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Abstract Background Heterologous prime-boost immunization protocols using different gene expression systems have proven to be successful tools in protecting against various diseases in experimental animal models. The main reason for using this approach is to exploit the ability of expression cassettes to prime or boost the immune system in different ways during vaccination procedures. The purpose of the project was to study the ability of recombinant vaccinia virus (VV) and bacterial plasmid, both carrying the NS1 gene from tick-borne encephalitis (TBE) virus under the control of different promoters, to protect mice against lethal challenge using a heterologous prime-boost vaccination protocol. Results The heterologous prime-boost vaccination protocol, using a VV recombinant and bacterial plasmid, both containing the NS1 TBE virus protein gene under the control of different promoters, achieved a high level of protection in mice against lethal challenge with a highly pathogenic TBE virus strain. No signs of pronounced TBE infection were detected in the surviving animals. Conclusion Heterologous prime-boost vaccination protocols using recombinant VV and bacterial plasmids could be used for the development of flavivirus vaccines.

Discrete systems of coupled linear mechanical oscillators with nonreciprocal interaction are a model for a variety of physical systems. In general, the presence of nonreciprocal interactions renders their dynamics nonconservative, but under certain conditions it remains conservative. In this paper we show which thermodynamic properties induced by nonreciprocity can be observed in conservative systems and which are specific to nonconservative systems. To this end, we formulate a criterion for identifying conservative systems and construct a measure to quantify the deviation from conservativity.

In the article "Point-of-care blood glucose testing: post-market performance assessment of the Accu-Chek Inform II hospital-use glucose meter," published in the Terapevticheskii Arkhiv journal, Vol. 95, No.12, 2023 (DOI: 10.26442/00403660.2023.12.202522), errors were made: the term "measurements at the place of treatment" was changed, as well as the section "Conflict of interest." At the request of the authors' team, errors in the conflict of interest and the wording of the term have been corrected, and the section "Information about the authors" has been updated. The publisher replaced the original version of the published article with the corrected one; the information on the website was also corrected. Correct text of the section "Conflict of interest": Conflict of interest. All authors are not employees or consultants of Roche Diagnostics and have not received any compensation from Roche Diagnostics. Correct wording of the term in Russian: "измерения по месту лечения". Changes were made to the title of the article in Russian: "Измерения глюкозы по месту лечения: пострегистрационное испытание госпитального глюкометра Акку-Чек Информ II", the text of the abstract, keywords, citation, in the text of the article, and abbreviations. Information of the place of work has been updated: Center for Laboratory Diagnostics of the Russian Children Clinical Hospital, a Branch of the Pirogov Russian National Research Medical University. The publisher apologizes to readers and authors for the errors and is confident that the correction of errors will ensure the correct perception and interpretation of the results of the study described in the text.

Background: A point-of-care glucose testing (POCT) is an essential component of care in patients with hyperglycemia and hypoglycemia in inpatient and outpatient settings. In Russian medical facilities (MFs), conventional glucose meters designed for self-monitoring by patients with diabetes are commonly used for POCT. These home-use meters have two serious disadvantages: the first is large measurement bias and the second - they can't be integrated into laboratory information systems, so measurement data have to be recorded into patient charts manually. Both factors may lead to medical errors. It is reasonable to use in the MFs specialized POCT glucose meters, as they are superior to conventional ones in accuracy and may be easily connected to laboratory information systems. With this in mind, physicians at the Russian Children's Clinical Hospital decided to substitute conventional meters with the Accu-Chek Inform II POCT meter, however, after preliminary performance assessment of the model., Aim: To test the Accu-Chek Inform II performance characteristics: accuracy, linearity, repeatability, and mean absolute relative difference (MARD)., Materials and Methods: Performance of the Accu-Chek Inform II was tested by comparing the results of parallel C GL measurements with the meter and reference laboratory analyzer in capillary blood samples. Overall, 99 parallel C GL measurements were made in 45 samples. Accuracy was evaluated according to the ISO 15197-2013 and POCT12-A3 criteria., Results: The Accu-Chek Inform II meter met the requirements of ISO 15197-2013 and POCT12-A3 and demonstrated high linearity (correlation coefficient, r =1,0), good repeatability (mean coefficient of variation, CV=1,38%) and acceptable MARD (4,9%)., Conclusion: The Accu-Chek Inform II POCT glucose meter may be efficiently and safely used in inpatient and outpatient MFs and particularly in pediatric clinics.

