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Hypoglycemic and hypolipidemic effects of Corchorus olitorius leaves as a food supplement on rats with alloxan-induced diabet

A. G. Nduka, A. O. Oluwatoyin, A. O. Temitope, O. Odutola

Department of Biochemistry, Benjamin S. Carson School of Medicine, Babcock University, Ilisan-Remo, Ogun State, Nigeria;

Received: 16 October 2019; Accepted: 15 May 2020

Corchorus olitorius is a species of shrub in the family Malvaceae known to have rich sources of chemical compound. The leaves of this plant are locally consumed as a vegetable and as a remedy in Africa, Middle East and Southern Asia. This study was designed to evaluate the hypoglycemic and hypolipidemic effects of C. olitorius leaves sample given for 14 days as a food supplement to rats with alloxan-induced diabet. Twenty one male albino rats weighing 150-200 g were randomly assigned into three groups: 1 – control rats; 2 –  diabetic rats; 3 – diabetic rats fed with C. olitorius leaves as supplement. Phytochemical analysis of C. olitorius leaves ethanolic extract revealed the presence of alkaloids, anthraquinone, cardiac glycosides, saponins, tannins, phenols, phlobatannins and flavonoids. Diabetic rats fed with C. olitorius leaves as a supplement were shown to have significantly (P < 0.01) decreased plasma glucose, LDL-cholesterol, total cholesterol and triglycerides levels when compared with untreated diabetic rats. The findings from this study indicated that C. olitorius leaf possesses hypoglycemic and hypolipidemic properties.

Cytotoxic action of maleimide derivative 1-(4-Cl-benzyl)-3-chloro-4-(CF(3)-phenylamino)-1H-pyrrole-2,5-dione toward mammalian tumor cells and its capability to interact with DNA

N. S. Finiuk1,2, I. I. Ivasechko1, O. Yu. Klyuchivska1,
H. M. Kuznietsova3, V. K. Rybalchenko3, R. S. Stoika1,2,3*

1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Ivan Franko National University of Lviv, Ukraine;
3Taras Shevchenko National University of Kyiv, Ukraine;

Received: 21 November 2019; Accepted: 15 May 2020

Development of chemical compounds capable to supress tumor progression is a perspective strategy of cancer treatment. Heterocyclic compounds possess a broad spectrum of biological activities, including anticancer one. According to the previous results of in silico modeling maleimide derivative 1-(4-Cl-benzil)-3-Cl-4-(CF3-phenylamino)-1Н-pyrrole-2,5-dione (MI-1) has a potential effect as an inhibitor of tyrosine protein kinases. The present study was aimed at in vitro evaluation of MI-1 cytotoxic effects toward tumor cells of various lines. The viability of tumor cells after  incubation with MI-1 was measured by means of 3,4,5-dymetyltiazol-2-yl-2,5-diphenyl-tetrazolium bromide (MTT) test. The MI-1 compound was shown to be toxic for a majority of studied tumor cell lines with IC50 value ranging from 0.8 to 62.2 μg/ml depending on the tissue origin of cells. The most prominent effect of MI-1 towards human cervix carcinoma (KB3-1 and KBC-1) cells with six times higher toxicity towards the multidrug resistant sub-line KBC-1 cells comparing with the action of Doxorubicin was demonstrated.  MI-1 inhibited the viability of human pancreatic, hepatocarcinoma, and colon carcinoma cells only in high doses, while human and rat glioblastoma cells were not sensitive to MI-1. Thus, the MI-1 anticancer activity dropped in the following rank of tumor cells: cervix > breast > pancreatic carcinoma > liver carcinoma > colon carcinoma > glioblastoma. Experiments on replacement of methyl green dye from DNA-methyl green complex showed that MI-1 intercalated into DNA molecule structure. The increase of the amount of the additional band of super-spiral DNA in the presence of MI-1 was revealed by means of DNA retardation at electrophoresis in the agarose gel and this effect was more pronounced than the effect of doxorubicin. The data presented indicate a new DNA-targeting mechanism of maleimide derivative 1-(4-Cl-benzil)-3-Cl-4-(CF3-phenylamino)-1Н-pyrrole-2,5-dione anticancer action.

