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Introduction. The use of thrombopoietin receptor agonists, especially eltrombopag, in the treatment of aplastic anemia (AA) patients who did not respond to the previous immunosuppressive therapy (IST), is accompanied by the development of a hematological response in 40–60 % of patients.Aim — to study the effi cacy of using eltrombopag in treatment programs for AA patients refractory to previous IST.Methods. The study included 20 AA patients who were treated at the National Research Center for Hematology from 2015 to 2020. These patients did not respond to the conducted IST (ATG + CsA). Eltrombopag was administered at a dose of 150 mg/day. The results of treatment were assessed at 3 and 6 months: the achievement of hematological improvement, partial and complete remission, as well as the identifi cation of possible clonal evolution were determined.Results. Eleven out of 20 (55 %) patients responded to treatment: 2 patients developed hematological improvement, 6 patients — partial remission, 3 patients — complete remission. All 11 patients responded to treatment within 12 months from the start of eltrombopag, but further positive dynamics of hematological parameters are possible. The median duration of treatment with eltrombopag was 11 (1–48) months. Most of the patients were treated with eltrombopag in combination with CsA. The duration of the course of treatment with eltrombopag depended on the response received (stable hematological improvement, remission, as well as the detection of clonal evolution) or its absence and the need for ATG or BMT. An aberrant karyotype was found in 2 AA patients who received eltrombopag: in one patient monosomy of chromosome 7 was detected 1 month after the start of treatment, in another patient, 37 months later, a clone with a derivative of chromosome 16 from t(1;16) and subclone with complex disorders of the karyotype without signs of myelodysplasia in the bone marrow.Conclusion. The inclusion of the TPO receptor agonist eltrombopag in the treatment program for AA patients allows for a stable hematological response and remission of AA for patients who have not responded to IST. The effectiveness of eltrombopag is determined by adherence to the treatment algorithm, the optimal duration of the course, and the dose of the drug used. There is still a need for long-term observation of the patient and control morphological and cytogenetic studies.

The paper proposes a methodological approach to the development of a model for assessing the investment attractiveness of innovative projects based on the provisions of fuzzy logic and set theory. Methods and models for evaluating the investment project effectiveness are described. The limitations of existing methods due to the uncertainty of the external environment are presented. An example of a quantitative assessment of a specific innovative project investment attractiveness is given.

Autoantibodies, immunoglobulins G (IgG) againstthe desmosomal proteins desmogleins 1 and 3, play a significant role in the pathogenesis of pemphigus vulgaris. The basic therapy for pemfigus includes systemic corticosteroids, but their use should be as briefas possible because of the severe side effects. In cases of corticosteroid-resistant pemfigus, adjuvant therapy, in particularextracorporeal methods, is used. The most effective and safest extracorporeal therapy is immunosorbtion. Immunosorbtion is based on the removal of pemphigus antibodies from the blood using an affinity sorbent during a therapeutic apheresis procedure. Existing immunosorbents are nonselective and increase the risk of infection. We designed an immunosorbent based on an agarose matrix, Affi-Gel 15, and human recombinant desmoglein 3, as aligand, for a selective removal of autoantibodies from pemphigus patients sera. It was shown on a pemphigus experimental modelin vivo(neonatal Balb/c mouse model) andin vitrothat the immunosorbent can effectively remove desmoglein 3-associated autoantibodies. The experimental results demonstrate that the solid-phase matrix immunosorbent Affi-Gel 15Dsg3 is a promising product for the development of pemphigus therapy.

The article summarizes information about the actinic keratosis. Presented data of the literature about the prevalence of the disease, the features of the clinical picture, diagnosis, differential diagnosis and treatment of dermatosis.

Methylenecarboxamide (glycine) morpholine oligomers (gMOs) with a modified backbone are new and promising nucleic acid analogues. In this work, a combination of circular dichroism spectroscopy, optical melting, and molecular dynamics simulations was used to investigate hybridization properties of gMOs, as well as the structure and dynamics of their tandem complexes with DNA and RNA. It was shown that the structure of nucleic acids in modified complexes is similar to that of the fully native analogues. The energies of binding and cooperative interactions at the helix-helix interface in the nick were determined experimentally and by computer simulation analysis. Here, we found for the first time, the possibility to determine and predict precisely the thermodynamic parameters of complementary complex formation using the original experimental and computer simulation approaches. It was shown that the use of simulation data in the explicit solvent and the molecular mechanics Poisson-Boltzmann (or generalized Born) surface area methods for the calculation of the hybridization enthalpy makes it possible to evaluate the thermal stability of DNA and gMO tandem duplexes with DNA or RNA with an unexpectedly high accuracy. We found that at high ionic strength and neutral pH, the observed thermal stability of the gMO/RNA tandem complex is similar to that of DNA/DNA and lower than that of gMO/DNA which is close to that of DNA/RNA.

