Background: Multi-year research into the synergistic effect of cyclophosphamide (CP) and complex composite double-stranded DNA preparation (DNAmix) has made it possible to develop the novel Karanahan technology for treating malignant tumors. After being applied, the technology results in eradication of cancer stem cells, induction of extensive apoptosis of committed cancer cells, and destruction of suppressor activity of tumor-associated stroma. The therapeutic efficacy of the Karanahan cancer treatment technology is independent of both tumor immunogenicity and the patient's immune status. Breast cancer is the most common human malignancy; its treatment remains extremely challenging in current clinical practice. It is suggested that in patients with advanced breast cancer, the Karanahan technology will ensure therapeutic eradication of cancer stem cells, substantially slowing down disease progression and extending the survival of palliative patients. The objective of this clinical trial is to examine the feasibility and practical applicability of the Karanahan technology in clinical practice for patients with advanced breast cancer and identify critical elements of the technology when transferred from animal models to humans. Methods: Clinical trials of feasibility of the Karanahan technology were conducted in patients with advanced stage IV breast cancer and palliative status. Patients were enrolled consecutively based on the decision of a local case conference. The officially registered trial protocol contained all the requisite provisions, involving the inclusion/non-inclusion criteria, conditions of therapeutic intervention, informed consent, and study endpoints. Results: According to the key characteristics of the Karanahan technology, the therapeutic success is independent of the molecular subtype of breast cancer. Presence of cancer stem cells in the tumor was shown to be important for all the subtypes of the disease. The findings indicate that the analyzed treatment regimen ensures a positive local therapeutic response. Partial tumor response to the Karanahan cancer treatment technology or disease stabilization was observed in eight patients. Karanahan treatment activates the adaptive antitumor immune response in 78% of patients. The median survival time of patients significantly increased from 6 months (historical control) to 20 months after Karanahan treatment. The study design was optimized, including the selection of the optimal dose and refinement of the elements of the intervention procedure. The accrual rate for patients meeting the inclusion criteria suggests that randomized large-scale clinical trials should be conducted at the major cancer treatment centers. Conclusions: The results of studying the feasibility and practical applicability of the novel antitumor Karanahan therapy indicate that measures can be prepared for developing a protocol and design for pilot and large-scale clinical trials of this approach. The critical elements of the experimental Karanahan technology for transferring it to clinical practice for treating advanced breast cancer have been identified.

BackgroundAt present, there are no universal markers of tumor stem cells known, including for B-lymphomas. Previously, we have shown that Epstein-Barr virus-induced B-cell lymphoma culture contains cells capable of internalizing TAMRA-labeled DNA. These cells form sphere-forming centers and are essential for the development of xenografts genetically identical to the initial culture.ObjectiveTo analyze the stem characteristics of cells that internalize DNA.MethodsSorting and RNA sequencing of two subpopulations (TAMRA + and TAMRA-) of Epstein-Barr virus-induced B-cell lymphoma culture and a series of quantitative real-time reverse transcription PCR were performed.ResultsTAMRA + cells were shown to have increased synthesis of mRNA of genes associated with the maintenance of a poorly differentiated state (SOX2, NANOG, POU5F1, CYP26A1), self-renewal (FZD5, FZD7, TCF3, LEF1) and epithelial-mesenchymal transition (MMP2, ITGB7). Transcriptomic analysis revealed that in TAMRA + cells, the synthesis of mitochondrial genes, as well as caspases and some apoptosis inhibitors, is reduced. TAMRA + cells possess clonogenic properties, increased level of synthesis of mRNA for key genes associated with self-renewal and poorly differentiated state maintenance.ConclusionsInternalization of the TAMRA-DNA probe is the marker of B-lymphoma cancer stem cells and can be used to detect tumor stem cells and develop new approaches to targeted treatment of B-lymphoma.

