Computer-aided Evaluation of Polyvalent Medications' Pharmacological Potential. Multiphytoadaptogen as a Case Study.

Many human diseases including cancer, degenerative and autoimmune disorders, diabetes and others are multifactorial. Pharmaceutical agents acting on a single target do not provide their efficient curation. Multitargeted drugs exhibiting pleiotropic pharmacological effects have certain advantages due to the normalization of the complex pathological processes of different etiology. Extracts of medicinal plants (EMP) containing multiple phytocomponents are widely used in traditional medicines for multifactorial disorders' treatment. Experimental studies of pharmacological potential for multicomponent compositions are quite expensive and time-consuming. In silico evaluation of EMP the pharmacological potential may provide the basis for selecting the most promising directions of testing and for identifying potential additive/synergistic effects. Multiphytoadaptogen (MPhA) containing 70 major phytocomponents of different chemical classes from 40 medicinal plant extracts has been studied in vitro, in vivo and in clinical researches. Antiproliferative and anti-tumor activities have been shown against some tumors as well as evidence-based therapeutic effects against age-related pathologies. In addition, the neuroprotective, antioxidant, antimutagenic, radioprotective, and immunomodulatory effects of MPhA were confirmed. Analysis of the PASS profiles of the biological activity of MPhA phytocomponents showed that most of the predicted anti-tumor and anti-metastatic effects were consistent with the results of laboratory and clinical studies. Antimutagenic, immunomodulatory, radioprotective, neuroprotective and anti-Parkinsonian effects were also predicted for most of the phytocomponents. Effects associated with positive effects on the male and female reproductive systems have been identified too. Thus, PASS and PharmaExpert can be used to evaluate the pharmacological potential of complex pharmaceutical compositions containing natural products.

[Quantitative targeted screening of proteins associated with lung adenocarcinoma cancer by the method of selected reaction monitoring]. / Napravlennyi kolichestvennyi skrining markerov adenokartsinomy legkogo metodom monitoringa vybrannykh reaktsii.

In the present study, we applied selected reaction monitoring (SRM) to a group of proteins that were previously reported to be associated with lung cancer (Novikova S.E. et al. (2017) Biomeditsinskaya khimiya, 63, 181-210. [1]). Measurements were performed on 59 plasma samples. These samples included: 23 samples of plasma of patients diagnosed with lung adenocarcinoma (LAC), 11 samples of plasma of patients diagnosed with squamous cell lung carcinoma (SqCC), 25 samples of donors with no previous history of oncological diseases, and one pooled sample from each of the above group. As a result of the SRM measurements 52 proteins were detected at least in one individual plasma sample. Statistical analysis showed that there were two groups confidently differentiated by the concentration value of 8 proteins wherein 5 proteins displayed increased level (P00738, P26639, P21926, P08603, P51149) in LAC group and 3 proteins (P51884, O15162, Q8N2K0) indicated diminishing the concentration level towards the control level. Data on protein concentrations obtained for LAC and SqCC did not distinguish the samples by statistical clustering analysis. These potential biomarkers can be used for further development of methods for early diagnostics of lung cancer.

Is myelin basic protein a potential biomarker of brain cancer?

Myelin basic protein is a potential biomarker for the central nervous system diseases in which the myelin sheath is destroyed. Using pseudo-selected reaction monitoring and the method of standard additions, we have measured the myelin basic protein level in the cerebrospinal fluid of patients with neurotrauma (n = 6), chronic neurodegenerative diseases (n = 2) and brain cancer (n = 5). Myelin basic protein was detected only in four out of five cerebrospinal fluid samples of patients with brain cancer. The cerebrospinal fluid myelin basic protein level ranged from 3.7 to 8.8 ng ml-1. We suggest that monitoring of myelin basic protein in cerebrospinal fluid can serve as a diagnostic test for the brain cancer.

Horizontal Transfer of Tamoxifen Resistance in MCF-7 Cell Derivates: Proteome Study.

Using estrogen-dependent MCF-7 breast cancer cells and tamoxifen-resistant MCF-7/T subline we have shown that their co-cultivation lead to increase in tamoxifen resistance in the parent MCF-7 cells. The proteome analysis of MCF-7/T cells and new-generated resistant cells revealed 21 common proteins differently expressed in both the resistant cell lines, among them - 6 proteins were associated with the drug or hormonal resistance. Both resistant lines were characterized with suppression of estrogen receptor and activation of SNAIL1-signaling - mesenchymal pathway playing an important role in the down-regulation of estrogen receptor and maintaining of the estrogen-independent phenotype.

[The connection of miR-21 and miR-155 with regulation of 15-HPGDH mRNA in human breast cancer cells]. / Sviaz' miR-21 i miR-155 s reguliatsiei mRNK 15-HPGD v kletkakh raka molochnoi zhelezy cheloveka.

