Detection of polyamines using immunobiosensor based on zinc oxide nanoparticles.

AIM: To analyze the performance of biosensor based on nanoparticles of zinc oxide for the detection of spermine and spermidine in solution and in cell culture. MATERIALS AND METHODS: Zinc oxide nanoparticles were used for preparing biosensor containing antibodies to spermine and spermidine. Polyamine concentration is solutions of spermine and spermidine as well as in lyophilisate of MCF-7 cells was measured by luminescence of the samples excited by laser beam at 380 nm. RESULTS: The minimum concentration for the detection of polyamines in model solutions is 10 ng/ml, and maximum one is 100 ng/ml. A higher level of luminescence intensity of nanoparticles was found during analysis the polyamines in MCF-7 lyophilisate allowing for detecting polyamines at concentrations from 100 cells/ml to 100,000 cells/ml. CONCLUSIONS: The proposed biosensor system for determining the level of biogenic polyamines in cell lyophilisate using the optical properties of zinc oxide nanoparticles is promising for further improvement of the methodology and its implementation for detection and measurement of polyamines in biological systems.

[Calpains: general characteristics and role in various states of the organism].

Calpains are a family of cytoplasmic calcium-dependent proteinases with papain-like activity. They participate in a variety of processes in the body: age changes, functioning of endothelium and pulmonary system, regulation of apoptosis and necrosis, development of various hypometabolic states, arterial hypertension, diabetes and chronic kidney disease, tumor growth. It is concluded that calpains, causing limited proteolysis of substrates, play an important role in a wide range of biological phenomena. Their activity is associated with the response to the calcium-dependent signaling and the effects of aging. Inhibition of calpains activity contributes to inhibition of endothelial dysfunction, cardiovascular disease, formation of structural and functional changes in the kidney tissue, has neuroprotective effect, preventing sarcopenia, reduces inflammatory reactions caused by hyperventilation of the lungs.

[Detection of the level of antibodies against bovine leucosis virus in the cow milk by immune sensor].

An immune sensor based on the surface plasmon resonance (SPR) was developed for express diagnostics of bovine leucosis. Sensor used for detection of the level of antibodies against bovine leukaemia virus (BLV) in the milk serum. It was shown that immune sensor analysis is more sensitive, rapid and simple in comparison with the traditional AGID test. It was stated that the developed immune sensor may be used for performance of screening of bovine leucosis at the farms and the minimal dilution of the milk serum should be 1:20.

[Thermobiosensors: features of design and function and prospects for their use]. / Termobiosensory: osoblyvosti konstruktsiï i funktsionuvannia ta perspektyva praktychnoho vykorystannia ïkh.

Thermobiosensors are biosensors, which utilize thermal sensors (thermistors, thermocouple) for measuring changes of heat arising during ferment, immune or metabolic reactions. Thermosensors apply to an estimation of a condition of ferment systems, interactions of immune components, kinetics of a biocatalysis, and also for application in biochemical diagnostics, at definition of quality of products, monitoring of an environment and control of biotechnological processes. In the review the data, concerning of the major principle of development and creation mini- and microthermobiosensors, are given on the basis of modern achievement in the field of technology of micromanufacturing. The various types mini-, micro- and multithermobiosensors, and also their application in separate areas of the biochemical and clinical analysis and monitoring of an environment are surveyed. The development and modern achievement in creation of separate variants thermosensors is in detail described on the basis of thermistors and thermocouples. The place thermosensors in general development of biosensor controls is allocated and the attention to prospect of their further improvement and application is given.

[Immobilization of immunoglobulins using lectins with an immune sensor method based on surface plasmon resonance]. / Immobilizatsiia imunohlobuliniv za dopomohoiu lektyniv z metoiu stvorennia imunnykh sensoriv na osnovi poverkhnevoho plazmonnoho rezonansu.

The effect of different lectins upon the response of immune sensor based on surface plasmon resonance (SPR) was investigated. Lectins affinity to carbohydrates of the IgG can be used to increase the density and orientation of IgG molecules at their immobilisation on the sensor surface. Conditions were elaborated for enhancement of immune sensor response in comparison with that one for bare or preliminary treated with dodecanthiol thin gold sensor surface. It was shown that human IgG revealed maximal affinity to wheat germ lectin and mouse monoclonal antibodies and rabbit IgG--to helix pomatia lectin. Pig antibodies, similar to human IgG, showed the greatest affinity to wheat germ lectin.

[Express diagnostics of bovine leucosis by immune sensor based on surface plasmon resonance]. / Ekspresna diahnostyka leikozu velykoï rohatoï khudoby za dopomohoiu imunnoho sensora na osnovi poverkhnevoho plazmonnoho rezonansu.

An immune sensor based on the surface plasmon resonance (SPR) was developed for express diagnostics of bovine leucosis. The sensor was used for detection of the level of antibodies against bovine leukaemia virus (BLV) in the blood serum. The industrially manufactured BLV antigen for screening test in the agar gel immunodiffusion (AGID) required the additional purification in order to be used in immune sensor analysis. It was shown that immune sensor analysis was more sensitive, rapid and simple in comparison with the traditional AGID test. It was stated that the developed immune sensor was capable to be used for performance of bovine leucosis screening at the farms and the minimal dilution of the serum should be 1:500.

