[Possibilities of an experimental damaging effect on the retinal pigment epithelium]. / Vozmozhnosti eksperimental'nogo modelirovaniya povrezhdayushchego vozdeistviya na pigmentnyi epitelii setchatki.

PURPOSE: To simulate the damaging effect on retinal pigment epithelium (RPE) in an experiment studying the effect of human neuronal precursors (NPs). MATERIAL AND METHODS: The study was carried out on 31 rabbits (31 eyes) of the Chinchilla breed, which were divided into 3 groups: the 1st group received a subretinal injection of balanced saline solution (BSS); the 2nd group - subretinal injection of BSS with vitrectomy, displacement of the injection bladder away from the injection site using a perfluororganic compound (PFOC) and laser coagulation; the 3rd group - subretinal injection of a culture of NPs using the same method as in the group 2. All rabbits were observed for 21 days using ophthalmoscopy, optical coherence tomography (OCT) and autofluorescence (AF). RESULTS: In the 1st group, 4 out of 5 rabbits were observed to have total retinal detachment and vitreoretinal proliferative processes in the early postoperative period after subretinal injection of the BSS. In the 2nd group, OCT and AF revealed atrophy of the outer and inner layers of the retina as well as disorganization of the photoreceptors-RPE-Bruch's membrane complex in the area of injection on the 21 day after the operation. In the 3rd group, the OCT data obtained during the 21 days of observation showed that a hyperreflective zone at the level of the RPE-Bruch's membrane complex corresponding to the NPs injection site was preserved, while there was a partial loss of the outer retinal layers - but of a smaller volume compared to the BSS injection. The suggested method of subretinal injection led to a reduced number of complications: in the 1st group, postoperative complications amounted to 80%, while in the 2nd and 3rd groups - 45%. CONCLUSION: The study proposes a new method for retinal injection of BSS, which can help reduce RPE degeneration patterns and possible postoperative complications, thus increasing research efficiency. Subretinal injection of a culture of neuronal precursors derived from human induced pluripotent stem cells (iPSCs) in an experiment can serve as a universal model for studying the survival and integration of stem cells.

[Morphological and functional indicators of retinal pigment epithelium and photoreceptor apparatus in inherited retinal diseases]. / Morfofunktsional'nye pokazateli retinal'nogo pigmentnogo epiteliya i fotoretseptornogo apparata pri nasledstvennykh zabolevaniyakh setchatki.

PURPOSE: To evaluate the relationship between the morphological and functional parameters of retinal pigment epithelium (RPE) and photoreceptors (PR) in inherited retinal diseases (IRD). MATERIAL AND METHODS: The study included 52 patients (104 eyes), 23 of them with Stargardt Disease (STGD), 19 with cone-rod dystrophy (CRD), 10 with retinitis pigmentosa/pigmentary abiotrophy (RP) of comparable disease durations. All patients underwent standard and additional ophthalmological examination: fundus autofluorescence (AF), spectral optical coherence tomography (OCT), computer perimetry (CP), electro-oculography (EOG), Ganzfeld electroretinography (gERG). RESULTS: Comparison of the groups of IRD patients and groups according to the degree of RPE damage with the control group revealed an increase in differences in the EOG and gERG indicators as the area and depth of damage to the RPE and PR progressed. The patterns of changes in RPE and PR, the frequency of their occurrence with IRD in this patient sample are described. A moderate correlation was found between the amount of RPE loss and EOG light rise, as well as between the defect of the ellipsoid zone and the amplitude of α- and ß-waves, the latency of ß-wave of the gERG. Some patients showed a mismatch between a small defect of the ellipsoid zone and RPE with significant damage to the visual field and reduction of the EOG and gERG indicators. The obtained electrophysiological indicators revealed pathological changes in RPE and PR, more significant and widespread in some cases than it was shown with visualization methods. Weak and moderate correlations between visual acuity, and RPE damage and light sensitivity index with loss of ellipsoid zone were calculated. CONCLUSIONS: Modern methods of retinal examination can help obtain complete and versatile picture of morphological and functional state of the retina in IDR that supplement each other. EOG and gERG have capability to determine the degree of RPE and PR functions impairment including those cases when morphological studies are not sufficiently informative.

[Possibilities of treating retinal diseases in patients with damaged retinal pigment epithelium]. / Vozmozhnosti lecheniia zabolevanii setchatki, soprovozhdaiushchikhsia povrezhdeniem retinal'nogo pigmentnogo épiteliia.