Ivanenko T., Hrekova T., Abramova T., Shiryaeva A., Vasilenko G. Influence of experimental metabolic syndrome on bioimpedance spectroscopy indicators of rat body. Journal of Education, Health and Sport. 2021;11(08):470-479. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2021.11.08.053 https://apcz.umk.pl/JEHS/article/view/JEHS.2021.11.08.053 https://zenodo.org/record/5559742 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2021; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 02.08.2021. Revised: 16.08.2021. Accepted: 31.08.2021. UDC: 616-008.9-092:616-092-055.1/.2]-073:577.352.2]-092.9 INFLUENCE OF EXPERIMENTAL METABOLIC SYNDROME ON BIOIMPEDANCE SPECTROSCOPY INDICATORS OF RAT BODY T. Ivanenko, T. Hrekova, T. Abramova, A. Shiryaeva, G. Vasilenko Zaporizhzhia State Medical University, Zaporozhzhia, Ukraine Ivanenkotv@i.ua Abstract The objective of this experiment was to establish sex-related differences in the parameters of bioimpedance spectroscopy of the Wistar rat’s bodies in the experimental metabolic syndrome (endocrine-salt model). Sexual dependencies of bioimpedance measurements in intact animals have been determined for the first time: males have higher amount of total fluid, but lower one of total fat. The intra/extracellular fluid balance in males is characterized with ratio 2:1, while in females is the other one - 1:1. For the first time the formation of the metabolic syndrome has already been determined on the seventh day in females which lead to decreasing in the percentage of total fat and to changing of the intra / extracellular fluid balance to ‘male’ type - 2:1. The last one should be considered as a sign of intracellular hyperhydration. In males the examined parameters have been being within the control values throughout 21 days of experiment. Key words: bioimpedance spectroscopy; sex-related differences; metabolic disorders; corticosteroids; rats.

BODIPY dyes are photostable neutral derivatives of 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene. These are widely used as chemosensors, laser materials, and molecular probes. At the same time, BODIPY dyes have small or moderate Stokes shifts like most other fluorophores. Large Stokes shifts are preferred for fluorophores because of higher sensitivity of such probes and sensors. The new boron containing BODIPY dye was designed and synthesized. We succeeded to perform an annulation of pyrrole ring with coumarin heterocyclic system and achieved a remarkable difference in absorption and emission maximum of obtained fluorophore up to 100 nm. This BODIPY dye was equipped with linker arm and was functionalized with a maleimide residue specifically reactive towards thiol groups of proteins. BODIPY residue equipped with a suitable targeting protein core can be used as a suitable imaging probe and agent for Boron Neutron Capture Therapy (BNCT). As the most abundant protein with a variety of physiological functions, human serum albumin (HSA) has been used extensively for the delivery and improvement of therapeutic molecules. Thiolactone chemistry provides a powerful tool to prepare albumin-based multimodal constructions. The released sulfhydryl groups of the homocysteine functional handle in thiolactone modified HSA were labeled with BODIPY dye to prepare a labeled albumin-BODIPY dye conjugate confirmed by MALDI-TOF-MS, UV-vis, and fluorescent emission spectra. Cytotoxicity of the resulting conjugate was investigated. This study is the basis for a novel BODIPY dye-albumin theranostic for BNCT. The results provide further impetus to develop derivatives of HSA for delivery of boron to cancer cells.

Natural and modified nucleoside polyphosphates are indispensable tools in modern biological investigations.1–4 Their applications include but are not limited to polymerase chain reactions (PCR), si...

In accordance with Russian Federal program of import substitution of foreign medicines quality of Russian drugs in the N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia, played vincristine-RONC (VC-RONC), which as a drug – the generic’s passed preclinical pharmacological and toxicological testing in comparison with foreign firms vincristine Teva of Israel (VC-Teva). The aim. The aim of present study was the comparative pathomorphological evaluation of the effect of VC-RONC and VC-Teva on the internal organs of rats. Materials and methods. Used 50 weinbrenner male rats, at 10 rats per group. VC-RONC and VC-Teva rats were administered intravenously 3 times daily at aquatoxicity total dose corresponding to the MTD and 1/2 MTD. Control rats in the same regime intravenously administered of 0,9 % sodium chloride solution. Rats were deduced from the experience of 3 and 30 days after the end of administration of the drugs. Conducted macroscopic and histological examination of internal organs by conventional methods, including fixation of the material in 10 % formalin and coloring sections with hematoxylin and eosin. The micropreparations of the internal organs was analyzed under light microscope at magnifications of 100, 400, 1000. Results. VC-RONC, as VC-Teva in cumulative doses of 0,5 and 0,25 mg/kg in 3 days after the end of introductions in the internal organs of rats cause similar slightly pronounced morphological changes: hypoplasia in the bone marrow and spleen, destructive changes in the testes, focal degenerative changes in the kidney and liver of individual rats. On the 30th day after the application of both drugs, some rats regardless of the dose occurred similar symptoms residual morphological changes in the bone marrow, testes, kidneys and liver. Conclusion. Based on the results of macroscopic and histological examination the conclusion about the similarity of the influence of VC-RONC and VC-Teva on the internal organs of rats was made.

We have studied the excision efficiency of human apurinic/apyrimidinic endonuclease 1 (APE1) and tyrosyl-DNA phosphodiesterase 1 (TDP1) on matched or mismatched bases located at the 3' end of DNA primers. We have used model DNA duplexes, which mimic DNA structures that occur during either replication (DNA with a 3' recessed end) or repair (DNA with a single-strand break). Both APE1 and TDP1 are more efficient in removing ribose-modified dNMP residues from mismatched pairs rather than canonical pairs. Thus, both of these enzymes may act as proofreading factors during the repair synthesis catalyzed by DNA polymerases including DNA polymerase β (Polβ). The design of new DNA polymerase inhibitors, which act as DNA or RNA chain terminators, is one of the main strategies in the development of antiviral agents. The excision efficacy of APE1 and TDP1 has also been studied for 3'-modified DNA duplexes that contain ddNMP or phosphorylated morpholino nucleosides (MorB) commonly used as terminators in the DNA synthesis. We have also investigated the insertion of ddNTP and morpholino nucleotides catalyzed by Polβ and human immunodeficiency virus reverse transcriptase. This experiment has pointed to MorCyt, cytosine-containing morpholino nucleoside, as a potential antiviral agent.

