Assessing cell lines with inducible depletion of cohesin and condensins components through analysis of metaphase chromosome morphology.

One of the most productive strategies for finding the functions of proteins is to study the consequences of loss of protein function. For this purpose, cells or organisms with a knockout of the gene encoding the protein of interest are obtained. However, many proteins perform important functions and cells or organisms could suddenly lose fitness when the function of a protein is lost. For such proteins, the most productive strategy is to use inducible protein degradation systems. A system of auxin-dependent protein degradation is often implemented. To use this system, it is sufficient to introduce a transgene encoding a plant-derived auxin-dependent ubiquitin ligase into mammalian cells and insert a sequence encoding a degron domain into the gene of interest. A crucial aspect of development of cell lines engineered for inducible protein depletion is the selection of cell clones with efficient auxin-dependent degradation of the protein of interest. To select clones induced by depletion of the architectural chromatin proteins RAD21 (a component of the cohesin complex) and SMC2 (a component of the condensin complex), we propose to use the morphology of metaphase chromosomes as a convenient functional test. In this work, we obtained a series of clones of human HAP1 cells carrying the necessary genetic constructs for inducible depletion of RAD21 and SMC2. The degradation efficiency of the protein of interest was assessed by flow cytometry, Western blotting and metaphase chromosome morphology test. Based on our tests, we showed that the clones we established with the SMC2 degron effectively and completely lose protein function when induced by auxin. However, none of the HAP1 clones we created with the RAD21 degron showed complete loss of RAD21 function upon induction of degradation by auxin. In addition, some clones showed evidence of loss of RAD21 function even in the absence of induction. The chromosome morphology test turned out to be a convenient and informative method for clone selection. The results of this test are in good agreement with flow cytometry analysis and Western blotting data.

[Public health risk threats associated with coronavirus infection in Moscow and the need to counteract].

PURPOSE OF THE STUDY: to determine the main threats to the health of the population of Moscow based on the analysis of the medical and demographic situation in Moscow and the Russian Federation in the context of the COVID-19 pandemic, in order to develop management decisions at the regional level. MATERIAL AND METHODS: Materials of the Federal State Statistics Service (Rosstat), as well as statistical collections of the Russian Research Institute of Health of the Ministry of Health of Russia for 2013-2022 were used. RESULTS: Coronavirus infection has changed the morbidity and mortality rates of the population of the Russian Federation and its constituent entities. In the Russian Federation mortality in 2019-2021 increased from 12.3 to 13.6‰, while in Moscow the increase in the indicator is significant - from 9.5 to 16.7‰. The incidence of COVID-19 in Moscow in 2020-2021 (over the two COVID years) increased from 6191.0 to 8976.0‰oo. The noted decrease in incidence rates across almost all classes of diseases during the COVID period (2019-2020) was associated with the lower medical check-ups and preventive activities, while during 2019-2021 there was an increase in disease incidence of the following classes: mental disorders, behavior disorders; diseases of the blood and blood-forming organs and certain disorders involving the immune mechanism, etc., indicating the need for rehabilitation of this group of patients. CONCLUSIONS: The need to counter the threats to the health of the population of Moscow caused by the coronavirus infection requires the development of management decisions aimed at strengthening preventive activities and rehabilitation of patients in the post-COVID period.

Design and testing of a high perveance sheet beam electron gun.

This paper presents the design, prototype engineering, and initial testing of a relatively low-voltage, high aspect ratio electron gun for versatile pulsed power applications. The sheet beam, diode electron gun is designed for (23-25) kV voltage and 6 µA/V1.5, as is required for x-band klystron, as a part of a compact Klylac delivering electron beam energy in the MeV range. The gun prototype was engineered, built, and tested using a Scandinova M1-0.5 modulator. Beam loss on the anode aperture is evaluated from transient waveforms and equivalent circuit modeling.

Growth Induction of Solid Ehrlich Ascitic Carcinoma in Mice after Proton Irradiation of Tumor Cells Ex Vivo.

This study presents data on the growth rate and frequency of induction of the solid form of Ehrlich's ascites carcinoma (EAC) in mice in the short and long term after inoculation of ascitic cells irradiated ex vivo with a proton beam in the dose range of 30-150 Gy. It was shown that the growth rate of solid tumors after inoculation of irradiated cells ex vivo coincided with the growth of tumors in the control group. The frequency of tumor induction in mice after inoculation of EAC cells irradiated at a dose of 30 Gy was 80%, 60 Gy-60%, 90 Gy-25%, and 120 Gy-10%; at irradiation at a dose of 150 Gy, no tumors appeared during the entire observation period. Thus, we determined the dose of proton radiation required to eliminate tumor cells and/or signaling factors that can lead to the induction of tumor growth of EAC in mice.

