Unfortunately, the demographic situation in modern Russia over the past 30 years leaves much to be desired and is one of the most discussed topics in the scientific community. According to current statistics, the male factor occupies up to 50% of the infertility structure today. And over the past 10 years, the number of diagnosed cases of male infertility has doubled. It is also worth mentioning the notorious events of recent years — the new coronavirus infection pandemic and a special military operation that are testing the demographic red lines to this day. From the viewpoint of medicine and demography, these events are associated with colossal reproductive losses and a decrease in already low birth rates for the state. One of the most relevant ways to raise the birth rate may be to expand the scope of assisted reproductive technologies (ART), including for male population. A promising ART procedure with a much more extensive list of indications for use than is commonly believed in our country is sperm cryopreservation, which has long ceased to be a "desperate method". This article presents possible indications for the use of sperm cryopreservation, as well as prospects and problems of introducing the method into the practice of healthcare in the Russian Federation. KEYWORDS: sperm cryopreservation, male infertility, cryotrauma, special military operation, ART. FOR CITATION: Faniev M.V., Prokopiev Ya.V., Kadyrov Z.A., Faustova K.V., Cherepnev G.V., Mazitova M.I., Antropova E.Yu. Problems and prospects of sperm cryopreservation in modern Russia. Russian Medical Inquiry. 2024;8(4):215–220 (in Russ.). DOI: 10.32364/2587-6821- 2024-8-4-5.

Hemorrhagic fever with renal syndrome (HFRS) is endemic in Tatarstan, where thousands of cases are registered annually. Puumala orthohantavirus is commonly detected in human case samples as well as in captured bank voles, the rodent hosts. The pathogenesis of HFRS is still not well described, although the cytokine storm hypothesis is largely accepted. In this study, we present a comprehensive analysis of a fatal HFRS case compared with twenty four non-fatal cases where activation of the humoral and cellular immune responses, pro-inflammatory cytokines and disturbed blood coagulation were detected using immunological, histological, genetic and clinical approaches. Multiple organ failure combined with disseminated intravascular coagulation syndrome and acute renal failure was the cause of death. Decreased Interleukin (IL)-7 and increased IL-18, chemokine (C-C motif) ligand (CCL)-5, stem cell growth factor (SCGF)-b and tumor necrosis factor-beta (TNF-β) serum levels were found, supporting the cytokine storm hypothesis of hantavirus pathogenesis.

Multiple sclerosis (MS) is an autoimmune neurodegenerative disease characterized by chronic brain inflammation. Leukocyte infiltration of brain tissue causes inflammation, demyelination, and the subsequent formation of sclerotic plaques, which are a hallmark of MS. Activation of proinflammatory cytokines is essential for regulation of lymphocyte migration across the blood–brain barrier. We demonstrate increased levels of many cytokines, including IL-2RA, CCL5, CCL11, MIF, CXCL1, CXCL10, IFNγ, SCF, and TRAIL, were upregulated in cerebrospinal fluid (CSF), whereas IL-17, CCL2, CCL3, CCL4, and IL-12(p40) were activated in MS serum. Interaction analysis of cytokines in CSF demonstrated a connection between IFNγ and CCL5 as well as MIF. Many cells can contribute to production of these cytokines including CD8 and Th1 lymphocytes and astrocytes. Therefore, we suggest that IFNγ released by Th1 lymphocytes can activate astrocytes, which then produce chemoattractants, including CCL5 and MIF. These chemokines promote an inflammatory milieu and interact with multiple chemokines including CCL27 and CXCL1. Of special note, upregulation of CCL27 was found in CSF of MS cases. This observation is the first to demonstrate CCL27 as a potential contributor of brain pathology in MS. Our data suggest that CCL27 may be involved in activation and migration of autoreactive encephalitogenic immune effectors in the brain. Further, our data support the role of Th1 lymphocytes in the pathogenesis of brain inflammation in MS, with several cytokines playing a central role.

Based on the active ingredient of the drug Ximedon (1,2-dihydro-4,6-dimethyl-1- N -(2-hydroxyethyl)pyrimidone-2, referred below to as pyrimidine ( I ), novel derivatives containing biogenic acids: succinic, L-ascorbic, para -aminobenzoic, nicotinic, and L-2-amino-4-(methylthio)butanoic (L-methionine) acids have been synthesized. The parameters of acute toxicity (LD 50 ) have been studied. The antitoxic effect of the compounds upon the injury by the hepatotropic poison carbon tetrachloride has been examined as the primary evaluation of their hepatoprotective properties. It has been found that, according to toxicological safety, the compounds synthesized belong to classes III and IV (moderately and little toxic compounds). The conjugates of pyrimidine ( I ) with ascorbic acid and methionine (LD 50 more than 5400 mg/kg) are least toxic. Pyrimidine ( I ) and its derivatives possess the antitoxic activity upon acute poisoning with carbon tetrachloride; the combined injection of carbon tetrachloride with pyrimidine ( I ) or its derivatives leads to an increase in the survival of animals and the normalization of the integral functional parameters, weight and body temperature, which decrease upon toxic injury. In addition, pyrimidine ( I ) and some of its derivatives (conjugates with L-ascorbic, succinic, para -aminobenzoic, and nicotinic acids) decrease the weight coefficients of the liver and kidneys (the organ-to-body-weight ratio) and the activity of transaminases, the markers of hepatic cytolysis, which increase upon toxic injury with carbon tetrachloride. The area of the pathological injury of the liver by steatosis and necrosis decreases by the action of pyrimidine ( I ) and its novel derivatives (conjugates with L-ascorbic, succinic, and nicotinic acids) two to three times. Advantages of pyrimidine ( I ) and its novel derivatives over the hepatoprotective drug Thiotriazolin have been revealed.