Morpholino derivatives of nucleosides are widely used in medicine and biology as monomers in the synthesis of DNA or siRNA analogues. They are considered to be promising agents for anticancer,1 ant...

Three novel uracil-based morpholine nucleosides each containing three different orthogonally reactive functional groups, namely, amino, carboxy and acetylene ones, were synthesized. The obtained monomers are intended for the synthesis of labeled nucleotide, nucleic acid or peptide mimics.

The article provides an overview of specialized food products intended for young children suffering from food allergies.

True (acantholytic) pemphigus is an autoimmune disease characterized by blisters and erosions in the skin and/or mucous tunics. True (acantholytic) pemphigus can be diagnosed based on an assessment of clinical manifestations and results of cytology, histology and immunology tests. The current diagnostics methods are characterized by certain advantages and shortcomings yet none of them has 100% sensitivity and specificity. To diagnose pemphigus, a complex of studies is needed taking into consideration the body of clinical symptoms and laboratory indices.

Objective : to assess the vitamin status of preschoolers according to the results of one-hour urinary vitamin excretion and the data of actual nutrition. Subjects and methods . A survey was conducted in winter 2016, covering 33 children (19 boys and 14 girls) aged from 2 to 7 years who went to a Yekaterinburg preschool educational institution. The amount of essential nutrients was calculated according to the data of actual nutrition during 5 days. The vitamin status was evaluated from the urinary excretion of thiamine, riboflavin, and 4-piridoxyl acid, by testing a morning fasting urine portion. Results. Determination of urinary excretion revealed deficiency of vitamin B2 in 24.2% of the examinees, and that of vitamins B1 and B6 in significantly more children (in 66.7–69.7%). According to the actual nutrition data, inadequate intakes of vitamins B2, A, and C were detected in 27.3–30.3% of the examinees, those of niacin and vitamin B1 in 54.5 and 69.7%, respectively. The proportion of the coincidental data obtained by two methods for assessing the vitamin status was 63.6% for vitamin B1 and 69.7% for vitamin B2. Conclusion . The findings suggest that the two methods to assess the supply of vitamins B1 and B2 are interchangeable and the children need additional vitamin intakes.

The paper gives the comparative characteristics of the mechanism of action of fermented milk drinks and probiotic foods. It deals with the results of investigations of the efficacy of probiotic microorganisms in infant nutrition. The issues of supplementing the Russian infant foods with probiotic Bifidobacterium BB-12 strains are considered. The paper presents the data that show that despite the considerable similarity of the mechanisms of action of probiotic foods and fermented mild drinks, their high nutritional value, and organoleptic properties, their final clinical effect may vary greatly. It also gives the results of a number of studies of the comparative clinical efficacy of fermented milk drinks and probiotic foods in 3–14-year-old children with acute enteric infections, which in combination with the data available in the literature suggest that there is a need for personalized recommendations on the use of the above foods in infant nutrition.

A convenient approach has been proposed to the synthesis of nicotinamide adenine dinucleotide (NAD+) mimetics, which comprise morpholino analogues of nucleosides. The approach is based on the use of ADP conjugates containing an amino group, which is tethered to the terminal phosphate through the aliphatic linker by the phosphodiester bond. We have synthesized four series of the NAD+ mimetics, which differ in the type of the modified nucleoside (2-aminomethylmorpholine (Mor) or 2-aminomethyl-4-carboxymethylmorpholine (MorGly) derivatives), in the linker length, and in the manner of the nucleoside attachment to the ADP derivative. We have studied the efficiency of NAD+ mimetics in the inhibition of the auto-poly(ADP-ribosyl)ation by the PARP 1 enzyme. The linker length, the mode of the attachment of the morpholino nucleoside analogue, and the nature of the heterocyclic base of the modified nucleoside were shown to inf luence the inhibition efficiency.

Bullous dermatoses are the diseases characterized by vesication and erosion of skin and/or mucos coats, production of autoantibodies against the structural components of epidermis and/or derma. The most frequent diseases of these disorders are pemphigus, bullous pemphigoid, dermatitis herpetiformis, beningn familial chronic pemphigus (Gougerot Hailey-Hailey disease).Diagnostics of bullous dermatoses is based on assessement of clinical signs, results of cytological and histological studies and immunological tests. However none of the current diagnostic methods allows to reveal bullous dermatosis severally and with absolute certainty. Consequently it is necessary to perform wide range of clinical and laboratory tests for early diagnosis and treatment order.

Network traffic monitoring on the basis of sequential and associative–sequential principles of current- information search is assessed in terms of efficiency. Associative–sequential search for information in network traffic permits increased productivity and faster decision making during searches in large information spaces.

The paper comparatively analyzes the nutritive value of fresh squeezed and reconstituted juices intended for infant feeding during the first year of fife. It presents laboratory data on the content of the trace elements copper and chromium in the fresh squeezed juices under the brand name of FrutoNyanya. Requirements for their raw materials, manufacture and technology, as well as a quality control system for baby juices and nectars are provided. Practical recommendations are given for the use of juices in the feeding of infants during the first year of fife.

