Pronounced Cytotoxic Effect Research Articles

Fabrication and Biocompatibility of Layer-by-layer Assembled Composite Graphene Oxide-polysaccharide Microcapsules

The present study is focused on the construction and characterization of the morphology and biocompatibility of polysaccharide multilayered microcapsules (PMC) composed of natural polyelectrolytes (chitosan/alginate/hyaluronic acid), and on the effect of graphene oxide (GO) incorporation in the polymer matrix. The insertion of GO in the polymer matrix is an innovative and still evolving strategy used to modify the properties of the polyelectrolyte microcapsules. We have fabricated a number of hybrid GO-polysaccharide multilayered capsules by layer-by-layer assembling technique onto a CaCO3 core, followed by core decomposition in mild conditions. Hybrid microcapsules with different composition were constructed by varying the number or localization of the incorporated GO-layers. It was found that the thickness of the hybrid microcapsules, evaluated by atomic force microscopy, decreases after incorporation of GO nanosheets in the polymer matrix. Analysis of the viability and proliferation of fibroblasts after incubation with hybrid PMC revealed pronounced concentration-dependent cytotoxic and antiproliferative effect. Based on the results, we can conclude that the hybrid multilayered microcapsules made of natural polysaccharides and graphene oxide could be used for biomedical applications.

Synthesis, structural characterization, and cytotoxic activity in tumor cells of Cu(II) and Co(II) complexes with o‐vanillin amino acids Schiff bases

Two new families of Cu(II) and Co(II) mononuclear complexes with mixed ligands, [Cu(VanTrpt)(bipy)]·MeOH (CuVanTrpt), [Cu(VanSer)(bipy)]·4H2O (CuVanSer), [Cu(VanTyr)(bipy)]·H2O (CuVanTyr), [Cu(VanThr)(bipy)]·MeOH (CuVanThr), [Co(VanTrpt)(bipy)(OH2)]·H2O·MeOH (CoVanTrpt), [Co(VanSer)(bipy)(OH2)]·MeOH (CoVanSer), [Co(VanTyr)(bipy)(OH2)]·3H2O·MeOH (CoVanTyr), and [Co(VanThr)(bipy)(OH2)] (CoVanThr), (VanX are Schiff base proligands resulted from the condensation of o‐vanillin with D,L‐tryptophan (VanTrpt), L‐serine (VanSer), L‐tyrosine (VanTyr), and L‐threonine (VanThr), respectively, and bipy is 2,2′‐bipyridine), have been obtained and characterized on the basis of elemental analysis, magnetic measurements, and spectral data (FT‐IR and UV–Vis–Nir spectroscopy). Crystal structures of CuVanTrpt, CuVanSer, and CuVanTyr have been solved by single‐crystal X‐Ray diffraction. Biological evaluation revealed that they are cytotoxic in HeLa (human carcinoma of the uterine cervix) and LSR‐SF‐SR (rat sarcoma induced by Rous sarcoma virus strain Schmidt‐Ruppin) cells, showing an enhancement of cytotoxicity dependent of concentration and treatment time. The most pronounced cytotoxic effect is expressed by CuVanSer and CoVanSer. Various cytopathological changes of both types of cancer cells were observed after treatment with Cu(II) and Co(II) complexes for 72 h at concentrations of 100 and 200 μg ml−1. LSR‐SF‐SR cells were found to be relatively more sensitive to the cytotoxic effect of the complexes investigated as compared to HeLa cells.

Effect of a novel thiazole derivative and its complexes with polymeric carriers on the processes of lipid peroxidation in lymphoma cells