Retinal diseases associated with damage to retinal pigment epithelium (PPE) are the most frequent causes of irreversible loss of vision in adults. Since there is no therapeutic treatment available that could repair RPE and its connections with the adjacent photoreceptors, the review focuses on various methods of surgical treatment. One of the most promising methods at present is the use of stem cells derivatives. Results of numerous experimental and clinical trials show that use of human induced pluripotent stem cells in the treatment of degenerative diseases of the retina can be considered effective and promising.

Optimisation of environmental remediation: how to select and use the reference levels.

A number of past industrial activities and accidents have resulted in the radioactive contamination of large areas at many sites around the world, giving rise to a need for remediation. According to the International Commission on Radiological Protection (ICRP) and International Atomic Energy Agency (IAEA), such situations should be managed as existing exposure situations (ExESs). Control of exposure to the public in ExESs is based on the application of appropriate reference levels (RLs) for residual doses. The implementation of this potentially fruitful concept for the optimisation of remediation in various regions is hampered by a lack of practical experience and relevant guidance. This paper suggests a generic methodology for the selection of numeric values of relevant RLs both in terms of residual annual effective dose and derived RLs (DRLs) based on an appropriate dose assessment. The value for an RL should be selected in the range of the annual residual effective dose of 1-20 mSv, depending on the prevailing circumstances for the exposure under consideration. Within this range, RL values should be chosen by the following assessment steps: (a) assessment of the projected dose, i.e. the dose to a representative person without remedial actions by means of a realistic model as opposed to a conservative model; (b) modelling of the residual dose to a representative person following application of feasible remedial actions; and (c) selection of an RL value between the projected and residual doses, taking account of the prevailing social and economic conditions. This paper also contains some recommendations for practical implementation of the selected RLs for the optimisation of public protection. The suggested methodology used for the selection of RLs (in terms of dose) and the calculation of DRLs (in terms of activity concentration in food, ambient dose rate, etc) has been illustrated by a retrospective analysis of post-Chernobyl monitoring and modelling data from the Bryansk region, Russia, 2001. From this example, it follows that analysis of real data leads to the selection of an RL from a relatively narrow annual dose range (in this case, about 2-3 mSv), from which relevant DRLs can be calculated and directly used for optimisation of the remediation programme.

[Rheumatoid atlanto-axial dislocation: a surgical approach]. / Atlantoaksial'nye dislokatsii revmatoidnogo geneza: khirurgicheskaia taktika.

Pathological processes in the craniovertebral region (CVR) are usually accompanied by dislocation complications leading to gross neurological disorders. One of the diseases that affect the CVR and lead to atlanto-axial dislocation (AAD) is rheumatoid arthritis. Errors in the diagnosis of rheumatoid disease and in the choice of a treatment approach may cause adverse outcomes. OBJECTIVE: To define the approach for surgical treatment of AAD associated with rheumatoid disease of the CVR, with allowance for the rigidity of deformity. MATERIAL AND METHODS: Five patients with rheumatoid AAD, 4 females and 1 male, aged 54 to 73 years underwent surgery. All dislocations were anterior ones. Three patients had mobile pannus-associated dislocation. In 2 cases, AAD was rigid and combined with odontoid invagination into the foramen magnum (FM). RESULTS: In all mobile AAD cases, decompression of the brainstem and restoration of the normal anatomical relationships in the CVR were achieved by dislocation correction and atlanto-axial fusion performed from the posterior approach, avoiding transoral interventions. In this case, spontaneous resorption of the pannus occurred within several months after surgery. In the postoperative period, all patients achieved significant regression of pain and neurological disorders. Complications in the form of wound infection developed in 1 case. CONCLUSION: A decision algorithm for choosing a surgical treatment option was based on the degree of deformity stability. Mobile AADs serve as an indication for indirect decompression using instrumental correction of dislocation and atlantoaxial fixation from the posterior approach. In the case of fixed AAD, posterior fixation is complemented by anterior decompression via the transoral approach.

[Assessment of Cyto- and Genotoxicity of Underground Waters from the Far Eastern Center on Radioactive Waste Treatment Site].