Extramedullary lesions in multiple myeloma occur on the extension of neoplastic plasma cells outside the cortical bone. On the local tumor growth the extramedullary mass are found most commonly in vertebrae, ribs, sternum and skull. Single or multiple malignant mass can be found in the skin, liver, breast, kidney and lymph nodes via hematogenous spread of the malignant plasma cells. Incidence of extramedullary lesions in multiple myeloma patients is 7 to 18% at MM diagnosis and up from 2.3 to 20% at relapse or progression. In the last decade, we noted the increase of extramedullary lesions diagnosis at the beginning of the disease that was associated with the wide use of modern x-ray diagnostic methods. Thus, in 2009 the International Myeloma Working Group (IMWG) recommended undergoing the magnetic resonance imaging scans of the spine and pelvis, especially in the absence of osteolytic lesions according to the results of x-ray study. The results of computer tomography allow more accurately find the localization and the sizes of extramedullary mass. Nowadays, it is widely discussed the advantages of positron emission tomography using not only at the beginning of the disease, but also during the treatment, assessing anti-tumour response. The application of standard therapy in case of extramedullary plasmocytomas at the beginning of MM is associated with the reduction of both overall and progression-free survival. Several articles indicate the high efficiency of the programs including bortezomib, there are also some reports concerning the successful application of lenalidomide and pomalidomide in the treatment of MM associated with presence of extramedullary plasmocytomas.

We report on the design, synthesis and molecular modeling study of conjugates of adenosine diphosphate (ADP) and morpholino nucleosides as potential selective inhibitors of poly(ADP-ribose)polymerases-1, 2 and 3. Sixteen dinucleoside pyrophosphates containing natural heterocyclic bases as well as 5-haloganeted pyrimidines, and mimicking a main substrate of these enzymes, nicotinamide adenine dinucleotide (NAD+)-molecule, have been synthesized in a high yield. Morpholino nucleosides have been tethered to the β-phosphate of ADP via a phosphoester or phosphoramide bond. Screening of the inhibiting properties of these derivatives on the autopoly(ADP-ribosyl)ation of PARP-1 and PARP-2 has shown that the effect depends upon the type of nucleobase as well as on the linkage between ADP and morpholino nucleoside. The 5-iodination of uracil and the introduction of the P−N bond in NAD+-mimetics have shown to increase inhibition properties. Structural modeling suggested that the P−N bond can stabilize the pyrophosphate group in active conformation due to the formation of an intramolecular hydrogen bond. The most active NAD+ analog against PARP-1 contained 5-iodouracil 2ʹ-aminomethylmorpholino nucleoside with IC50 126 ± 6 μM, while in the case of PARP-2 it was adenine 2ʹ-aminomethylmorpholino nucleoside (IC50 63 ± 10 μM). In silico analysis revealed that thymine and uracil-based NAD+ analogs were recognized as the NAD+-analog that targets the nicotinamide binding site. On the contrary, the adenine 2ʹ-aminomethylmorpholino nucleoside-based NAD+ analogs were predicted to identify as PAR-analogs that target the acceptor binding site of PARP-2, representing a novel molecular mechanism for selective PARP inhibition. This discovery opens a new avenue for the rational design of PARP-1/2 specific inhibitors.

The development of new derivatives and analogues of nucleic acids for the purposes of molecular biology, biotechnology, gene diagnostics, and medicine has been a hotspot for the last two decades. Methylenecarboxamide (glycine) morpholine oligomer analogues (gM) seem to be promising therapeutic candidates because of the ability to form sequence specific complexes with DNA and RNA. In this paper we describe new approaches to the determination of thermodynamic parameters for hybridization of tandem oligonucleotide complexes with the complementary template. It makes possible to determine changes in enthalpy and entropy corresponding to the binding of an individual oligomer with the template, and to the formation of cooperative contact at the helix-helix interface of two neighboring duplex fragments (in the nick). We have experimentally analyzed the series of model tandem complexes of different length at various oligomer concentrations, ionic strength, and pH. The analysis of thermodynamic parameters of complex formation for native and modified oligomers revealed higher Gibbs free energy values of hybridization and cooperative interaction of morpholine-containing complexes at the helix-helix interface under standard conditions (1 M NaCl, pH 7.2). Further comparative analysis of the hybridization properties of modified oligomers at ionic strength and pH allows us to determine the charge state of the morpholine backbone and the thermodynamic origin of the effects observed. It was found that the decrease in pH to 5.5 led to the protonation of internal morpholine nitrogens. The obtained results prove the veracity of the proposed model and the possibility to evaluate thermodynamic parameters of short native and modified oligomers with high accuracy.

The fundamental roles of nucleoside triphosphates and nucleotide cofactors such as NAD(+) in biochemistry are well known. In recent decades, continuing research has revealed the key role of 5'-capped RNA and 5',5'-dinucleoside polyphosphates in the regulation of vitally important physiological processes. Last but not least, the commercial potential of nucleoside triphosphate synthesis can hardly be overestimated. Nevertheless, despite decades of investigation and the obvious topicality of the research on the chemical synthesis of the nucleotide compounds containing phosphate anhydride linkages, none of the existing procedures can be considered an up-to-date "gold standard". However, there are a number of fruitful synthetic approaches to forming phosphate anhydride linkages in satisfactory yield. These are summarized in this concise review, organized by the type of active phosphorous intermediate and reagents used.