[Treatment of ventral hernias in patients with morbid obesity]. / Taktika lecheniya ventral'nykh gryzh u patsientov s morbidnym ozhireniem.

The review is devoted to the treatment of ventral hernias in patients with morbid obesity. This issue is important due to significant number of such patients and no unambiguous clinical recommendations. The advantages of simultaneous surgery (with bariatric intervention) are obvious, i.e. lower risk of postoperative hernia incarceration and no need for re-hospitalization with another intervention. High risk of bariatric population makes it necessary to minimize surgery time and surgical trauma. A staged approach with reducing body weight surgically or conservatively before hernia repair is often chosen. Hernia repair should be performed using laparoscopic or robotic techniques with obligatory use of mesh implants. Panniculectomy or abdominoplasty as the main surgery is a valid option. Currently, it is necessary to develop clear criteria for selecting patients with morbid obesity for staged and simultaneous treatment of ventral hernias.

Tribotronic and electrochemical properties of platinum-nanofluid interfaces formed by aqueous suspensions of 5 and 40 nm TiO2 nanoparticles.

Nanoparticles (NPs) can be highly beneficial as additives to lubricating fluids, and the tribotronic response of charged NPs tuned by external fields represents an area of great technological potential. Tribotronic response, however, is expected to be highly size dependent, which represents a significant design challenge. To explore this issue, quartz crystal microbalance and cyclic voltammetry were employed to characterize nanotribological and electrochemical behavior of platinum-nanofluid interfaces formed by aqueous suspensions of different-sized negatively charged titanium dioxide (TiO2) NPs. Suspensions of 5, 40, and 100 nm NPs were all observed to reduced interfacial frictional drag forces upon introduction into pure water in zero field conditions, with reductions for the 40 nm NPs about twice those of 5 nm particles at comparable concentrations. Suspensions of 100 nm NPs produced even greater reductions, but rapidly precipitated from the suspension when left unstirred. NPs were also driven to and from Pt electrode surfaces by applying external electric fields with varying amplitudes and modulation frequencies. For electric fields of sufficient amplitude and duration, the 40 nm TiO2 nanosuspension exhibited tribological properties consistent with a reversible electrophoretic deposition of the NPs, accompanied by changes in the electrochemical attributes and increasing interfacial drag. The 5 nm NP properties were consistent with progressive reductions in interfacial drag forces at the NP-suspension interface linked to field-induced increases in concentration.

Universal Maximum Strength of Solid Metals and Alloys.

Interstitial electron density ρ_{o} is offered as a direct metric for maximum strength in metals, arising from universal properties derived from an electron gas. ρ_{o} sets the exchange-correlation parameter r_{s} in density-functional theory. It holds also for maximum shear strength τ_{max} in polycrystals [M. Chandross and N. Argibay, Phys. Rev. Lett. 124, 125501 (2020)PRLTAO0031-900710.1103/PhysRevLett.124.125501]. Elastic moduli and τ_{max} for polycrystalline (amorphous) metals are linear with ρ_{o} and melting T_{m} (glass-transition T_{g}) temperature. ρ_{o} or r_{s}, even with rule-of-mixture estimate, predicts relative strength for rapid, reliable selection of high-strength alloys with ductility, as confirmed for elements to steels to complex solid solutions, and validated experimentally.

[The problems of rural health care and prospects of development].

Further reforming of the rural health care should be proceeded with special attention to availability of medical care in countryside.The purpose of the study is to develop on the basis of analysis of accounting data and research results recommendations for improving organizational forms of medical care to country dwellers. The exploration of peopleware and main performance indicators of medical organizations in rural areas was implemented.The statistical, analytical and sociological methods were applied. The reporting forms 47, 30, 12 of Federal statistical monitoring are analyzed.Results. The provision of population with medical personnel in rural municipalities in 2010-2018 increased from 12.5 to 14.5 per 10 thousand of rural population. The provision with paramedical personnel made up to 52.3%oo and during the same years indicator decreased from 55.4 to 52.3%oo. During the analysis period, 4241 feldsher obstetric posts were reduced and in 2018 their number made up to 33,350. The number of feldshers in rural areas decreased on 18.5%. In 2005-2018 number of central district hospitals and district hospitals decreased in 2.5 times. At that, provision of beds in municipalities of rural areas decreased from 49.6 to 38.8%oo. There significant winding up of district hospitals and their restructuring into branches of central district hospitals occurred. In 2018, remained only 47 out of them equipped with 1549 beds. Unfortunately, the reporting forms of the Ministry of Health of the Russian Federation do not account branches of central district hospitals. This information is to be included in accountability of the Ministry of Health.It is necessary to expand scope of authority of local government bodies in resolving medical and social problems in rural territories.