A complex (dusty) plasma system is well known as a paradigmatic model for studying the kinetics of solid-liquid phase transitions in inactive condensed matter. At the same time, under certain conditions a complex plasma system can also display characteristics of an active medium with the micron-sized particles converting energy of the ambient environment into motility and thereby becoming active. We present a detailed analysis of the experimental complex plasmas system that shows evidence of a non-equilibrium stationary coexistence between a cold crystalline and a hot fluid state in the structure due to the conversion of plasma energy into the motion energy of microparticles in the central region of the system. The plasma mediated non-reciprocal interaction between the dust particles is the underlying mechanism for the enormous heating of the central subsystem, and it acts as a micro-scale energy source that keeps the central subsystem in the molten state. Accurate multiscale simulations of the system based on combined molecular dynamics and particle-in-cell approaches show that strong structural nonuniformity of the system under the action of electostatic trap makes development of instabilities a local process. We present both experimental tests conducted with a complex plasmas system in a DC glow discharge plasma and a detailed theoretical analysis., (© 2022. The Author(s).)

Binary and ternary composites (CM) based on M-type hexaferrite (HF), polymer matrix (PVDF) and carbon nanomaterials (quasi-one-dimensional carbon nanotubes-CNT and quasi-two-dimensional carbon nanoflakes-CNF) were prepared and investigated for establishing the impact of the different nanosized carbon on magnetic and electrodynamic properties. The ratio between HF and PVDF in HF + PVDF composite was fixed (85 wt% HF and 15 wt% PVDF). The concentration of CNT and CNF in CM was fixed (5 wt% from total HF + PVDF weight). The phase composition and microstructural features were investigated using XRD and SEM, respectively. It was observed that CM contains single-phase HF, γ- and β-PVDF and carbon nanomaterials. Thus, we produced composites that consist of mixed different phases (organic insulator matrix-PDVF; functional magnetic fillers-HF and highly electroconductive additives-CNT/CNF) in the required ratio. VSM data demonstrate that the main contribution in main magnetic characteristics belongs to magnetic fillers (HF). The principal difference in magnetic and electrodynamic properties was shown for CNT- and CNF-based composites. That confirms that the shape of nanosized carbon nanomaterials impact on physical properties of the ternary composited-based magnetic fillers in polymer dielectric matrix.

Preclinical studies of human cellular and tissue-based products (HCT/Ps) for transplantation therapy of type 1 diabetes mellitus (T1DM) necessarily involve animal models, particularly mouse models of diabetes induced by streptozotocin (STZ). These models should mimic the clinical and metabolic manifestations of T1DM in humans (face validity) and be similar to T1DM in terms of the pathogenetic mechanism (construct validity). Furthermore, since HCT/Ps contain human cells, modeling of diabetes in immune-deficient animals is obligatory. Here we describe the most simplified diabetes model in Nude mice. Diabetes was induced in 31 males by a single intraperitoneal injection of STZ in normal saline at a medium-to-high dose of 150 mg/kg body weight. Fourteen control animals received only saline. Non-fasting plasma glucose (PG) levels were measured periodically for 50 days. All STZ-treated mice survived beyond 50 days. By day 15 after STZ administration, 22 of 31 (71%) mice developed stable diabetes based on the following criteria: (1) non-fasting PG ≥ 15 mmol/L on consecutive measurements up until day 50; (2) no diabetes remission. The mean non-fasting PG in mice with stable diabetes over the period of 35 days was equal to 25.7 mmol/L. On day 50, mean plasma insulin concentration, mean pancreatic insulin content, and the average number of β-cells in pancreatic islets were 2.6, 8.4, and 50 times lower, respectively, than in the control animals. We consider that our Nude mouse model of diabetes meets face validity and construct validity criteria and can be used in preclinical studies of HCT/Ps., (Copyright ® 2020 National Research University Higher School of Economics.)

Electrothermal elements are used in various energy harvesters, coolers, and radiation detectors. The optimal operation of these elements relies on mastering two competing boundary conditions: the maximization of the electrothermal response and the blockade of lattice (phonon) thermal conduction. In this work, we propose and demonstrate that efficient electrothermal operation and phonon blocking can be achieved in solid-state thermionic junctions, paving the way for new phonon-engineered high-efficiency refrigerators and sensors. Our experimental demonstration uses semiconductor-superconductor (Sm-S) junctions where the electrothermal response arises from the superconducting energy gap and the phonon blocking results from the acoustic transmission bottleneck at the junction. We demonstrate a cooling platform where a silicon chip, suspended only from the Sm-S junctions, is cooled by ~40% from the bath temperature. We also show how the observed effect can be used in radiation detectors and multistage electronic refrigerators suitable for cooling of quantum technology devices., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Currently, many DNA vaccines against infectious diseases are in clinical trials; however, their efficacy needs to be improved. The potency of DNA immunogen can be optimized by targeting technologies. In the current study, to increase the efficacy of NS1 encoded by plasmid, proteasome targeting was applied. NS1 variants with or without translocation sequence and with ornithine decarboxylase as a signal of proteasomal degradation were tested for expression, localization, protein turnover, proteasomal degradation and protection properties. Deletion of translocation signal abrogated presentation of NS1 on the cell surface and increased proteasomal processing of NS1. Fusion with ornithine decarboxylase led to an increase of protein turnover and the proteasome degradation rate of NS1. Immunization with NS1 variants with increased proteasome processing protected mice from viral challenge only partially; however, the survival time of infected mice was prolonged in these groups. These data can give a presupposition for formulation of specific immune therapy for infected individuals.

