A Comparative Kidney Transcriptome Analysis of Bicarbonate-Loaded insrr-Null Mice.

The maintenance of plasma pH is critical for life in all organisms. The kidney plays a critical role in acid-base regulation in vertebrates by controlling the plasma concentration of bicarbonate. The receptor tyrosine kinase IRR (insulin receptor-related receptor) is expressed in renal ß-intercalated cells and is involved in alkali sensing due to its ability to autophosphorylate under alkalization of extracellular medium (pH > 7.9). In mice with a knockout of the insrr gene, which encodes for IRR, urinary bicarbonate secretion in response to alkali loading is impaired. The specific regulatory mechanisms in the kidney that are under the control of IRR remain unknown. To address this issue, we analyzed and compared the kidney transcriptomes of wild-type and insrr knockout mice under basal or bicarbonate-loaded conditions. Transcriptomic analyses revealed a differential regulation of a number of genes in the kidney. Using TaqMan real-time PCR, we confirmed different expressions of the slc26a4, rps7, slc5a2, aqp6, plcd1, gapdh, rny3, kcnk5, slc6a6 and atp6v1g3 genes in IRR knockout mice. Also, we found that the expression of the kcnk5 gene is increased in wild-type mice after bicarbonate loading but not in knockout mice. Gene set enrichment analysis between the IRR knockout and wild-type samples identified that insrr knockout causes alterations in expression of genes related mostly to the ATP metabolic and electron transport chain processes.

Complete and partial time-delay signature suppression in a laser array.

We model dynamics of a quantum dot based micropillar laser array subject to the time-delayed optical feedback. The global coupling provided by the feedback generates a rich set of various instabilities including chaotic regimes with strong time-delay signature in the autocorrelation function. We demonstrate that the dispersion of the array coupling phases leads to effective suppression of the time-delay signature due to the dispersion of the system's internal timescales. We find that the transition to the complete suppression of the time-delay signature appears via a chimera state where highly correlated and non-correlated laser outputs coexist. The degree of correlation in the chimera state depends on the coupling phase dispersion.

Cytotoxic Activity of Macrophages as a Tumor Malignancy Factor.

An experimental cell-based model was developed to study antibody-independent cytotoxic activity of macrophages that allows selection of derivative tumor cells resistant to this activity and clarification of the mechanisms responsible for this selection. Cytotoxic macrophages showing antibody-independent cytotoxic activity were obtained. Derivatives of PC3 and H1299 cells resistant to cytotoxic macrophage activity were generated. The main characteristics of the obtained derivative cells were studied. The proposed experimental model can be used as a tool for studying the mechanisms of the development of tumor cells resistance to antibody-independent cytotoxic activity of macrophages.

The Mechanism of the Development of Macrophage Tolerance in Tumor Microenvironment.

Bacteria forming the resident microbiome of the tumor are an integral component of its microenvironment. The interaction of the tumor microbiome with the tumor or tumor stromal cells is not well understood. We hypothesized that bacteria in the tumor microenvironment induce macrophage tolerance. Macrophage tolerance is a phenomenon of macrophage inability to respond to a repetitive inflammatory stimulus, which leads to a loss of cytotoxic activity. We studied the development of macrophage tolerance under the influence of bacteria and cytokines of the tumor microenvironment in vitro. It was found that the macrophage tolerance in the tumor stroma can develop in response to bacterial cell wall components and inflammatory factors. The acquired tolerance is inability of macrophages to produce proinflammatory cytokines TNFα, IL-1ß, and MCP-1 and activation of the production of immunosuppressive IL-10.

[Analysis of the development of acute coronary syndrome during the COVID-19 pandemic].

The study compares two groups of patients with acute coronary syndrome, depending on the presence of COVID-19 in the anamnesis. The comparison groups analyzed the risk factors for cardiovascular diseases, including smoking, heredity, gender differences, age, diabetes, and obesity. The results of biochemical blood tests were analyzed. It was found that patients with acute coronary syndrome who underwent COVID-19 were less likely to have risk factors for cardiovascular diseases. Patients with acute coronary syndrome who underwent COVID-19 were found to have lower blood glucose, cholesterol, very-low-density lipoprotein, and triglycerides. As a result of the study, it was revealed that COVID-19 is an independent risk factor for the development of acute coronary syndrome.

Tracking photon jumps with repeated quantum non-demolition parity measurements.