ТiО(2) hepatotoxicity under long-term administration to rats

O. V. Tsymbalyuk, S. P. Veselsky, A. M. Naumenko, T. L. Davydovska,
I. S. Voiteshenko, I. I. Сhyzh, V. A. Skryshevsky

Institute of High Technologies, Taras Shevchenko National University of Kyiv, Ukraine;

Received: 30 March 2020; Accepted: 15 May 2020

Titanium dioxide (ТіО2) powder which is used as a white dye was  considered to be an inert material for a long time despite its accumulation  in liver tissues after penetration into  organism. The aim of the study was to estimate biochemical markers of liver functioning in blood plasma and ATPase activity of erythrocyte plasma membrane under the oral administration of ТіО2 nanoparticles suspension (0.1 mg/kg, daily) to Wistar rats for 30 and 100 days. A significant increase of alanine aminotransferase and aspartate aminotransferase activity as well as of direct, indirect and bound bilirubin content, a decrease of connjugated (taurocholic, taurochenodeoxycholic, taurodeoxycholic, glycocholic, glycochenodeoxycholic, and glycodeoxycholic) and free (glycodeoxycholic and deoxycholic) bile acids concentration with concomitant increase of free cholic acid concentration in blood plasma of rats under ТіО2 administration were revealed, indicating a significant impairment of pigment exchange in the liver of rats. Under ТіО2 administration a substantial  inhibition of erythrocyte plasma membrane Мg2+-dependent ouabain-sensitive Na+,K+-ATPase and ouabain-insensitive ATPase was observed. These results presume the disturbance of transplasmalema ion-transporting processes and cells ionic homeostasis  induced by ТіО2.

Both maternal and newborn IgMs inhibit influenza virus-induced hemagglutination in vitro

A. P. Pogribna1*, M. Y. Grom2, I. V. Sokol3,
V. Berestoviy3, D. O. Govsieiev4

1Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
2Research Institute “Zhyttya”, Kyiv, Ukraine;
3Bogomolets National Medical University, Kyiv, Ukraine;
4Kyiv City Maternity Hospital No 5, Ukraine;

Received: 16 October 2019; Accepted: 15 May 2020

Most serum immunoglobulins M (IgMs) are “natural IgMs”, which are produced apparently spontaneously without exogenous antigenic or microbial stimuli. The IgMs are the first immunoglobulins expressed in the human fetus, and the maternal IgM do not cross the placenta in the normal conditions. We currently lack a clear understanding of the molecular basis for immunological differences or identities of IgM repertoires between adults and neonates, so we have tried to apply a simple and illustrative method to compare the properties of such IgM antibodies. This study was undertaken to compare the abilities of pairs of maternal and newborn highly-purified total serum IgM antibodies to block influenza virus agglutinins. We collected ten pairs of serum samples from cord blood of apparently healthy newborns and venous blood of their mothers. The highly purified total IgM antibodies were obtained by sequential salt fractionation and affinity chromatography. The effect of IgM antibodies on virus hemagglutinin interaction with erythrocytes was evaluated by hemagglutination reaction using influenza virus vaccine. According to the titer of influenza hemagglutinins, the IgM antibodies of newborns decreased hemagglutination of erythrocytes two to four times more efficiently compared to IgM antibodies of their mothers (8 out of 10 cases). Our results demonstrate that serum IgMs of newborns are able to interfere with influenza virus agglutinins even more efficiently than those of adults. These data may be useful for better understanding of immune system development in newborns.

Adaptor protein Ruk/CIN85 affects redox balance in breast cancer cells

I. R. Horak*, N. V. Latyshko, O. O. Hudkova, T. O. Kishko,
O. V. Khudiakova, D. S. Gerashchenko, T. D. Skaterna,
I. P. Krysiuk, S. G. Shandrenko, L. B. Drobot

