Category Archives: Uncategorized
Edaravone reduces the markers of oxidative stress and neuroinflammation in neocortex of rats with acute intracerebral hemorrhage and type 2 diabetes mellitus
V. L. Holubiev*, A. E. Lievykh, V. A. Tkachenko,
Yu. V. Kharchenko, V. I. Zhyliuk
Department of Pharmacology, Dnipro State Medical University, Dnipro, Ukraine;
*e-mail: 209@dmu.edu.ua
Received: 05 August 2024; Revised: 22 October 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Type 2 diabetes mellitus (T2DM) is associated with a higher incidence of hemorrhagic stroke in a severe form. The aim of this study was to estimate the markers of oxidative stress and neuroinflammation in the brain of rats with acute intracerebral hemorrhage (ICH) and T2DM after treatment with edaravone. T2DM was induced by a single intraperitoneal injection of nicotinamide/streptozotocin, ICH – by stereotactic microinjection of bacterial collagenase. Rats were randomized into four groups: 1 – intact control; 2 – T2DM; 3 – T2DM+ICH; 4 – T2DM+ICH+edaravone 6 mg/kg/day. Edaravone (a drug to treat neural injury after acute cerebral ischemic stroke) was administered intraperitoneally for 10 days starting from the 60th day after diabetes mellitus induction and 30 min after ICH induction. Brain homogenates were assessed for the content of advanced glycation end products (AGEs) and advanced oxidation protein products (AOPPs). The levels of TNF-α and 8-hydroxy-2′-deoxyguanosine (8-OHdG) were measured with ELISA. The increased content of 8-OHdG and TNF-α in brain homogenates of animals of T2DM group compared to the control was shown. It was revealed that in brain homogenates of animals of T2DM+ICH group the content of these markers significantly exceeds that for T2DM group, and in addition, an elevated AOPPs level was observed. Our results demonstrated that edaravone prevented the elevation of TNF-α level, reduced oxidative DNA damage by decreasing 8-OHdG content, and attenuated the formation of AGEs and AOPPs in the brains of experimental animals. These findings suggest that edaravone may have therapeutic potential in diabetic patients with acute ICH.
Plasma gelsolin and matrix metalloproteinase-3 levels as diagnostic markers for psoriatic arthritis
Y. A. Zamzam1*, T. F. Mansour2, R. M. Salem3,
H. A. A. Hanout3, R. A. Mostafa1
1Department of Clinical Pathology, Faculty of Medicine, Tanta University, Egypt;
2Department of Internal Medicine (Rheumatology Unit), Faculty of Medicine, Tanta University, Egypt;
3Department of Rheumatology and Rehabilitation, Faculty of medicine, Tanta University, Egypt;
*e-mail: yosrazamzam@yahoo.com
Received: 18 June 2024; Revised: 10 August 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Recent studies have revealed a high prevalence of undiagnosed psoriatic arthritis (PsA) in patients with psoriasis. Diagnosis of psoriatic arthritis has proven challenging because the symptoms of the disease are nonspecific, rheumatoid factor is not detectable, and acute phase reactant levels may be normal. Therefore, identifying soluble biomarkers for diagnosing PsA in psoriasis patients may help in early diagnosis and proper management. The aim of the work was to evaluate plasma gelsolin and matrix metalloproteinase-3 (MMP-3) levels as potential markers for PsA. This case-control study included 25 healthy controls and 50 psoriasis patients, who were divided into 25 patients with psoriasis only and 25 patients with psoriatic arthritis. Plasma levels of gelsolin and MMP-3 were measured using ELISA. It was shown that patients with PsA had significantly lower gelsolin and significantly higher MMP-3 plasma levels compared to patients with psoriasis only. For detecting PsA, gelsolin and MMP-3 had sensitivity of 96% and specificity of 92 and 80% for each, respectively. Gelsolin level negatively while MMP-3 level positively correlated with such parameters as disease activity for psoriatic arthritis, composite psoriatic disease activity index, and inflammatory markers including high-sensitivity C-reactive protein and erythrocyte sedimentation rate. It was concluded that plasma gelsolin and MMP-3 levels could serve as potential biomarkers for diagnosing PsA and monitoring the disease progression in PsA patients.