[Case report of impacted mandibular third molar extraction using marked surgical bur]. / Udalenie retinirovannogo tret'ego molyara nizhnei chelyusti s ispol'zovaniem markirovannogo khirurgicheskogo bora.

Third molar extraction is a very common manipulation in dental practice, which involve risks of injury of the important adjacent anatomical structures. In order to prevent such complications, the authors suggest using marked surgical burs providing maximal control of the tooth dissection.

[Extra-intracranial bypass surgery using the orifice of maxillary artery in a patient with middle cerebral artery aneurysm. Case report and literature review]. / Pervyi opyt vypolneniya ekstra-intrakranial'nogo shuntirovaniya s ispol'zovaniem ust'ya verkhnechelyustnoi arterii u patsienta s anevrizmoi srednei mozgovoi arterii. Opisanie klinicheskogo sluchaya i obzor literatury.

OBJECTIVE: To present the technique of extra-intracranial bypass surgery using the orifice of maxillary artery bypass, to evaluate the advantages and disadvantages of this and alternative revascularization options. MATERIAL AND METHODS: Radial artery graft harvesting was performed at the 1st stage. Simultaneously, the second team of surgeons performed a combined (submandibular and anterior) access to the donor artery (mandibular segment of maxillary artery behind the ramus of the mandible). Craniotomy and mobilization of potential recipient arteries (M2-M3 segments of the middle cerebral artery) were performed at the 2nd stage. Distal anastomosis in end-to-side fashion was formed with M3 segment of the middle cerebral artery. At the 3rd stage, radial artery was passed through a subcutaneous tunnel in zygomatic region. The orifice of maxillary artery was resected together with distal external carotid artery (ECA) and orifice of superficial temporal artery. After transposition of ECA and orifice of maxillary artery, proximal end-to-end anastomosis was performed with radial artery. After that, the main surgical stage was performed, i.e. exclusion of M3 segment of the middle cerebral artery together with aneurysm. RESULTS: Harvesting of mandibular segment of the maxillary artery as a donor vessel reduces the length of bypass graft to 12-14 cm since this branch is localized close to the skull base. You can also form optimal proximal end-to-end anastomosis for intracranial redirecting blood flow maxillary artery. CONCLUSION: The described method makes it possible to form anastomosis with a short bypass graft and reduce the risk of thrombosis. This procedure is effective for cerebral bypass in patients with skull base tumors, complex aneurysms, and occlusive-stenotic lesions of carotid arteries.

[Models and sculptural casts in forensic medicine and medical criminology]. / Mulyazhi i skul'pturnye slepki v sudebnoi meditsine i meditsinskoi kriminalistike.

The study objective is to present facts from the history of casting art and biographical information about the casting artists S.P. Fiveysky and M.A. Kurbatov, the founders of the casting craft in Russia. The attributes of the technology for making death masks, their features, advantages, and disadvantages as objects that capture appearance are described. The role of models and death masks in forensic practice, forensic science, and pedagogical activity is addressed.

[COVID-19-associated mucormycosis in ophthalmological practice in the Volgograd region]. / COVID-19-assotsiirovannyi mukormikoz v oftal'mologicheskoi praktike Volgogradskogo regiona.

The problem of fungal infections in the era of COVID-19 has acquired special significance. This infection, directly or indirectly, through the use of glucocorticoids and antibiotics in its treatment, as well as poorer self-management of chronic diseases, has led to a wide spread of risk factors for fungal diseases among people who have had a novel coronavirus infection. The article presents two cases of COVID-19-associated mycosis, more related to mucormycosis, which were diagnosed by ophthalmologists in the Volgograd region. In the first case, the severe course of rhino-orbito-cerebral form of mucormycosis required a number of surgical interventions and prolonged treatment in the intensive care unit. In the second case, the patient asked for help without signs of aggravation of the general condition, but with irreversible local manifestations. In both cases, the eyeball was removed. Morphological examination revealed aseptal ribbon hyphae of different diameters, branching mainly at right angles, more typical for fungi of the Mucorales family. Due to the severe consequences of the disease, clinicians, including ophthalmologists, need to be especially alertness in patients with the described symptoms and risk factors in the post-COVID period.