The interaction of viral proteins with host cell components plays an important role in antiviral immune response. One of the key steps of antiviral defense is the formation of immunoproteasomes. The effect of nonstructural protein 1 (NS1) of tick-borne encephalitis virus on the immunoproteasome formation was studied. It was shown that cell expression of NS1 does not reduce the efficacy of the immunoproteasome generation in response to interferon-γ stimulation and even increases the content of the immunoproteasome subunits without the interferon-γ treatment. Thus, NS1 of tick-borne encephalitis virus activates, rather than blocks the mechanisms of immune defense in the cell.

The individual portable systems of control of glucose level in blood commonly known as glucometers permit to patients with diabetes mellitus to independently correct pharmaceutical therapy. The effectiveness of this correction depends on accuracy of control of glucose level. The evaluation was implemented concerning minimal admissible accuracy and clinical accuracy of control of glucose level of devices Contour TC, Satellite Express and One Touch Select according standards expounded in GOST 15197-2011 and international standard ISO 15197-2013. It is demonstrated that Contour TC and One Touch Select meet the requirements of these standards in part of accuracy while Satellite Express does not.

The control of electronic and thermal transport through material interfaces is crucial for numerous micro and nanoelectronics applications and quantum devices. Here we report on the engineering of the electro-thermal properties of semiconductor-superconductor (Sm-S) electronic cooler junctions by a nanoscale insulating tunnel barrier introduced between the Sm and S electrodes. Unexpectedly, such an interface barrier does not increase the junction resistance but strongly reduces the detrimental sub-gap leakage current. These features are key to achieving high cooling power tunnel junction refrigerators, and we demonstrate unparalleled performance in silicon-based Sm-S electron cooler devices with orders of magnitudes improvement in the cooling power in comparison to previous works. By adapting the junctions in strain-engineered silicon coolers we also demonstrate efficient electron temperature reduction from 300 mK to below 100 mK. Investigations on junctions with different interface quality indicate that the previously unexplained sub-gap leakage current is strongly influenced by the Sm-S interface states. These states often dictate the junction electrical resistance through the well-known Fermi level pinning effect and, therefore, superconductivity could be generally used to probe and optimize metal-semiconductor contact behaviour.

Tick-borne encephalitis virus (TBEV) is one of the most prevalent and medically important tick-borne arboviruses in Eurasia. There are overlapping foci of two flaviviruses: TBEV and Omsk hemorrhagic fever virus (OHFV) in Russia. Inactivated vaccines exist only against TBE. There are no antiviral drugs for treatment of both diseases. Optimal animal models are necessary to study efficacy of novel vaccines and treatment preparations against TBE and relative flaviviruses. The models for TBE and OHF using subcutaneous inoculation were tested in Cercopithecus aethiops and Macaca fascicularis monkeys with or without prior immunization with inactivated TBE vaccine. No visible clinical signs or severe pathomorphological lesions were observed in any monkey infected with TBEV or OHFV. C. aethiops challenged with OHFV showed massive hemolytic syndrome and thrombocytopenia. Infectious virus or viral RNA was revealed in visceral organs and CNS of C. aethiops infected with both viruses; however, viremia was low. Inactivated TBE vaccines induced high antibody titers against both viruses and expressed booster after challenge. The protective efficacy against TBE was shown by the absence of virus in spleen, lymph nodes and CNS of immunized animals after challenge. Despite the absence of expressed hemolytic syndrome in immunized C. aethiops TBE vaccine did not prevent the reproduction of OHFV in CNS and visceral organs. Subcutaneous inoculation of M. fascicularis with two TBEV strains led to a febrile disease with well expressed viremia, fever, and virus reproduction in spleen, lymph nodes and CNS. The optimal terms for estimation of the viral titers in CNS were defined as 8-16 days post infection. We characterized two animal models similar to humans in their susceptibility to tick-borne flaviviruses and found the most optimal scheme for evaluation of efficacy of preventive and therapeutic preparations. We also identified M. fascicularis to be more susceptible to TBEV than C. aethiops.