Purification and physico-chemical properties of Bacillus atrophaeus protease with elastolytic and fibrinogenolytic activity
O. V. Gudzenko1*, L. D. Varbanets1, V. O. Chernyshenko2,
Y. M. Stohnii2, A. M. Ostapchuk3, V. O. Ivanytsia3
1Institute of Microbiology and Virology named after D. K. Zabolotny,
National Academy of Sciences of Ukraine, Kyiv;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Odesa I.I. Mechnikov National University, Odesa, Ukraine;
*e-mail: alena.gudzenko81@gmail.com
Received: 18 September 2024; Revised: 21 October 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Microbial proteases, among which proteases capable of cleaving elastin, fibrin, fibrinogen, and collagen, have been a matter of interest to researchers due to their significant biotechnological potential along with low production cost. We previously showed that Bacillus atrophaeus 08 synthesizes an extracellular protease complex that exhibits high elastolytic, fibrinogenolytic, fibrinolytic activity, and minor caseinolytic and collagenase activity. The aim of the work was to isolate and purify the Bacillus atrophaeus 08 protease from the culture liquid supernatant and to study the physicochemical properties and substrate specificity of enzyme preparation. Precipitation with ammonium sulfate of 90% saturation, gel-permeation and ion-exchange chromatography were used in the experiment. According to the data obtained, the yield of the purified enzyme with a molecular weight of about 30 kDa was 6%, its elastase activity increased 30 times (420 U/mg protein), and fibrinogenolytic activity 31.8 times (350 U/mg protein). In addition, it also exhibited fibrinolytic (35.3 U/mg protein), minor caseinolytic activity (1.2 U/mg protein) and no collagenase activity. The optimum of elastin hydrolysis was at 37°C, pH 3.0 and 9.0-10.0, the optimum for fibrinogen hydrolysis was 12°C, pH 4.0. SDS-PAAG electrophoresis showed that the Bβ-chain of fibrinogen was almost not cleaved even after 1 h of incubation with the enzyme, while the Aα-chain disappeared already at the 30th min with the production of fragments with M.W. of about 30-45 kDa. The activity of the studied enzyme preparation towards fibrin was much lower than towards fibrinogen.
Generation of the MCF-7 cell sublines with CRISPR/Cas9 mediated disruption of estrogen receptor alfa (ESR1) expression
L. O. Savinska1, S. A. Kvitchenko1,2, S. S. Palchevskyi1,
I. V. Kroupskaya1, A. V. Mazov1, O. M. Garifulin1, V. V. Filonenko1*
1Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
2ESC “Institute of Biology and Medicine”, Taras Shevchenko National Univercity of Kyiv, Ukraine;
*e-mail: filonenko@imbg.org.ua
Received: 28 October 2024; Revised: 05 November 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Supported by the literature, our initial hypothesis was that Estrogen Receptor alfa (ESR1) may function as a master regulator by influencing the expression of epithelial-to-mesenchymal transition (EMT)-related genes in cancer cells. To explore this further, we used the CRISPR/Cas9 gene editing system to create MCF-7 sublines with down-regulated ESR1 expression and analyzed its impact on EMT initiation. By applying two distinct types of gRNA for gene editing, we established six MCF-7 cell sublines with either nearly complete or partial down-regulation of the ESR1 isoforms. Unexpectedly, the data obtained revealed no discernible impact of ESR1 down-regulation on EMT manifestation as Western blot and Real-Time qPCR analysis of selected clones revealed no changes in EMT markers expression. We suggested that those of the ESR1 isoforms, the expression of which was not affected by gene editing, could be crucial for the initiation of EMT. The obtained cell models will be used further to evaluate the activity of ESR1 isoforms.
Potential of isothiocyanate sulforaphane from broccoli to combat obesity and type 2 diabetes: involvement of NRF2 regulatory pathway
M. V. Ivanochko1, M. M. Bayliak1, V. I. Lushchak1,2*
1Department of Biochemistry and Biotechnology,
Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
2Research and Development University, Ivano-Frankivsk, Ukraine;
*e-mail: volodymyr.lushchak@pnu.edu.ua
Received: 03 September 2024; Revised: 11 November 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Biologically active food components are now considered to be remedies for the prevention and treatment of metabolic disorders of different etiology. The aim of this review was to analyze the current data on the application of isothiocyanate sulforaphane, found in broccoli and other cruciferous plants, for the treatment of T2DM, obesity, and their comorbidities with the presentation of established molecular, particularly dependent on NF-E2-related factor-2 (Nrf2), and signaling mechanisms of therapeutic effects.
Biochemical and cellular mechanisms of immunogenic cell death
M. Klishch, N. Skorokhyd, R. Panchuk, R. Stoika*
Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, Ukraine;
*e-mail: stoika@cellbiol.lviv.ua; stoika.rostyslav@gmail.com
Received: 26 April 2024; Revised: 11 September 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Immunogenic cell death (ICD) is a mode of programmed cell death that leads to the activation of anticancer immune response and determines the long-term success of anticancer therapies. Here, we provide a review of the known molecular and cellular mechanisms of ICD. Usually, solid tumor experimental models have been used in ICD studies. However, ascites tumor models may possess some advantages over them. The results of our investigation on the approbation of murine Nemeth-Kellner lymphoma as an experimental ascites tumor model for ICD studies are presented.