Category Archives: Uncategorized
Long-term hypocholesterolemic effect of amidated alginate in rats
M. Marounek1, Z. Volek1, T. Taubner1, D. Dušková1, L. Kalachniuk2
1Institute of Animal Science, Prague, Czech Republic;
e-mail: marounek.milan@vuzv.cz;
2National University of Life and Environmental Sciences of Ukraine, Kyiv;
e-mail: kalachnyuk_liliya@nubip.edu.ua; lilkalachnyuk@gmail.com
The effect of octadecylamide of alginic acid on blood serum and hepatic cholesterol, and the faecal output of fat and sterols was examined in female rats fed diets containing cholesterol and palm fat at 10 and 50 g/kg, respectively for 10 weeks. Amidated alginate, supplied at 10 and 20 g/kg, significantly decreased serum cholesterol from 5.25 to 2.99 and 2.39 µmol/ml, respectively, and decreased hepatic cholesterol from 30.7 to 12.3 and 9.4 µmol/g, respectively. Amidated alginate increased the faecal output of fat and at higher dosing significantly decreased faecal output of bile acids. Faecal output of bile acids and hepatic cholesterol significantly correlated (r = 0.791; P < 0.001). The results of the present experiment showed that hypocholesterolemic effect of amidated alginate persisted within 10 weeks of feeding.
The effect of intranasally administered TLR3 agonist larifan on metabolic profile of microglial cells in rat with C6 glioma
Y. Hurmach1, M. Rudyk1, V. Svyatetska1, N. Senchylo1,
O. Skachkova2, D. Pjanova3, K. Vaivode3, L. Skivka1
1ESC Institute of Biology and Medicine, Taras Shevchenko National University of Kyiv, Ukraine;
2Research Laboratory of Experimental Oncology, National Cancer Institute, Kyiv, Ukraine;
3Latvian Biomedical Research and Study Centre, Riga, Latvia;
e-mail: jhurmach@gmail.com
Glioma-associated microglia/macrophages (GAM) represent an attractive therapeutic target for the development of the alternative methodology in the treatment of gliomas. This study was aimed to investigate the effect of intranasally administered TLR3 agonist Larifan on microglial cell metabolic profile in rats with C6 glioma. Our results demonstrate progressive generation microglial cell population with immunosuppressive and pro-inflammatory properties in C6 glioma-bearing brain. Intranasally delivered TLR3 agonist is capable to abrogate the creation of this pro-tumoral immune infiltrates, probably, through the effect on myeloid-derived suppressor cells, and can be considered as a promising agent for glioma therapy aimed the GAM re-education.
The effects of PDK4 inhibition on AMPK protein levels and PGC-1α gene expression following endurance training in skeletal muscle of Wistar rats
S. Aminizadeh1, Y. Masoumi-Ardakani1, B. Shahouzehi2
1Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Iran;
e-mail: soheilaminizadeh@gmail.com; ymab125@yahoo.com;
2Cardiovascular Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Iran;
e-mail: bshahouzehi@gmail.com
There are regulatory networks in cells which surveil the physiological and environmental states. These cellular regulations are conducted through gene expression modulation. Skeletal muscle is able to adapt shortly and produce ATP at different conditions. AMPK (AMP-activated protein kinase) and PGC-1α (peroxisome proliferator-activated receptor-gamma coactivator-1alpha) are important regulators of cellular energy homeostasis. We designed this study to examine the effects of interactions between endurance training and PDK4 (pyruvate dehydrogenase kinase 4) inhibition on AMPK and PGC-1α expression in rat skeletal muscle. Thirty-two male Wistar rats were randomly selected and divided into 4 groups (n = 8); Group 1 control which did not receive any treatment, Group 2 received dichloroacetic acid (DCA) (150 mg/kg/day), Group 3 (endurance training group), Group 4 which received DCA and performed endurance training. AMPK protein expression, PDK4 and PGC-1α gene expression were measured by western blotting and real-time PCR, respectively. Our data showed that PDK4 inhibition caused AMPK protein elevation. Endurance training (group 2) and PDK4 inhibition (group 4) induce significant enhancement of PGC-1α gene expression compared to control group. The group which received DCA showed significant elevation of PDK4 gene expression compared to control group (P = 0.001), also other two groups (groups 2 & 3) showed significant elevation of PDK4 gene expression compared to control (P = 0.006). It seems that the combination of endurance training and PDK4 inhibition by up-regulation of PGC-1α expression, effectively improves energy state and performance in skeletal muscle.
