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Breaking the genetic code – a new revolutionary stage in the development of molecular biology: 1968 Nobel Prize laureates M. W. Nirenberg, H. G. Khorana, R. W. Holley

O. P. Matyshevska*, V. M. Danilova, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: matysh@yahoo.com

Received: 28 October 2021; Accepted: 12 November 2021

This review presents the life stories of M. Nirenberg, H. Khorana, and R. Holley, winners of the 1968 Nobel Prize in Physiology or Medicine, the history of the discoveries made by these scientists, and the methodological approaches used in their works. Owing to the M. Nirenberg and H. Khorana research, the nucleotide compositions of all mRNA triplet codons were decoded. H. Khorana was the first scientist to experimentally prove the direct link between the nucleotide sequence of DNA and the amino acid sequence of the synthesized protein and to obtain a synthetic gene. R. Holley was the first to completely decode the sequence of transport RNA, determine its secondary structure and role in protein synthesis on the ribosome. The Nobel Prize awarded to the scientists was a recognition of their contribution in understanding the mechanisms of coding and reading genetic information and marked a breakthrough moment in the development of molecular biology.

Differential impact of the temperature stress and soil drought on lipoxygenase activity in winter rye plants

L. M. Babenko, K. O. Romanenko*, I. V. Kosakivska

M.G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: katerynaromanenko4@gmail.com

Received: 5 July 2021; Accepted: 12 November 2021

Lipoxygenase cascade is a source of physiologically active compounds, the presence of which is considered not only as a sign of damage but also as a trigger of adaptive responses to stress. The aim of the study was to determine the effects of short-term (2 h) heat (40°C) and cold (4°C) temperature stress and moderate soil drought on lipoxygenase (LOX) activity in 14-day-old winter rye (Secale cereale L. ‘Boguslavka’) plants. The shoots were found to have both membrane-bound 9-LOX1 and 9-LOX2 and soluble 13-LOX activity, the roots – membrane-bound 9-LOX activity. After heat stress, the activity of 9-LOX1 and 9-LOX2 in the shoots increased by 3 and 2 times, respectively, the activity of 9-LOX in the roots – by 2 times, and 13-LOX activity in the shoots decreased by 1.5 times. After the cold stress, the activity of 9-LOX1 and 9-LOX2 in the shoots raised by almost 1.5 times, the activity of 9-LOX in the roots – by 1.2 times. Moderate soil drought caused enhancement in the activity of both membrane-bound isoforms of 9-LOX in the shoots by 1.5–2 times and in the roots – by 3 times. The established fluctuations indicate that molecular forms of LOX with different localization are differentially involved in the winter rye response to temperature stress and moderate soil drought.

Yeast concentration in the diet defines Drosophila metabolism of both parental and offspring generations

O. M. Strilbytska1*, N. P. Stefanyshyn1,
U. V. Semaniuk1, O. V. Lushchak1,2*

1Department of Biochemistry and Biotechnology, Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
2Research and Development University, Ivano-Frankivsk, Ukraine;
*e-mail: olya_b08@ukr.net or oleh.lushchak@pnu.edu.ua

Received: 18 April 2021; Accepted: 12 November 2021

Parental dietary nutrients epigenetically influence offspring metabolism. Our analysis revealed unforeseen patterns in how enzymes of the main metabolic pathways respond to protein content in the diet. We reared parental flies Drosophila melanogaster on four types of diet with different dry yeast concentrations ranging from 0.25% to 15%. The subsequent generation was fed by the same diet, so the only variable in the experiments was the yeast concentration in the parental diet. We showed that protein restriction in the parental diet led to higher lactate dehydrogenase (LDH) activity in parents, and this effect was inherited in their progeny. The transgenerational effect of parental dietary yeast on malate dehydrogenase (MDH) activity was found only in males. An elevated level of dietary yeast was sufficient to enhance alanine transaminase ( ALT) and aspartate transaminase (AST) activity in parents, however, did not affect ALT activity and decreased AST  in their offspring. A low yeast parental diet was shown to cause higher urea content in F1 males. It is concluded that parental dietary yeast plays a critical role in metabolic health that can be inherited through generation.