Breast cancer is the most frequent cancer and the leading cause of cancer-related deaths in women worldwide. We determined the expression of COX2, COX1, 15-HPGDH mRNA and miRNAs (miR-21, miR-155) in three estrogen positive human breast cancer cell lines (MCF-7, BT-474, ZR-75-1). According to the results of three independent experiments the amount of COX1 and COX2 mRNA was significantly higher in the ZR-75-1 than in MCF-7 and BT-474 cells. Levels of total 15-HPGDH; functional 15-HPGDH mRNA in BT-474 cell line were lower than in MCF-7 and ZR-75-1 ones. The synthesis of 15-HPGDH enzyme in BT-474 line was blocked at the nuclear immature pre-mRNA processing level. miR-155 expression level was significantly lower than miR-21 in breast cancer cell lines. Correlations between the dysregulation of miR-21, miR-155 and 15-HPGDH, COX-1, COX-2 mRNA were identified. Expression of miR-21 was high in MCF-7, ZR-75-1 and BT-474 cell lines. Our results show that miR-21 and miR-155 regulate activity of several genes in cancer cells, their effect on the individual genes was in some cases cumulative. Based on our results, we concluded that miR-21, miR-155 suppress the work of tumor suppressor gene 15-HPGDH and induce potential oncogene COX-2 that promotes cell malignancy and metastasis of breast cancer.

[Search for potential gastric cancer biomarkers using low molecular weight blood plasma proteome profiling by mass spectrometry].

Gastric cancer, one of the most widespread malignant tumors, still lacks reliable serum/plasma biomarkers of its early detection. In this study we have developed, unified, and tested a new methodology for search of gastric cancer biomarkers based on profiling of low molecular weight proteome (LMWP) (1-17 kDa). This approach included three main components: sample pre-fractionation, matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), data analysis by a bioinformatics software package. Applicability and perspectives of the developed approach for detection of potential gastric cancer markers during LMWP analysis have been demonstrated using 69 plasma samples from patients with gastric cancer (stages I-IV) and 238 control samples. The study revealed peptides/polypeptides, which may be potentially used for detection of this pathology.

[Prostate cancer detection using mass-spectrometric profiling of low-molecular weight blood proteome].

Currently, there are no reliable biomarkers of blood plasma for early detection of prostate cancer (PC), a one of the most common malignancies in men. This study developed, universalized and tested a new standardized methodology of detection of prostate cancer biomarkers--profiling of low-molecular weight proteome of blood plasma (1-17 kDa). This approach includes three main components: a preliminary sample preparation, time-of-flight mass spectrometry with an matrix-assisted laser desorption/ionization ALDI-TOF-MS), and the processing of data using bioinformatics software package. The potentials and prospects of the approach developed for identification of potential PC markers are demonstrated. 46 samples of blood plasma of PC patients and 26 controls were screened. This evaluation identified peptides/polypeptides that have the potential to be used for the detection of disease.

[Production of active oxygen by neutrophils in patients with cancer of the esophagus and stomach before and after surgical interventions].

The state of the oxygen-depended antimicrobial activity of neutrophils on oncologic patients was studied. Production of active oxygen by non-activated and activated neutrophils of peripheral blood from 19 patients (the average age of 56 years, 16 males and 3 females) with esophageal cancer and cancer of the stomach and metastasis to the esophagus was recorded by chemiluminescence before and after surgical interventions (transpleural gastrectomy, Lewis type operation) on the 1st, 3rd and 7th days after the operations. The oxygen-depended antimicrobial activity of the neutrophils was investigated by their ability to produce active oxygen when incubated with opsonized zymosan and N-formyl-methionyl-leucyl-phenylalanine. Luminol chemiluminescence recorded the whole pool of active oxygen and showed the summary activity of myeloperoxidase and NADPH-oxidase, while luceginin chemiluminescence measured formation of superoxide anion radical (*O(-)2) and evaluated the activity of NADPH-oxidase. The results showed that the procedure provided estimation of the functional state of the neutrophil leukocytes and their ability to produce active oxygen. Impairment of the myeloperoxidase- and NADPH-depended production of active oxygen by non-activated and activated neutrophils observed before operations resulted in altered synthesis of active oxygen after the surgery. This is one of the leading factors of the infection development in the end. Detection of the myeloperoxidase- and NADPH-depended production of active oxygen could be of help in revealing the groups of risk among patients at the stage of the preoperative examination and serve as a prognostic factor of the development of severe infectious complications during the postoperative period.

[Lipoxins: study of various biosynthesis pathways]. / Lipoksiny. Issledovanie razlichnykh putei biosinteza.

Lipoxins A4 and B4 were obtained by using soybean lipoxygenase and blood cells as a source of enzymatic activity. The conditions facilitating lipoxin biosynthesis from arachidonic acid catalyzed by soybean 15-lipoxygenase were selected. A comparative analysis of lipoxin biosynthesis with the use of cell suspensions containing only granulocytes and of mixed suspensions (platelets + granulocytes and platelets + total fraction of blood leucocytes) was carried out.

[Arachidonic acid metabolism in the neutrophilic granulocytes in lymphogranulomatosis]. / Obmen arakhidonovoi kisloty v neitrofil'nykh granulotsitakh pri limfogranulematoze.