[Application of chemiluminescent technique for determination of Daphnia magna sensitivity to various types of toxic agents]. / Zastosuvannia khemiliuminestsentnoho metodu dlia vyznachennia chutlyvosti Daphnia magna do riznykh typiv toksychnykh rechovyn.

The character and intensity of activated chemiluminescence of Daphnia living medium are investigated depending on the effect on them of various concentration different in nature toxic substances, such, as potassium bichromate, methomyl and tween-80. It is shown, that the toxic influence of these agents can be defined by registration of stimulating effect of Daphnia's exometabolites on the process of chemiluminescence. The sensitivity of the method makes: for potassium chromate--0.005 mg/l, methomyl--0.0013 mg/l and tween-80--0.1 mg/l.

[Application of lectins for determination of antibodies to glycoproteins of leukemia virus in bovine blood serum]. / Vykorystannia lektyniv dlia vyiavlennia antytil do hlikoproteïniv virusu leikozu v syrovattsi krovi velykoï rohatoï khudoby.

The possibility of using different lectins for the determination of level of specific antibodies to leukosis virus virus in bovine blood serum whit the help of the surface plasmon resonance was investigated. The modification of transducer surface with lectins provides for immobilisation of glycosilated proteins of leukaemia virus. It allows to make selective determination of antibodies to glicosilated protein, the quantity of which more high in blood serum of ill animals in the comparison with the immunised ones. Moreover, the treatment of surface with FGA-P and PLA lectins increases the sensitivity of immune sensor to the specific antibodies in the comparison with the non-treated one.

[Enzymatic sensor for determing triglycerides]. / Fermentnyi sensor dlia vyznachennia rivnia tryatsylglitseroliv.

An enzymatic sensor based on ion selective field effect transistors was developed for the determination of triacylglyceroles in the range of concentrations from 0.1 to 2 mM. It was show that the sensitivity of the sensor was equal to 0.1 mM and time of one measurement was 2 min. The optimal environment for the triacylglyceroles registration is 1 mM sodium phosphate buffer (pH 7.3).

[Immunosensors: sources of origin, achievements and perspectives]. / Imunosensory: vytoky vynyknennia, dosiagnennia ta perspektyvy.

The analysis of the recent data in the literature and results of investigations in the field of the development and study of function efficiency of different types of immune sensors, that are performed at the Department of Biochemistry Sensory and Regulatory Systems of the A. V. Palladin Institute of Biochemistry of Ukrainian National Academy of Sciences are presented. Sources of origination and perspectives of the devlopment of biosensors are discussed as well. The paper also gives an overview of main research projects at the Department, mainly in the filed of biosensors. They include development of the scientific bases for the creation of a new generation of chemo- and biosensors for their application in medicine and ecology. Multi-immune, multi-enzyme and combined multi-parametrical sensors can provide express analyses in laboratory and field conditions with the purpose to perform immune chemical diagnostics of diabet, kidney diseases, immune defficiencies, autoimmune, allergic, pre-infarction and pre-tumor states as well as to control total toxicity of the environment and identification of main types of toxic elements in it. The investigations are based on the latest achivements in the field of physics, chemistry, information technology and electronics with the use of different types of planar electrodes, ion sensitive field effect transistors (ISFETs), semiconductor capacitive structures, termistors, optrodes, piezocrystalls and application of such methods and effects as laser correlation spectroscopy, chemiluminescence, fluorescence, surface plasmon resonance, photoluminescence of porous silicon, interferometry, evanescent wave technique, nonemmiting energy transfer and holography.

[Development of enzyme biosensor based on trypsin and conductometric thin-film electrodes for protein and artificial substrates determination]. / Enzimosensor na osnovi tripsinu ta konduktometrichnykh planarnykh elektrodiv dlia viznacheniia vmistu bilkiv peptidnikh substrativ u vodnikh rozchinakh

Two analytical systems based on the soluble and immobilized trypsin and conductometric thin-film electrodes were developed to measure concentration of artificial substrate and protein in solution. It was shown that these systems allow one to determine concentrations of Ha-benzoyl-L-arginin-ethyl-ester in the range of 0.1-1.0 mM and concentrations of HSA: 0.1-2.0 mg/ml with soluble and 0.1-0.8 mg/ml with immobilized trypsin.

[The effect of hypoxic hypoxia on the physicochemical and functional properties of hemoglobin in rats]. / Vplyv hipoksiinoï hipoksiï na fizyko-khemichni i funktsiini vlastyvosty hemohl'obinu shchuriv.

The effect of intermittent altitude chamber hypoxia is established to cause an increase in solvability of laboratory rat hemoglobin. Results of immunochemical and fluorescent analysis of the samples of hemoglobin and its component are presented and discussed. They prove that changes in solvability of hemoglobin are determined by the conformational reconstructions of the respiration protein as a result of formation of the complexes with internally erythrocytic metabolites.