Category Archives: Uncategorized
Comparative cytotoxic activity of carboplatin and β-cryptoxanthin in free and liposomal forms against breast cancer cell line
M. W. Shafaa*, N. S. Elkholy
Physics Department, Medical Biophysics Division, Faculty of Science,
Helwan University, Cairo, Egypt;
*e-mail: medhatwi@hotmail.com
Received: 1 June 2020; Accepted: 17 May 2021
The study of the effectiveness of the synthetic and natural anticarcinogenic compounds in liposomal form is urgent for their possible use in therapy. In this work, the alkylating agent carboplatin and the representative of carotenoids β-cryptoxanthin were used. The aim of the research was to study the toxicity of these compounds in free and liposomal forms against breast cancer MCF-7 cell line. According to DSC and FTIR data, when carboplatin or β-cryptoxanthin were added to liposomal bilayers, a single peak was observed indicating their mutual mixing. Integration of β-cryptoxanthin into bilayer was found to be more proper for the creation of PE acyl chains ordered and cooperative state. It was found that MCF-7 cells sensitivity was much higher to the free β-cryptoxanthin than to the free carboplatin with IC50 42 and 235 μg/ml, respectively. The IC50 values for β-cryptoxanthin loaded into liposomes and for free carboplatin were similar. At the same time, no cytotoxic effect of carboplatin-loaded liposomes was observed. The data obtained allow proposing a possible antitumor treatment regimen where carboplatin is replaced by free β-cryptoxanthin or its liposomal form to increase the effectiveness of breast cancer therapy.
The effect of CO donor hemin on the antioxidant and osmoprotective systems state in Arabidopsis of a wild-type and mutants defective in jasmonate signaling under salt stress
M. A. Shkliarevskyi1, Yu. E. Kolupaev1,2*, T. O. Yastreb1,
Yu. V. Karpets1, A. P. Dmitriev3
1Dokuchaev Kharkiv National Agrarian University, Ukraine;
2Karazin Kharkiv National University, Ukraine;
3Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: plant.biology.knau@gmail.com
Received: 6 December 2020; Accepted: 17 May 2021
The role of the gasotransmitter carbon monoxide (CO) in signaling and adaptive processes in plants has been studied insufficiently. There are indirect data indicating jasmonate signaling participation in realization of CO effects, but the possible connection between carbon monoxide and jasmonate signaling during plant adaptation to salt stress remains unclear. We studied the carbon monoxide donor hemin effect on the Arabidopsis of a wild-type (Col-0) and defective in jasmonate signaling coi1 and jin1 mutants response to the salt stress. Arabidopsis thaliana 4-week-old plants were grown on a modified Hoagland’s medium. Plants were incubated for 24 h in usual or 2 µM hemin containing culture medium, then transferred to 150 mM NaCl containing media and incubated for 24 h before the medium was replaced with the usual one. It was shown that salt stress caused water deficiency and superoxide dismutase and catalase activity decrease in the plants of all three genotypes. Treatment with 2 μM hemin stabilized the levels of catalase activity and photosynthetic pigments and increased guaiacol peroxidase activity in a wild-type, but not in coi1 and jin1 mutant plants after stress induction. Treated with hemin wild-type Arabidopsis plants accumulated more proline and sugars in response to stress than treated coi1 and jin1 mutants. It was concluded that jasmonate signaling can be involved in adaptive processes induced by exogenous carbon monoxide.
