Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry最新文献

Arachidonic acid (AA) is widely distributed in many organs and can be metabolized in several ways into small lipid molecules: eicosanoides that perform a physiological and/or pathophysiological role. Among them, epoxyeicosatrienoic acids (EET) and hydroxyeicosatetraenoic acids (HETE), which are produced by cytochrome P450 (CYP) enzymes, have attracted much attention. AA metabolites play an important role in such cellular processes as proliferation, differentiation, and apoptosis and are also involved in the development of organs and tissues as well as in the pathogenetic processes underlying the development of various diseases; in particular, AA metabolites formed with the participation of CYP are involved in normal and pathological processes associated with the functioning of the cardiovascular system. This review focuses on the formation and cardiovascular effects of CYP-generated AA metabolites and summarizes current literature data on the physiological and pathophysiological significance of the CYP-dependent AA metabolic pathway and its relevance to the cardiovascular system in both health and disease.

A study is performed of the effect elevated concentrations of antibodies have on the immune response in patients with autoimmune thyroiditis (AIT) living in Arkhangelsk. Results from immunological examinations of 108 individuals with AIT are presented. The immunological study included examining hemograms, the contents of phenotypes of lymphocytes, cytokines, immunoglobulins, and antibodies of thyroid peroxidase (TPO), double-stranded DNA, and nucleoproteins. It was estanlished that a rise in the concentration of antibodies (AB) to TPO, ds-DNA, to RNP in patients with AIT is associated with elevated contents of mature T lymphocytes (CD3+), cytotoxic T lymphocytes (CD8+), cells labeled for programmed cell death (CD95+), and a drop in concentrations of IL-10. An increase in the concentration of cytotoxic lymphocytes and cells labeled for apoptosis and a simultaneous drop in the concentration of regulatory IL-10 is not beneficial in terms of prognostic value for the accumulation of autoantigens and autoantibodies.

Tyrosine protein kinase HER2 plays an important role in carcinogenesis. In breast cancer (BC), the HER2 gene is amplified in approximately 20% of cases. Trastuzumab is used to treat BC with HER2 gene amplification. Trastuzumab is not effective in treating tumors with low HER2 expression, but trastuzumab deruxtecan was recently found to significantly improve prognosis in these patients. Nonetheless, there are still difficulties with accurate diagnosis of HER2-low BC, especially at the preoperative stage. In this study, when comparing the results between core needle biopsies and resection specimens using three immunohistochemistry scores (0, 1+, and 3+) of the HER2 protein level, we observed only moderate agreement (66.2%, κ = 0.486) in patients who did not undergo neoadjuvant therapy (n = 71). Other miRNA- or protein-coding genes regulated by HER2 signaling pathways may be additional markers for the scoring of the HER2 level. We measured levels of HER2-regulated miR-378a, -205, and -21 and mRNA levels of their target genes TRPS1, ITGA2, BCL6, and PTEN in BC surgical specimens. For estrogen receptor-positive BC (n = 64), we confirmed that the expression of miR-21, miR-378a, TRPS1, PTEN, and BCL6 is associated with the HER2 protein level. Moreover, the expression profile of miR-378a, BCL6, and PTEN differed between HER2 1+ and HER2 3+ tumors. TRPS1 expression was significantly higher in HER2 3+ tumors compared with HER2 0 tumors, but there was no difference in the amount of TRPS1 mRNA between HER2 1+ and HER2 3+ tumors. Thus, an analysis of the expression of miR-21, miR-378a, TRPS1, PTEN, and BCL6 may help to distinguish between HER2-negative, HER2-positive, and HER2-low BCs.