Pemphigus vulgaris is a chronic autoimmune bullous disease characterized by the formation of blisters on the skin and/ or mucous tunics as a result of acantholysis. To search for new molecular and biological targets, study pathogenetic mechanisms of the disease development and develop new methods of treatment, it is urgent to create an experimental model of pemphigus in laboratory animals reproducing clinical, histological and immunological signs of pemphigus. Goal of the study. To develop an experimental model of pemphigus by injecting IgG produced from the blood serum taken from patients with pemphigus to neonatal mice of the BALB/c inbred line. Results. Accumulated IgG products taken from patients with pemphigus (main groups) and healthy volunteers (control group) were injected intraperitoneally to neonatal mice of the BALB/с in the doses of 10-30 mg per mouse. Clinical, histological and immunomorphological signs of pemphigus were revealed in the mice from the main group, which received intraperitoneal injections of IgG taken from patients with pemphigus in the dose of 30 mg per mouse. No signs of pemphigus were observed in the mice from the control group, which received injections of IgG taken from healthy people. This study confirms the role of pemphigus autoantibodies in the pathogenesis of pemphigus vulgaris and shows that passive transmission of antibodies to laboratory animals is possible.

Nutrition is of particular importance in young children who are characterized by exceptionally high growth rates in conjunction with scare resources of nutrients in their body. The physiological features of infants suggest that this active life period in the child requires adequate attitude towards his nutrition that largely differs from that during the first year of life. The paper considers the issues of organizing the nutrition of children aged 1 to 3 years, by using commercial infant dairy products — new combined products — sterilized milk- and fruit-milk-based desserts and milk cocktails for infant feeding.

The Far-Eastern wildcat (Prionailurus bengalensis euptilurus) is a rare and poorly investigated nondomestic felid species. An attempt of freezing and cryopreserving Far-Eastern wildcat spermatozoa in CaniPlus Freeze (CPF) medium is reported. Sperm was collected by electroejaculation from five adult Far-Eastern wildcat captive-born males. Epididymal spermatozoa from five adult randomly bred domestic cat males were used as a reference. The viability of frozen-thawed spermatozoa evaluated by double staining with SYBR Green I and PI followed by the subsequent confocal laser scanning microscopy (CLSM) was 38.2% ± 3.0% for the domestic cat and 38.0% ± 10.2% for the Far-Eastern wildcat. The motility of frozen-thawed spermatozoa was 30.8% ± 9.8% for the domestic cat and 33.7% ± 15.1% for the Far-Eastern wildcat. Sperm morphology was assessed by light microscopy. The total percentage of normal spermatozoa after freezing and thawing was 51.9 ± 5.9 for the domestic cat and 55.0% ± 6.4% for the Far-Eastern wildcat. Defects of flagella were the most frequently observed abnormalities in both species (32.2% ± 4.8% and 30.8% ± 4.4% of all reported anomalies for the domestic cat and Far-Eastern wildcat, respectively). Domestic cat epididymal and Far-Eastern ejaculatory spermatozoa fertilized in vitro-matured oocytes of the domestic cat (30.0% ± 5.5% and 35.5% ± 15.0%, respectively). Taken together, these results suggest that the freezing of Far-Eastern wildcat spermatozoa with CPF medium is a suitable method for Felidae cryopreservation.

To determine the prevalence of amp1q21 and its relationship to the clinical manifestations of multiple myeloma (MM).In December 2009 to March 2016, a total 134 patients aged 30 to 81 years (median 57 years) underwent a pretreatment FISH-study of bone marrow (BM) with centromeric and locus-specific DNA probes to identify amp1q21, t(11;14), t(4;14), t(14;16), t(14;20), t(6;14), trisomies of chromosomes 5, 9, 15, del13q14, del17p13/TP53, and t(8q24)/cMYC. Induction therapy with bortezomib-containing cycles was performed. Autologous stem cell transplantation was carried out in 48 patients. The median follow-up of patients was 19.3 months (3.2-77.4 months). Disease progression was diagnosed in 69 (51.5%) patients; 12 patients also underwent FISH study during disease progression.At the onset of MM, amp1q21 was detected in 53 (39.6%) patients. The overall 5-year survival rate in patients with amp1q21 was almost 2 times lower than that in those without amp1q21 (43.5 and 79.4%, respectively; p=0.07). The overall 5-year survival rate in patients with one extra copy of 1q21 (only 3 copies) was 67.3%, that in those with 2 or more extra copies of 1q21 (only 4-7 copies) was 20.9% (p=0.0016). Nine (75%) of the 12 patients examined during disease progression were found to have amp1q21: 2 cases were detected in the period of progression to have amp1q21 in its absence at disease onset; 7 cases had amp1q21 both at MM onset and progression; however, the number of copies of 1q21 was unchanged.Аmp1q21 is one of the most common chromosomal abnormalities in patients with new-onset MM and may appear in the course of disease progression. The presence of аmp1q21 is an important prognostic factor and must have to be included in the diagnostic study both at disease onset and progression.Цель исследования. Определить распространенность amp1q21 и ее связь с клиническими проявлениями множественной миеломы (ММ). Материалы и методы. C декабря 2009 г. по март 2016 г. 134 пациентам в возрасте от 30 лет до 81 года (медиана 57 лет) до начала лечения выполнено FISH-исследование костного мозга (КМ) с центромерными и локус-специфичными ДНК-зондами для выявления amp1q21, t(11;14), t(4;14), t(14;16), t(14;20), t(6;14), трисомий хромосом 5, 9, 15, del13q14, del17p13/TP53, t(8q24)/сMYC. Индукционная терапии выполнялась курсами, содержащими бортезомиб. Трансплантация аутологичных стволовых клеток выполнена 48 больным. Медиана наблюдения за больными составила 19,3 мес (3,2-77,4 мес). Прогрессирование заболевания диагностировано у 69 (51,5%) пациентов, 12 пациентам FISH-исследование также проведено в период прогрессирования заболевания. Результаты. В дебюте ММ amp1q21 обнаружена у 53 (39,6%) больных. Общая 5-летняя выживаемость у больных с amp1q21 почти в 2 раза ниже, чем у больных без нее, - 43,5 и 79,4% соответственно (р=0,07). Общая 5-летняя выживаемость у больных с одной дополнительной копией 1q21 (всего 3 копии) составила 67,3%, с 2 дополнительными копиями и более 1q21 (всего 4-7 копий) - 20,9% (р=0,0016). Из 12 пациентов, обследованных в период прогрессирования заболевания, amp1q21 выявлена у 9 (75%): в 2 случаях amp1q21 выявлена в период прогрессирования в отсутствие в дебюте; в 7 случаях amp1q21 обнаружена и в дебюте, и в период прогрессирования ММ, однако количество копий 1q21 не изменилось. Заключение. Аmp1q21 - одно из наиболее распространенных хромосомных нарушений у больных с впервые выявленной ММ и может появляться в ходе прогрессирования заболевания. Наличие аmp1q21 является важным прогностическим фактором и, вероятно, должна включаться в диагностические исследования как в дебюте, так и в период прогрессирования заболевания.