Despite the end of the COVID-19 pandemic, there still remain risks of new aggressive strains of coronavirus. As the human population increases progressively, it is mandatory to ensure both preventive measures and an immediate response to emerging infectious threats. Another essential component for rapidly restraining a new possible pandemic is the development of new anticoronaviral therapeutics. In the present study, the anticoronaviral capabilities of Gc protein-derived macrophage-activating factor (GcMAF) are characterized. It is demonstrated that the administration of GcMAF to Syrian hamsters infected with SARS-CoV-2 within the first phase of infection (six days postinfection) is accompanied by (i) a statistically significant reduction in the viral load of the lung tissue and (ii) the switching of the inflammatory status of the lung tissue to a neutral one in terms of mRNA expression levels of the groups of pro/anti-inflammatory cytokines and chemokines. The potential mechanism for this antiviral action and the containment of the inflammatory response by the drug associated with the engagement of terminal N-acetylgalactosamine GcMAF and C-type lectin domain containing 10A expressed at the surface of lung-infiltrating macrophages and pneumocytes, which simultaneously express angiotensin-converting enzyme 2, is discussed.

In this work, we describe the design and flight performance of the REFOS instrument (from Russian REntgenovskiy FOto Spektrometr; in English “X-ray Photo Spectrometer”). REFOS is a soft X-ray spectrophotometer that registers full-disk integrated solar spectra (“Sun as a star”). It operates on board the Impulse-1 nanosatellite, which was launched on 24 June 2023. REFOS has a 1.2 – 30 keV spectral range, a 0.123 keV nominal full width at half-maximum (FWHM) resolution at 5.9 keV, and a cadence of 16 s. We illustrate the instrument flight performance using the spectra of the X5.0 flare that occurred on 31 December 2023 at 21:55 UT. For this flare, REFOS registered a meaningful signal in all of its energy bins. Based on a comparison between the GOES and REFOS fluxes, we corrected the REFOS spectral sensitivity. Additionally, we assessed the quality of the calibration based on the shape of the continuum. The observed continuum allows diagnosing the plasma temperature, and the observed spectral lines allow studying abundances of the Mg, Si, S, Ar, Ca, Fe, and Ni.

Vitamin D3 transporter (DBP) is a multifunctional protein. Site-specific deglycosylation results in its conversion to group-specific component protein-derived macrophage activating factor (GcMAF), which is capable of activating macrophages. It has been shown that depending on precursor conversion conditions, the resulting GcMAF activates mouse peritoneal macrophages towards synthesis of either pro- (IL-1β, TNF-α—M1 phenotype) or anti-inflammatory (TGF-β, IL-10—M2 phenotype) cytokines. The condition for the transition of the direction of the inflammatory response of macrophages when exposed to GcMAF is the initial glycosylated state of the population of DBP molecules and the associated effective deglycosylation of DBP by β-galactosidase. In vivo experiments with GcMAF exhibiting anti-inflammatory properties on models of induced arthritis in mice and cystitis in rats indicate a significant anti-inflammatory effect of the macrophage activator. The feasibility of unidirectional induction of anti-inflammatory properties of macrophages allows creation of combined therapeutic platforms where M2 macrophages are among the key therapeutic components.

The association of coronal mass ejections (CMEs) with flares is related to the question of whether reconnection is necessary for the CME eruption. Indeed, if reconnection happens during a CME eruption, the plasma is heated, which can be observed as a flare. In this work, we study the CME-flare association using data obtained with the Mg xii spectroheliograph on board the Complex Orbital Observations Near-Earth of Activity on the Sun (CORONAS-F) satellite. This instrument is sensitive only to the emission of plasma with a temperature greater than 4 MK, which makes it a convenient tool for detection of flaring activity. During our analysis, we first searched for CMEs detected during the Mg xii observations by the Large Angle and Spectroscopic Coronagraph (LASCO). Then, we visually checked the Mg xii images for flaring activity. We found that during the Mg xii observations (2001 – 2003), 198 CMEs were detected by LASCO. One hundred sixty of them (81

(1) Background: Multi-year research into the synergistic effect of cyclophosphamide and complex composite double-stranded DNA preparation (DNAmix) has made it possible to develop the novel Karanahan technology for treating malignant tumors, which is based on the eradication of cancer stem cells; (2) Methods: Pilot clinical trials for Karanahan technology were conducted in 12 patients with advanced stage IV breast cancer; (3) Results: The findings indicate that the analyzed treatment regimen ensures a local positive therapeutic response, significantly increases patient survival time, and activates the adaptive antitumor immune response. Adjustment of the basic regimen of Karanahan technology has been developed, which consists of increasing the dose of the DNAmix complex composite double stranded DNA preparation and its lymphotropic administration to the main lymph depots simultaneously with intratumoral injection; (4) Conclusions: Karanahan technology has a promising therapeutic potential in the treatment of inoperable stage IV breast cancer.

