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Urine neutrophil gelatinase-associated lipocalin as an early biochemical marker of microalbuminuria in predicting early kidney damage in patients with type 2 diabetes mellitus
P. J. Padmini, V. Ashok*
Shri Sathya Sai Medical College and Research Institute, Sri Balaji Vidyapeeth Deemed to be University, Tamilnadu, India;
*e-mail: dr.ashokmbbs1986@gmail.com
Received: 08 July 2021; Accepted: 12 November 2021
Diabetic nephropathy is one of the microvascular complications of diabetes mellitus. The study was done to evaluate the diagnostic value of neutrophil gelatinase-associated lipocalin (NGAL) urine level as a biomarker for the early detection of nephropathy in type 2 diabetic patients. This study was performed on 150 patients with type 2 diabetes mellitus classified into three equal groups according to their urine albumin/creatinine ratio (ACR), including patients with normoalbuminuria (ACR < 30 mg/g creatinine), microalbuminuria (ACR = 30–300 mg/g creatinine) and macroalbuminuria (ACR > 300 mg/g creatinine). Fifty apparently healthy subjects were selected as a control group. Urine NGAL was estimated by a particle-enhanced turbidimetric immunoassay using Hitachi 917 analyzer. The serum level of glucose and creatinine was also estimated. The results showed that NGAL urine level was significantly elevated in diabetes patient groups with microalbuminuria and macroalbuminuria when compared to the control group and diabetes patient group with normoalbuminuria. The levels of urine NGAL correlated positively with microalbuminuria in patients with diabetes. Receiver operating characteristic curves revealed that urine NGAL had a higher diagnostic value for diabetic nephropathy early detection compared to ACR and microalbuminuria in patients with diabetes mellitus.
Extracellular vesicles produced by mouse breast adenocarcinoma 4T1 cells with up- or down-regulation of adaptor protein Ruk/CIN85 differentially modulate the biological properties of 4T1 WT cells
A. Yu. Zhyvolozhnyi1,2*, I. R. Horak1, D. S. Geraschenko1, M. O. Gomozkova3,
O. O. Hudkova1, S. J. Vainio2, A. A. Samoylenko2, L. B. Drobot1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Faculty of Biochemistry and Molecular Medicine, University of Oulu, Finland;
3Brigham Young University-Idaho, Rexburg, USA;
*e-mail: ppndl2@gmail.com
Received: 02 November 2021; Accepted: 12 November 2021
Extracellular vesicles (EVs) are secreted by most cell types under both physiological and pathological conditions and were proposed to be actively involved in intercellular communication. The mode of EVs action is dependent on their cargos composition. EVs play an important role in tumor initiation, recurrence, metastasis and therapeutic resistance. EVs marker proteins Alix and Tsg101 and cortactin are the binding partners of adaptor protein Ruk/CIN85. The present study aims to analyze the regulatory effects of EVs produced by 4T1 cells with overexpression (RukUp) or down-regulation (RukDown) of adaptor protein Ruk/CIN85 on proliferation rate, migration and invasion activity of parental 4T1 WT cells. EVs from conditioned medium of 4T1 RukUp or RukDown cells were isolated by differential centrifugation followed by further purification using Exo-spin™ kit (Cell Guidance Systems). The number and size of EVs were characterized by NTA (Malvern Panalytical NanoSight NM300) instrument. The content of marker proteins and Ruk/CIN85 in isolated EVs was analyzed by Western-blotting. The viability, migration and invasion activity of 4T1 WT cells were studied using MTT-test, scratch-test and Boyden chamber assay, respectively. It was demonstrated for the first time that adaptor protein Ruk/CIN85 is a constitutive component of EVs produced by 4T1 cells. It was also shown that EVs produced by 4T1 cells with different levels of Ruk/CIN85 expression are characterized by a specific profile of the content of its multiple molecular forms. It turned out that the ability of EVs to modulate the proliferative activity, motility and invasiveness of 4T1 WT cells was tightly correlated with the biological properties of 4T1 cells that produce EVs (highly aggressive 4T1 RukUp cells or weakly invasive 4T1 RukDown cells). Our data suggest that adaptor protein Ruk/CIN85 is not only a constitutive component of cargos composition of EVs produced by tumor cells but, depending on its content in EVs, plays an active role in the control of carcinogenesis.
