Category Archives: Uncategorized

Exometabolites of endospore-forming bacteria of Bacillus genus identified by genomic-metabolomic profiling

А. M. Ostapchuk, М. D. Shtenikov*, V. О. Ivanytsia

Odesa I. I. Mechnykov National University, Ukraine;
*e-mail: shtenikovnik@gmail.com

Received: 30 March 2020; Accepted: 13 November 2020

The set of unique bioactive metabolites produced by marine bacilli is already known but the metabolomic of these bacteria  is underinvestigated. The aim of this work was to carry out  the comparative analysis of metabolomic and genomic traits of Bacillus velezensis ONU 553, Bacillus pumilus ONU 554, Bacillus subtilis ONU 559 strains isolated from Black Sea bottom sediments. Organic extracts of each strain were analyzed using high-resolution liquid chromatography-mass spectrometry. General annotation of genomes was performed using PATRIC, search for secretory signals in the primary structure of selected proteins with using Signal IP analysis.  The search of biosynthetic gene clusters was performed using antiSMASH, PRISM 3 and BiG-SCAPEs, the reconstruction of metabolites- with PRISM 3 and TransATor analyzes. The study allowed to found and identified 90, 33 and 43 metabolites in the strains Bacillus velezensis ONU 553, Bacillus pumilus ONU 554 and  Bacillus subtilis ONU 559 respectively. The compounds found in metabolome were subdivided into two groups: those which are known members of the genus Bacillus and those new to both genus and prokaryotes in general. Among the secondary metabolites of studied strains the variants of the nonribosomal peptide class  surfactins (anhteisoC16-surfactin, surfactin B2-me ester), gageostatins, fengycins and amicoumacins, and the secreted protease inhibiting pentapeptide GPFPI were identified. The biosynthetic clusters of lipopeptides of the pumilacidin subgroup and amicoumacin antibiotic AI-77A were identified for the first time with the use of  bioinformatic tools. The data obtained replenish the  understanding of the marine bacilli biosynthetic potential.

Honeybee chitosan-melanin complex: isolation and investigation of antimicrobial activity

M. Lootsik1, N. Manko1, O. Gromyko2,
S. Tistechok2, M. Lutsyk (Jr.)3, R. Stoika1*

1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Ivan Franko National University of Lviv, Ukraine;
3Danylo Halytsky Lviv National Medical University, Ukraine;
*e-mail: stoika.rostyslav@gmail.com

Received: 4 May 2020; Accepted: 13 November 2020

Antimicrobial activity of marine crustaceans chitosans is well studied and is widely used in medicine, while chitosans of insects are poorly investigated in this aspect, though they might also be of practical significance. The aim of this study was to isolate and purify chitosan-melanin complex (CMC) from the honeybee­ corpses and to estimate its antimicrobial activity. Antibacterial activity of CMC was evaluated by MTT test, antifungal activity towards Candida albicans was estimated by calculating colony forming units (CFU method). The modified method of CMC isolation and purification was described which differs from the known analogs in deacetylation of chitin-melanin complex by its hydrolysis in 40% NaOH without previous melanin elimination and in further purification of CMC by differential solubilization at distinct pH values. The anti-microbial activity of CMC was characterized by prevalence of candidacidal effect, IC50 towards laboratory strain of C. albicans was 50 μg/ml. The ranking of studied bacteria sensitivity to the CMC action decreased as: E. coli > St. aureus > Ps. aeruginosa. It is suggested that CMC isolated from the honeybee corpses might be a perspective constituent of medicinal compositions for treatment of lesions caused by C. albicans infection.

Diazepinone effect on liver tissue respiration and serum lipid content in rats with a rotenone model of Parkinson’s disease

L. Ya. Shtanova1,2*, P. I. Yanchuk1, S. P. Vesеlsky1,
O. V. Tsymbalyuk1, T. V. Vovkun2, V. S. Moskvina2, O. V. Shablykina2,
S. L. Bogza2, V. N. Baban1, A. A. Kravchenko3, V. P. Khilya2

1Institute of High Technologies, Taras Shevchenko National University of Kyiv, Ukraine;
2Taras Shevchenko National University of Kyiv, Ukraine;
3Chuiko Institute of Surface Chemistry, National Academy of Sciences, Kyiv;
*e-mail: shtanova@ukr.net

