Immune Response of Mice Against Babesia canis Antigens is Enhanced When Antigen is Coupled to Gold Nanoparticles.

PURPOSE: The aim of this study was to isolate Babesia canis soluble antigens and to investigate the effect of their conjugates with gold nanoparticles on the immunogenicity in laboratory animals. METHODS: A procedure was developed for isolating and purifying B. canis antigens. The isolated culture antigen of B. canis 495 was coupled to gold nanoparticles, and the conjugate was used to immunize laboratory mice. RESULTS: Western blotting showed that the resultant antiserum specifically recognized the proteins of the B. canis strains isolated from naturally infected dogs. The antibody titer, the respiratory activity of peritoneal macrophages, the proliferative activity of splenocytes, and the production of cytokines were maximal when the animals were immunized with the antigen-nanoparticle conjugate emulsified in complete Freund's adjuvant. Without adjuvant, the babesial antigen was weakly immunogenic. CONCLUSION: Therefore, the use of gold nanoparticles as an antigen carrier induced a broad immune response involving both cellular and humoral responses. The antibodies raised by the proposed procedure are potentially effective at immunodetection of Babesia canis infections in dogs.

Insitu magnesium calcium phosphate cements formation: From one pot powders precursors synthesis to in vitro investigations.

Calcium phosphate cements are of great interest for researchers and their applications in medical practice expanded. Nevertheless, they have a number of drawbacks including the insufficient level of mechanical properties and low degradation rate. Struvite (MgNH4PO4) -based cements, which grew in popularity in recent years, despite their neutral pH and acceptable mechanical performance, release undesirable NH4 + ions during their resorption. This issue could be avoided by replacement of ammonia ions in the cement liquid with sodium, however, such cements have a pH values of 9-10, leading to cytotoxicity. Thus, the main goal of this investigation is to optimize the composition of cements to achieve the combination of desirable properties: neutral pH, sufficient mechanical properties, and the absence of cytotoxicity, applying Na2HPO4-based cement liquid. For this purpose, cement powders precursors in the CaO-MgO-P2O5 system were synthesized by one-pot process in a wide composition range, and their properties were investigated. The optimal performance was observed for the cements with (Ca + Mg)/P ratio of 1.67, which are characterized by newberyite phase formation during setting reaction, pH values close to 7, sufficient compressive strength up to 22 ± 3 MPa (for 20 mol.% of Mg), dense microstructure and adequate matrix properties of the surface. This set of features make those materials promising candidates for medical applications.

The biological acoustic sensor to record the interactions of the microbial cells with the phage antibodies in conducting suspensions.

The acoustic biological sensor for the analysis of the bacterial cells in conducting suspension was developed. The sensor represented the two channel delay line based on the piezoelectric plate of Y-X lithium niobate thick of 0.2mm. Two pairs of the interdigital transducers (IDT) for the excitation and reception of shear horizontal acoustic wave of zero order (SH0) in each channel were deposited by the method of photolithography. One channel of the delay line was electrically shorted by the deposition of thin aluminum film between IDTs. The second channel remained as electrically open. The liquid container with the volume of 5ml was fixed on the plate surface between IDTs by the glue, which did not cause the additional insertion loss. For the first time the influence of the conductivity of the cell suspension on the registration of the specific and nonspecific interactions of the bacterial cells with phage-antibodies (phage-Abs) was studied by means of the developed sensor. The dependencies of the change in insertion loss and phase of the output signal on the conductivity of the buffer solution at specific/nonspecific interactions for the electrically open and shorted channels of the delay line were obtained. It was shown that the sensor successfully registered the interactions of microbial cells with phage-Abs in the range of the conductivity of 2-20 µS/cm on the model samples A. brasilense Sp245 - specific phage-Abs. The sensor in the time regime of the operation fast reacted on the specific/nonspecific interaction and the time of the stabilization of the output parameters did not exceed 10min.

Electro-acoustic sensor for the real-time identification of the bacteriophages.

The electro-acoustical sensor for the real-time identification of the bacteriophage FAl-SR65 is described. The polyclonal antibodies specific to the bacteriophage FAl-SR65 were for the first time obtained, and the possibility of their application to the bacteriophage detection by using the electro-acoustical method of analysis was also studied for the first time. The electro-acoustical sensor was based on the lateral electric field excited piezoelectric resonator. The frequency dependences of the real and imaginary parts of the electrical impedance of such a resonator, loaded with the virus-antibody suspension, significantly differed from those measured for the same resonator with the same virus suspension but without antibodies (control). The limit for the reliable bacteriophage detection during their interaction with the antibodies (sensitivity level) was determined to be 106 phages ml-1. The obtained results demonstrate the possibility to record the bacteriophage-antibody interactions and can be used as the basis for the development of the biological sensor for the quantitative virus detection.