Quantum error correction is required for a practical quantum computer because of the fragile nature of quantum information. In quantum error correction, information is redundantly stored in a large quantum state space and one or more observables must be monitored to reveal the occurrence of an error, without disturbing the information encoded in an unknown quantum state. Such observables, typically multi-quantum-bit parities, must correspond to a special symmetry property inherent in the encoding scheme. Measurements of these observables, or error syndromes, must also be performed in a quantum non-demolition way (projecting without further perturbing the state) and more quickly than errors occur. Previously, quantum non-demolition measurements of quantum jumps between states of well-defined energy have been performed in systems such as trapped ions, electrons, cavity quantum electrodynamics, nitrogen-vacancy centres and superconducting quantum bits. So far, however, no fast and repeated monitoring of an error syndrome has been achieved. Here we track the quantum jumps of a possible error syndrome, namely the photon number parity of a microwave cavity, by mapping this property onto an ancilla quantum bit, whose only role is to facilitate quantum state manipulation and measurement. This quantity is just the error syndrome required in a recently proposed scheme for a hardware-efficient protected quantum memory using Schrödinger cat states (quantum superpositions of different coherent states of light) in a harmonic oscillator. We demonstrate the projective nature of this measurement onto a region of state space with well-defined parity by observing the collapse of a coherent state onto even or odd cat states. The measurement is fast compared with the cavity lifetime, has a high single-shot fidelity and has a 99.8 per cent probability per single measurement of leaving the parity unchanged. In combination with the deterministic encoding of quantum information in cat states realized earlier, the quantum non-demolition parity tracking that we demonstrate represents an important step towards implementing an active system that extends the lifetime of a quantum bit.

[Parvovirus B19 infection in HIV-infected patients].

Here we provide a review of the literature and a description of our own clinical case. The patient was a 32-year-old woman who had been infected with HIV for 6 years without antiretroviral therapy. The test results showed CD4 87 cells/l, viral load 3750 copies/ml. Normochromic normocytic anemia and reticulocytopenia developed soon. In the myelogram, all erythroblasts were 0.5%. The viral load of parvovirus B19 DNA according to PCR was more than 9 million IU/ml. Pure red cell aplasia associated with parvovirus B19 was diagnosed. We started antiretroviral therapy with efavirenz, lamevudine and tenofovir. In addition to blood transfusions, we administered intravenous donor immunoglobulin with a dose increase from 5000 mg to 20 000 mg per day. After discontinuing of intravenous immunoglobulins, the laboratory test results were stable over the next 5 months: hemoglobin was more than 115 g/L, reticulocytes more than 3%, in the myelogram all erythroblasts were 21%. However, the elimination of parvovirus B19 wasnt achieved. The maximum decrease in viral load for parvovirus B19 was down to 720 IU/ml. A typical feature of the case was the lack of pure red cell aplasia of the bone marrow with the existing viral load of parvovirus B19. HIV infection progressed: 44 cells/l, viral load not determined. The case ended lethally.

[Methods of Evaluating the Efficiency of CRISPR/Cas Genome Editing].

The CRISPR/Cas system is currently widely used for genome editing. The procedure of genome editing includes two necessary steps: (i) searching for the most effective guide RNA, and (ii) analyzing clones for presence of the desired mutation. This review presents the methods used to assess the efficiency of the CRISPR/Cas system and to confirm mutation in the target locus and discusses their advantages and disadvantages. It aims to provide information that could help researchers to choose a technique most appropriate for their specific tasks and available resources.

Optimization of Heterologous Expression of Insulin Receptor-Related Receptor Ectodomain.

In this paper, we present an approach to optimize the heterologous expression of the receptor tyrosine kinase IRR, which further simplifies the purification of the IRR from the medium and increases the final yield. The approach proposed by us can find application in the biotechnological production of other large-scale recombinant proteins produced for medical purposes.

[Pure red cell aplasia of the bone marrow in combination with thymoma. A literature review and own data].

Eight patients were observed with a rare combination of thymoma and pure red cell aplasia of bone marrow (PRCA), of which seven women were between 44 to 68 years old. The diagnosis of PRCA was established before the detection of thymoma in 1 patient, simultaneously in 3, after - in 4. Seven patients underwent timomectomy. The weight of removed thymomas was from 200 to 780 grams. Morphological type A thymoma variant (spindle cell) was installed in 2 patients, type B1 - in 2, type B2 - in 2, type B3 - in 2. Complete remissions were obtained using cyclophosphamide and cyclosporin in 5 patients, lasting from 6 months to 7 years. The results of immunological studies with the identification of non - hemolytic antibodies to the proteolytic antigen (Pr1d) on the erythrocyte membrane in 4 patients are presented. Of these, two studied patients simultaneously detected antibodies to the Pr1d antigen and the interspecific antigen of mammalian erythroblasts (IAME). It is shown that the lifespan of red blood cells are not changed. The direct Coombs test was negative in 5 patients, but with the help of aggregate hemaglutination test and enzyme immunoassay, antibodies were detected on the surface of erythrocytes. The pathogenesis of this combination of diseases remains unclear and needs to be elucidated.