Background. Many types of tumors are sensitive to changes in prooxidant-antioxidant balance. Thus, further studies on reactive oxygen species inducing antitumor drugs that generate oxidative stress-dependent cytotoxic effects are promising. Our previous works showed that thiazole derivatives in combination with polymeric carriers have a pronounced cytotoxic effect on tumor, while not being cytotoxic against pseudo-normal cells in vitro. It was found that thiazole derivatives in complex with PEG-based polymeric carriers affected the antioxidant system of lymphoma cells in vitro. The aim of this work was to study the in vitro effect of the complex of thiazole derivative N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) in combination with polymeric carriers poly(VEP-co-GMA)-graft-mPEG (Th1), poly(PEGMA) (Th3) and poly(PEGMA-co-DMM) (Th5) on the level of lipid peroxidation products in NK/Ly cells. Materials and Methods. The experiments were conducted on white wild-type male mice with a grafted NK/Ly lymphoma. Ascites tumor cells were inoculated into mice intraperitoneally. Abdominal drainage with ascites of anesthetized mice was performed with a sterile syringe on the 7th–10th days after inoculation. Investigated compounds BF1, polymeric carriers Th1, Th3, Th5 and combination of BF1 + Th1 (Th2), BF1 + Th3 (Th4) and BF1 + Th5 (Th6) at a final concentration of 10 μM were added to the lymphoma samples and incubated for 10 minutes. The level of lipid peroxidation products, such as lipid hydroperoxides and thiobarbituric acid-positive products) were determined according to the techniques described below. Results. All applied complexes based on thiazole derivative BF1 and PEG-based polymeric carriers at a concentration of 10 μM increased the activity of lipid hydroperoxi­des in the lymphoma cells by 29–36% compared to control. Complexes Th2 and Th6 increased the significance of BF1 influence on lymphoma cells from P <0.05 to P <0.01. Among all of the studied complexes, Th4 and Th6 significantly increased the level of TBA-positive products, while Th2 and BF1 did not change the content of the secondary products of lipid peroxidation. None of the unconjugated polymeric carriers affected the level of lipid peroxidation products. Conclusions. Thus, based on the results of this work, thiazole derivative BF1 in complex with polymeric carriers increases the level of primary and secondary products of lipid peroxidation in lymphoma cells. Polymeric carriers enhanced the effect of thiazole derivative on the studied parameters, so complexes of thiazole derivatives and PEG-containing polymeric carriers should be taken into consideration and further investigated as potential antitumor agents.

Biochemistry and toxicology of proteins and peptides purified from the venom of Vipera berus berus.

The isolation and characterization of individual snake venom components is important for a deeper understanding of the pathophysiology of envenomation and for improving the therapeutic procedures of patients. It also opens possibilities for the discovery of novel toxins that might be useful as tools for understanding cellular and molecular processes. The variable venom composition, toxicological and immunological properties of the common vipers (Vipera berus berus) have been reviewed. The combination of venom gland transcriptomics, bottom-up and top-down proteomics enabled comparison of common viper venom proteomes from multiple individuals. V. b. berus venom contains proteins and peptides belonging to 10–15 toxin families: snake venom metalloproteinase, phospholipases A2 (PLA2), snake venom serine proteinase, aspartic protease, L-amino acid oxidase (LAAO), hyaluronidase, 5′-nucleotidase, glutaminyl-peptide cyclotransferase, disintegrin, C-type lectin (snaclec), nerve growth factor, Kunitz type serine protease inhibitor, snake venom vascular endothelial growth factor, cysteine-rich secretory protein, bradykinin potentiating peptide, natriuretic peptides. PLA2 and LAAO from V. b. berus venom produce more pronounced cytotoxic effects in cancer cells than normal cells, via induction of apoptosis, cell cycle arrest and suppression of proliferation. Proteomic data of V. b. berus venoms from different parts of Russia and Slovakian Republic have been compared with analogous data for Vipera nikolskii venom. Proteomic studies demonstrated quantitative differences in the composition of V. b. berus venom from different geographical regions. Differences in the venom composition of V. berus were mainly driven by the age, sex, habitat and diet of the snakes. The venom variability of V. berus results in a loss of antivenom efficacy against snakebites. The effectiveness of antibodies is discussed. This review presents an overview with a special focus on different toxins that have been isolated and characterized from the venoms of V. b. berus. Their main biochemical properties and toxic actions are described.

Study of the laser radiation effect in combination with doxorubicin on the survival of MCF7 and MCF7DOX culture cells