Major stages of the pathogenesis of true acantholytic pemphigus are interpreted in terms of autoimmune pathology; therefore, treatment of this group of diseases remains pathogenetic and is aimed at suppressing the synthesis of autoantibodies against keratinocyte proteins. Treatment with the use of systemic glucocorticosteroids is currently the main method of treatment for true acantholytic pemphigus. To reduce the course dose of glucocorticosteroids, decrease the risk of adverse effect development and achieve long-term remission, further studies of disease pathogenesis and development of new treatment methods for reducing the doses of glucocorticosteroids are of great importance.

An efficient solid-phase-supported peptide synthesis (SPPS) of morpholinoglycine oligonucleotide (MorGly) mimics has been developed. The proposed strategy includes a novel specially designed labile linker group containing the oxalyl residue and the 2-aminomethylmorpholino nucleoside analogues as first subunits.

С целью анализа современных данных о методах определения пролиферативной активности, а также для выбора оптимального метода для работы с тканями с низкой ожидаемой пролиферативной активностью исследовали новые публикации в отрасли. Установили, что подсчет митотического индекса имеет низкую чувствительность и специфичность, а такие методы как иммуноферментный анализ, иммуноблоттинг, иммуногистохимическое исследование, полимеразная цепная реакция отличаются высокой чувствительностью и информативностью, но только иммуногистохимические исследования обеспечивают визуализацию с возможностью топической диагностики. Это свидетельствует, что, несмотря на преимущества и отличия каждого маркера пролиферации, определение бромдексиуридина позволяет проводить динамическую оценку и определение пролиферативной активности в тканях с низкой пролиферативной активностью благодаря его кумулятивному эффекту., Proliferation is an increasing of the body's tissues by cell multiplication by division. It may refer to the processes of different nature. Thus, cell proliferation underlies regenerative tissue neoplasms; proliferation observed in various hyperplasias; finally cell proliferation underlying tumor growth [Mitchison, 2003, Sulić at al., 2005]. Therefore, the identification and determining of the activity of proliferative processes is an important task of experimental and practical medicine.The aim of this review was to analyze the contemporary scientific literature data about methods of determining of the proliferative activity, as well as the choice of an optimal method for its assessing in tissues with expected low proliferative activity.There are many approaches to assess the proliferative activity. A number of proteins involved in cell cycle regulation has been opened last decades. They may serve as selectable markers of proliferating cells either in vivo, or in vitro. Determining of the proliferative activity is possible with using of different methods, such as counting of the mitotic index, enzyme immunoassay, immunoblotting, immunohistochemical analysis, PCR techniques. Consider some of them.Mitotic index. Determining of the mitotic index is based on counting of the percentage of dividing cells of the total number of cells analyzed. The test tissue is treated with colchicine or its derivatives - substances that have properties to prevent the formation of microtubules, thereby preventing divergence of individual chromosomes in anaphase. After processing tissue the cell cycle stops in it, thereby enabling the identification of mitotic cells at the time of application of the drug [Прохорова и др., 2003]. The mitotic index (MI) calculated using the formula:MI = (P+M+A+T)/N * 100,where (Р+М+А+Т) is amount of cells at the stages of prophase, metaphase, anaphase and telophase, respectively, and N – the whole amount of cells analyzed.The main disadvantages of this method are:¾ low specificity;¾ inability to assess the proliferative activity in the dynamics;¾ inability to detect the proliferative activity in tissues with expected low proliferative activity.Such methods as enzyme immunoassay, immunohistochemical analysis, immunoblotting, PCR techniques provide high sensitivity, specificity, informativity, but immunohistochemical techniques also provides visibility with the possibility of topical diagnosis. These techniques use monoclonal antibodies produced against specific antigens associated with cell proliferation. For today, there are methods of identifying of such proteins as cyclins, Ki-67, bromodeoxyuridine. We would like to elaborate on some of them.Proliferating cell nuclear antigen (PCNA) is most commonly identified cyclin; it is an auxiliary protein of DNA polymerase delta. This molecule can be detected in human and animal paraffin-embedded or frozen tissue [Takahashi et al., 1993]. PCNA does not require pre-injection into the tissue under study, as confirmed by the positive results on archival samples, but it has some significant drawbacks:¾ anti-PCNA immunohistochemistry may give a weak signal in non-proliferating tissues [Takahashi et al., 1993];¾ the intensity of the signal depends on the method of fixing and amount of pre-heat treatment [Sasaki et al., 1992, Takahashi et al., 1993];¾ it does not allow evaluating the proliferative activity characteristics and dynamics in in tissues with expected low proliferative activity.All steps in the application of this methodology held in vitro. The manufacturer recommends a 30-minute incubation period at room temperature. Formalin-fixed and paraffin-embedded tissue sections require unmasking of the antigen with high temperature treatment in10 mMcitrate buffer (pH 6.0) before immunostaining.Ki-67 antigen, also known as MKI67 - is another endogenous antigen [Scholzen, Gerdes, 2003], a nuclear protein that is bound and may be required for cell proliferation [Bullwinkel et al., 2003]. Furthermore, it is associated with transcription. Inactivation of the Ki-67 antigen leads to inhibition of the synthesis of ribosomal RNA [Bullwinkel et al., 2003, Rahmanzadeh, 2007]. During the interphase Ki-67 can be detected only in the nucleus, whereas