Metabolism of arachidonic acid in neutrophils was studied in vitro in blood samples obtained from 36 patients with Hodgkin's disease of various stage and histology and 32 healthy donors. The basic metabolites were: leucotriene B4, such products of its omega-oxidation as 20-hydroxy-leucotriene B4 (20-OH-LTB4) and 20-carboxy-leucotriene B4 (20-COOH-LTB4), and 5-hydroxyeicosatetraenic acid. Their profile proved identical in patients with Hodgkin's disease and healthy donors. Most patients with Hodgkin's disease showed a decrease in leucotriene B4 and 5-hydroxyeicosatetraenic acid and an increase in the level of omega oxidation of leucotriene B4 as assessed by omega catabolite/leucotriene B4 ratio. The levels of 5-hydroxyeicosatetraenic acid and leucotriene B4 omega-oxidation were found to depend upon histology and stage of Hodgkin's disease. They were nearly normal in patients with stage III, mixed cellular disease, B-symptoms and signs of biologic activity of tumor. A direct correlation was established between the level of leucotriene B4 omega oxidation in neutrophils and that of ceruloplasmin in blood serum of patients with various stages of Hodgkin's disease.

[Fatty acid composition of phospholipids of erythrocyte membranes and risk of breast cancer]. / Sostav zhirnykh kislot fosfolipidov membran éritrotsitov i risk raka molochnoi zhelezy.

Five fatty acids, two saturated--palmitic (C16:0), stearic (C18:0), and three unsaturated--oleic (C18:1), linoleic (C18:2) and arachidonic (C20:4) were assayed in red cell membranes of breast cancer patients and healthy controls. No significant difference in the levels of saturated fatty acids was established between the two groups, although mean values were somewhat lower in cancer patients. Mean levels of unsaturated fatty acids were also lower in breast cancer patients, with the difference being significant for arachidonic acid in postmenopausal women. There was no significant difference in polyunsaturated/saturated acid ratio between patients and controls in either cycling or postmenopausal group. In controls, the levels of the fatty acids checked, particularly, those of arachidonic acid were higher in menopause than in premenopause. However, no difference in the fatty acid levels was observed between pre- and postmenopausal cancer patients.

[Congenital disorders of tyrosine metabolism and their correction in children with tumors]. / Vrozhdennye narusheniia obmena tirozina i ikh korrektsiia pri opukholiakh u detei.

The paper deals with the evaluation of quantitative indexes of urine excretion of tyrosine and its metabolites--highly carcinogenic p-hydroxyphenyllactic acid and homogentisic acid--in infants suffering various tumors and their parents. Also, the effect of ascorbic acid (100 mg/kg body weight daily) on tyrosine metabolism was studied. The sick children and their parents showed increased urine excretion of tyrosine and p-hydroxyphenyllactic acid matched by a decrease in that of homogentisic acid. The said changes suggest inhibition of p-hydroxyphenylpyruvate dioxygenase (CE 1.13.11.27). High doses of ascorbic acid were shown to arrest or significantly inhibit the excretion of carcinogenic metabolite of tyrosine in patients with hemoblastoses and nephroblastoma.

[Determination of retinol in human blood plasma using high pressure liquid chromatography]. / Opredelenie retinola v plazme krovi cheloveka metodom zhidkostnoi khromatografii vysokogo davleniia.

A quantitative procedure for estimation of retinol in human blood plasma using high pressure liquid chromatography involved separation of the retinoids on an reversed phase analytical column. C19-retinoid aldehyde was used as an internal standard. The procedure enabled to detect up to 0.015 microgram of retinol/ml of blood plasma and may be used for routine clinical analyses.

[Determination of carminomycin and its basic metabolite 13-dihydrocarminomycin by a highly efficient liquid chromatographic method with a fluorescent detector]. / Opredelenie karminomitsina i ego osnovnogo metabolita 13-digidrokarminomitsina metodom vysokoéffektivnoi zhidkostnoi khromatografii s fliuorestsentnym detektorom.

Carminomycin, an antitumor antibiotic, and 13-dihydrocarminomycin, its main metabolite, were determined in the blood plasma of patients with highly efficient liquid chromatography and a fluorescent detector (lambda ex=492 nm, lambda em=538 nm). The liquid chromatograph manufactured by Spectro-Physics, model SP-8000, with Particyl Column 10/25 was used for the determination. The composition of the mobile phase of acetonitrile--0.1 M H3PO4 was 93:7 by the volume. The mobility speed was 2 ml/min. The retention time of carminomycin, rubomycin (used as the national standard) and dihydrocarminomycin was 6.8, 8 and 10.2 minutes, respectively. All three substances showed symmetrical peaks with high resolution. The early phase kinetics of carminomycin after its intravenous injection to the patients in a dose of 24 mg (18 mg/m2) is satisfactorily described by the diexponential equation. The pharmacokinetic parameters of carminomycin corresponding to the open two-compartmental system are presented. Highly efficient liquid chromatography may be used for the pharmacokinetic description of biotransformation of carminomycin and some other antibiotics of the anthracycline group.