The use of lasers in oncology is constantly evolving. In recent years, the mechanisms of the effect of laser radiation, particularly of the infrared spectrum, on tumour cells in combination with various chemotherapeutic agents have been studied.
 The aim. To study the effect of laser radiation with a wavelength of 660 and 810 nm on the survival of tumour cell cultures MCF7 and MCF7DOX in the presence of doxorubicin.
 Materials and methods. Tumour cells of MCF7 and MCF7DOX cultures were cultured in DMEM medium (Biowest, France) with 10 % fetal calf serum (FST) (Biowest, France) and 40 μg / ml gentamicin (Sigma, USA). Doxorubicin was added to the cells to a final concentration of 1 μg / ml. Cells were irradiated with a laser (Photonica-Plus, Ukraine) with a wavelength of 660 and 810 nm (irradiation time – 5 min, power density – 50 mV/cm2, irradiation dose 15 J/cm2). The results were recorded using a multiwell spectrophotometer (Labsystems Multiskan PLUS, Finland). Photomicrographs of cells were taken using a Carl Zeiss microscope, Germany.
 Results. Cell survival assessment and morphological characteristics in micropreparations indicate the antitumor efficacy of the combined effects of laser irradiation and doxorubicin. The most pronounced cytotoxic effect on MCF7-DOX culture cells was caused by doxorubicin exposure for 90 min in combination with infrared laser irradiation (λ=810 nm).
 Conclusions. Infrared laser light synergises with the toxic effects of doxorubicin and creates favourable conditions for apoptosis of tumour cells, as evidenced by cytomorphological data.

Antiproliferative Effects of the Aptamer d(GGGT)4 and Its Analogues with an Abasic-Site Mimic Loop on Different Cancer Cells.

In this paper, we study the T30923 antiproliferative potential and the contribution of its loop residues in six different human cancer cell lines by preparing five T30923 variants using the single residue replacement approach of loop thymidine with an abasic site mimic (S). G-rich oligonucleotides (GRO) show interesting anticancer properties because of their capability to adopt G-quadruplex structures (G4s), such as the G4 HIV-1 integrase inhibitor T30923. Considering the multi-targeted effects of G4-aptamers and the limited number of cancer cell lines tested, particularly for T30923, it should be important to find a suitable tumor line, in addition to considering that the effects also strictly depend on G4s. CD, NMR and non-denaturating polyacrylamide gel electrophoresis data clearly show that all modified ODNs closely resemble the dimeric structure of parallel G4s’ parent aptamer, keeping the resistance in biological environments substantially unchanged, as shown by nuclease stability assay. The antiproliferative effects of T30923 and its variants are tried in vitro by MTT assays, showing interesting cytotoxic activity, depending on time and dose, for all G4s, especially in MDA-MB-231 cells with a reduction in cell viability approximately up to 30%. Among all derivatives, QS12 results are the most promising, showing more pronounced cytotoxic effects both in MDA-MB-231 and Hela cells, with a decrease in cell viability from 70% to 60%. In summary, the single loop residue S substitution approach may be useful for designing antiproliferative G4s, considering that most of them, characterized by single residue loops, may be able to bind different targets in several cancer cell pathways. Generally, this approach could be of benefit by revealing some minimal functional structures, stimulating further studies aimed at the development of novel anticancer drugs.

The Inhibitory Response to PI3K/AKT Pathway Inhibitors MK-2206 and Buparlisib Is Related to Genetic Differences in Pancreatic Ductal Adenocarcinoma Cell Lines.

The aberrant activation of the phosphoinositide 3-kinase (PI3K)/ protein kinase B (AKT) pathway is common in pancreatic ductal adenocarcinomas (PDAC). The application of inhibitors against PI3K and AKT has been considered as a therapeutic option. We investigated PDAC cell lines exposed to increasing concentrations of MK-2206 (an AKT1/2/3 inhibitor) and Buparlisib (a pan-PI3K inhibitor). Cell proliferation, metabolic activity, biomass, and apoptosis/necrosis were evaluated. Further, whole-exome sequencing (WES) and RNA sequencing (RNA-seq) were performed to analyze the recurrent aberrations and expression profiles of the inhibitor target genes and the genes frequently mutated in PDAC (Kirsten rat sarcoma virus (KRAS), Tumor protein p53 (TP53)). MK-2206 and Buparlisib demonstrated pronounced cytotoxic effects and limited cell-line-specific effects in cell death induction. WES revealed two sequence variants within the direct target genes (PIK3CA c.1143C > G in Colo357 and PIK3CD c.2480C > G in Capan-1), but a direct link to the Buparlisib response was not observed. RNA-seq demonstrated that the expression level of the inhibitor target genes did not affect the efficacy of the corresponding inhibitors. Moreover, increased resistance to MK-2206 was observed in the analyzed cell lines carrying a KRAS variant. Further, increased resistance to both inhibitors was observed in SU.86.86 carrying two TP53 missense variants. Additionally, the presence of the PIK3CA c.1143C > G in KRAS-variant-carrying cell lines was observed to correlate with increased sensitivity to Buparlisib. In conclusion, the present study reveals the distinct antitumor effects of PI3K/AKT pathway inhibitors against PDAC cell lines. Aberrations in specific target genes, as well as KRAS and TP53, individually or together, affect the efficacy of the two PI3K/AKT pathway inhibitors.