Immunoproteasomal processing of mycobacterial antigens is necessary to control the infection and to protect the organism from development of active form of tuberculosis. Here we investigate the activation of immunoproteasome subunit genes transcription in peritoneal monocytes of C57Bl/6 mice infected with vaccine M. bovis BCG and virulent strain M. tuberculosis H37Rv. The level of transcription of LMP2, LMP7, MECL1 subunits didn't increase for one and two days after a single infection. Two rounds of infection with BCG strain M. bovis led to enhancement of the only LMP7 subunit gene transcription. However after subsequent infection of monocytes with vaccine followed by virulent strain infection the dramatic rise of all immunoproteasomal subunit genes transcription was observed. Activation of transcription of the gene coding the PA28alpha subunit of regulatory complex PA28 was observed only after a single infection of monocytes with strain M. bovis BCG. Thus, vaccination with strain M. bovis BCG promotes effective activation of immunoproteasomal genes in case of subsequent contact with virulent strain M. tuberculosis H37Rv.

Hepatocellular carcinoma (HCC) is among the most frequent malignancies in humans. HCC therapy is not efficient and the usual outcome is poor. In this regard, novel approaches to treat and prevent HCC are urgently needed. The Alpha-fetoprotein (AFP) is a serological marker of HCC. Recently it has been shown, that the DNA vaccines expressing AFP are capable in generating immune response against AFP. However, both, the immunization procedures and DNA vaccines used before were complex and not always very effective. We have shown that DNA vaccine encoding HIV-1 reverse transcriptase (RT) with fused ornithine decarboxylase (ODC) degradation signal induced a strong Th-1 immune response against RT in mice. Using this approach we designed a set of novel DNA vaccines bearing AFP and ODC degradation signal. Results obtained on transfected cells demonstrated efficient expression and fast proteasomal degradation of the recombinant AFP. The anti-tumor immune response stimulation was shown in immunized animals and most importantly a notable retardation of tumor growth was observed as a result of protective vaccination.

A system of equations describing motion of dust particles in gas discharge plasma is formulated. This system is developed for a monolayer of dust particles with an account of dust particle charge fluctuations and features of the discharge near-electrode layer. Molecular dynamics simulation of the dust particles system is performed. A mechanism of dust particle average kinetic energy increase is suggested on the basis of theoretical analysis of the simulation results. It is shown that heating of dust particles' vertical motion is initiated by forced oscillations caused by the dust particles' charge fluctuations. The process of energy transfer from vertical to horizontal motion is based on the phenomenon of the parametric resonance. The combination of parametric and forced resonances explains the abnormally high values of the dust particles' kinetic energy. Estimates of frequency, amplitude, and kinetic energy of dust particles are close to the experimental values.

Mouse ornithine decarboxylase (ODC) degrades in proteasome in an ubiquitin-independent manner with an averagehalf-life of 2 h. The 37 amino acid long C-terminal fragment known as a degradation signal (degron) is responsible for the effective degradation of ODC. Recently, amino acids being critical for degradation in the ODC-degron have been mapped. Mutations of Cys441 and Ala442 led to protein stabilization, while a substitution of other amino acids composing ODC-degron had almost no effect on the protein turnover; whereas insertions or deletions in region between Ala442 and ODC C-terminus diminished greatly rate of protein degradation, e.g. positioning of the key amino acids from the C-terminus was shown to be crucial. Using these data we introduced both key amino acids into the alfa-fetoprotein with truncated exportation signal (deltaAFP), at the same distance from the C-terminus as they being in the ODC (deltaAFPCAG and deltaAFPLCAG). Removal of N-terminal exportation signal prevented secretion of modified proteins. Using in silico approach we demonstrated no significant difference in hydrophobicity or secondary structure between C-terminus of deltaAFP and mutated proteins. The degradation kinetics of deltaAFP, deltaAFPCAG, deltaAFPLCAG in cyloheximide-chase and proteasome inhibition assay (using MG132) was identical. Obtained results suggest that introduced substitutions are insufficient for effective recognition of mutated deltaAFP by26S proteasome. We assume thatadditional amino aci ds composing ODC-degron or their combine action could also affect degradation. Besides that, one cannot exclude that conformation of the mutated deltaAFP limits its C-terminus accessibility to proteasome.

We demonstrate quantum-limited electronic refrigeration of a metallic island in a low-temperature microcircuit. We show that matching the impedance of the circuit enables refrigeration at a distance, of about 50 microm in our case, through superconducting leads with a cooling power determined by the quantum of thermal conductance. In a reference sample with a mismatched circuit this effect is absent. Our results are consistent with the concept of electromagnetic heat transport. We observe and analyze the crossover between electromagnetic and quasiparticle heat flux in a superconductor.

We study quasiparticle energy relaxation at subkelvin temperatures by injecting hot electrons into an Al island and measuring the energy flux from quasiparticles into phonons both in the superconducting and in the normal state. The data show strong reduction of the flux at low temperatures in the superconducting state, in qualitative agreement with the theory for clean superconductors. However, quantitatively the energy flux exceeds the theoretical predictions both in the superconducting and in the normal state, suggesting an enhanced or additional relaxation process.