Hepatoprotective properties of a new pyrimidine derivative - L-ascorbate 1-(2-hydroxyethyl)-4,6-dimethyl-1,2-dihydropyrimidine-2-one, synthesized on the basis Xymedon, were assessed in white rats exposed to CCl4. The compound under study administered prior to exposure to CCl4 reduced the deviation of biochemical parameters from reference values and severity of structural and morphological changes in liver, when compared to the control. Hepatoprotective properties of the studied compound were more pronounced than those of Xymedon.

A comparative study of the physical adsorption of RNAse A and RNAse Bacillus pumilis onto a negatively charged surface of mica, a hydrophobic surface of pyrolytic graphite and a surface of lipid layers of dipalmitoylphosphatidylcholine (DPPC) was performed by atomic force microscopy. It was found that microbial RNAse, unlike RNAse A, 1) is adsorbed onto the negatively charged surface of mica in the form of monomers and dimers; 2) exhibits enhanced tropism to the hydrophobic surface of pyrolytic graphite; 3) modifies morphotopography and thickness of the lipid bilayer of DPPC. The detected surface-dependent differences in adsorption of RNAses are consistent with the features of their structure and cytotoxic properties.

Transmembrane potential of mitochondria is a sensitive biomarker of metabolic activity of cells. Here, we studied mitochondrial potential psi m in Yarrowia lipolytica yeast cells treated with two fullerene C-60 derivatives: bis-nitroxide methanofullerene and 3-phospho-pentafullerene acid. Transmembrane mitochondrial potential was measured by vital ratiometric cationic fluorochrome JC-1 using flow cytometry. The fullerene C-60 derivatives tested in a concentration of 10 mu g/ml developed cytoprotective effect in the yeast cells challenged either with non-ionic detergent tween-80, or Tris-buffer, pH 9.0. Treatment with bis-nitroxide methanofullerene resulted in a 6-fold increase in proportion of cells with high psi m, while 3-phospho-mentafullerene acid evoked a 1,5-fold increase in this subset compared to the stressed cells. Hence, both fullerene derivatives counteract psi m dissipation in challenged cells.

We developed a rapid method to study Yarrowia lipolytica single yeast cell mitochondrial responses based on flow cytometry and potential-sensitive ratiometric fluorochrome JC-1. The proposed model has several advantages as it allows to measure mitochondrial membrane potential in intact cells (without isolation of mitochondria), has an internal positive control (uncoupler of oxidative phosphorylation CCCP), and produces a normalized measurement of the transmembrane mitochondrial potential, taking into account the heterogeneity of cell size and number of mitochondria in a single cell. The similarity of the mitochondrial complex I structure in mammals and Yarrowia lipolytica opens prospects for the model implementation in search for human bioenergetic response modifiers.

We developed a rapid method to study Yarrowia lipolytica single yeast cell mitochondrial responses based on flow cytometry and potential-sensitive ratiometric fluorochrome JC-1. The proposed model has several advantages as it allows to measure mitochondrial membrane potential in intact cells (without isolation of mitochondria), has an internal positive control (uncoupler of oxidative phosphorylation CCCP), and produces a normalized measurement of the transmembrane mitochondrial potential, taking into account the heterogeneity of cell size and number of mitochondria in a single cell. The similarity of the mitochondrial complex I structure in mammals and Yarrowia lipolytica opens prospects for the model implementation in search for human bioenergetic response modifiers.

The aim of our work was to assess long-term results of autologous peripheral blood hematopoietic stem cell application in patients with peripheral arterial diseases. Peripheral blood hematopoietic stem cells mobilized by granulocyte colony-stimulating factor were transplanted intramuscularly to 30 patients with peripheral arterial diseases (Ilb stage by Pokrovsky). Standart tredmill test, ankle-brachial index estimation and ankle-brachial index restoration time estimation after loading were performed on 0, 3, 6, 12 and 60 months after transplantation. Immunohistochemical study of injured gastrocnemius muscle biopsies taken before peripheral blood hematopoietic stem cells transplantation and 3 months after the procedure was performed. Peripheral blood hematopoietic stem cells transplantation increases capillary density (22,4%, p = 0,0005). Ankle-brachial index increased by 18,1% on month 6 after transplantation without a tendency to change on month 12. 60 months after transplantation initial to hematopoietic stem cells transplantation ankle-brachial index rates were marked. Painless walking distance was increasing at all times of observation progressively, on month 60 no walking distance limitation was marked by most patients. Ankle-brachial index restoring time shows positive trend of the functional state of limb during the first year after transplantation, 60 months after transplantation it showed no «walking reserve» limitation in most patients. 5-years survival was 79%, death causes were stroke, cardiac pathology (3 cases), lung cancer. So, peripheral blood hematopoietic stem cells transplantation allows eliminating peripheral arterial diseases symptoms and preserving limb in long-term period. Autologous transplantation of peripheral blood hematopoietic stem cells has no complications and is safe for therapy of patients with peripheral arterial diseases II stage.