Synthesis of RNA by in vitro transcription has many applications in the production of vaccines, anticancer and gene therapy medicines. Despite several advantages of mRNA molecules over DNA in terms of transient transgene expression, rapid degradation of RNA is the limiting factor for the efficacy of mRNA-based therapeutics. Translational efficiency and stability of mRNA depends on the sequence of 5′ and 3′ untranslated regions, cap structure, coding sequence (CDS), and the poly(A) tail. So far, the role of the poly(A) tail sequence in mRNA stability and translational efficiency remains poorly investigated. There are few studies evaluating the influence of poly(A) tails containing certain non-A nucleotides or segments of non-A nucleotides on the efficacy of mRNA therapeutics. In our work, we conducted a comprehensive assessment of the effects of mRNA poly(A) tail on luciferase expression in cell lines HEK293 and DC2.4 and in BALB/c mice. Our results showed that mRNAs with a poly(A) tail consisting of 100 adenines separated by a segment of 10 nucleotides (50A-GCAUAUGACU-50A) have the best translational efficiency both in vitro (in cell lines) and in vivo (in BALB/c mice). Nonetheless, the results were inconsistent and depended both on cell lines and on the coding sequence (NanoLuc or firefly luciferase). Thus, effects of a mRNA poly(A) tail on luciferase expression may depend on the sequence of the gene in question and on specific features of the expression of RNA-binding proteins in cell lines.

Cardiovascular diseases (CVDs) are one of the world’s main causes of mortality. The role of matrix metalloproteinases (MMPs) as markers of CVD complications has been actively discussed in recent years. It has been proven that a high level of MMPs contributes to the development of unstable plaque, aggravating a patient’s prognosis. Being an independent risk factor, obesity also results in complications of cardiovascular diseases. Studies on the effect an increase in the body mass index and adipose tissue hormones on the level of MMPs have begun to appear. Results from clinical and experimental studies over the last 5 years devoted to studying matrix metalloproteinases are presented here. The role of MMPs in the pathogenesis of atherosclerosis is reflected. MMPs are considered biomarkers of the development of adverse cardiovascular events. Results from studies on the effect obesity has on the level of MMPs are also presented here. A search was performed for articles in the PubMed, Google Scholar, and Elibrary databases for the period from 2018 to 2023 using the keywords metalloproteinases and atherosclerosis, metalloproteinases and obesity, and the connection between atherosclerosis and obesity separately for each type of MMP.

Hepatitis is a disease that attacks the organs of the human liver that causes inflammation that makes the liver function. Telomere length and telomerase activity in chronic liver disease and hepatocarcinoma correlate with disease severity. Telomeres are protected by her six shelterin proteins or telosome, namely TERF1/TRF1, TERF2/TRF2, POT1, TPP1, RAP1, and TIN2. TRF2 protein has an important role in regulating the molecular events that maintain telomere integrity. The objective of the research was to determine the concentration of TERF2 in leukocytes in patients with hepatitis. The research design method is descriptive with a case-control study involving 62 subjects. Blood samples were taken from patients with hepatitis from Klinik Pratama Prof. Qomariyah, which amounted to 32 people, including 8 males and 24 females (40–60 years). As a control, employees of YARSI University with the same age range, amounting to 24 people, including 20 men and 4 women. Measurements of TERF2 used the ELISA method. TERF2 protein concentrations in patients with hepatitis leukocytes were significantly higher (p < 0.05) compared to controls. Similarly, TERF2 levels between male patients with male controls and between female patients with female controls also showed significant differences in sufficient concentration (p < 0.05). Instead, the concentrations of TERF2 between patients with hepatitis men and women did not have a difference. The concentration of TERF2 in leucocyte patients with hepatitis showed a concentration higher than in healthy people, both men and women.

Dyslipidemia is a well-known risk factor for the development of cardiovascular diseases and atherosclerosis. The effects of combined pretreatment with atorvastatin and fenofibrate (Tricor) were studied in a mouse model of acute lipemia induced by a general lipase inhibitor, poloxamer 407 (P-407, 250 mg/kg). This lipemia is characterized by significantly increased serum levels of triglycerides (TG), low-density lipoprotein (LDL) cholesterol, together with decreased concentration of high-density lipoprotein (HDL) cholesterol. Atorvastatin pretreatment had a hypolipidemic effect, decreasing concentrations of LDL cholesterol and increasing HDL cholesterol. Pretreatment of mice with fenofibrate decreased TG level, increasing HDL cholesterol. Combined pretreatment with atorvastatin and fenofibrate decreased TG and total cholesterol. Elevation of the serum cystatin C level was found in control and lipemic mice pretreated with atorvastatin, fenofibrate, or both. Liver expression of lysosomal acid lipase increased in atorvastatin- or/and fenofibrate-pretreated groups of lipemic mice. It was concluded that increased expression of lysosomal acid lipase is related to the removal of lipid droplets from hepatocytes, thus preventing acute lipemia. Lastly, cystatin C may be a “theranostic” biomarker for hypolipidemic drugs.