Received: 5 March 2020; Accepted: 13 November 2020

Parkinson’s disease (PD) is a chronic and progressive age-related neurodegenerative disorder. Accumulation of α-synuclein aggregates, oxidative stress, mitochondrial dysfunction and lipid metabolism  disturbance are thought to be the key violations at PD pathogenesis. Despite long-time  research the causes of PD occurrence are not yet clear. We investigated the influence of diazepinon, a new derivative of benzodiazepine, on liver tissue respiration (LTR), serum lipid content and  behavioral parameters of rats with modeled PD induced by intraperitoneal injections of 2.0 mg/kg rotenone (ROT) within 28 days. LTR was assessed using the polarograph LP-9. Blood samples for biochemical analysis were collected from the inferior vena cava. The behavioral parameters of rats were studied by the open field test. We showed  that in rats with ROT – induced PD the coefficient of liver oxygen consumption was decreased by 33.5% (P < 0.001), the serum content of phospholipids, cholesterol, cholesterol esters, free fatty acids and triglycerides was reduced by 21.4% (P < 0.001), 28.8% (P < 0.001), 26.8% (P < 0.001), 30.3% (P < 0.01) and 41.5% (P < 0.001) respectively and the motor disorders were detected. Diazepinone application resulted in a full recovery of  LTR,  serum concentration of phospholipids, partial recovery of serum free fatty acids and triglycerides content and significant improvement of motor behavior. However  diazepinone did not affect the reduced concentration of cholesterol and cholesterol esters in the serum of rats with simulated PD.

The content of pro-inflammatory cytokines IL-1β, IL-6, IL-17A and TNFα in the blood of patients with type 2 diabetes after therapy with metformin

K. P. Zak*, O. V. Furmanova, V. V. Popova, Ya. A. Sayenko

SI “V.P. Komisarenko Institute of Endocrinology and Metabolism of National Academia of Medical Sciences of Ukraine”, Kyiv;
*e-mail: kpzak2017@gmail.com

Received: 28 February 2020; Accepted: 13 November 2020

Currently the world society is extremely worried about the global increase in the number of patients with diabetes on our planet. Annually 4 million people die of this disease, and the cost of its treatment reaches trillions of dollars. A new highly effective oral antidiabetic drug  metformin (1,1-dimethylbiguanide hydrochloride) is one of the most common hypoglycemic remedies  currently prescribed for the first-line treatment of patients with type 2 diabetes (T2D). However, the mechanism of its curative effect is still not clear. The results of our study showed  that metformin treatment of patients with newly diagnosed T2D was followed by  pronounced normalization of  the increased levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-17A and TNFα),  inflammation indexes and lymphocyte’s immunophenotype. The obtained data confirm the existing hypothesis about the inflammatory nature of T2D and indicate that the immune system, in particular proinflammatory cytokines, plays a significant role in the mechanism of the curative effect of metformin at  T2D.

Oxidative/antioxidant balance and matrix metalloproteinases level in the knee cartilage of rats under experimental osteoarthritis and probiotic administration

O. Korotkyi*, K. Dvorshchenko, L. Kot,
T. Vovk, M. Tymoshenko, L. Ostapchenko

ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: korotkyi@gmail.com

Received: 28 June 2020; Accepted: 13 November 2020

The aim of this work was to investigate the effect of the poly-strain probiotic on oxidative-antioxidant balance and the level of matrix metalloproteinases (MMPs) in rat knee cartilage under experimental osteoarthritis. Osteoarthritis was induced by a single injection of monoiodoacetate into the knee joint of rats. Probiotic was administered daily for 14 days. Knee cartilages homogenate was used to evaluate  the content of reactive oxygen species (superoxide anion and hydrogen peroxide), products of lipids peroxidation (diene conjugates, TBA-active compound, Shiff bases), to determine superoxide dismutase and catalase activity and activity of glutathione-dependent  antioxidant enzymes, the level of reduced and oxidized glutathione. The level of MMPs -1, -2, -3, -8 expression was estimated by ELISA. Osteoarthritis was found to cause a significant increase in the reactive oxygen species level, lipid peroxidation products content, superoxide dismutase and catalase activity, level of all studied MMPs, and also depletion of glutathione-dependent antioxidant system and the decrease in the ratio between reduced and oxidized glutathione.The administration of the probiotic was followed by the tendency for the restoration of the parameters to the values of the control group. Thus, the administration of the probiotic to rats with osteoarthritis may be considered as an anti-inflammatory and antioxidant agent for further clinical trials.