The aim of our work was to assess long-term results of autologous peripheral blood hematopoietic stem cell application in patients with peripheral arterial diseases. Peripheral blood hematopoietic stem cells mobilized by granulocyte colony-stimulating factor were transplanted intramuscularly to 30 patients with peripheral arterial diseases (IIb stage by Pokrovsky). Standart tredmill test, ankle-brachial index estimation and ankle-brachial index restoration time estimation after loading were performed on 0, 3, 6, 12 and 60 months after transplantation. Immunohistochemical study of injured gastrocnemius muscle biopsies taken before peripheral blood hematopoietic stem cells transplantation and 3 months after the procedure was performed. Peripheral blood hematopoietic stem cells transplantation increases capillary density (22,4%, p = 0,0005). Ankle- brachial index increased by 18,1% on month 6 after transplantation without a tendency to change on month 12. 60 months after transplantation initial to hematopoietic stem cells transplantation ankle-brachial index rates were marked. Painless walking distance was increasing at all times of observation progressively, on month 60 no walking distance limitation was marked by most patients. Ankle-brachial index restoring time shows positive trend of the functional state of limb during the first year after transplantation, 60 months after transplantation it showed no «walking reserve» limitation in most patients. 5-years survival was 79%, death causes were stroke, cardiac pathology (3 cases), lung cancer. So, peripheral blood hematopoietic stem cells transplantation allows eliminating peripheral arterial diseases symptoms and preserving limb in long-term period. Autologous transplantation of peripheral blood hematopoietic stem cells has no complications and is safe for therapy of patients with peripheral arterial diseases II stage.

The exact mechanism by which cytotoxic ribonucleases reach the cytosol of tumor cells remains unclear. The interaction of ribonucleases with a lipid bilayer is involved in the translocation of ribonucleases across the endosomal membrane. Here, we aimed to study the hydropathy character of toxic antitumor ribonucleases (bovine seminal ribonuclease and binase) and two non-toxic ribonucleases (bovine pancreatic ribonuclease and human pancreatic ribonuclease) by sliding-window hydrophobicity analysis. Comparative hydropathy plot analysis of the non-toxic pancreatic ribonucleases and their toxic variants was also performed. The data obtained indicate that some cytotoxic ribonucleases have a hydrophobic segment, which is sterically available for the hydrophobic interaction with a tumor cell membrane and endosomal membrane. After dissociation, subunits of dimeric ribonucleases are probably capable of thermodynamically favorable interaction with the interfacial region of a lipid bilayer. Remarkably the hydrophobic segment is not identified in the amino acid sequences of non-toxic ribonucleases. The paper describes the hydrophobic properties of toxic RNases that are essential for both the model of a lipid-protein interaction and the cytotoxicity mechanism unraveling.

Preliminary incubation of peritoneal macrophages with binase in a concentration of 100 mkg/ml (and more) results in the reduction of the macrophages' death by necrosis induced by hydrogen peroxide. In this case the cells mostly enter apoptosis. The protection effect of binase may be a consequence of a stabilizing effect of binase on cytoplasmic membrane. The possibility of the lipid bilayer stabilization is indicated by the change in DPPC phase transition parameters in the presence of binase, which have been determined in a model experiment by differential scanning calorimetry. However, the influence of binase on the transmission of H2O2-induced pro-apoptotic signals should not be ruled out.

Изучено влияние двух концентраций (40 и 400 мкг/мл) биназы (РНКазы Bacillus intermedius) на индукцию активных форм кислорода в моноцитах и гранулоцитах венозной крови человека методом лазерной проточной цитофлуориметрии. Установлено, что биназа избирательно действует на E. coli- и форболмиристатацетат-зависимый окислительный взрыв в гранулоцитах и не влияет на эти процессы в моноцитах. Профиль ответа гранулоцитов зависит от используемого индуктора окислительного взрыва и концентрации биназы. В условиях окислительного взрыва, стимулированного E. coli, биназа концентрационно зависимо увеличивает долю клеток, продуцирующих АФК, и интенсивность генерации АФК на клетку. В гранулоцитах, in vitro стимулированных ФMA, наблюдается тенденция к разнонаправленному концентрационно зависимому действию биназы на интенсивность продукции АФК на клетку.