[Immunodetection of bacteriophages by a piezoelectric resonator with lateral electric field].

It has been demonstrated that electroacoustic analysis with polyclonal antibodies can be used for bacteriophage detection. The frequency dependences of the real and imaginary parts of electrical impedance of a resonator with a viral suspension with antibodies were shown to be essentially different from the dependences of a resonator with control viral suspension without antibodies. It was shown that ΦAl-Sp59b bacteriophages were detected with the use of antibodies in the presence of foreign virus particles. The ΦAl-Sp59b bacteriophage content in the analyzed suspension was ~1010­106 phages/mL; the time of analysis was no more than 5 min. The optimally informative parameter for obtaining reliable information was the change in the real or imaginary part of electrical impedance at a fixed frequency near the resonance upon the addition of specific antibodies to the analyzed suspension. It was demonstrated that the interaction between bacteriophages and antibodies can be recorded, offering good prospects for the development of a biological sensor for liquid-phase identification and virus detection.

[The analysis of biochemical markers of MCF-7 cells apoptosis induced by recombinant analog of lactaptin].

Earlier we isolated and characterized human milk pro-apoptotic peptide - lactaptin and generated its engineered analog - RL2. It was shown that both lactaptin and RL2 are capable to induce apoptotic death of MCF-7 cells. In this report we have analyzed biochemical markers of RL2 induced MCF-7 apoptosis. The activation of initiator and effector caspases as well as mitochondrial membrane potential and cytoplasm membrane changes were analyzed using flow cytometry and Western-blot methods. We have found that RL2 induced apoptotic death of MCF-7 cells was accompanied by PS exposure on the plasma membrane surface. It also was shown that RL2 has induced dissipation of mitochondrial membrane potential and resulted in activation of initiator caspases 8, 9 and effector caspase 7.

Altered juvenile hormone metabolism, reproduction and stress response in Drosophila adults with genetic ablation of the corpus allatum cells.

Juvenile hormone (JH), which controls many developmental and physiological processes in Drosophila melanogaster, is synthesized de novo in the specialized endocrine glands, corpus allatum (CA). The present study concerns JH metabolism, reproduction and stress resistance in Drosophila with genetic ablation of a part of CA cells. The correlated regulation of JH biosynthesis and degradation in Drosophila adults has been found: ablation of CA cells led to (1) a dramatic decrease in activity of the key regulatory enzyme of JH biosynthesis, juvenile hormone acid methyl transferase and (2) a considerable increase in JH-hydrolyzing activity. It has been also shown that ablation of CA cells caused three significant physiological changes: (1) an increase in the intensity of response of JH degradation system to heat stress; (2) a disturbance of reproduction; (3) a decrease in stress resistance. Pharmacological rise of JH level rescued JH-hydrolyzing activity, fecundity and stress resistance in CA-ablated females. Pronouncedly, all the physiological effects caused by CA ablation were significant in females but not in males indicating a sexual dimorphism of JH physiological roles in Drosophila adults.

[Extracorporeal methods of blood purification in cardiovascular diseases]. / Ekstrakorporal'nye metody ochishcheniia krovi pri serdechno-sosudistykh zabolevaniiakh.

Increasing sensitivity to drugs, elimination of endotoxicosis, recovery of normal homeostatic control after hemocarboperfusion help to break off the vicious circles associated with severe cardiovascular disorders, and improve the effect of subsequent medication. Hemocarboperfusion added to combined intensive care and resuscitation procedures was associated with a 82.9% success rate.

[Complex immunochemical study of plasma proteins and soluble antigens of leukocytes in patients with ischemic heart disease during hemoperfusion therapy]. / Kompleksnoe immunokhimicheskoe issledovanie belkov plazmy krovi i rastvorimykh antigenov leikotsitov u bol'nykh ishemicheskoi bolezn'iu serdtsa pri gemoperfuzionnoi terapii.

An immunochemical analysis of 16 individual plasma proteins and 8 leukocyte soluble antigens (LSA) was performed over time in 11 coronary patients after hemosorption. The concentration of plasma lipoproteins gradually returned to normal within 2-3 weeks. Leukocyte dysfunction in the patients manifested itself in change of the specific LSA concentration in hemolysates as compared to that in donors. Shortly after hemosorption a selective 2-8-fold increase in the LSA concentration in the hemolysates was noted. Response of the leukocyte system was observed in all the cases and recorded in repetition of the procedures. Individual LSA function is under study.