in mitosis most of the protein moves to the surface of the chromosomes. Ki-67 is present during all active phases of the cell cycle (G1, S, G2 and mitosis), but it is absent in resting cells (G0) [Scholzen, Gerdes, 2003]. This causes the possible variability of nuclear staining intensity. For detection in paraffin-embedded sections as well as in the case of PCNA thermal pre-treatment is required. Ki-67 can be detected in the tissue without its prior administration. However, it like as PCNA has several drawbacks:¾ does not assess the features of the proliferative activity in the dynamics;¾ does not assess the features of the proliferative activity in tissues with expected low proliferative activity.Cell proliferation index Ki-67 stronger correlated with the index of cell proliferation of bromodeoxyuridine than with PCNA, since the antigen Ki-67 has shorter half-life (1.5 - 2 hours), as compared with PCNA [Bromley et al., 1996], [Bologna-Molina et al., 2013]. All stages of the usage of this methodology held in vitro. The manufacturer recommends a 30-minute incubation period at room temperature or overnight at 40 C. The prior proteolytic processing of formalin-fixed and paraffin-embedded tissue sections is required. The antibodies react with the core antigen of humans and other mammals. Ki-67 protein initially was identified using prototype monoclonal antibodies, which were obtained by immunizing of mice with the nuclei of cells of Hodgkin's lymphoma [Gerdes et al., 1983]. However, currently MIB-1 monoclonal antibodies are exist; they are directed against another epitope of the same antigen. MIB-1 are used in clinical trials to determine the Ki-67 proliferative index. The main advantage of MIB-1 antibodies compared with the original Ki-67 antibodies (and the reason why they substantially replaced the original antibodies in clinical usage) is the possibility of using in fixed paraffin-embedded tissue samples after heat processing, unlike the original antibodies [Bánkfalvi, 2000].Bromodeoxyuridine (BrdU) is an exogenous proliferation marker; it is synthetic nucleoside analogue of thymidine that is used for studying of the DNA replication [Lehner et al., 2011]. BrdU can be transmitted to daughter cells during the replication [Kee et al., 2002] and defined at least in 2 years after its application [Eriksson et al., 1998]. Enzymes (helicase, topoisomerase) and DNA-binding proteins unwind the DNA and hold the matrix in the untwisted state and rotate the DNA molecule. The correctness of the replication ensures with an exact match of complementary base pairs and DNA polymerase activity, able to detect and correct errors. Method’s principle is based on the detection of BrdU that is capable to replace thymidine during replication, incorporating into new DNA [Lehner et al., 2011]. Anti-BrdU-antibody binds with BrdU, which has been exogenously administered into the newly synthesized DNA, enables the visualization of cells in which DNA replication occurs. The substance can get into the DNA only in S-phase of the cell cycle, thus eliminating the variability of staining [Lehner et al., 2011, Teruaki et al., 2011].Features of this method are:¾ it needs pre-injection into the tissue under study, but the exact timing and dosage of BrdU allows for evaluation of the proliferative activity in the dynamics;¾ prolonged administration of BrdU can be used to assess the proliferative activity in tissues with expected low proliferative activity, since it will be detected in each molecule of the newly synthesized DNA after administration, and transmitted to daughter cells.BrdU incorporated into the core is highly stable antigen, which gives a strong and reliable signal regardless of the method of fixation and further processing of tissues. Research can be carried out on small rodents. Important qualities of this method are:¾ low toxicity of the drug (approved for use in humans) [Takao et al., 1985, Takamitsu et al., 2006];¾ minimally invasive administration, thus reducing the risk of death in experimental animals;¾ the initial stage of the method is performed in vivo [Lehner et al., 2011].BrdU does not affect the biochemical constants, neuroautonomic indicators and endocrine balance [Hoshino et al., 1985]. This is an important condition for maintaining of indicators of the simulated pathology. The second investigation phase conducted in vitro on conventionally made tissue sections, which can also be used to perform other histochemical studies. For successful immunohistochemical detection of incorporated bromodeoxyuridine the DNA denaturation is required [Konishi et al., 2011].For the integrated assessment of the methods above and decision of the most appropriate method for studying of the proliferative activity in the dynamics and in in tissues with expected low proliferative activity we have compiled a table.Conclusions. Bromodeoxyuridine is a versatile and highly effective marker of DNA replication and cell proliferation, allowing carrying out a dynamic observation of the experiment, including tissues with expected low proliferative activity. It can be widely used in chronic experiments on small rodents, in the simulation of various pathologies and studying the effect of pharmaceuticals on the activity of proliferation., З метою аналізу сучасних даних про методи визначення проліферативної активності, а також для вибору оптимального методу для роботи із тканинами з низькою очікуваною проліферативною активністю дослідили новітні публікації в галузі. Встановили, що підрахунок мітотичного індексу має низьку чутливість і специфічність, а такі методи як імуноферментний аналіз, імуноблотинг, імуногістохімічне дослідження, полімеразна ланцюгова реакція мають високу чутливість та інформативність, але лише імуногістохімічні дослідження дають змогу візуалізації з можливістю топічної діагностики. Це свідчить, що, незважаючи на переваги і відмінності кожного маркера проліферації, визначення бромдезоксіуридину дає можливість здійснювати динамічне оцінювання та визначення проліферативної активності у тканинах із низькою проліферативною активністю через його кумулятивний ефект.