Administration of platelet-rich plasma or concentrated bone marrow aspirate after mechanically induced ischemia improves biochemical parameters in skeletal muscle
А. Т. Pidlisetsky1, G. V. Kosiakova2, T. M. Goridko2, A. G. Berdyschev2,
O. F. Meged2, S. I. Savosko3*, О. V. Dolgopolov4
1Lviv Regional Hospital for War Veterans and Repressed named after Yu. Lypa, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Bogomolets National Medical University, Kyiv, Ukraine;
4SI “Institute of Traumatology and Orthopedics of NAMS of Ukraine”, Kyiv;
*e-mail: savosko_s@ukr.net
Received: 16 September 2020; Accepted: 17 May 2021
Ischemia is known to cause structural and functional damage to skeletal muscle. In this research platelet-rich plasma (PRP) and cell bone marrow aspirate (CBMA) which are considered to be promising in regenerative medicine were used to improve the animal muscles condition after ischemia. Mechanically induced ischemia (MII) of the rabbit skeletal muscles was caused by applying a tourniquet to the hind limb for 6 hours. At 6 h after tourniquet was removed the rabbit PRP or CBMA solutions were injected into the tibia muscle and animals were divided into experimental groups (n = 15): 1 – ischemia + saline, 2 – ischemia + PRP, 3 – ischemia + CBMA. Intact animals were the control group (n = 5). Animals were removed from the experiment on 5, 15, and 30th days of reperfusion after ischemia. A progressive hypotrophy, muscle fibers destruction and leukocyte infiltration was observed after tibia muscle MII. The ischemic state was accompanied by catalase and superoxide dismutase activation, persistent NO2– overproduction and a significant decrease in glutathione peroxidase activity in the muscle homogenates. It was shown that after PRP or CBMA local injection into the ischemic muscle catalase activity remained at the elevated level, SOD activity approached the control level and the most pronounced effects were NO2– level decrease and glutathioperoxidase activity restoration. After CBMA injection angiogenesis activation and the increase in the number of nuclei in the muscle fibrils were observed. The obtained data allow to expand the range of biochemical parameters for assessing the muscles condition after ischemia and indicate a positive effect of platelet-rich plasma and cell bone marrow aspirate injection on skeletal muscle regeneration.
Lipid peroxidation and DNA fragmentation in fresh and cryopreserved spermatozoa of men at different spermatogenesis state
T. O. Yurchuk*, O. V. Pavlovich, G. O. Gapon,
A. Y. Pugovkin, M. P. Petrushko
Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Department of Cryobiology of Reproductive System, Kharkiv;
*e-mail: taisiya.yur@gmail.com
Received: 11 July 2021; Accepted: 17 May 2021
Cryopreservation of spermatozoa is widely used in the treatment of infertility by assisted reproductive technologies. However, the cryopreservation causes an oxidative stress which can induce pathological changes in the male gametes. The aim of the research was to evaluate lipid peroxidation (LPO) and DNA fragmentation as well as correlation between these parameters in the fresh and cryopreserved spermatozoa of men with normozoospermia or oligoasthenoteratozoospermia (OAT) and spermatozoa derived from the epididymis of men with azoospermia. The level of malondialdehyde (MDA) in a TBA test, superoxide dismutase (SOD) activity and total antioxidant activity (AOA) were assessed. DNA integrity was estimated by acridine orange staining technique. It was shown that MDA level and SOD activity were significantly higher in the fresh spermatozoa of oligoasthenoteratozoospermic group compared with the fresh spermatozoa of normozoospermic group. After cryopreservation the MDA level increased in all groups and was the highest in OAT group where the greatest AOA decline was detected. DNA fragmentation frequency was 2.6 and 4.1 times higher in the fresh and cryopreserved OAT spermatozoa respectively as compared with the fresh normozoospermic ones (7.2%). DNA fragmentation was found to be the lowest in the fresh (6.2%) and cryopreserved (5.8%) epididymal spermatozoa. After cryopreservation SOD activity in epididymal spermatozoa was lower than in normozoaspermic. In spermatozoa of the studied groups the MDA level positively correlated with DNA fragmentation (0.79 Pearson’s correlation coefficient) both before and after cryopreservation. It is suggested that due to the detected low DNA fragmentation and LPO level in epididymal spermatozoa their use could be an alternative approach for infertility treatment by assisted reproductive technologies.