[Machine learning for resting state fMRI-based preoperative mapping: comparison with task-based fMRI and direct cortical stimulation]. / Primenenie mashinnogo obucheniya dlya predoperatsionnogo kartirovaniya po dannym funktsional'noi magnitno-rezonansnoi tomografii pokoya: sravnenie so stimul-zavisimoi funktsional'noi magnitno-rezonansnoi tomografiei i pryamoi elektricheskoi stimulyatsiei kory.

OBJECTIVE: To develop a system for preoperative prediction of individual activations of motor and speech areas in patients with brain gliomas using resting state fMRI (rsfMRI), task-based fMRI (tb-fMRI), direct cortical stimulation and machine learning methods. MATERIAL AND METHODS: Thirty-three patients with gliomas (19 females and 14 males aged 19 - 540) underwent DCS-assisted resection of tumor (19 ones with lesion of motor zones and 14 patients with lesions of speech areas). Awake craniotomy was performed in 14 cases. Preoperative mapping was performed according to special MRI protocol (T1, tb-fMRI, rs-fMRI).Machine learning system was built on open source data from The Human Connectome Project. MR data of 200 healthy subjects from this database were used for system pre-training. Further, this system was trained on the data of our patients with gliomas. RESULTS: In DCS, we obtained 332 stimulations including 173 with positive response. According to comparison of functional activations between rs-fMRI and tb-fMRI, there were more positive DCS responses predicted by rs-fMRI (132 vs 112). Non-response stimulation sites (negative) prevailed in tb-fMRI activations (69 vs 44). CONCLUSION: The developed method with machine learning based on resting state fMRI showed greater sensitivity compared to classical task-based fMRI after verification with DCS: 0.72 versus 0.66 (p<0.05) for identifying the speech zones and 0.79 versus 0.62 (p<0.05) for motor areas.

[Comparison of resting state and task-based functional MRI in preoperative mapping in patients with brain gliomas]. / Sravnenie funktsional'noi magnitno-rezonansnoi tomografii v sostoyanii pokoya i stimul-zavisimoi funktsional'noi magnitno-rezonansnoi tomografii v predoperatsionnom kartirovanii u patsientov s gliomami golovnogo mozga.

OBJECTIVE: To analyze and compare the results of cerebral cortex mapping with task-based (tb-fMRI) and resting-state functional MRI in patients with glioma of eloquent cortical areas. MATERIAL AND METHODS: There were 55 patients (24 men and 31 women aged 24 - 74 years, median 39) with glial tumors. In 26 patients, the tumor was located in motor areas. Twenty-nine patients had lesions of Broca and Wernicke's areas. All patients underwent preoperative tb-fMRI and rs-fMRI. Then, resection of tumor was carried out in all cases. RESULTS: Comparison of fMRI and rs-fMRI activation maps was assessed by calculating the Dice coefficient for inclusive speech and motor cortex masks and exclusive masks without brainstem, cerebellum, subcortical nuclei. Inclusive Dice coefficient for motor cortex ranged from 0.11 to 0.50, for speech cortex - from 0.006 to 0.240 (p<0.05). In case of exclusive masks, this value ranged from 0.15 to 0.55 for motor cortex and from 0.004 to 0.205 for speech cortex (p<0.05). CONCLUSION: When comparing the results of cortical mapping in patients with glial tumors, the use of hemispheric exclusive and inclusive masks did not significantly increase activation maps matching. Probably, low degree of correspondence was associated with different genesis of activations, as well as with high variability of speech cortex.

[The medical support of large-scale international events].

The medical support is one of the most important components of effectiveness of international sports events. The purpose of the study is to analyze results of implementation of organizational approaches to medical support of international sports competitions at the World Skills Championship 2019 in Kazan, previously developed and successfully approved on the model of the 2013 World's Students Games (Kazan). The medical support plan included preparation of regulatory base, resources of medical infrastructure at sports venues, including personnel training and measures to eliminate the medical and sanitary consequences of emergencies. At the objects of the Championship, 21 medical aid posts, 23 mobile medical teams functioned, and 22 ambulance teams were on duty. The estimated number of medical staff involved in work at the Championship venues was 120 people, or 8.9 people per 100 contestants. There were 1883 registered medical requests. The largest percentage of requests fell on staff (29.6%), volunteers (20.4%), contestants (18.1%) and visitors (10.7%). Among the accredited persons, the most requested medical assistance was for respiratory diseases (8.5 ‰), diseases of the nervous system (8.4 ‰) and injuries (7.2 ‰). The application of developed approaches to medical support of international sports competitions for the World Skills World Championship allowed to organize provision of medical care to client groups at high level.