To study whether the genotype of a mycobacterial strain might affect the ability of macrophages (MP) to produce the key cytokines, we determined the synthesis of interferon-gamma (IFN-gamma), tumor necrosis factor-gamma (TNF-gamma), and interleukin-6 (IL-6) in the infection of murine MP with M. tuberculosis H37Rv and M. bovis BCG, the strains that belong to the same genetic group (M. tuberculosis complex) but are opposite in virulent properties. MP infection with a virulent and attenuated M. tuberculosis complex strain was shown to differently affect the synthesis of IFN-gamma, TNFaalpha-, and IL-6. MP infection with M. tuberculosis H37Rv activated the generation of IFN-gamma and that with M. bovis BCG substantially increased the levels of TFN-alpha and IL-6. The findings suggest that this model may be used to investigate the specific features of mycobacterial strains of various genotypic clusters with eukaryotic cells.

Phenomenon of antibody dependent tick-borne encephalitis virus (TBEV) infectivity enhancement in the human monocytic cell culture J-96 has been studied. Three specific commercial sera that are generally used for therapy and prevention of TBE in humans were used. Results showed that enhancement of TBEV infectivity was markedly apparent following use of 2 out of 3 studied sera. Data obtained in the study prove that phenomenon of antibody dependent TBEV infectivity enhancement in monocytes cell culture may exist, but conditions for its realization in the organism are very unlikely due to the narrow range of antibodies' concentration and quantity of viral particles. A the same time, the fact of possible aggravation of TBE course after use of specific antisera for treatment and prevention of TBE in humans should not be ignored.

Infection with many encephalitic viruses is associated with the induction of the proinflammatory cytokine interleukin (IL)-6. In some situations, induction of high levels of this cytokine is associated with a protective response, but in others it can be linked to tissue damage and disease. In the studies reported here, levels of serum IL-6 and virus-specific antibodies were measured on admission to hospital and correlated with clinical outcomes. Only some patients demonstrated raised levels of serum IL-6, and there was no correlation between high levels of this cytokine and either gender or the severity of clinical disease. A statistically significant association between raised IL-6 and age was observed, with all individuals below the age of 26 showing normal levels of serum IL-6, regardless of clinical presentation. Furthermore, not all patients had detectable levels of virus-specific serum immunoglobulin G (IgG) antibodies, but an inverse and statistically significant correlation between raised IL-6 levels and IgG titre was observed. Consequently, serum levels of IL-6 cannot be used as a reliable indicator of disease outcome.

Tissue precursors and genesis of female reproductive tract carcinoma vis-a-vis its carcinomatous and sarcomatous patterns remain unknown. To determine the clonal origin of 17 female reproductive tract carcinomas, such molecular, genetic and immunohistochemical techniques as PCR-SSCP and/or denaturing gel electrophoresis for K-ras, p53 and PTEN genes; D17S786, CHRNB1, TP53, BAT26 and BAT40 microsatellites and immunostaining for p53 protein were used. Carcinomatous and sarcomatous components were studied separately. Eight tumors were assumed to be monoclonal (combination or conversion tumors), while one--of an obscure origin. Our results suggest that carcinosarcomas were characterized by chromosomal instability. Moreover, it was shown that it is necessary to combine immunohistochemical techniques with a battery of methods including genetic ones to determine clonal origin of immunologically--stained carcinosarcomas.

Ex vivo production of IFN-gamma, TNF-alpha, and IL-6 by mouse peritoneal macrophages was studied during successive infection with the vaccine strain M. bovis BCG and virulent strain M. tuberculosis H37Rv. The increase in the concentrations of TNF-alpha and IL-6 did not depend on the sequence of macrophage infection with the vaccine or virulent strain, but was related to the presence of the vaccine strain M. bovis BCG in the medium. IFN-gamma production depended on infection of macrophages with the virulent strain M. tuberculosis H37Rv. The concentration of IFN-gamma was maximum during primary infection with the virulent strain and did not increase after successive infection with the virulent and vaccine strain.

We use a hysteretic Josephson junction as an on-chip detector of the third moment of shot noise of a tunnel junction. The detectable bandwidth is determined by the plasma frequency of the detector, which is about 50 GHz in the present experiment. The third moment of shot noise results in a measurable change of the switching rate when reversing polarity of the current through the noise source. We analyze the observed asymmetry assuming adiabatic response of the detector.

Potentially immunoactive regions of the NS1 nonstructural protein of the tick-borne encephalitis virus that can stimulate the antibody formation in vivo and protect animals from this disease were chosen on the basis of theoretical calculations. Eleven 16- to 27-aa peptides containing the chosen regions were synthesized. The ability of the free peptides (without any high-molecular-mass carrier) to stimulate the production of antipeptide antibodies in mice of three lines and ensure the formation of protective immunity was studied. Most of these peptides were shown to exhibit the immunogenic activity in a free state. Five fragments that can protect mice from the infection by a lethal dose of tick-borne encephalitis virus were found.