This study has been completed in the frames of activities on the environment assessment in the vicinity of the Far Eastern center (FEC) on radioactive waste treatment (a branch of Fokino, Sysoev Bay). Underground waters collected at the FEC technical site were surveyed both with instrumental techniques and bioassays. Concentrations of some chemicals (ranged to the third hazard category) in the samples collected are over the permitted limits. Activities of 137Cs and 90Sr in waters amount up to 3.8 and 16.2 Bq/l, correspondingly. The integral pollution index is over 1 in all the samples and could amount up to 165. The Allium-test application allows the detection of the sample points where underground waters have an enhanced mutagenic potential. Dependencies between biological effects and pollution levels are analyzed. The findings obtained could be used for the monitoring optimized and decision making on rehabilitation measures to decrease negative influence of the enterprise on the environment.

Radiation safety during remediation of the SevRAO facilities: 10 years of regulatory experience.

In compliance with the fundamentals of the government's policy in the field of nuclear and radiation safety approved by the President of the Russian Federation, Russia has developed a national program for decommissioning of its nuclear legacy. Under this program, the State Atomic Energy Corporation 'Rosatom' is carrying out remediation of a Site for Temporary Storage of spent nuclear fuel (SNF) and radioactive waste (RW) at Andreeva Bay located in Northwest Russia. The short term plan includes implementation of the most critical stage of remediation, which involves the recovery of SNF from what have historically been poorly maintained storage facilities. SNF and RW are stored in non-standard conditions in tanks designed in some cases for other purposes. It is planned to transport recovered SNF to PA 'Mayak' in the southern Urals. This article analyses the current state of the radiation safety supervision of workers and the public in terms of the regulatory preparedness to implement effective supervision of radiation safety during radiation-hazardous operations. It presents the results of long-term radiation monitoring, which serve as informative indicators of the effectiveness of the site remediation and describes the evolving radiation situation. The state of radiation protection and health care service support for emergency preparedness is characterized by the need to further study the issues of the regulator-operator interactions to prevent and mitigate consequences of a radiological accident at the facility. Having in mind the continuing intensification of practical management activities related to SNF and RW in the whole of northwest Russia, it is reasonable to coordinate the activities of the supervision bodies within a strategic master plan. Arrangements for this master plan are discussed, including a proposed programme of actions to enhance the regulatory supervision in order to support accelerated mitigation of threats related to the nuclear legacy in the area.

[Genetic Cell Reprogramming: A New Technology for Basic Research and Applied Usage].

Gene function disclosure and the development of modern technologies of genetic manipulations offered the possibility of genetic reprogramming application to alter cell specialization. With the involvement of a gene set that encodes the transcription factors responsible for the pluripotent state, any cell of an adult body could be reprogrammed into the embryonal.state and pluripotency could be induced in this cell. Such reprogrammed cells were called induced pluripotent stem cells (iPSCs), and they are capable of again passing through all developmental stages. This provides new possibilities for studies of the basic mechanisms of developmental biology, the formation of specific cell types, and the whole body. In culture, iPSCs could be maintained permanently in a nontransformed state and permit genetic manipulations while maintaining their pluripotent properties. Such a unique combination of their properties makes them an attractive tool for studies of various pathologies and for the delineation of treatment approaches. This review discusses the basic and applied aspects of iPSCs biology.

[RADIATION SAFETY DURING REMEDIATION OF THE "SEVRAO" FACILITIES].

Within a framework of national program on elimination of nuclear legacy, State Corporation "Rosatom" is working on rehabilitation at the temporary waste storage facility at Andreeva Bay (Northwest Center for radioactive waste "SEVRAO"--the branch of "RosRAO"), located in the North-West of Russia. In the article there is presented an analysis of the current state of supervision for radiation safety of personnel and population in the context of readiness of the regulator to the implementation of an effective oversight of radiation safety in the process of radiation-hazardous work. Presented in the article results of radiation-hygienic monitoring are an informative indicator of the effectiveness of realized rehabilitation measures and characterize the radiation environment in the surveillance zone as a normal, without the tendency to its deterioration.

[RADIATION HYGIENIC MONITORING AT THE AREA OF THE LOCATION OF THE FAR EASTERN CENTER FOR RADIOACTIVE WASTE MANAGEMENT (FEC "DALRAO"--BRANCH OF FSUE "ROSRAO")].