Production of recombinant SARS-COV-2 proteins and diphtheria toxoid CRM197-based fusion
O. I. Krynina1, S. I. Romaniuk1, O. B. Gorbatiuk1,2,
O. H. Korchynskyi1,3,4, А. V. Rebriiev1, Ya. S. Kulyk1,
Ye. O. Kozadaieva1, A. A. Siromolot1,5, M. M. Guzyk1,
D. V. Kolybo1*, S. V. Komisarenko1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2State Institute of Genetic and Regenerative Medicine, National Academy of Medical Sciences of Ukraine, Kyiv;
3Centre for Innovative Research in Medical and Natural Sciences, Faculty of Medicine, University of Rzeszow, Rzeszow, Poland;
4S. Gzhytskyi National University of Veterinary Medicine and Biotechnologies, Lviv, Ukraine;
5ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: kolibo@biochem.kiev.ua
Received: 10 October 2021; Accepted: 12 November 2021
The quickly emerged global COVID-19 pandemic raised a desperate need in the development of protecting vaccines targeting this disease. Therefore, a generation of effective producers of recombinant SARS-CoV-2 proteins became an urgent task. Its resolving contributes to the study of functional SARS-CoV-2 properties, as well as will allow developing the domestic COVID-19 vaccine in Ukraine, thus playing an important strategic role in tackling the pandemics. The aim of the study was to generate prokaryotic and eukaryotic producers of recombinant SARS-CoV-2 proteins and to isolate nucleocapsid (N) protein, receptor-binding domain (RBD) of spike (S) protein, as well as RBD fused to the carrier – diphtheria toxoid CRM197. For this purpose, appropriate genetic constructs, in particular, replication deficient recombinant AdvC5-based adenoviral vectors expressing the SARS-CoV-2 proteins and CRM197-fused conjugate were created through methods of molecular biology and genetic engineering. Restriction analysis and/or DNA sequencing confirmed that we created the correct constructs. Immobilized metal affinity chromatography was used to purify the recombinant proteins. Compliance of their properties was confirmed by the results from polyacrylamide gel electrophoresis, Western blotting, immunoenzymatic assay and MALDI-TOF mass spectrometry. As a result, we generated E. coli Rosetta (DE3) bacterial strain and HEK293 cell line producing recombinant SARS-CoV-2 proteins and CRM197-based fusion. In addition, pure N protein, RBD of S protein and RBD-CRM197 fusion protein were isolated. The obtained recombinant SARS-CoV-2 proteins can be used to study immunogenic and antigenic properties of the SARS-CoV-2 proteins. Cells producing recombinant SARS-CoV-2 proteins and RBD-CRM197 fusion protein are able to provide cheap and safe synthesis of the antigenic substances for domestic development and production of immunodiagnostics for COVID-19 and COVID-19 vaccines in Ukraine.
Biochemical and molecular-physiological aspects of the nitric oxide action in the utera
H. V. Danylovych, Yu. V. Danylovych
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: danylovych@biochem.kiev.ua
Received: 18 May 2021; Accepted: 12 November 2021
The sources of the nitric oxide (NO) formation in the uterus and the dynamics of changes in its content in different periods of organ functioning in human and animals are analyzed. The biochemical mechanisms of NO action on the myometrium contractile activity, the significance of NO in the physiological processes during pregnancy and labor, the importance of mitochondria as a reliable NO source in the smooth muscle and the possible ways of NO influence on Ca2+ transport and bioenergetic processes in mitochondria are considered. The authors’ data concerning ionic and membrane mechanisms of NO action on Ca2+-homeostasis of uterine myocytes, identification of nitric oxide in uterine smooth muscle mitochondria, biochemical characteristics of the NO-synthase reaction and the possible role of NO in the regulation of Ca2+ transport in these subcellular structures and in the electron transport chain functioning are presented and discussed.
Nobel Prize laureate Kary Mullis and the polymerase chain reaction (PCR)
V. M. Danilova*, O. P. Matyshevska, S. V. Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: valdan@biochem.kiev.ua
Received: 11 May 2021; Accepted: 22 September 2021
The article highlights the major life and career milestones and the extraordinary personality of 1993 Nobel Prize laureate in Chemistry Kary B. Mullis. The background of Mullis’ invention of the polyme-rase chain reaction (PCR), a revolutionary and monumental method of molecular biology and genetics of the 20th century, is described. The PCR technique is based on multiple selective copying of a particular segment of DNA with the help of enzymes in vitro. Under these conditions, only the target region is copied, and only if it is present in the studied sample. The invention of the PCR method has been one of the most outstanding events in molecular biology in recent decades.
