of the ATPase cycle. The 40-kDa actin-binding protein affected the strong actin-myosin binding. We suggest that the 40-kDa actin-binding protein is involved in regulation of the actin-myosin interaction in the smooth muscle of the mussel.

[Forensic medical assessment of the mechanisms of development of gunshot lesions caused by elastic destructive agents].

This paper reports the results of analysis of the data obtained in experimental studies and practical expert assessments of body injuries inflicted by rubber balls for traumatic weapons. The causes accounting for the polymorphism of such injuries and the mechanisms of their development were elucidated by means of damage simulation taking into consideration the physical and dynamic properties of elastic ball-type destructive agents and the morphological structure of different anatomical regions of the human body. The results of the study may be of interest for differential diagnostics of gunshot lesions caused by elastic destructive agents.

Investigation of nanostructured oxides: synthesis, structure and properties.

The process of forming three-component nanocrystalline fibers and powders of zirconia, yttria and alumina is studied depending on the component ratio and heat treatment temperature. It has been found that in the investigated system at 500-600 degrees C a nanocrystalline triple solid solution is formed, which exists up to 1200 degrees C. Beyond the above temperature, the triple solid solution decomposes into individual components. Specific regularities of changes in the crystalline structure and size of nanograins of oxides of triple solid solutions in the ZrO2(Y2O3)-Al2O3 system are established depending on the composition and thermal action. The structure--crystallite size--physical-chemical property relationship is also considered. The proposed synthesis method enables preparing nanocrystalline fibers and powders with a high degree of dispersion and reactive activity, whose use in composite materials and ceramics improves their service properties.

[Combined treatment of recurrent ovarian cancer using citoreductive surgery and intraoperative hypertermical introabdominal hyper fusion].

This article analyses results of combined treatment of 40 female patients' age 28-73 (average 55) years with recurrent ovarian cancer who were treated in the Lviv state oncologist regional medically-diagnostic center from 2008 to 2010 year, using citoreductial interferes and introoperational pyrexias introabdominal hyper fusion.

[The application of the videographic images as samples for the comparative craniofacial study].

The results of analysis of 181 forensic medical examinations for craniofacial personality identification are presented. Video snapshots of materials used in the practical forensic medical work and specimens prepared for experimental stdies are shown to be of a higher informative value than conventional photographs. The use of life-time video snapshots for the purpose of comparison significantly improves the efficacy of craniofacial identification by providing much wider opportunities for studying the signs of descriptive portraits, comparing skull measurements and life-time video snapshots obtained at different head rotations and camera angles, and making use of specific characteristics of the stomatological status.

[Mitochondrial genome and human mitochondrial diseases].

Today there are described more than 400 point mutations and more than hundred of structural rearrangements of mitochondrial DNA associated with characteristic neuromuscular and other mitochondrial syndromes, from lethal in the neonatal period of life to the disease with late onset. The defects of oxidative phosphorylation are the main reasons of mitochondrial disease development. Phenotypic diversity and phenomenon of heteroplasmy are the hallmark of mitochondrial human diseases. It is necessary to assess the amount of mutant mtDNA accurately, since the level of heteroplasmy largely determines the phenotypic manifestation. In spite of better understanding of the processes of phenotypic expression, currently there are no adequate treatments for mitochondrial diseases.

[The genetic history of long-term Russian resident populations of polar northeastern Siberia based on mitochondrial DNA variability].

The mtDNA variation has been studied in representatives of the Russkoe Ust'e (n = 30), Kolyma (n = 31), and Markovo (n = 26) ethnic subgroups originating from Russian military men, hunters, and fishers who married local Yukaghir women and settled at the Arctic Ocean coast and on the Anadyr' River more than 350 years ago. The mtDNA haplotypes characteristic of indigenous Siberian peoples have been demonstrated to form the basis of the mitochondrial gene pool of long-term Russian resident populations of the region. Only one of 30 identified haplotypes belonging to 11 haplogroups (H2a) is characteristic of European populations. The C and D haplogroups are the most diverse. The analysis has revealed the characteristics of the population structure of the long-term Russian resident populations and allowed them to be interpreted in terms of recent historical and environmental processes.

[Role of natural selection in evolution of mitochondrial haplogroups from Northeastern Eurasia].

The role of natural selection in the evolution of human populations from Northeastern Eurasia was studied. Selection for the regions-specific haplogroup C was demonstrated.

[Expression of early developmental genes in vole Microtus rossiaemeridionalis].

The expression of genes Sox2, Klf4, Myc, Sall4, Gata6, Foxa2, Hnf4a, Cdx2, Esrrb, Hand1 in cultivated cells, embryos and organs of adult voles Microtus rossiaemeridionalis was studied. High resemblance of the expression patterns of these genes in the organs of adult voles, mice and humans was demonstrated. It was established that genes Gata6, Foxa2 and Hnf4a were specifically expressed in vole extraembryonic endoderm cells, while Cdx2 and Handl genes, in trophoblast stem cells. This shows that these genes can be used markers for corresponding vole cell lines. Indirect confirmation pointing to the fact that Oct4 gene is a marker gene for epiblast cells both in the vole and mouse was obtained.