Linear immunogenic peptides corresponding to amino acid sequences from the NS1 non-structural protein from tick-borne encephalitis virus (strain Sophyin) were predicted using established algorithms and synthesized. Of the 12 peptides predicted, 11 were able to induce peptide-specific antibodies in BALB/c mice but only 1 of these 11 was able to induce antibodies, which reacted with the native protein in a radio-immune precipitation assay. This peptide corresponds to amino acids 37--55, and forms one of the predicted structurally conserved alpha helices of the virus NS1 protein. It was able to protect 60% of animals against lethal challenge with the homologous highly pathogenic tick-borne encephalitis virus strain, and adoptive transfer experiments indicated the involvement of the antibodies induced by this peptide in its protective activity in mice.

To assess immunogenetic markers of insulin dependence (IMID), 117 patients with diabetes mellitus of the adults (DMA) were examined. Of them, 97 patients (77 females and 20 males, 41 and 10 obese, respectively) were insulin dependent and 20 (8 males and 12 females) had classic diabetes mellitus (DM) type 2. All the analysed immunogenetic markers (genotype SS/SS, alleles 04*, 0301 in HLA-DQ*A2, haplotype A1*0301-B1*0302) were associated most often with insulin-dependent DMA. The following conclusions were made: both DMA variants can be definitely differentiated only by clinico-anamnestic screening; there were no differences in distribution of the alleles 0301 in DQ*A1 and 0302 in DQ*B1 in different variants of DMA; alleles 04* and 0301 DQ*A1 significantly more frequently occur in DM type 1 than DM type 2; 0301 allele was more frequent in patients with insulin-dependent DM type 2 vs DM type 2; an expectant genetic marker of insulin-dependence (haplotype 0301/0302) occurs significantly more frequently in insulin-dependent DMA than in healthy subjects and DM type 2.

A vaccinia virus recombinant containing the non-structural tick-borne encephalitis virus (TBEV) NS1 gene was developed. The recombinant expressed native dimeric form of the NS1 protein in infected cells and protected mice against lethal infection with TBEV.

Aim: To test IL-6 and IL-2 serum concentrations as indirect prognostic indicators in tick-borne encephalitis (TBE) course., Material and Methods: 13 TBE patients' sera were examined for IL-2 and IL-6., Results: IL-6 was not detected in patients with TBE fever but was detected in meningoencephalytic TBE., Conclusion: TBE patients demonstrate a positive correlation between the disease severity and serum IL-6. This criterion is proposed as a prognostic factor of a TBE course.

Six peptide fragments of the envelope protein E of the tick-borne encephalitis virus involving the predicted T-helper epitopes were synthesized. Their ability to induce antibodies without conjugation with any high-molecular-mass carrier was studied in mice of three lines. Five of six synthesized peptides exhibited immunogenic properties, which differed in dependence on the haplotype of immunized mice. The peptide binding to the antiviral antibodies was studied, and two peptides were revealed that demonstrated a high ability to recognize the viral antibodies in the horse and human sera. These peptides are promising for the development of diagnostic agents for the tick-borne encephalitis virus.

Three intramuscular injections (50 micrograms each) with bacterial plasmid pMV45 carrying nonstructural gene of NS1 protein of tick-borne encephalitis (TBE) virus protected 88% Balb/c mice from lethal challenge with the virus. Antibodies to NS1 nonstructural protein were detected in the sera of vaccinated mice after the challenge. Absence of antibodies to E structural protein indicated absence of manifest infectious process in mice vaccinated with plasmid and challenged with a lethal dose of TBE virus.

Antiviral activity of phosprenyl was studied in BALB/c mice infected with tick-borne encephalitis (TBE) virus. Up to 60% animals infected with TBE virus survived after 1-3 intramuscular injections of phosprenyl. The mortality in the untreated group infected with the virus was 100%. Direct antiviral effect of phosprenyl was studied in sensitive SPEV cells infected with TBE virus. The titer of the virus decreased 10-fold in the cells treated with the drug vs. untreated control cells. Phosprenyl stimulates some interleukins: gamma-interferon, tumor necrosis factor-alpha, and interleukin-6. The stimulating effect of the drug manifests in intact animals and in those infected with TBE virus and treated with phosprenyl. The prospects of further trials of the drug as a therapeutic and prophylactic agent in TBE are discussed.

The fusion motif of tick-borne encephalitis virus E protein has been predicted to be located within its conserved region (98-120). Results are presented to demonstrate that non-neutralizing monoclonal antibody which recognizes a synthetic peptide corresponding to residues 98-113 of the E protein sequence can block the fusion of the virus particles with artificial membranes.