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

Received: 25 February 2020; Accepted: 15 May 2020

Excessive reactive oxygen species (ROS) production may lead to damage of cellular proteins, lipids and DNA, and cause cell death. Our previous findings demonstrated that increased level of adaptor protein Ruk/CIN85 contributes to breast cancer cells malignancy. The aim of this study was to investigate the role of Ruk/CIN85 in the maintaining of the redox balance in cancer cells. Mouse breast adenocarcinoma 4T1 cells with different levels of Ruk/CIN85 expression were used as a model in this study. Activities of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), aldehyde dehydrogenase (ALDH) and formaldehyde dehydrogenase (FALDH), as well as H2O2 and aldehydes content were measured using fluorometric assays. Gene expression correlations between Ruk/CIN85 and antioxidant enzymes in breast cancer samples were analyzed using ist.medisapiens transcriptomic database. It was demonstrated that Ruk/CIN85-overexpressing 4T1 cells were characterized by increased production of H2O2 and reduced activities of CAT, GPx and SOD. Overexpression of Ruk/CIN85 resulted in decreased content of aldehydes together with increased activity of ALDH, while in Ruk/CIN85-knocked down 4T1 cells, activities of ALDH and FALDH were decreased. The data of transcriptomic analysis revealed the correlations between SH3KBP1 expression and CAT, GPX4, ALDH1A1, ALDH1L1, ALDH2, GSR, SOD1 in human breast carcinomas samples. The obtained results indicate that adaptor protein Ruk/CIN85 affects redox balance in mouse breast adenocarcinoma 4T1 cells.

mPTP opening differently affects electron transport chain and oxidative phosphorylation at succinate and NAD-dependent substrates oxidation in permeabilized rat hepatocytes

H. M. Mazur, V. M. Merlavsky, B. O. Manko, V. V. Manko*

Ivan Franko National University of Lviv, Ukraine;

Received: 10 October 2019; Accepted: 15 May 2020

Mitochondrial Ca2+ overload may trigger the opening of mitochondrial permeability transition pore (mPTP) and its prolonged activation leads to cell death. ATP synthase is considered as a possible molecular component of the pore. The aim of this study was to investigate the state of oxidative phosphorylation at Ca2+-induced activation of mPTP in permeabilized hepatocytes. Hepatocytes were isolated by two-stage Seglen method. Permeabilization was performed using digitonin. Oxygen consumption rate was measured with Clark electrode. Oxidative phosphorylation was determined as the ratio of the ADP-stimulated respiration and substrate-stimulated respiration rates (ADP/S). It was established that increasing of Ca2+ concentration in the medium inhibited oligomycin effects and suppressed ADP- and FCCP-stimulated respiration upon succinate or glutamate, pyruvate and malate mixture oxidation. The mPTP inhibitor cyclosporin A did not directly affect respiration and oxidative phosphorylation after elevation of Ca2+ concentration and mPTP activation. When cyclosporine A was added before increasing Ca2+ concentration, the electron transport chain function (FCCP-stimulated respiration) was not impaired while the partial disruption of oxidative phosphorylation (ADP-stimulated respiration) was observed only upon succinate oxidation. The results obtained showed that inhibition of oxidative phosphorylation was the primary event in mPTP activation, possibly due to the involvement of ATP synthase in pore opening. In the case of NAD-dependent substrates oxidation that effect was stronger and faster than at succinate oxidation, due to the lower mitochondria energization.

Vitamin D(3) regulates hepatic VEGF-A and apelin expression in experimental type 1 diabetes

D. O. Labudzynskyi1*, I. O. Shymanskyi1, O. O. Lisakovska1,
A. O. Mazanova1, L. V. Natrus2, M. M. Veliky1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Bogomolets National Medical University, Kyiv, Ukraine;

Received: 09 July 2019; Accepted: 15 May 2020

The deficiency of vitamin D is associated with the risk of various chronic diseases, including diabetes mellitus and its complications. Given the strong genomic action of vitamin D hormone-active form, its deficiency can lead to dysfunction of cytokine signaling pathways, including those dependent on vascular endothelial growth factors (VEGFs) and apelin. The present study was carried out to define the link between VEGF-A and apelin expression in liver, hepatocytes viability and vitamin D status at experimental type 1 diabetes in mice. We established that chronic hyperglycemia at streptozotocin-induced diabetes was accompanied by a 2.2-fold decrease in 25OHD content in the serum and increased hepatocytes apoptosis and necrosis. Vitamin D deficiency correlated with increased apelin and VEGF-A (8- and 1.6-fold respectively) expression. Almost complete restoration of circulatory 25OHD content in serum was achieved at vitamin D3 treatment (800 IU/kg, per os, for 2 months) followed by reduced apelin and VEGF-A expression in liver and the decline of hepatocytes apoptosis. We conclude that vitamin D3 can be involved in cell survival, angiogenesis and fibrogenesis by modulating  VEGF-A and apelin dependent regulatory systems in diabetic liver.