Alginate Encapsulation to Enhance Biopreservation Scope and Success: A Multidisciplinary Review of Current Ideas and Applications in Cryopreservation and Non-Freezing Storage.

BACKGROUND: The development of encapsulation technologies has played an important role in improving cryopreservation outcomes for many cell and tissue types over the past 20 years. Alginate encapsulation cryopreservation (AECryo) has been incorporated into a range of applications in biotechnology, species conservation and clinical therapies, using cells from many different phyla, including higher plants, animal and human cells. This review describes the background to the origins of AECryo, the development of AECryo in higher plant tissues, broadening to current applications in algal conservation, the roles for AECryo in preserving phytodiversity, fungal species and in animal and human cells. OBJECTIVE: The main aims are to provide information resources on AECryo in different areas of biology and to stimulate new ideas for wider applications and future improvement. The translation of this useful biopreservation strategy into new opportunities for cell cryopreservation and storage at non-freezing temperatures are also discussed.

Coinheritance of HbD-Punjab/ß+-thalassemia (IVSI+5 G-C) in patient with Gilbert's syndrome.

Thalassemia and qualitative hemoglobinopathy are hereditary disorders of Hb synthesis that lead to change in the Hb conformation or a decrease in the synthesis of structurally normal Hb, and consequently, to erythron pathology. Many variants of Hb are unstable or have altered affinity for oxygen, and, in heterozygous form can be associated with clinical and hematological manifestations (hemolytic anemia, hypochromic microcytic anemia, erythrocytosis). HbD-Punjab [ß121 (GH4) Glu → Gln; HBB: C.364G> C] is variant of Hb carrying the amino acid substitution in the 121 position of ß-globin chain. In all cases reported so far, patients with HbD-Punjab/ß+-thalassemia (IVSI+5 G-C) combination experienced typical thalassemia with hypochromic microcytosis. HbD-Punjab was detected by electrophoresis from 37 to 94% of total Hb. The article describes rare clinical case of the cohabitation of HbD-Punjab/ß+-thalassemia (IVSI+5 G-C) in a patient with homozygous variant of Gilbert's syndrome observed in AS Loginov Moscow Clinical Scientific Center.

Perfusion bioreactor-based cryopreservation of 3D human mesenchymal stromal cell tissue grafts.

The generation of an off-the-shelf in vitro engineered living tissue graft will likely require cryopreservation. However, the efficient addition and removal of cryoprotective agents (CPA) to cells throughout the volume of a three-dimensional (3D) tissue graft remains a significant challenge. In this work, we assessed whether a perfusion bioreactor-based method could be used to improve the viability of cryopreserved mesenchymal stromal cell- (MSC) based tissue constructs as compared to using conventional diffusion-based methods. The bioreactor was first used to saturate 3D constructs with CPA under perfused flow. Following cryopreservation, the bioreactor was then also used for the efficient removal of the CPA from the 3D tissues. We demonstrate that addition and removal of CPA under perfused flow significantly increased the viability of MSC within cryopreserved 3D tissue constructs as compared to conventional diffusion-based methods.

[Behavioral Phenotype of Mice with Alkali Sensor IRR Gene Knockout].

Receptor-like tyrosine kinase IRR (the insulin receptor-related receptor) can be activated by extra- cellular alkaline media. IRR is found in organs that come in contact with liquids of extremal pH, and also in specific cells of the nervous systems where its function is not known. In this study, we analyzed the phenotype of IRR knockout mice in a series of behavioral tests. In control experi- ments, null-mutation littermate mice were analyzed. In the "Social interaction" test, the knockout animals showed a reduced number of social contacts. No statistically significant differences in im- mobility time were revealed in the "Forced swim" test, yet the number of animals that showed pro- longed immobility time, was higher in the group of knockout mice. In the "Resident-intruder" test, wild-type mice demonstrated their typical aggressive behavior whereas 7 out of 16 knockout animals stayed inert and, in contrast, attacked by the intruder. The obtained data suggest that the IRR gene inactivation results in disturbances of the aggressive-defensive behavior typical of the parental mouse strain.

Closed Vitrification System as A Platform for Cryopreservation of Tissue Engineered Constructs.