Aims. We aim to improve the existing techniques to probe the nanoflare hypothesis for the coronal heating problem. For this purpose, we propose using the solar extreme ultraviolet (EUV) emission variability registered with modern space-based imagers. Methods. We followed a novel model-based approach. As a starting point, we used the EBTEL 0d hydrodynamic model. We integrated the arising system of stochastic differential equations to calculate the covariance matrix for plasma parameters. We then employed a Taylor expansion technique to relate model parameters with observable EUV intensity variation statistics. Results. We found that in the high-frequency approximation, the variability of the EUV emission is defined by the dimensionless factor ϖ , which is inversely proportional to the frequency. We calculated the factor ϖ throughout the solar disk and found that it does not exceed 0.01, except for the finite number of compact regions. The distribution of ϖ follows the power law with an index of ≈ − 2.6. To validate our approach, we used it to probe the temperature of the coronal plasma. We show that the line-of-sight temperature distribution is close to homogeneous with a mode of ≈1.25 MK, which is in perfect agreement with the results of the spectroscopic diagnostics.

Macrophages are the immune cells of high-immunological plasticity, which can exert both pro- and anti-inflammatory activity, as well as repolarize their phenotype to the opposite or neutral one. In this regard, M2 macrophages of the tumor-associated stroma (TAS) are a promising therapeutic target in treating malignant neoplasms. Using FACS assay, we have estimated the CD11b+/Ly-6G+/Ly-6C+ fraction of macrophages from the peritoneum and TAS in intact healthy mice and those with developed Lewis carcinoma, both untreated and treated according to Karanahan technology in combination with group-specific macrophage activator (GcMAF-RF). As well, the pattern of pro- and anti-inflammatory cytokines mRNA expression in different groups of experimental and tumor-bearing animals was assessed. It was found that: (i) exposure of intact mice to GcMAF-RF results in the increased number of CD11b+/Ly-6C+ peritoneal macrophages and, at the same time, the expression pattern of cytokines in peritoneal macrophages switches from that characteristic of the mixed M1/M2 phenotype to that characteristic of the neutral M0 one; (ii) combination of Karanahan technology and GcMAF-RF treatment results in M0/M1 repolarization of TAS macrophages; (iii) in tumor-bearing mice, the response of peritoneal macrophages to such a treatment is associated with the induction of anti-inflammatory reaction, which is opposite to that in TAS macrophages.

In this part of the study, the first component of the concept of “natural genome reconstruction” is being proven. It was shown with mouse and human model organisms that CD34+ hematopoietic bone marrow progenitors take up fragments of extracellular double-stranded DNA through a natural mechanism. It is known that the process of internalization of extracellular DNA fragments involves glycocalyx structures, which include glycoproteins/protein glycans, glycosylphosphatidylinositol-anchored proteins and scavenger receptors. The bioinformatic analysis conducted indicates that the main surface marker proteins of hematopoietic stem cells belong to the indicated groups of factors and contain specific DNA binding sites, including a heparin-binding domain and clusters of positively charged amino acid residues. A direct interaction of CD34 and CD84 (SLAMF5) glycoproteins, markers of hematopoietic stem cells, with double-stranded DNA fragments was demonstrated using an electrophoretic mobility shift assay system. In cells negative for CD34, which also internalize fragments, concatemerization of the fragments delivered into the cell occurs. In this case, up to five oligonucleotide monomers containing 9 telomeric TTAGGG repeats are stitched together into one structure. Extracellular fragments delivered to hematopoietic stem cells initiate division of the original hematopoietic stem cell in such a way that one of the daughter cells becomes committed to terminal differentiation, and the second retains its low-differentiated status. After treatment of bone marrow cells with hDNAgr, the number of CD34+ cells in the colonies increases to 3 % (humans as the model organism). At the same time, treatment with hDNAgr induces proliferation of blood stem cells and their immediate descendants and stimulates colony formation (mouse, rat and humans as the model organisms). Most often, the granulocyte-macrophage lineage of hematopoiesis is activated as a result of processing extracellular double-stranded DNA. The commitment process is manifested by the appearance and repair of pangenomic single-strand breaks. The transition time in the direction of differentiation (the time it takes for pangenomic single-strand breaks to appear and to be repaired) is about 7 days. It is assumed that at the moment of initiation of pangenomic single-strand breaks, a “recombinogenic situation” ensues in the cell and molecular repair and recombination mechanisms are activated. In all experiments with individual molecules, recombinant human angiogenin was used as a comparison factor. In all other experiments, one of the experimental groups consisted of hematopoietic stem cells treated with angiogenin.