Intensification ofactivities in the field of spent nuclear fuel (SNF) and radioactive waste (RW) management in the Far East region of Russia assumes an increase of the environmental load on the territories adjacent to the enterprise and settlements. To ensure radiation safety during works on SNF and radioactive waste management in the standard mode of operation and during the rehabilitation works in the contaminated territories, there is need for the optimization of the existing system of radiation-hygienic monitoring, aimed at the implementation of complex dynamic observation of parameters of radiation-hygienic situation and radiation amount of the population living in the vicinity of the Far Eastern Center for Radioactive Waste Management (FEC "DALRAO"). To solve this problem there is required a significant amount of total and enough structured information on the character of the formation of the radiation situation, the potential ways of the spread of man-made pollution to the surrounding area, determining the radiation load on the population living in the vicinity of the object. In this paper there are presented the results of field studies of the radiation situation at the plant FEC "DALRAO", which were obtained during the course of expedition trips in 2009-2012.

Characterization of Rabaptin-5 γ isoform.

Rab GTPases are key regulators of intracellular membrane traffic acting through their effector molecules. Rabaptin-5 is a Rab5 effector in early endosome fusion and connects Rab5- and Rab4-positive membrane compartments owing to its ability to interact with Rab4 GTPase. Recent studies showed that Rabaptin-5 transcript is subjected to extensive alternative splicing, thus resulting in expression of Rabaptin-5 isoforms mostly bearing short deletions in the polypeptide chain. As interactions of a Rab GTPase with different effectors lead to different responses, functional characterization of Rabaptin-5 isoforms becomes an attractive issue. Indeed, it was shown that Rab GTPase effector properties of Rabaptin-5 and its α and δ isoforms are different. This work focused on another Rabaptin-5 isoform, Rabaptin-5γ. Despite its ability to interact with Rab5, endogenously produced Rabaptin-5γ was absent from early endosomes. Rather, it was found to be tightly associated with trans-Golgi network and partially localized to a Rab4-positive membrane compartment. The revealed intracellular localization of Rabaptin-5γ indicates that it is not involved in Rab5-driven events; rather, it functions in other membrane transport steps. Our study signifies the role of alternative splicing in determination of functional activities of Rab effector molecules.

Molecular barriers to processes of genetic reprogramming and cell transformation.

Genetic reprogramming by ectopic expression of transcription factor genes induces the pluripotent state in somatic cells. This technology provides an opportunity to establish pluripotent stem cells for each person, as well as to get better understanding of epigenetic mechanisms controlling cell state. Interestingly, some of the molecular processes that accompany somatic cell reprogramming in vitro are also characteristic for tumor manifestation. Thus, similar "molecular barriers" that control the stability of epigenetic state exist for both processes of pluripotency induction and malignant transformation. The reprogramming of tumor cells is interesting in two aspects: first, it will determine the contribution of epigenetic changes in carcinogenesis; second, it gives an approach to evaluate tumor stem cells that are supposed to form the entire cell mass of the tumor. This review discusses the key stages of genetic reprogramming, the similarity and difference between the reprogramming process and malignant transformation.

[Fleas of small mammals from the northeastern Russian Far East].

Results of the study of fleas (Siphonaptera) collected from small mammals (insectivores and rodents) in the Russian Far East (Magadan Province, and Khabarovsk and Kamchatka Territories) are represented. Fourteen flea species were revealed in 17 species of small mammals.

[The methods of assessment of health risk from exposure to radon and radon daughters].

The critical analysis of existing models of the relationship dose-effect (RDE) for radon exposure on human health has been performed. Conclusion about the necessity and possibility of improving these models has been made. A new improved version ofthe RDE has been developed. A technique for assessing the human health risk of exposure to radon, including the method for estimating of exposure doses of radon, an improved model of RDE, proper methodology risk assessment has been described. Methodology is proposed for the use in the territory of Russia.

[Radio-ecological situation in the area of JSC "Priargunsky Production Mining and Chemical Association"].

In order to assess the radioecological situation created in the area of the location of diversified uranium mining enterprise "Priargunsky Production Mining and Chemical Association" (PIMCU) there was investigated the radioactivity of a number of the compartments of environment, both at the industrial site and beyond it, as well as the volume activity of radon inside the ground and working premises. Radioecological situation in the vicinity of the uranium mines was performed in comparison with the background (fixed reference, control) district, where there is no uranium mining. Performed studies have shown the significant excess content of 226Ra, 232Th, 210Pb, 222Rn in soil, water open water bodies and local foods near uranium mines compared to areas outside the zone of influence of uranium mining that allows to make a conclusion about the significant technogenic pollution of local areas of the plant and adjoining territory.