Предложена модель выбора базовых функций при автоматизированной идентификации временных рядов, алгоритм и программное средство идентификации временных рядов на основе ассоциативно-мажоритарного подхода, позволяющие идентифицировать вид базовых зависимостей динамических процессов. Задача идентификации вида базовых функций решается с использованием классической теории распознавания образов. Идентификация производится путем сравнения исходного образа с эталонами, хранящимися в едином пространстве образов. Пространство образов представляет собой область памяти устройства (ассоциативной памяти), на котором производится идентификация. Особенностью предложенной модели является оперативность сравнения исходных образов с образами эталонов за счет использования единого признакового пространства и возможности сравнения всех образов за один такт. Для повышения оперативности идентификации в алгоритме выбора базовых функций также использован ассоциативно-мажоритарный подход к хранению и поиску идентификационных данных в электронной памяти. Предложенный алгоритм и программное средство выбора базовых функций являются универсальными, так как позволяют идентифицировать вид базовой зависимости в любом динамическом процессе независимо от специфики исследуемой предметной области. Автоматизированная идентификация вида базовых функций сокращает время построения прогнозных моделей и позволяет оперативно прогнозировать дальнейшие варианты протекания процесса.

The authors disclosed the participation of apoptosis proteins in the development of acantholysis in patients with pemphigus. In this connection, studies of the PERP protein passing apoptosis signals and regulating desmosomal functions in keratinocytes are of interest. There is no information about any studies aimed at the PERP protein expression in patients with pemphigus in available literature. Goal. To assess the PERP protein expression in the skin of patients with pemphigus. Materials and methods. There was a study of 22 patients with pemphigus, a patient with bullous pemphigoid and ten healthy people. The PERP protein expression was studied in the biopsy materials obtained from lesion foci and apparently healthy skin of the patients as well as healthy people using the indirect immunofluorescence method. Results. The PERP protein expression was revealed in patients with pemphigus on areas of apparently intact skin, in lesion foci in the patient with bullous pemphigoid and skin of healthy volunteers in the membrane of keratinocytes from all epidermal layers.The PERP protein expression in the blister operculum in lesion foci in patients with pemphigus was absent. Conclusion. Substantial differences in the PERP protein expression in the blister operculum and apparently intact skin of patients with pemphigus were revealed.

Goal. To determine the nucleotide protein-coding PERP gene sequence and assess the relation between the revealed mutations/polymorphisms and development of true acantholytic pemphigus as well as particular features of its course. Materials and methods. The protein-coding PERP gene DNA sequence was studied by the sequence analysis method in 18 patients with true acantholytic pemphigus. Results. Two polymorphisms were discovered in patients with true acantholytic pemphigus in Exon 3 of the PERP gene for the first time: rs648802 (non-synonymous) and rs648396 (synonymous). The incidence of wild type genotypes in the revealed polymorphisms (С/С genotype rs648802 and Т/Т genotype rs648396) in healthy volunteers reliably exceeded that in patients (p = 0.049). Patients with true acantholytic pemphigus are characterized by a higher incidence rate of mutant heterozygous genotypes С/G rs648802 and Т/C rs648396 (p = 0.09). Mutant heterozygous genotypes of the polymorphisms (G/G genotype rs648802 and С/С genotype rs648396) were revealed in patients with the earlier onset of the disease (41-60 years) (p = 0.025) more often while heterozygous genotypes (С/G genotype rs648802 and T/С genotype rs648396) were revealed when the disease developed at the age of 61 or older more often (p = 0.01). Conclusion. Identification of the polymorphous genotype by the sequence method or other molecular methods (e.g. PCR) can be used to forecast the terms when true acantholytic pemphigus can emerge in genetically inclined patients. However, it should be noted that it is necessary to specify the preliminary results obtained based on a greater sample of patients with true acantholytic pemphigus.

A versatile strategy for the synthesis of $$\hbox {NAD}^{+}$$ mimetics was developed, involving an efficient pyrophosphate linkage formation in key conjugates containing a functional amino group which acts as useful reactive anchor for further derivatization. These $$\hbox {NAD}^{+}$$ mimetics consist of ADP conjugated through a diphosphate chain to an extended aliphatic linker bearing an aromatic acid residue. A number of conjugates containing aromatic carboxylic acids were found to inhibit poly(ADP-ribose) synthesis catalyzed by poly(ADP-ribose) polymerase-1 (PARP-1). A new class of potential PARP-1 inhibitors mimicking $$\hbox {NAD}^{+}$$ , a substrate in the PARP-1 catalyzed reaction, was proposed.