BACKGROUND: Cryopreservation of mesenchymal stromal cells (MSCs) and MSCs-based tissue engineered constructs (TECs) is a promising strategy for regenerative medicine. OBJECTIVE: To examine vitrification system consisting of multicomponent vitreous solution, closed type container, human adult MSCs and two-step exposure procedure as a platform for cryopreservation of MSCs-based TECs. MATERIALS AND METHODS: Vitrification properties of solutions were studied by visual analysis and calorimetry. Viability (trypan blue, MTT-test), metabolic activity (Alamar Blue assay) and adhesion of cells were assessed both after exposure with vitreous solutions and following rapid cooling-thawing in standard cryovials. RESULTS: The feasibility of the vitrification system was tested on MSCs suspensions (S-MSCs) and alginate encapsulated MSCs (AE-MSCs). The minimal concentrations of cryoprotectants, which allowed avoiding ice formation during rapid cooling and rewarming comprised 10 % for dimethylsulfoxide, 20 % for ethylene glycol, 20 % for 1.2-propanediol and 0.5 M sucrose. To achieve viability and metabolic activity rates of AE-MSCs comparable to S-MSCs after vitrification the extension of the exposure time within the same vitreous solution was sufficient. After vitrification both S-MSCs and AE-MSCs retained the capacity to osteogenic and adipogenic differentiation. CONCLUSION: Data demonstrate that this vitrification system can be used as a platform for development of effective protocols for cryopreservation of MSCs-based TECs.

Structural and functional analyses of the sixth site of neurexin alternative splicing.

In this study, we found the sixth site of alternative splicing (SS6) of neurexin 1a from the rat brain. This site is located between the fifth LNS and the third EGF-like domains. The insertion in the SS6 site corresponds to the 9-residue peptide VALMKADLQ, which is conserved among animals. We demonstrated that the SS6 insertion regulates tissue-specific expression of neurexin 1α.

[Actual sanitary, epidemiological and hygienic aspects of a dietitian's activities in stationary medical institutions].

The article shows the relevance of the main areas of dietitians' training to sanitary and epidemiological and hygienic issues of organization of clinical nutrition in stationary medical institutions (MIs) at training and refresher courses on dietetics. The attention is focused on the new legislative, policy and regulatory instruments, sanitary and epidemiological and hygienic requirements, providing high quality, safety and efficacy of nutritional therapy in MIs. The role of dietitian in the organization of clinical nutrition is highlighted. There were set out rights and responsibilities of a dietitian as a representative of MI under inspections by Rospotrebnadzor bodies; the demands, put forward by these bodies to the tested object, and actions, taken by them.

[CONTROVERSIAL ISSUES OF THE ANESTHESIOLOGICAL MAINTENANCE OF DELIVERY IN OBESE PATIENTS].

The review represents debatable issues of the anesthesia management of the surgical delivery and postoperative period in patients with obese: the choice of anesthesia methods, anesthesia as a risk factor of maternal and perinatal morbidity and mortality and methods of intraoperative pain management. Authors made an attempt to reveal preventable risk factors of complications in obese patients: professional skills, technical equipment, standards compliance, period of time from the decision till delivery itself selection of an adequate dose of local anesthetic during spinal anesthesia, necessity of monitoring and correction of intra-abdominal hypertension. The article discusses the variety of problems, and in case offurther researches they will help to decrease frequency of the anesthetic complications, that determine obstetric and perinatal outcomes in obese patients.

Long-term evolution of broken wakefields in finite-radius plasmas.

A novel effect of fast heating and charging a finite-radius plasma is discovered in the context of plasma wakefield acceleration. As the plasma wave breaks, most of its energy is transferred to plasma electrons. The electrons gain substantial transverse momentum and escape the plasma radially, which gives rise to a strong charge-separation electric field and azimuthal magnetic field around the plasma. The slowly varying field structure is preserved for hundreds of wakefield periods and contains (together with hot electrons) up to 80% of the initial wakefield energy.

A sugar pretreatment as a new approach to the Me2SO- and xeno-free cryopreservation of human mesenchymal stromal cells.

Experimental and clinical applications of mesenchymal stromal cells (MSCs) require the development of new approaches and improvements of existing methods of cell cryopreservation. Cryoprotective solutions for effective cell preservation usually contain 10% dimethyl sulfoxide (Me2SO) and fetal bovine serum (FBS) limiting clinical application of MSCs. We have developed a novel approach to the cryopreservation of human MSCs comprising inclusion of sugars into incubation medium for 24 hrs prior to cryopreservation (pretreatment) with their obligatory presence in cryoprotective solution. Such combined application of mannitol, lactose, sucrose, trehalose or raffinose on cultivation and cryopreservation stages resulted in a significant increase of MSCs viability. Optimal concentration of sugars for cell pretreatment was 200 mM, as an additive in cryoprotective solution - 300 mM. Highest cell viability and metabolic activity assessed by Alamar Blue test were achieved with sucrose, trehalose and raffinose. Using these sugars about 50% of pretreated cells after cryopreservation in the absence of Me2SO and FBS preserved their survival and metabolic activity. During following recultivation cryopreserved MSCs were able to attachment, proliferation and multilineage differentiation towards osteogenic and adipogenic lineages. The data obtained indicate that the approach included pretreatment of cells with sugars combined with their presence in cryoprotective solution is feasible for future regenerative medicine projects.