Generation of CRISPR/Cas9 modified human iPSC line with correction of heterozygous mutation in exon 6 of the CaSR gene.

The calcium-sensing receptor (CaSR) gene encodes a cell membrane G protein-coupled receptor (GPCR) which has a key role in maintaining the extracellular Ca2+ homeostasis. We aimed at correcting the compound heterozygous mutation in the 6th [c.1656delA, p.I554SfsX73] and 7th [c.2217 T > A, p.C739X] exons of the CASR gene which the original patient-derived iPSC line had. The mutation is associated with neonatal severe primary hyperparathyroidism of the patient. We generated and characterized a CRISP/Cas9-edited hiPSC line with the restored sequence in the sixth exon of the CASR gene, bearing only heterozygous mutation in the 7th exon. The results showed that the new genetically modified cell line has karyotype without abnormalities, typical hiPSCs morphology, characteristic expression of pluripotency markers, and ability to develop into three germ layers, and differentiates in chondrogenic, adipogenic, osteogenic directions. This new cell line will complement the existing pool of CaSR-mutated cell lines, a valuable resource for in-depth understanding of neonatal severe primary hyperparathyroidism. This will allow further exploration of the application of pharmacological drugs in the context of personalized medicine to correct Ca-homeostasis disorders.

[Some physiological and biochemical indicators of underyearling Laxman's shrews (Sorex cecutiens Laxmann) and even-toothed shrews (Sorex isodon Turov) under conditions of different population densities].

Based on the results of a study conducted in 2006-2010 in the Buyunda River basin (a feeder of the Kolyma River), the influence of the population density of common shrews (Sorex) on some of the physiological and biochemical parameters (glycogen and lipids in the liver, the relative weight of the spleen, the white and brown adipose tissue cellularity of bone and brain tissue) was investigated. The content of energy reserve substances was correlated with the number of animals (fat deposits had a negative correlation; the glycogen content in the liver had a direct correlation). For the rest of the physiological-biochemical parameters, no significant correlation with the population density was detected, although for the content of brown fat and cellularity of bone marrow tissue in Sorex isodon, as well as the relative weight of the spleen in both species of shrews, a trend was observed. We suggest that the identified physiological changes indicate irregular feeding of animals in years with higher population densities.

[Regulatory supervision and radiation survey in the area of location of former military technical bases].

Activities related to the rehabilitation of areas and facilities of the temporary storage of spent nuclear fuel and radioactive waste (SNF and RW) at Andreeva Bay and Gremikha on the Kola Peninsula and in the Primorsky Krai in the Russian Far East is an important component of the regulatory functions of the Federal Medical biological Agency (FMBA of Russia). Technical support to the FMBA of Russia in this activity is provided by A.L Burnazyan Federal Medical Biophysical Center Main research interests include evaluation of radiological threats to determine the priority directions of regulation, a detailed analysis of the radiation situation at areas, territories and in vicinity of temporary waste storage facilities, radiation control and environmental monitoring, the development of digital maps and geoinformation systems, project expertise in the field of rehabilitation of PVC including the management of SNF and RW Implementation of these natural, practical and theoretical works is completed by development a set of regulatory documents ensuring adherence to radiation safety for the stuff population and the environment, and the also documents governing the management of SNF and RW waste in the territories of PVC.

In vitro histogenesis of human embryonic stem cells into retina components.

We developed a protocol of in vitro differentiation of human embryonic stem cells into three-dimensional structures histologically and molecularly similar to the developing retina.

[Worldwide experience and recent trends in gene therapy of ischaemic diseases].

Fifteen odd years have passed since the first application of a gene-therapeutic modality in clinical practice for treatment of lower-limb chronic ischaemia. Over this time, vast experience has been gained worldwide, with not less than one thousand patients treated by gene-based therapies, thus making it possible to generalise the published findings of these clinical trials. Resulting from such an analysis, it should be recognized that the least dangerous gene therapeutic modalities available so far are plasmid ones, with the most efficient being those containing the gene of vascular endothelial growth factor VEGF(165). The most convincing results were obtained while treating chronic ischemia of the lower extremities, whereas gene-based therapy used for treatment of coronary artery disease failed to have yielded, as of yet, clear cut positive results.