Complexation-induced nanoarchitectonics of sulfonate cailx[4]resorcinol substituted at the upper rim by N-methyl-d-glucamine fragments: Morphological transition and in vitro anticancer activity

Complexes of various nitrogen-containing ligands with transition metals are known for their anticancer activity. At the same time, the shape of aggregates of these complexes can have a significant effect on cytotoxicity and selectivity of action in tumor cells, however, this issue has received insufficient attention in the literature. In this work, the selective binding ability of sulfonate calix[4]resorcinol functionalized at the upper rim by N-methyl-D-glucamine fragments towards copper (II) ion (Cu2+) was demonstrated. N-methyl-D-glucamine calix[4]resorcinol weakly interacts with Zn2+, but effectively binds Cu2+ with the formation of a 2:1 complex. The complexation of calix[4]resorcinol with copper leads to a significant change in the shape of aggregates from rounded to rod-like particles, which is confirmed by TEM images. The rod-like aggregates of calix[4]resorcinol–copper(II) showed a pronounced cytotoxic effect against HuTu 80 cell line, while not affecting Chang liver cells.

Synthesis of 1,4,2,6-dithiadiazinane 1,1-dioxide and study of its cytotoxic activity

Cyclothiomethylation of sulfamide using paraformaldehyde or bis(dimethylamino)methane, hydrogen sulfide or its sodium salts catalyzed by rare-earth salts affords 1,4,2,6-dithiadi azinane 1,1-dioxide. The heterocycle was found to exhibit a pronounced cytotoxic effect against suspension tumor cell lines (Jurkat, HL60, K562, and U937).

Microwave-assisted synthesis of Co(II), Ni(II), Ti(III), and Ti (IV) complexes of iminochromeno-pyrido-pyrimidine and in vitro antiproliferative effect against H460, MCF-7, and HCT116

The purpose of this paper is the synthesis of new metal complexes and their biological application. Four new transition metal complexes (2–5) were synthesized using conventional as well as microwave methods from 7-iminochromeno[4,3-d]pyrido[1,2-a]pyrimidin-6(7H)-one (1) and corresponding metal salts. Ligand 1 and metal complexes 2–5 were characterized by IR, 1H-NMR, 13 C-NMR, Mass spectroscopy and elemental analysis. The free ligand and the metal complexes have been tested for in vitro antiproliferative activity. The evaluation of the antiproliferative effects of compounds (1–5) on three tumor cell lines (H460, MCF-7, and HCT116) shows that all tested compounds have demonstrated moderate to pronounced antiproliferative and/or cytotoxic effect. The most active compounds were ligand 1 and metal complexes 3 and 5.

Microplastics and nanoplastics: Size, surface and dispersant – What causes the effect?

There is increasing evidence that humans are exposed to microplastic particles through contaminated food. Although suitable analytical methods are still lacking, it is likely that these contaminations also contain a nanoplastics fraction. It is known from nanotoxicology that particles may acquire altered toxicological properties with decreasing particle sizes. Particles can also have different surface modalities and functionalizations. Moreover, nano- and microplastics as materials with probably a relatively low toxicity are often applied at high concentrations in in vitro tests, and therefore the solvating agent, namely the dispersant in which the particles are supplied may have a major impact on the outcome. This might be misinterpreted as particle effect. Therefore, it is crucial to determine what causes the effect – size, surface or dispersant? In this study this question was investigated by applying established in vitro models for the intestinal barrier (differentiated Caco-2 monoculture and mucus- and M-cell co-culture) and hepatocytes (differentiated HepaRG cells), mimicking the oral route of particle uptake. A complex set of nine different polystyrene micro- and nanoparticles was used to elucidate the effect of particle size, surface modification and dispersant. Uptake and transport as well as biochemical endpoints were measured, complemented by particle characterization. The results show that indeed some dispersants can cause a more pronounced cytotoxic effect than the particles themselves. Surface modification and particle size show a clear influence on the uptake and cytotoxicity of nano- and microplastic particles.

The Effect of Fatty Acids on Ciprofloxacin Cytotoxic Activity in Prostate Cancer Cell Lines-Does Lipid Component Enhance Anticancer Ciprofloxacin Potential?