Prediction of pEC50(M) and molecular docking study for the selective inhibition of arachidonate 5-lipoxygenase

N. R. Das1, P. G. R. Achary2*

1Department of CSIT, Siksha ‘O’ Anusandhan deemed to be University, Bhubaneswar, Odisha, India;
2Department of Chemistry, Faculty of Engineering and Technology (ITER), Siksha ‘O’ Anusandhan deemed to be University, Bhubaneswar, Odisha, India;
*e-mail: pgrachary@soa.ac.in

Received: 28 April 2021; Accepted: 12 November 2021

Arachidonate 5-lipoxygenase (ALOX5) is considered a prime target for drug discovery in the area of liver fibrosis, rheumatoid arthritis, atherosclerosis, cancer and asthma. To date, the lead rate in the discovery of drugs that inhibit ALOX5 for the treatment of the above diseases is not satisfactory. So, the development of powerful and effective ALOX5-targeted drugs is desired. In this regard, Quantitative Structure-Activity Relationship (QSAR) and molecular docking can have a major role in screening and designing drugs. In this work, 3D-QSAR models were proposed, which were built using the techniques like Multiple Linear Regression (MLR), and Partial Least Squares (PLS) for the pEC50(M) taking a diverse dataset of 112 molecules. The technique of the ‘Index of Ideality of Correlation (IIC)’ was also investigated to generate an optimal descriptor derived from the SMILES molecular structure. The effect of the number and nature of descriptors on the model were analyzed. The models can be helpful in providing better directions for the development of novel drug targets for 5-lipoxygenase. A significant improvement in the stability of the model was observed by the incorporation of the optimal descriptor. The molecular docking results showed that the ALOX5 receptor was well inhibited by the 112 ligands showing the least binding energy (-10.8 Kcal/mol).  In order to validate the binding mode of the ligands docked with AutoDock Vina software, the top-scored compounds were re-docked using DockThor online docking server. The results obtained from docking suggest that the ligands with IDs 18, 20, 24, 30 and 44 are some of the potential inhibitors for ALOX5.

Indicators of mineral metabolism and dental status of young rats born from female with methionine-induced hyperhomocysteinemia

O. Kutelmakh1*, R. Lesyk2,3, Yu. Chumakova4,
S. Bondar1, V. Dronenko1, L. Shkilniak1

1Vinnytsia National Pirogov Memorial Medical University, Vinnytsya, Ukraine;
2Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
3University of Information Technology and Management in Rzeszow, Rzeszow, Poland;
4International Humanitarian University, Odessa, Ukraine;
*e-mail: ahzelo1968vin@i.ua

Received: 12 July 2021; Accepted: 12 November 2021

We assume that the violation of homocysteine (Hcy) metabolism and vitamin D level in mothers’ utero causes various abnormalities in offspring, including hard-woven teeth and periodontium impairments. The aim of this study was to evaluate the biochemical parameters of blood and alveolar bone homogenates and assess dental status of offspring born from female rats under long-term methionine hyperhomocysteinemia (HHcy). Clinical, biochemical and morphometric studies were performed on 30 female rats and 281 infantile and early juvenile rats. Serum Hcy levels were determined by competitive immunoassay with electrochemiluminescent detection, 25-hydroxyvitamin D (25OHD) level was estimated  by ELISA. No violation of Hcy serum level was found in rats born from females with long-term methionine-induced hyperhomocysteinaemia. Hyperhomocysteinaemic diet caused by long-term use of methionine reduced the level of total and ionized calcium and phosphates in the blood of pregnant females and their offspring. Elevated levels of homocysteine and decreased vitamin D levels in pregnant females caused a delay in the complete physiological eruption of permanent molars in their offspring. Biochemical changes in the blood, jawbones and the presence of early carious processes in the teeth of young rats, in our opinion, are the result of metabolic disorders in utero caused by hyperhomocysteinaemia and vitamin D deficiency in their mothers.

Properties of alpha-L-fucosidase for serum of patients with hepatocellular cancer and cytotoxicity on some cancer cell lines

Z. M. A. A. Hamodat

University of Mosul, College of Science, Chemistry Department, Mosul – Iraq;
e-mail: zahraahamodat@uomosul.edu.iq