Disturbance of the transmembrane phosphatidylserine asymmetry in hepatocytes as an apoptosis marker under the action of xenobiotics on rats
O. A. Nakonechna, L. A. Babijchuk, A. I. Bezrodna
Kharkiv National Medical University, Kharkiv, Ukraine;
e-mail: bezrodnaya.ai@gmail.com
It has been reported that unfavorable chemical environmental factors affect the functional state of liver, activate free radical processes against the background of the reduced antioxidant activity, change physico-chemical properties and membrane phospholipid composition of hepatocytes. The aim of our research was to estimate phosphatidylserine distribution in the phospholipid bilayer of hepatocyte membranes and apoptosis stages in hepatocytes of rats under the influence of surfactants: ethyleneglycol (EG), polyethyleneglycol 400, (PEG-400) and polypropyleneglycol (PPG) at a dose of 1/10 DL50. It was found in the subacute toxicological experiment on rats that the investigated xenobiotics EG, PEG-400 and PPG at a dose of 1/10 DL50 caused phosphatidylserine translocation to the outer membrane in the phospholipid bilayer of hepatocytes. This is a specific signal for macrophages aiming at recognition and elimination of apoptotic cells. Analysis of cell death modes under the influence of the investigated xenobiotics at a dose of 1/10 DL50 revealed that the intake of xenobiotics was associated with an increase in the amount of apoptotic/necrotic hepatocytes.
Influence of C(60) fullerene on the ischemia-reperfusion injury in the skeletal muscle of rat limb: mechanokinetic and biochemical analysis
D. O. Zavodovskyi1, S. Yu. Zay2, T. Yu. Matvienko1, Yu. I. Prylutskyy1,
N. Y. Nurishchenko1, S. S. Paradizova3, L. L. Bezuh3, U. Ritter4, P. Scharff4
1ESC Institute of Biology and Medicine, Taras Shevchenko National University of Kyiv, Ukraine;
2Lesya Ukrainka Eastern European National University, Lutsk, Ukraine;
3SI The Territorial Medical Association of the Ministry of Internal Affairs of Ukraine in Kyiv;
4Institute of Chemistry and Biotechnology, Technical University of Ilmenau, Ilmenau, Germany;
e-mail: Lab@univ.kiev.ua
Influence of the pristine C60 fullerene aqueous colloidal solution (C60FAS) on the ischemia-reperfusion injury in the skeletal muscle of rat limb was studied. Skeletal muscle damage effects were induced by 3 h lasting vascular ischemia. The impact of C60FAS was studied after its intramuscular injection immediately after 1 h of reperfusion at different doses, namely: 1, 2 and 3 mg/kg of body weight. Changes in the mechanokinetic parameters of ischemic skeletal muscle contraction at different modes of functioning and biochemical parameters of blood were used as markers of ischemic injury, and analyzed in detail under action of C60FAS. The obtained results indicate on great promise of use of C60FAS to reduce the consequences of ischemic muscle trauma.
Immunogenicity assay of KatG protein from Mycobacterium tuberculosis in mice: preliminary screening of TB vaccine
P. Purkan1, R. Budiyanto1, R. Akbar1, S. P. A. Wahyuningsih2, W. Retnowati3
1Biochemistry Division, Chemistry Department, Faculty of Sciences and Technology, Airlangga University, Campus C, Jl. Mulyorejo-Surabaya, Indonesia;
2Biology Department, Faculty of Sciences and Technology, Airlangga University, Campus C, Jl. Mulyorejo-Surabaya, Indonesia;
3Microbiology Department, Faculty of Medicine, Airlangga University, Campus C, Jl. Moestopo-Surabaya, Indonesia;
e-mail: purkan@fst.unair.ac.id
The tuberculosis (TB) disease is still widely found even though BCG vaccine given to many people. Ineffectiveness of the BCG vaccine is one of causes that make the difficulties in preventing TB transmission. Objective of the research was to determine the immunogenicity of KatG protein of M. tuberculosis clinical isolate L19 in mice. The KatG protein as antigen was prepared by expression of the katG gene of M. tuberculosis clinical isolate L19 in Escherichia coli BL21 using pColdII-DNA vector. After purification by affinity chromatography, the KatG was vaccinated to mice to detect its immunogenicity. The expression of katG in E. coli BL21 could result in KatG protein with molecular weight 80 kDa in sodium dodecyl sulfate gel electrophoresis (SDS-PAGE). The pure KatG protein could significantly stimulate the immune response of mice by triggering the antibodies production of IgG1, IgG2a, IgG2b, IgG2c, IgG3, and IgM. The highest antibody level was obtained when the mice were vaccinated by KatG L19 with the dose of 45 μg/ml. Of the antibodies, the IgG2c isotype was dominantly produced in the blood serum. The KatG protein exhibited a high immunogenicity in mice, so it is possible to develop as a vaccine candidate for TB. A clinical test should be performed in a future to ensure its safety as a therapeutic protein.