Indicators of lipid metabolism disorders in the blood serum of adolescents with metabolic syndrome

L. A. Strashok1,2, O. V. Buznytska1,2*, О. М. Meshkova3

1Kharkіv Medical Academy of Postgraduate Education, Ukraine;
2V.N. Karazin Kharkіv National University, Ukraine;
3Bogomolets National Medical University, Kyiv, Ukraine;
*e-mail: ebuznickaa@ukr.net; elena.buznytska@gmail.com

Received: 16 April 2018; Accepted:13 November 2020

Atherogenic dyslipidemia is one of the main and early indication of adolescentrs with metabolic syndrome. Because there is not enough information about the nature of dyslipidemia in adolescents and the possibilities of its correction and prevention, the study of this problem is relevant. The goal of the study was to estimate the indexes of lipid metabolism in blood serum of adolescents with metabolic syndrome. We examined 200 obese patients aged 14–18 years, who were divided into two groups with and without metabolic syndrome. The control group consisted of 30 adolescents of similar age with normal body weight. The lipid profile of blood was studied by standardized methods using Cormay Multi semi-automatic photometer­. The level of β-lipoproteins in blood serum was determined by the turbidimetric method. Elevated levels of triglycerides, low and very low density lipoprotein cholesterol, β-lipoproteins and a tendency to decreased high density lipoprotein level cholesterol were found in the  blood serum of adolescents with metabolic syndrome  as the signs of atherogenic dyslipidemia. Reliable correlations both between the indexes of lipid profile and between atherogenic dyslipidemia indicators and anthropometric indexes were established, indicating an increased risk of lipid metabolic disturbances in individuals with abdominal obesity.

Autoantibodies to myelin basic protein and histone H1 as immune biomarkers of neuropsychological disorders in patients with multiple sclerosis

S. Ya. Kyryliuk1, T. I. Nehrych1, N. K. Svyrydova2,
Ye. O. Trufanov2, R. S. Stoika3, Yu. Ya. Kit3

1Danylo Halytsky Lviv National Medical University, Ukraine;
2Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine;
3Institute of Cell Biology National Academy of Sciences of Ukraine, Lviv;
e-mail: sinitska@ukr.net

Received: 15 April 2020; Accepted: 13 November 2020

Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system with different disorders of neurological and higher cortical functions. It is important to identify biomarkers that can control the dynamics of neuropsychological changes and predict the progression of this process. The aim of the study was to investigate the pathogenic and clinical significance of serum autoantibodies to the myelin basic protein (MBP) and histone H1 in the occurrence of neurological and neuropsychological disorders in patients with MS. Fifty-five patients diagnosed for MS were examined. A general clinical and neurological examination, determination of cognitive status, depression level and the content of autoantibodies to histone H1 and MBP in the blood serum were conducted. Blood serum samples of 20 healthy volunteers were used in control.  The serum of patients with MS was shown to contain antibodies of IgG class to MBP and histone H1. The level of anti-histone H1 IgG-antibodies in blood serum of MS patients was found to be higher compared with the level of anti-MBP IgG-antibodies (P < 0.05). Increased levels of anti-MBP antibodies correlated with the severity of trunk ataxia, impaired conceptualization, and mood. High level of anti-histone H1 antibodies correlated with the severity of paresis, trunk ataxia, impaired conceptualization, semantic language, and mood. Determination of the level of anti-histone H1 antibodies in blood serum of patients with MS might serve as a biomarker of inflammatory and, probably, of the neurodegenerative processes of this disease and determine the dynamics of clinical course of the MS. Anti-MBP antibodies play an important role in the pathogenesis of the MS and are an additional marker of the severity of the clinical course of neurological and some neuropsychological disorders.

Activation of the PI3K/AKT/MTOR/P70S6K1 signaling cascade in peripheral blood mononuclear cells in patients with type 2 diabetes

T. S. Vatseba1*, L. K. Sokolova2, V. M. Pushkarev2,
O. I. Kovzun2, B. B. Guda2, V. V. Pushkarev2,
M. D. Tronko2, N. V. Skrypnyk1, L. M. Zaiats1

1Ivano-Frankivsk National Medical University, Ivano-Frankivsk, Ukraine;
2SI “V.P. Komisarenko Institute of Endocrinology and Metabolism of NAMS of Ukraine”, Kyiv;
*e-mail: tamara.vatseba@gmail.com