Olfactory dysfunction is an early marker of COVID-19 infection. However, individuals may develop chronic olfactory impairment for more than six months in 1-10 % of cases. The study's objective is to evaluate the efficacy and safety of intranasal immunotherapy using bioactive substances produced by M2 macrophages for the treatment of people with long-term post-COVID-19 hyposmia. Seven individuals with long-term persistent hyposmia (7 to 24 months), associated with PCR-confirmed coronavirus infection were evaluated for olfactory function at baseline, one, and six to twelve months after therapy. The intranasal inhalation of M2 macrophage conditioned medum (one time per day for 28-30 days) was well tolerated. Furthermore, olfactometry demonstrated that the patients restored their capacity to perceive (Kruskal-Wallis H test 14.123, p = 0.0009) and recognize odours (H =11.674, p = 0.0029). In addition, the subjective evaluation of smell significantly improved (H = 11.935, p = 0.0026). At the 6- to 12-month follow-up, the majority of patients (5/7) reported extremely high levels of satisfaction with the outcomes, and the remaining two patients also felt generally positive about the therapy's success. Overall, our study showed that the use of intranasal inhalations as a method of delivering bioactive factors and the conditioned medium of M2 macrophages as a therapeutic agent are both safe, well tolerated and, according to preliminary data, clinically effective in the treatment of patients with long-term postCOVID-19 hyposmia.

It is well-established that double-stranded RNA (dsRNA) exhibits noticeable radioprotective and radiotherapeutic effects. The experiments conducted in this study directly demonstrated that dsRNA was delivered into the cell in its native form and that it induced hematopoietic progenitor proliferation. The 68 bp synthetic dsRNA labeled with 6-carboxyfluorescein (FAM) was internalized into mouse hematopoietic progenitors, c-Kit+ (a marker of long-term hematopoietic stem cells) cells and CD34+ (a marker of short-term hematopoietic stem cells and multipotent progenitors) cells. Treating bone marrow cells with dsRNA stimulated the growth of colonies, mainly cells of the granulocyte–macrophage lineage. A total of 0.8% of Krebs-2 cells internalized FAM-dsRNA and were simultaneously CD34+ cells. dsRNA in its native state was delivered into the cell, where it was present without any signs of processing. dsRNA binding to a cell was independent of cell charge. dsRNA internalization was related to the receptor-mediated process that requires energy from ATP. Synthetic dsRNA did not degrade in the bloodstream for at least 2 h. Hematopoietic precursors that had captured dsRNA reinfused into the bloodstream and populated the bone marrow and spleen. This study, for the first time, directly proved that synthetic dsRNA is internalized into a eukaryotic cell via a natural mechanism.