Astrocyte specific proteins content in the different parts of the rat and mongolian gerbil brain during ontogenesis
Y. P. Kovalchuk*, H. N. Shiyntum, G. A. Ushakovа
Oles Honchar Dniprо National University, Ukraine;
*e-mail: yulka.5868152@ukr.net
Received: 28 October 2021; Accepted: 17 May 2021
Astrocyte-specific proteins are used as markers of astrocytes, particularly in the case of age-related brain dysfunctions and neurodegenerative disorders. However, the data on the content of these proteins in different parts of the brain during ontogenesis are insufficient. In this research, the content of astrocyte specific Ca2+-binding protein S100B and glial fibrillary acidic protein (GFAP) in cerebellum, thalamus, and hippocampus of Wistar rats and Mongolian gerbils during ontogenesis were determined. Animals were divided by age into four groups (n = 6-10): 1 – newborn (1-day-old); 2 – 30-day-old; 3 – 90-day-old; 4 – 180-day-old. Fractions of soluble and filamentous proteins from the different parts of the brain were obtained by differential centrifugation and solubilization with 4M urea. The quantitative levels of S100B and GFAP were determined with ELISA. It was revealed that the content of S100B protein increased significantly by day 180 in all studied parts of the rat and gerbil brain. The content of GFAP both soluble and filamentous forms in the brain of 1 day-old animals was low, but raised significantly after 30 days in all studied parts of gerbil brain, and increased gradually up to 180 days in the rat brain. The magnitude of the increase in the content of the studied astrocyte-specific proteins was shown to be different and to depend on the brain part, postnatal development stage of the animals and their species.
Regulation of ionized calcium concentration in mitochondria matrix in the absence of exogenous Са(2+)
A. V. Sylenko*, S. G. Shlykov, L. G. Babich,
О. Yu. Chunikhin, S. O. Kosterin
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: sylenkoanna@ukr.net
Received: 15 September 2021; Accepted: 17 May 2021
Functional activity of mitochondria depends on ionized calcium content in its matrix. Thus, investigation of the ways to influence Ca2+ transport and accumulation in these organelles is an urgent issue. The aim of this work was to evaluate ionized calcium concentration in the mitochondria matrix in the absence of exogenous Ca2+ and in the presence of Mg2+ and ATP in incubation media. The concentration of ionized calcium in mitochondria isolated from the myometrium of non-pregnant rats was measured with the fluorescent probe Fluo-4 AM, mitochondria hydrodynamic diameter was assessed using dynamic light scattering. The ATP concentration-dependent increase in ionized calcium concentration in mitochondria matrix in the absence of exogenous Са2+ and the high level of the total Са2+ accumulation after further addition of exogenous Са2+ were registered. The effect of ATP on Са2+ concentration did not depend on the blockage of mitochondrial PTP with cyclosporine A or on H+-ATPase/ATP-synthase inhibition with oligomycin, instead the addition of 10mM theophylline and 30 mM NaHCO3 to the Mg2+-containing medium led to Са2+ concentration elevation in mitochondria matrix. It was shown that low Ca2+ concentration in the matrix correlated with the increase in mitochondria size, whereas high Ca2+ concentration correlated with relatively smaller size of the organelles. Activation of the soluble adenylyl cyclase by NaHCO3 with simultaneous inhibition of phosphodiesterase by theophylline was accompanied by the decline in Са2+ probe normalized fluorescence in mitochondria matrix under conditions of exogenous Ca2+ introduction. It was suggested that the soluble adenylyl cyclase may be involved in regulation of Са2+ concentration in mitochondria matrix.