The synthetic peptide with the conservative 98-113 sequence of protein E of tick-borne encephalitis virus was studied in order to elucidate its role in the functioning of flaviviruses. The peptide was shown to inhibit the in vitro infection of macrophages with the virus. An antibody that specifically binds this peptide was found among the set of monoclonal antibodies produced against protein E. This antibody was found to prevent penetration of the virus into liposomes. A correlation was found between our results and data on the spatial structure of protein E and its interspecies homology. The protein E 98-113 sequence of the tick-borne encephalitis virus was found to be the fusion site of the viral envelope with a cellular membrane.

Many-year routine use of EIA as an in vitro test demonstrated it as a highly reproducible and technological test for assessing the efficacy of vaccine against tick-borne encephalitis and its semiproducts at the intermediate stages of vaccine production. The reproducibility of mouse protection test is notably inferior to that of EIA.

The humoral immune response to flaviviruses is mainly directed to the major envelope protein, E, and a glycosylated non-structural protein, NS1. Cell-mediated immune responses, however, appear to be directed mainly against non-structural proteins. Experiments described here show that a defective recombinant adenovirus (Rad51) containing the gene encoding the NS1 protein of tick-borne encephalitis virus can induce a strong protective immune response against several pathogenic tick-borne flaviviruses in an experimental animal model, and can enhance the efficacy of conventional vaccine preparations. A protective immune response against a lethal virus challenge can also be induced by the passive transfer of antibodies, B cells or T cells from animals vaccinated with Rad51. Raised levels of non-neutralizing antibodies and cytokines associated with a T helper cell-type 1 immune response are also observed. These data demonstrate the importance of non-structural viral proteins in the protective immune response against flaviviruses and support the use of non-structural viral proteins as vaccine components.

Recombinant adenovirus expressing NS1 nonstructural protein of trick-borne encephalitis (TBE) virus (Rad 51) protected mice from many strains of TBE and Omsk hemorhagic fever (OHF) viruses, but virtually did not protect them from Negishi virus. During combined use of whole-virion inactivated TBE vaccine and Rad 51 the recombinant adenovirus notably potentiated the protective effect of the traditional vaccine. The results of adaptive transfer of immunological material from mice infected with Rad 51 showed that both the vaccinated animals' sera and the pool of T and B cells partially protected the recipient mice from lethal TBE infection. NS1 protein expressed by adenovirus increased the level of the key interleukins (IL) interferon, tumor necrosis factor, IL-1 beta, IL-2, and, probably, IL-4. Vaccination of mice with Rad 51 resulted in the appearance of antibodies to NS1 protein in rather high titers. The prospects of using Rad 51 as a vaccine against TBE are discussed.

Dendritic cells (DC) from mice infected with the murine retrovirus Rauscher leukaemia virus (RLV) are poor stimulators of allogeneic and syngeneic T cells and express lower, but still significant, levels of MHC class II. In this paper we further investigated the mechanism of the dysfunction of DC. DC from infected animals did not cause anergy of T cells during coculture for 3 or 6 days. They did not release a substantial amount of soluble factors which could suppress T cell responses. The low T cell responses on stimulation using RLV-infected DC could be overcome by the addition of control DC. Pretreatment of these control DC with monoclonal antibody against MHC class II molecules completely blocked their ability to restore stimulation of T cells in the presence of infected DC. However, antibody against MHC class I or mismatched MHC class II molecules did not prevent restoration of function. The reduced labeling of surface MHC class II molecules previously reported was shown to reflect a loss in total class II molecules within the cells; MHC class I levels were unaltered by exposure to the virus. In DC from RLV-infected mice biosynthesis of MHC class II was decreased by around 50% at the transcriptional level in comparison with beta-actin. Thus, the down-regulation of surface class II molecules observed in DC following RLV infection is a consequence of a specific block in its biosynthesis and the failure of DC to stimulate T cells may be a direct consequence of the reduced class II levels. Since reduced stimulation by DC is also seen in HIV-1 infection in humans we speculate that a similar mechanism might operate in retroviral infection in man.

Although inactivated viral vaccines have been dramatically successful in controlling many of the world's most devastating diseases, they frequently need several injections to ensure high levels of protection, and thus their efficacy is reduced in many situations. We have developed several rapid vaccination protocols for two commercial vaccine preparations against tick-borne encephalitis virus and studied their efficacy in an experimental murine model. Vaccination protocols as brief as two doses given over two days elicit efficient protection against challenge with potentially fatal doses of virus and this protection is afforded as soon as 5 or as long as 100 days after the first vaccination. The very rapid induction of protection and the poor antibody responses observed would suggest that cell-mediated immune responses are the most important mechanisms for the protection elicited by conventional inactivated vaccines against tick-borne encephalitis.