Effects of alpha-ketoglutarate on lifespan and functional aging of Drosophila melanogaster flies
M. P. Lylyk1, M. M. Bayliak1, H. V. Shmihel1,
J. M. Storey2, K. B. Storey2, V. I. Lushchak1
1Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
2Institute of Biochemistry, Carleton University, Ottawa, Canada;
e-mail: lushchak@pu.if.ua; bayliak@ukr.net
The effects of an alpha-ketoglutarate-supplemented diet on lifespan and functional senescence were evaluated in the Canton S strain of Drosophila melanogaster. The results suggest that effects of dietary alpha-ketoglutarate (AKG) are dose- and gender-dependent. In males, diets containing 1-10 mM AKG did not affect mean and maximum lifespans, except that an increased maximum lifespan observed at 10 mM AKG. Diet with 20 mM AKG shortened median lifespan and had no effect on maximum lifespan of males. In females, diets with low concentrations of AKG (1 and 5 mM) did not affect lifespan, whereas diets supplemented with 10 and 20 mM AKG increased both median and maximum lifespans. At a lifespan-prolonging concentration (10 mM), AKG decreased fecundity, increased cold resistance and had no effect on climbing activity or resistance to oxidative stress in flies of either gender at middle (24 days) and old (40 days) ages. Moreover, middle-aged AKG-fed females but not males were more resistant to heat stress that was accompanied by higher levels of HSP90 protein as compared with controls. Middle-aged flies on AKG-supplemented diets showed elevated oxidative stress and had higher total protein and triacylglycerol levels as compared with controls. Hence, anti-aging effects of AKG do not seem to be related to preventing oxidative stress development but involve metabolic rearrangement and synthesis of specific protective proteins, which aid to resist destructive processes with age.
Specificity and sensitivity of the new test for serological evaluation of tuberculosis using MPT83-MPT63 fusion antigen
A. A. Siromolot1,2, T. O. Chudina2, I. S. Danilova3,
O. M. Rekalova4, D. V. Kolybo1,2, S. V. Komisarenko2
1ESC Institute of Biology and Medicine, Taras Shevchenko National University of Kyiv, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3NSC Institute of Experimental and Clinical Veterinary Medicine, Kharkiv, Ukraine;
4SI National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky National Academy of Medical Sciences of Ukraine, Kyiv;
e-mail: saa0205@ukr.net
The aim of this work was to characterize the experimental test-system based on MPT83-MPT63 fusion antigen with reference commercial diagnostics and investigate the basic characteristics of enzyme-linked immunosorbent assay (ELISA) tests such as specificity and sensitivity. In addition, we investigated the correlation of biochemical and immunological parameters of blood samples of patients with tuberculosis, which was correctly diagnosed by those test-systems. It was shown that the developed test-system match the existing ones regarding the criteria of reliability and the basic requirements of ELISA kits. Recommendations for the testing of serum samples and indications for the use of the proposed serological diagnostic methods has been suggested.
Plasminogen modulates formation of reactive oxygen species in human platelets
A. A. Tykhomyrov, D. D. Zhernosekov, M. M. Guzyk, V. V. Korsa, T. V. Grinenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: artem_tykhomyrov@ukr.net
Reactive oxygen species (ROS) are considered to be important signalling molecules controlling many platelet functions. ROS production has been shown to be augmented by platelet activation, however, plasminogen (Pg) has not been studied in the context of modulating intraplatelet ROS levels. The aim of this study was to investigate the ability of different Pg forms to affect platelet metabolic activity/survival and intracellular ROS production in resting and activated platelets. Platelets isolated from donor plasma were pre-treated with Glu- or Lys-Pg (1.2 µM) and activated by thrombin (1.0 NIH unit/ml) or collagen (1.25 mg/ml). MTT assay was adapted to estimate total mitochondrial dehydrogenase activity, while intracellular ROS levels were monitored with the use of H2DCF-DA probe by flow cytometry. Lys-Pg was shown to slightly, but significantly, mitigate MTT reduction (P < 0.05 vs. control platelets). Two-fold elevation in metabolic activity of platelets stimulated by thrombin as compared to untreated cells was observed. However, this activation was less exhibited in the case of platelets pre-incubated with either Glu- of Lys-Pg, with a predominant effect of Lys-Pg. Unlike thrombin, collagen treatment dramatically suppressed metabolic activity of platelets by 60% compared to control (P < 0.05). Glu- or Lys-Pg pre-incubation had no effects on the activity of collagen-stimulated platelets. Two subpopulations of platelets were observed with distinct characteristics of intracellular ROS formation. Elevated ROS production was demonstrated in these populations of both thrombin- and collagen-treated platelets. Pg (Lys-form to greater extent) enhanced intracellular ROS generation in thrombin-stimulated platelets. These findings suggest that augmented ROS generation within platelets pre-treated with Pg followed by their stimulation may result in down-regulation of their survival and functional activity. This study adds to our understanding one more possible mechanism of Pg impact on the platelet function.