Received: 17 April 2020; Accepted: 13 November 2020

Modern research shows that patients with diabetes mellitus have an increased risk of cancer. PI3K/Akt/mTOR/p70S6K1 signaling pathway plays an important role in the pathogenesis of cancer and diabetes. The aim of this study was to determine the state of РІ3K/Akt/mTORC1/p70S6K signaling cascade activity in peripheral mononuclear blood cells (PBMC) of patients with type 2 diabetes (T2D) relatively to the insulin and insulin-like growth factor (IGF-1) concentrations in blood plasma. Enzyme-linked immunosorbent assay was used to examine the levels of insulin and IGF-1 in blood plasma as well as the content of phosphorylated forms of Akt (Ser473), PRAS40 (Thr246), and p70S6K (Thr389) in PMBC. It was shown that in the blood plasma of patients with T2D the levels of insulin and IGF-1 were increased. Phosphorylation and activation of Akt by the mTORC2 protein kinase complex was not observed. At the same time, the relative degree of phosphorylation of mTORC1 inhibitor, PRAS40, and its substrate, p70S6K, was higher in PMBC of T2D patients in comparison with control values. These data suggest that phosphoinositide-dependent protein kinase 1 (PDK1) and, possibly, mitogen-activated protein kinase (MAPK) could mediate the effects of IGF-1 on Akt activation under type 2 diabetes.

Protective effects of hexane fraction of Costus afer leaves against sodium arsenite-induced hepatotoxicity and nephrotoxicity in male albino wistar rats

G. N. Anyasor*, O. O. Aramide, O. S. Shokunbi

Department of Biochemistry, School of Basic Medical Sciences,
Benjamin S. Carson (Snr.) College of Health and Medical Sciences, Babcock University,
Ilishan-Remo, Ogun State, Nigeria;
*e-mail: anyasorg@babcock.edu.ng

Received: 13 July 2020; Accepted: 13 November 2020

Arsenite is a toxic metallic pollutant known to cause hepatotoxic and nephrotoxic injuries. Costus afer Ker Gawl. is an indigenous medicinal plant used as therapy for numerous tissue disorders. Thus, this study investigated the protective potential of C. afer hexane leaf fraction (CALHF) on sodium arsenite-induced hepatic and renal injuries in albino rats. Twenty-five male albino rats were randomly distributed into five groups of five rats each. Group 1: rats administered orally with 0.5 ml of 0.9% saline; Group 2: untreated rats induced with 5 mg/kg body weight (b.w.) sodium arsenite (i.p.); Group 3: rats induced with sodium arsenite and treated with 10 mg/kg b.w. silymarin (hepatoprotective drug); Group 4 and 5: rats induced with sodium arsenite and treated with 100 and 200 mg/kg b.w. CALHF, respectively. CALHF was orally administered daily, while sodium arsenite was administered every 48 hours for 14 days. Thereafter, rats were sacrificed, blood was collected to estimate hepatic and nephrotic functions. Hepatic and renal function tests showed that 100 and 200 mg/kg CALHF and 10 mg/kg silymarin treated animals had significantly reduced (P < 0.05) plasma alanine aminotransferase, aspartate aminotransferase, creatinine and urea levels, when compared with those of untreated animals. C. afer hexane leaf fraction exhibited hepatoprotective and nephroprotective effects against sodium arsenite induced toxicity in rats.

Effect of glutamic acid and cysteine on oxidative stress markers in rats

N. O. Salyha

Institute of Animal Biology, National Academy of Agrarian Sciences of Ukraine, Lviv;
е-mail: ynosyt@yahoo.com

Received: 7 May 2020; Accepted: 13 November 2020

Epinephrine (EPI) surges is known to be associated with stress induction and raising risk of heart strokes. The search for effective, nontoxic substances with antioxidative effects has been intensified in recent years. We focused our attention on two amino acids: L-glutamic acid (Glu) and L-cysteine (Cys). Our goal was to compare the effects of Glu, Cys and Glu in combination with Cys intraperitoneal administration on the antioxidant system indicators and the content of lipid peroxidation products in myocardium and spleen tissues of rats subjected to experimental EPI-induced stress. Rats were divided into five groups: EPI, EPI/Glu, EPI/Glu/Cys, EPI/Cys and control. The reduced glutathione (GSH) and TBA-active products level, glutathione peroxidase (GPx,), glutathione-S-transferase (GST), glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G6PDH) activity in tissues were measured. Our results indicate that epinephrine-induced stress increased the content of the lipid peroxidation products in myocardium and reduced the level of GSH in myocardium and spleen tissues of rats. Increasing of GPx activity in spleen only stressed animals were observed, while significantly lowered the GPx activity in groups of rats treated with amino acids (Glu, Glu/Cys, Cys). The obtained results suggest that the GR activity was significantly inhibited by stress in all investigated groups in spleen and epinephrine-induced rats and EPI/Cys groups of rats in myocardium. In rats treated with amino acids (particularly, Glu and Glu/Cys groups), we observed no significant difference in studied parameters. Our results indicate that application of Glu, Cys alone or in combination can increase GSH content in both studied tissues and activity of some antioxidative enzymes, and thus partially mitigated of epinephrine-induced stress in rats.