Flaviviruses elicit a humoral immune response to two virus-encoded, membrane-associated glycoproteins. One is the major virion surface envelope protein (E), which is recognized by antibody, whereas the other is a secreted, heavily glycosylated non-structural protein (NS1). Inoculation with either protein can give rise to a protective immune response, as can the passive transfer of E and NS1 monospecific monoclonal antibodies. Experiments reported here demonstrate that the secreted form of NS1, whether from cells infected with tick-borne encephalitis virus (TBEV) or from cells infected with a defective recombinant adenovirus containing the NS1 gene, occurs chiefly as a pentamer or hexamer and occasionally as a decamer or dodecamer. Intracellular forms of this protein however occur only as dimers. The higher M(r) forms secreted from the cell are exquisitely sensitive to detergent, suggesting they are held together by hydrophobic bonds. Both intracellular and extracellular forms of the dimer can be dissociated by heat, but at different temperatures. Unlike similar proteins from mosquito-borne viruses. NS1 from TBEV-infected cells cannot be dissociated at ambient temperatures by extremes of pH. Studies on the antigenic structure of this protein show it to have several highly conserved epitopes, confirming similar earlier conclusions from amino acid sequence analyses.

The authors examined 110 males and females admitted to hospital within 6 hours since the onset of new macrofocal anterior myocardial infarction running without complications. All the patients underwent transcardiac galvanization (TG) in the initial disease hours. Precardiac mapping, tetrapolar rheography and clinical observation provided evidence on TG course action on the necrosis mass, central hemodynamics and clinical appearance of the disease in the acute and postinfarction periods. TG promoted faster and more marked improvement of cardiac muscle contractility, attaining higher functional level of inotropic myocardial activity up to the disease month 6. This was associated with alleviation of circulatory disorders both at the hospital and postinfarction stage. The response to course TG results from cardioprotective action of constant electric current on ischemic myocardium which limits the zone of the lesion thus reducing early fatal cases number and preventing cardiogenic shock.

During the IF2-catalysed formation of the 30S initiation complex, the GTP requirement and its subsequent hydrolysis during 70S complex formation are considered to be essential for translation initiation in Escherichia coli. In order to clarify the role of certain amino acid residues believed to be crucial for the GTP hydrolytic activity of E. coli IF2, we have introduced seven single amino acid substitutions into its GTP-binding site (Gly for Val-400; Thr for Pro-446; Gly, Glu, Gln for His-448; and Asn, Glu for Asp-501). These mutated IF2 proteins were expressed in vivo in physiological quantities and tested for their ability to maintain the growth of an E. coli strain from which the functional chromosomal copy of the infB gene has been deleted. Only one of the mutated proteins (Asp-501 to Glu) was able to sustain cell viability and several displayed a dominant negative effect. These results emphasize that the amino acid residues we substituted are essential for the IF2 functions and demonstrate the importance of GTP hydrolysis in translation initiation. These findings are discussed in relation to a previously proposed theoretical model for the IF2 G-domain.

Exponentially growing rat islet cells (RINr) and hamster islet cells (HIT T-15) were incubated in presence of tolbutamide (10-1000 microM), gliclazide (0.1-10 microM) or glibenclamide (0.01-10 microM) for 15 hrs. Accumulation of insulin in culture medium was estimated by RIA. Effects of sulfonylureas (SU) on cell proliferation were assessed by 3H-thymidine (3H-T) incorporation into cellular DNA. All of SUs used stimulated insulin production in RIN and HIT cell cultures (with an exception of tolbutamide, which markedly suppressed insulin secretion in HIT cells at 1000 microM). 3H-T incorporation into RIN cells was elevated only in presence of gliclazide (10 microM), whereas tolbutamide at 1000 M significantly inhibited RIN cell proliferation. Gliclazide (0.1 microM) and glibenclamide (0.01-10 microM) enhanced 3H-T incorporation into HIT cells. Further detailed investigations of mechanisms of SU effects on islet cell reproduction will be of use for designing optimal strategy of hypoglycemizing therapy of diabetes mellitus.

The binding of tick-borne encephalitis virus to porcine kidney embryo cells was studied. Anti-idiotypic antibodies against TBE virus E protein precipitated gp110 kDa which is predicted to be a cellular receptor for TBE virus.

We have investigated the effects of Pro-Met-Asp-Phe-NH2 (PMAP) on insulin and glucagon release from human fetal pancreatic microfragments in vitro. Four batches of precultured microfragments were incubated for 24 hrs in medium containing 5.5 mM glucose, 17 mM glucose, 1 microM PMAP or 1 microM PMAP plus 17 mM glucose. PMAP significantly enhanced both basal and glucose-stimulated insulin release (2.2- and 4.1-fold, respectively). Glucagon secretion was markedly inhibited by glucose (17 mM). PMAP neither affected the basal glucagon release nor potentiated the inhibitory action of glucose on glucagon release. Hence, PMAR selectively regulates insulin production in human fetal islet tissue without affecting glucagon production. Our results suggest that the substances similar or related to PMAP may prove to be of clinical value in drug correction of diabetes mellitus.