Osteoprotegerin, sclerostin, and osteocalcin serum levels in thyroid disorder patients
F. F. Rija1, S. Z. Hussein2*, M. A. Abdalla3
1Department of Biology, Tikrit University – College of Sciences, Tikrit, Iraq;
2Clinical Biochemistry Unit, Salah Aldeen Health Directorate, Tikrit, Iraq;
3Department of Human Anatomy, Tikrit University College of Medicine, Tikrit, Iraq;
*e-mail: drmohammadahmad68@gmail.com
Received: 15 February 2021; Accepted: 22 September 2021
The maintenance of thyroid hormone homeostasis is essential in the regulation of skeletal system development, bone tissue mineralization and fracture risk prevention. This study was carried out to evaluate the serum level of osteoprotegerin (OPG), sclerostin (SOST), and osteocalcin (OC) in hypo- and hyperthyroidism patients. A total number of examined patients was 90 individuals, 30 of them were normal as a control group, the rest 60 individuals were arranged in two groups: 30 patients with hypothyroidism and 30 patients with hyperthyroidism. Serum assay tests for OPG, SOST, OC, thyroid stimulating hormone (TSH), total triiodothyronine (T3), and total thyroxine (T4) were used. The present study reported high levels of SOST, T3, and T4 and low levels of OPG, OC, and TSH in patients with hyperthyroidism compared to control. Whereas in patients with hypothyroidism only SOST and TSH levels were increased, but OPG, OC, T3, and T4 levels were low compared to control. The data obtained indicate the correlation on osteoprotegerin, sclerostin, and osteocalcin levels with thyroid hormones disturbances.
The impact of genetic factors on thyroid hormones metabolism in patients with diabetic kidney disease
N. O. Abramova1*, N. V. Pashkovska1, M. V. Vlasenko2
1Bukovinian State Medical University, Chernivtsi, Ukraine;
2Vinnitsa National Pirogov Memorial Medical University, Ukraine;
*e-mail: natalloka84@gmail.com
Received: 29 January 2021; Accepted: 22 September 2021
One out of eleven adults in the world has diabetes mellitus. 25% of them develop diabetic kidney disease. Thyroid hormones are involved in the regulation of almost all physiological processes in the body, including renal function. The aim of the research was to study the dependance of biochemical markers of renal function in patients with diabetic kidney disease on C/Т polymоrphism in the DIO1 gene. To assess the dependence of biochemical markers of renal function on the C/T polymorphism in the DIO1 gene, the following groups has been formed: 19 patients with CC genotype, 69 individuals – with CT and 14 ones- with TT genotypes. Content of urea and cleatinine in plasma, eGFR, as well as microalbumine and creatinine content in urine were sagnificantly higher in patients with TT genotype than in group of patients with CC genotype and control group (P < 0.05). Presence of the T allele in genotype is associated with violation of thyroid hormones metabolism with the development of nonthyroidal illness syndrome. Carriers of T allele with diabetic kidney disease had significantly worse biochemical indices of renal function, that indicates the dependence of these markers on DIO1 polymorphism.
Green synthesis of silver nanoparticles using aqueous extract of hot chili pepper fruits and its antimicrobial activity against Pseudomonas aeruginosa
O. E. Smirnov1,2, V. Ye. Kalynovskyi1, Yu. M. Yumyna1, P. P. Zelena1,
M. A. Skoryk3, V. M. Dzhagan4, N. Yu. Taran1
1ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
2Institute of Plant Physiology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
3G.V. Kurdyumov Institute for Metal Physics, National Academy of Sciences of Ukraine, Kyiv;
4V. Lashkaryov Institute of Semiconductors Physics, National Academy of Sciences of Ukraine, Kyiv;
e-mail: plantaphys@gmail.com
Received: 09 April 2021; Accepted: 22 September 2021
Green synthesis of different nanoparticles using the plants aqueous extracts has several advantages over other methods due to the environmentally favorable nature of plants. Moreover, such approach is also cost effective. This work describes the biosynthesis of silver nanoparticles (Ag-NPs) with the use of the aqueous extract of dry pericarps of hot chili peppers (Capsicum sp. cv. Teja (S-17) and cv. Carolina Reaper) with different levels of pungency and their antibacterial effect on the antibiotic resistant Pseudomonas aeruginosa. Phytochemical screening of pericarp tissues showed great distinction in contents of phenolic compounds and capsaicinoids as potential reducing agents wich correlated with total antiradical activity as analyzed by reduction of DPPH radicals. The biosynthesized Ag-NPs were characterized by UV-vis spectrophotometry and scanning electron microscopy (SEM). The average size of the nanoparticles in both samples was less than 25 nm. Іnitial concentration of both samples of Ag-NPs inhibited P. aeruginosa growth with equal efficiency.