In this work, we study where heating takes place during coronal mass ejections (CMEs). For this purpose, we have used the data of the Mg xii spectroheliograph on board the Complex Orbital Observations Near-Earth of Activity on the Sun (CORONAS)-F satellite. This instrument obtained images of the solar corona in the Mg xii 8.42 Å line, which emits only at temperatures higher than 4 MK. After analyzing the Mg xii data archive from 2001 to 2003, we found ten high-temperature eruptive events. Each of them was associated with a CME and nine were associated with a flare. The eruptive structures had temperatures higher than 4 MK and a characteristic size of 100 – 200 Mm. The events were observed by the Mg xii spectroheliograph for 10 min to 3 h. In the Mg xii images, the peak intensity of the eruptive structures was 0.2 – 14.4

Solar extreme ultraviolet (EUV) imaging instruments usually have a channel centered at 304 Å to observe the strong He ii 303.8 Å line, which is valuable for studying the dynamics of chromospheric and transition-region structures. In off-limb regions where He ii is weak, however, the coronal Si xi 303.3 Å line becomes significant and provides a background haze that reduces the contrast of He ii structures such as jets and macrospicules, complicating the interpretation of the observations. Generally, the separation of this background would require spectroscopic observations. In this article, we take an alternate approach by reconstructing the differential emission measure (DEM) of the quiescent corona to obtain synthetic radial emission profiles in the Si xi 303.3 Å line and show that at altitudes above 20 Mm it makes the major contribution to the background. We also find the silicon abundance to be significantly, by around 80

A biotechnology for personalized ex vivo gene therapy based on molecular genomic balancing of hematopoietic stem cell (HSC) chromatin with nucleosome monomers of human genomic DNA (hDNAnmr) has been developed and implemented in the clinic to change (to "correct") mutant chromosome loci genomes of dominant HSC clones that form mono- and oligoclonal hematopoiesis during aging and major (oncological, cardiovascular, neurodegenerative and autoimmune) fatal immune-mediated diseases of civilization. A fundamentally new biotechnological approach has been applied to the delivery of genetic material into eukaryotic stem and progenitor cells by establishing an artificial "recombinogenic situation" in them to induce homologous recombination (equivalent replacement) of mutant DNA regions with healthy hDNAnmr. In experimental preclinical trials, the effectiveness of genomic balancing technology has been proven to reduce the risk of sudden death in old animals and to increase the lifespan of outbred mice by 30% and Wistar rats by 57%. The improvement in their quality of life, compared with the control, is explained by an increase in the telomeric regions of the HSCs and HPCs chromosomes by 1.5-2 times. The potential of the technology to slow down the hereditary neurodegenerative diseases on the model of amyotrophic lateral sclerosis is shown. The effectiveness of this technology in clinical practice is presented on the example of a terminal patient with stage 4 neuroendocrine cancer. This technology used in the treatment of a number of oncological, neurodegenerative, autoimmune and hereditary diseases with clonal hematopoiesis is able to arrest the progression of the disease, prevent its recurrence, prolong the active life of a person, increase the average life expectancy and prevent sudden death.

Group-specific component macrophage-activating factor (GcMAF) is the vitamin D3-binding protein (DBP) deglycosylated at Thr420. The protein is believed to exhibit a wide range of therapeutic properties associated with the activation of macrophagal immunity. An original method for GcMAF production, DBP conversion to GcMAF, and the analysis of the activating potency of GcMAF was developed in this study. Data unveiling the molecular causes of macrophage activation were obtained. GcMAF was found to interact with three CLEC10A derivatives having molecular weights of 29 kDa, 63 kDa, and 65 kDa. GcMAF interacts with high-molecular-weight derivatives via Ca2+-dependent receptor engagement. Binding to the 65 kDa or 63 kDa derivative determines the pro- and anti-inflammatory direction of cytokine mRNA expression: 65 kDa—pro-inflammatory (TNF-α, IL-1β) and 63 kDa—anti-inflammatory (TGF-β, IL-10). No Ca2+ ions are required for the interaction with the canonical 29 kDa CLEC10A. Both forms, DBP protein and GcMAF, bind to the 29 kDa CLEC10A. This interaction is characterized by the stochastic mRNA synthesis of the analyzed cytokines. Ex vivo experiments have demonstrated that when there is an excess of GcMAF ligand, CLEC10A forms aggregate, and the mRNA synthesis of analyzed cytokines is inhibited. A schematic diagram of the presumable mechanism of interaction between the CLEC10A derivatives and GcMAF is provided. The principles and elements of standardizing the GcMAF preparation are elaborated.