Evaluation of the α-casein (CSN1S1) locus as a potential target for a site-specific transgene integration.

Transgenic animals are an important tool in biotechnology, including the production of recombinant proteins in the milk. Traditionally, expression constructs are based on hybrid vectors bearing mammary gland specific regulatory elements from the α-casein (Csn1s1), ß-casein (Csn2), whey acidic protein (WAP), or ß-lactoglobulin (BLG) genes. Overexpression from the randomly integrated vectors typically provides high levels of expression, but has drawbacks due to unpredictable genome localization. CRISPR-Cas9 targeted transgene integration into the endogenous casein locus could alleviate the need for extensive animal screening to achieve high and reproducible expression levels. We decided to evaluate such a "precise" integration approach, placing the human granulocyte-macrophage colony-stimulating factor (hGMCSF) gene under control of the mouse endogenous alpha-S1-casein (Csn1s1) promoter. We designed two types of transgene integrations: a knock-in in the second exon of the Csn1s1 (INS-GM) and a full-size Csn1s1 replacement with hGMCSF (REP-GM) which was never tested before. The INS-GM approach demonstrated low transgene expression and milk protein levels (0.4% of Csn2 transcripts; 2-11 µg/ml hGMCSF). This was probably caused by the absence of the 3'-polyadenylation signal in the hGMCSF transgene. REP-GM animals displayed high transgene expression, reaching and slightly exceeding the level of the endogenous Csn1s1 (30-40% of Csn2 transcripts), but yielded less hGMCSF protein than expected (0.2-0.5 mg/ml vs 25 mg/ml of Csn1s1), indicating that translation of the protein is not optimal. Homozygous inserts leading to the Csn1s1 knock-out did not have any long standing effects on the animals' health. Thus, in our experimental design, site-specific transgene integration into the casein locus did not provide any significant advantage over the overexpression approach.

Development of a two-channel system for monitoring the mirror elements of the Millimetron space observatory.

The paper presents the results of the development of an onboard two-stage telescope monitoring (control) system of the Millimetron space observatory. A 3D laser scanner is used for the preliminary inspection of the reflector elements, and an image analyzer based on the Foucault knife method is used for the final one. The results of the calculation and modeling of the monitoring system for tuning the telescope elements into space are presented. The signals formed in the system are calculated, including using data on the actual shape of the manufactured mirror elements. Estimates of the expected measurement accuracy are given, and the test results of the developed system are presented.

Balamuthia spinosa n. sp. (Amoebozoa, Discosea) from the brackish-water sediments of Nivå Bay (Baltic Sea, The Sound) - a novel potential vector of Legionella pneumophila in the environment.

We have found a new free-living amoeba species named Balamuthia spinosa n. sp. (Amoebozoa, Discosea) in the bottom sediments of the brackish-water Nivå Bay (Baltic Sea, The Sound). This species resembles members of the genus Stygamoeba morphologically and was (mis)identified as belonging to this genus during the initial investigation. However, SSU rRNA gene data show that it robustly groups with Balamuthia mandrillaris sequence among Acanthopodida and represents a new species of the genus Balamuthia. Fragments of Legionella pneumophila genome were found among the NGS contigs obtained from B. spinosa n. sp., suggesting that this species may be a vector of Legionella in the environment. We discuss a remarkable morphological and ultrastructural similarity between the genus Balamuthia and the genus Stygamoeba. In addition, our phylogenetic analysis based on the SSU rRNA gene sequences revealed a close relationship between the genera Stygamoeba and Vermistella. It is one more confirmation of the order Stygamoebida, which was formed basing on the morphological evidence. The position of these branches close to Thecamoebida clade is congruent with current phylogenomic data.

Generation of two iPSC lines from healthy donor with a heterozygous mutation in the VPS13B gene.

The human induced pluripotent stem cell (iPSC) lines, iCS-MAF1-1 and iCS-MAF1-11, were generated from fibroblasts. The donor has a heterozygous mutation in the VPS13B gene, which manifests in her child as Cohen syndrome. It is a Golgi pathology, characterized by postnatal microcephaly and delayed growth and mental development. However, the process underlying pathological changes leading to the onset of the disease is still unknown. The use of iPSC will allow describing the early stages of neurogenesis, which is undoubtedly relevant for identifying key stages of development, at which phenotypic manifestations of mutations in the VPS13B gene are found.