When developing vaccines and immunotherapeutic and enzymatic drugs, it is usually necessary to obtain a functional protein preparation, which can later be used for immunization or analytical studies in the process of creating a drug. Production in E. coli bacterial cells is the fastest and cheapest way to obtain such proteins. However, recombinant proteins, when produced in E. coli, often form insoluble and functionally inactive aggregates. The purpose of this work is to create a set of expression vectors for rapid screening of optimal conditions for the production of recombinant proteins prone to aggregation in E. coli in soluble form. The work used modern genetic engineering methods, including optimization and de novo synthesis of nucleotide sequences encoding helper polypeptides. For the production and chromatographic purification of proteins, the standard strain BL21(DE3) and the affinity chromatography method using a metal chelate sorbent were used. As a result, a set of nine vectors with helper polypeptides was obtained to increase solubility and increase product yield during the production of recombinant proteins in E. coli cells. The efficiency of obtaining protein drugs prone to molecular aggregation using this set of vectors has been demonstrated by the exampe of three cytokines: interleukin-31 (IL-31), interleukin-33 (IL-33), and transforming growth factor beta (TGF-beta1). The resulting set allows for rapid selection of a suitable helper polypeptide as well as optimization of conditions for obtaining a soluble form of recombinant proteins in E. coli during the pharmaceutical development of therapeutic drugs.

The development of effective delivery systems for antitumor drugs with increased specificity and controlled release is one of the challenges for biomedical chemistry. This paper presents studies of non-covalent interactions of human serum albumin, which may be a promising carrier for drug delivery, with antitumor antibiotic doxorubicin and folic acid, which have the potential of a guide ligand. Intermolecular interactions of doxorubicin and folic acid with human serum albumin were studied by spectroscopic methods at various pH levels and temperatures. The binding strength of doxorubicin and folic acid responded differently to pH changes. The affinity of the drug to protein increased with the transition from acidic to alkaline conditions, while pH 7.4 was optimal for binding for folic acid. In the case of the triple system, it was found that there was no significant effect of albumin complexation with folic acid on non-covalent interaction with doxorubicin. As expected, binding to these active compounds altered the conformation of the protein. At the same time, this change was minimal in physiological pH for folic acid and in alkaline for doxorubicin. Additionally, the therapeutic properties of doxorubicin, non-covalently bound to human serum albumin, were shown to be preserved in vitro.

Melanoma is one of the most aggressive types of cancer. Melanoma morbidity is increasing every year and mortality rate is high besides target therapy. Calorie restriction is lifestyle approach that can alter the activity of the pathways regulating key processes of melanomagenesis. The signaling cascades modulated by fasting includes JAK/STAT. Female С57Bl/6 mice were used for the investigation of the calorie restriction effect on the tumor growth and development. Control group is the mice on ad libitum diet, CR group is the mice on 30% calorie restricted diet. B16 melanoma cell transplantation has been carried out in the mice of the both groups after 3 months of ad libitum and CR regimes. During 15 days after melanoma cell implantation the formation of solid tumor has been occurred. Then the animals were euthanized and average mouse weight and tumor mass in both groups were evaluated. JAK/STAT gene expression was assessed by means of real-time PCR. The activity of glutathione peroxidase and glutathione-S-transferase and glutathione level were registered spectrophotometrically. 30% calorie restriction in mice with B16 melanoma decreased of tumor mass by 1.8 times (р = 0.049) as compared with the tumor mass of control group animals. STAT1 expression increased by 1.9 times, STAT3, STAT5b, STAT6, JAK1 and JAK2 expression decreased by 2.8, 3.8, 2.6, 3 and 4.5 times accordingly. Activity of glutathione peroxidase, glutathione-S-transferase and glutathione level decreased by 1.6, 3 and 1.5 times accordingly compare with control group. The received data can become pathogenical basis for the creation of new adjuvant method of anticancer therapy.