To overcome immune tolerance to cancer, the immune system needs to be exposed to a multi-target action intervention. Here, we investigated the activating effect of CpG oligodeoxynucleotides (ODNs), mesyl phosphoramidate CpG ODNs, anti-OX40 antibodies, and OX40 RNA aptamers on major populations of immunocompetent cells ex vivo. Comparative analysis of the antitumor effects of in situ vaccination with CpG ODNs and anti-OX40 antibodies, as well as several other combinations, such as mesyl phosphoramidate CpG ODNs and OX40 RNA aptamers, was conducted. Antibodies against programmed death 1 (PD1) checkpoint inhibitors or their corresponding PD1 DNA aptamers were also added to vaccination regimens for analytical purposes. Four scenarios were considered: a weakly immunogenic Krebs-2 carcinoma grafted in CBA mice; a moderately immunogenic Lewis carcinoma grafted in C57Black/6 mice; and an immunogenic A20 B cell lymphoma or an Ehrlich carcinoma grafted in BALB/c mice. Adding anti-PD1 antibodies (CpG+αOX40+αPD1) to in situ vaccinations boosts the antitumor effect. When to be used instead of antibodies, aptamers also possess antitumor activity, although this effect was less pronounced. The strongest effect across all the tumors was observed in highly immunogenic A20 B cell lymphoma and Ehrlich carcinoma.

We studied angiogenin production by human macrophages and evaluated the role of this factor in the macrophage-mediated regulation of fibroblasts. All macrophage subtypes, and especially the efferocytosis-polarized macrophages, M2(LS), actively produced angiogenin. Exogenous recombinant angiogenin dose-dependently enhanced the proliferation and differentiation of dermal fibroblasts. The addition of the angiogenin inhibitor to fibroblasts cultures suppressed the stimulating effect of exogenous angiogenin or M2(LS) conditioned media. These findings indicate the involvement of angiogenin in the macrophage-mediated paracrine regulation of skin fibroblasts.

Background: Double-stranded fragmented extracellular DNA is a participant, inducer, and indicator of various processes occurring in the organism. When investigating the properties of extracellular DNA, the question regarding the specificity of exposure to DNA from different sources has always been raised. The aim of this study was to perform comparative assessment of biological properties of double-stranded DNA obtained from the human placenta, porcine placenta and salmon sperm.Methods: The intensity of leukocyte-stimulating effect of different dsDNA was assessed in mice after cyclophosphamide-induced cytoreduction. The stimulatory effect of different dsDNA on maturation and functions of human dendritic cells and the intensity of cytokine production by human whole blood cells was analyzed ex vivo. The oxidation level of the dsDNA was also compared.Results: Human placental DNA exhibited the strongest leukocyte-stimulating effect. DNA extracted from human and porcine placenta exhibited similar stimulatory action on maturation of dendritic cells, allostimulatory capacity, and ability of dendritic cells to induce generation of cytotoxic CD8+CD107a+ T cells in the mixed leukocyte reaction. DNA extracted from salmon sperm stimulated the maturation of dendritic cells, while having no effect on their allostimulatory capacity. DNA extracted from human and porcine placenta was shown to exhibit a stimulatory effect on cytokine secretion by human whole blood cells. The observed differences between the DNA preparations can be caused by the total methylation level and are not related to differences in oxidation level of DNA molecules.Conclusions: Human placental DNA exhibited the maximum combination of all biological effects.

This paper describes in detail a compact (2U format) telescope operating in the extreme ultraviolet range for studying the solar corona at a wavelength of 17.14 nm. The telescope objective has been built according to the Ritchey-Chrétien scheme with the following parameters: effective focal length of 381.3 mm, field of view of 2×2∘, and angular resolution of 11 in. Reflective multilayer Al/Be coatings were used, having 55% reflectance and a 0.4 nm spectral bandwidth. The wavefront rms error is 30 nm. An interferometric technique for controlling the shapes of the substrates and for adjusting the optical system and detector of the telescope assembly is described in detail.