Received: 07 July 2021; Accepted: 12 November 2021

Alpha-L-fucosidase (FUCA) degrades many fucosylated glycans and has long been recognized as a tumor marker associated with the early detection of some cancers. This study aimed to purify and characterize alpha-L-fucosidase from the serum of patients with hepatocellular cancer and estimate its toxic effect against hepatocellular carcinoma HepG2, prostate cancer PC3 cell lines and the standard hepatocyte WRL-68 cell line. SDS-Page Electrophoresis technique was used to determine the purity of the purified alpha-L fucosidase and estimate its molecular weight. Three purification steps were used for FUCA purification: precipitation with 65% ammonium sulfate saturation, DEAE-cellulose ion exchange, Sephadex G-75 gel filtration. The procedure resulted in 54% recovery of the enzyme with 27.5-fold purification and 14 U/mg specific activity. It was demonstrated that FUCA purified from the serum of HCC patients showed a more toxic effect on HepG2 cells (IC50 of 65.74 µg/ml) than on PC3 prostate cancer cells (IC50 of 111.5 g/ml) and less toxic effect against standard hepatocyte WRL-68cells (IC50 of 214.5 µg/ml). We can conclude that the inhibitory effect of the purified FUCA on hepatocellular carcinoma is more than its effect on prostate cancer cells. Also, the purified FUCA may be used in studies on anticancer drug development in liver cancer.

Growth hormone, growth hormone receptor and insulin-like growth factor serum levels in patients with obesity and food addiction

O. Avsar1*, S. Sancak2, I. Koroglu3, E. Avci4

1Hitit University, Department of Molecular Biology and Genetics, Corum, Turkey;
2Fatih Sultan Mehmet Education and Research Hospital, Department of Endocrinology, Istanbul, Turkey;
3Arapgir Ali Özge State Hospital, Department of Internal Medicine, Malatya, Turkey;
4Health Sciences University, Department of Biochemistry, Ankara, Turkey;
*e-mail: orcunavsar@hitit.edu.tr

Received: 27 May 2021; Accepted: 12 November 2021

Obesity is a public health problem that increasingly becomes widespread and causes various complications. Food addiction is a hedonic eating behavior characterized by overconsumption of palatable foods (i.e., foods involve a high amount of salt, sugar and fat). Disturbances in the growth hormone signaling pathway were shown to be associated with increased food intake and adiposity. The study aimed to determine the growth hormone (GH), growth hormone receptor (GHR), insulin, and insulin-like growth factor 1 (IGF-1) serum levels in individuals with obesity and food addiction. The present study involved 30 adults with obesity (23 females and 7 males) and 10 healthy adults (5 females and 5 males). 18 obese adults were diagnosed with food addiction, whereas only 2 individuals with food addiction were in the control group. GH, GHR, IGF-1 and insulin values were analyzed with ELISA kits. It was revealed that the obese subjects had significantly lower serum IGF-1 levels compared to healthy individuals (144.55±22.69 ng/ml vs 338.70±61.90 ng/ml, P < 0.001)). No significant differences in the GH, GHR and insulin levels between obese and control groups were detected (P > 0.05). No significant differences between the group with food addiction and the group without food addiction in terms of gender, age, weight, BMI, GH, GHR, insulin and IGF-1 levels were observed. Our study demonstrates that normal IGF-1 levels may be protective for the development of obesity. The serum levels of GH, GHR, insulin, IGF-1 are not associated with food addiction and, therefore, can not be used as novel markers of food addiction.

Serum levels of alpha-melanocyte stimulating hormone, vitamin D, calcium, phosphorus and magnesium in COVID-19 patients

S. Z. Hussein1, M. A. Abdalla2*

1Clinical Biochemistry Unit, Salah Aldeen Health Directorate, Tikrit, Iraq;
2Department of Human Anatomy, Tikrit University College of Medicine, Tikrit, Iraq;
*e-mail: dr.mohammad68@tu.edu.iq

Received: 04 April 2021; Accepted: 12 November 2021

The COVID-19 pandemic occurred and quickly spread throughout the world. To improve the state of COVID-19 patients, it is important to identify the possible clinical differential diagnostic markers and their correlation with the severity of SARS-CoV-2 infection. In this study, the serum level of alpha-melanocyte stimulating hormone (alpha-MSH), vitamin D, calcium, phosphorus and magnesium in the serum of COVID-19 patients were analyzed. Blood samples were collected from 60 patients who attended Isolated Hospital in Tikrit City/Iraq from September to December 2020 and diagnosed by RT-PCR as COVID-19 positive and from 30 healthy individuals. It was shown that COVID-19 patients revealed high serum levels of α-MSH as compared with healthy individuals but low serum levels of vitamin D, calcium, and magnesium which may be recommended as supplements for those patients to increase the innate immune response.

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.