Presenting thematic ‘Molecular and clinical studies of hemostasis’ Issue

V. Chernyshenko, S. Komisarenko

Study of blood coagulation system became even more requested in recent year since the COVID-19 pandemia began. 2020 that was planned for the celebration of 75th Anniversary of Ukrainian hemostasiology and for publishing of this Issue turned out as the year of world-wide splash of the infection which has thrombosis as the most severe complication that can cause the death of patients. The opening review written by Dr Sandor Vari is summarizing the most recent findings on the pathogenesis and treatment of COVID-19, focusing on markers of endothelial dysfunction and thrombosis during this pathology
The nature and mechanisms of blood coagulation was the object of scientific interest worldwide since Hippocrates introduced the term “thrombus” to describe the blood clot. Ukrainian school of hemostasiology was founded by academician Volodymyr Belitser (1906-1988), remarkable scientist who was one of the most important contributors to the oxidative phosphorylation theory. In 1944 Volodymyr Belitser became the first Head of the laboratory of enzymes, later reorganized into Protein Structure and Functions department, that was focused on the blood circulation and clotting.
This Issue of The Ukrainian Biochemical Journal ‘Molecular and clinical studies of hemostasis’ can be assumed as the peculiar tribute to the school of Belitser presenting articles of Professors Leonid Medved, Yevgen Makogonenko and Eduard Lugovskoi which were the Heads of Protein Structure and Functions department in 1987-1998, 1998-2001 and 2009-2019 respectively, and two experimental works by scientists of Palladin Institute of Biochemistry of NAS of Ukraine focused on blood coagulation and fibrinolysis mechanisms. Tight connection between scientific explorations and medical use of knowledge in the field of hemostasiology is underlined by works of Vinnytsia and Ivano-Frankivsk medical Universities that are also presented in this Issue.
We are presenting ‘Molecular and clinical studies of hemostasis’ – the Thematic Issue of the Ukrainian biochemical journal – with a great hope and expectance that molecular studies and clinical approvals presenting in our collection one day will allow to overwhelm the deadly thrombotic complications of SARS-CoV-2 infection. Possibly the International project initiated by the RECOOP HST Association (that also guided the edition of this Issue) will have a chance to contribute to it.

COVID-19 infection: disease mechanism, vascular dysfunction, immune responses, markers, multiorgan failure, treatments, and vaccination

Vari S. G.

International Research and Innovation in Medicine Program Cedars-Sinai Medical Center,  Los Angeles, CA, USA;

The new SARS-CoV-2 virus is a great danger for the worldwide population since there is no known pre-immunity, no specific treatment, and no vaccine. Still, the testing and tracing are the best tools to isolate the infected and prevent the spread of COVID-19. The major goals are to save lives, reduce the mortality rate, increase the survival rate of those are in severe or critical conditions, reduce the hospital stay and accelera­te the recovery. This review summarizes the findings on the novel coronavirus that causes COVID-19 and outlines information about symptoms, testing, disease mechanism, vascular dysfunction, immune responses, treatments, and vaccination. At this time no vaccine is available to prevent COVID-19. A literature review reveals  that more research is necessary to investigate the interactions between respiratory viruses, human coronaviruses, and the new SARS-CoV-2 virus in the infected population to guide the design of COVID-19 specific therapeutics and vaccines. Almost every government on earth has realized that  daily life cannot return to normal until citizens have built up antibodies to safeguard them from the virus. Scientists and manufacturers worldwide are accelerating COVID-19 vaccine research, and pharmaceutical companies are already investing in the large-scale production of vaccines. A synopsis of the most recent sources presented in this review was the starting point for several COVID-19 research projects in the Regional Cooperation for Health, Science and Technology (RECOOP HST) Association managed by Cedars – Sinai Medical Center to gain a better understanding of the COVID-19 disease.

To the 70th birthday of the corresponding member of the NAS of Ukraine, Prof. R. S. Stoika