Double Nobel prize winner: Frederick Sanger – the father of genomics
T. V. Danylova1*, S. V. Komisarenko2
1National University of Life and Environmental Sciences of Ukraine, Kyiv;
*e-mail: danilova_tv@ukr.net,
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail:svk@biochem.kiev.ua
Received: 02 February 2021; Accepted: 23 April 2021
This paper aims to outline briefly the main stages of Frederick Sanger’s scientific activity – the only person to have won two Nobel Prizes in Chemistry (1958, 1980). His work on the structure of proteins, especially that of insulin, and the determination of base sequences in nucleic acids made an immense impact on the development of biochemistry and especially on the development of a new scientific field – molecular biology. His methods for determining the primary structure of proteins and nucleic acids helped biochemists and molecular biologists to determine the structure of many proteins and nucleic acids and laid the basis for genetic engineering.
Inclusion of enterosorbents in anti-inflammatory therapy improve treatment effectiveness in copd patients during exacerbations
S. V. Kucher*, O. V. Lototska
I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: kuchersv@tdmu.edu.ua
Received: 29 January 2021; Accepted: 23 April 2021
Inflammation plays a key role in the development of chronic obstructive pulmonary disease (COPD), and manifests as increased levels of inflammation markers such as cytokines. The purpose of this study was to examine the effect of enterosorbents on IL-1β, TNF-α and IL-10 levels in COPD patients with exacerbations, while taking into account their ages. Patients were divided into two age groups: age group 1 (40-59 years) and age group 2 (>60 years). Levels of IL-1β, TNF-α and IL-10 were determined by enzyme-linked immunosorbent assay. Levels of pro-inflammatory and anti-inflammatory cytokines were increased in both age groups of COPD patients during exacerbations compared to the healthy control group. Pro-inflammatory cytokines (IL-1β, TNF-α) increased more in age group 2. Basic therapy (BT) had positive impacts. Addition to BT of the enterosorbents Enterosgel and Carboline decreased serum levels of IL-1β, TNF-α and IL-10 in both age groups more than after BT alone. Greater changes were observed in age group 2. We found that addition of the enterosorbents Carboline or Enterosgel to basic therapy of COPD patients during exacerbations decreased levels of IL-1β, TNF-α and IL-10 in patients of both age groups (40-59 years and >60 years).
Clinical and immunological peculiarities of parasitic infections in children with digestive system disorders in Western Ukraine
K. T. Hlushko*, H. A. Pavlyshyn, K. V. Kozak
Department of Pediatrics No 2, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: glushko_kt@tdmu.edu.ua
Received: 29 January 2021; Accepted: 23 April 2021
Diseases of the digestive system such as gastroduodenal disorders (GDD) and hepatobiliary diseases (HBD) are fairly common among children. At the same time, intestinal parasites are widespread gastrointestinal tract infectious agents. We examined 108 children with GDD (n = 54) and HBD (n = 54) (mean age 11.8 ± 4.3 years) who were treated in a children’s hospital. The following were performed: blood test for specific Ig to Ascaris lumbricoides, Toxocara canis; feces analysis for helminth eggs, Giardia cysts; and pinworm test. The serum level of interleukin-4 (IL-4) was measured in 97 of 108 children. Overall parasitic co-infections were found in 60.2% of children: giardiasis in 30.6%, toxocariasis in 17.6%, ascariasis seropositivity in 13.9% and enterobiasis in 8.3% of cases. Parasitic infections (PIs) occurred in 72.2% of the HBD group and in 48.2% of the GDD group (P = 0.01). Among the PIs only ascariasis was found more often in the HBD group (22.2%) compared with the GDD group (5.6%) (P = 0.01). The mean age of children who were positive for PI (PI+) was 9.9 ± 4.6 years; while in the PI negative (PI–) group the mean age was 12.8 ± 3.3 years (P < 0.001). In GDD the IL-4 level in PI+ group was 9.3 ± 0.9 pg/ml in the versus 6.9 ± 1.8 pg/ml in the PI‒ group (P = 0.02). Children with HBD and PI had higher IL-4 levels (14.2 ± 18.8 pg/ml) compared to those without PI (7.5 ± 2.4 pg/ml) (P = 0.03). The differing impact on the course and immune response in children depending on co-infection with PIs indicates the need for additional testing for these infections.