Simple SummaryMost prostate cancers are initially hormone-dependent but later gain a hormone-independent phenotype associated with changes in lipid metabolism, including enhanced absorption of extracellular fatty acids. The aim of our study was to assess the effect of ciprofloxacin conjugates with fatty acids on different type of prostate cancer (LNCaP and DU-145) and normal (RWPE-1) cells, as well as their influence on cell lipid metabolism by proteomic analysis. All tested conjugates exhibited cytotoxic potential, the most powerful for oleic, elaidic and docosahexaenoic acids. The hormone-independent DU145 line was more sensitive to derivatives than the hormone-dependent LNCaP line. These results are consistent with previously observed pronounced cytotoxic effect of conjugates on a hormone-insensitive PC3 line. Tested derivatives decreased intensity of proteins involved in prostate cancer lipid metabolism. Our findings confirm the involvement of lipid metabolism in prostate carcinogenesis indicating a target for fatty acids as drug carriers.Purpose: To assess cytotoxic effect of ciprofloxacin conjugates with fatty acids on prostate cancer cells (LNCaP and DU-145) with different hormone sensitivity, based on previous promising results from the PC3 cells. Methods: Cytotoxicity were estimated using MTT and LDH tests, whereas its mechanisms were estimated by apoptosis and IL-6 assays. The intensity of proteins involved in lipid metabolism was determined using ML-CS assay. Results: The hormone insensitive DU-145 cells were more vulnerable than the hormone sensitive LNCaP cells. The IC50 values for oleic (4), elaidic (5) and docosahexaenoic acid (8) conjugates were 20.2 µM, 17.8 µM and 16.5 µM, respectively, in DU-145 cells, whereas in LNCaP cells IC50 exceeded 20 µM. The strong conjugate cytotoxicity was confirmed in the LDH test, the highest (70.8%) for compound (5) and 64.2% for compound (8) in DU-145 cells. This effect was weaker for LNCaP cells (around 60%). The cytotoxic effect of unconjugated ciprofloxacin and fatty acids was weaker. The early apoptosis was predominant in LNCaP while in DU-145 cells both early and late apoptosis was induced. The tested conjugates decreased IL-6 release in both cancer cell lines by almost 50%. Proteomic analysis indicated influence of the ciprofloxacin conjugates on lipid metabolic proteins in prostatic cancer. Conclusion: Our findings suggested the cytotoxic potential of ciprofloxacin conjugates with reduction in proteins involved in prostate cancer progress.

Experimental Study of the In Vitro and In Vivo Functional Activity of NKG2D Chimeric Antigen Receptor

Aim. To study antitumor cytotoxic effect of CAR-T NKG2D and CAR-T anti-CD19 in vitro and in vivo in order to compare antitumor activity of chimeric antigen receptors (CAR) with different structural and functional properties. Materials & Methods. CAR constructions were produced by molecular cloning. CAR-T cell populations were obtained by transduction of healthy donor T-lymphocytes with recombinant lentiviral particles coding CAR NKG2D or CD19 target antigen CAR sequences. CAR-T cell proportion was assessed by FusionRed fluorescence and EGFR membrane receptor imaging. Specific in vitro cytotoxic activity of CAR-T effector cells was analyzed by Real-Time Cytotoxicity Assay (RTCA) during co-cultivation with HeLa_CD19 target cell line using xCELLigence. Interferon-Y (IFN-y) synthesis in vitro and in vivo along with the degree of cytotoxic effect were analyzed by immunoassay of culture medium of co-cultivated effector cells and target cells as well as isolated auto-plasma from the peripheral blood of mice. To assess the in vivo functional activity, CAR-T cell populations were infused into immunodeficient NSG-SGM3 mice (10 000 000 cells/mouse) 12 days after HeLa_CD19 cell injection and confirmation of engraftment and tumor growth. Upon euthanasia, tumors were removed and fixed in paraffin to prepare histological sections. CAR-T cell tumor infiltration was assessed by CD3 antigen immunohistochemical staining. Results. The highest ligand (molecules MICA, ULBP1/2/3/4/5/6) expression levels were detected in HeLa cell line. The obtained NKG2D CAR-T cells showed a considerable cytotoxic activity against HeLa_CD19 target line (cell index [CI] = 1.27), which was, however, twice as low as that of CAR-T anti-CD19 (CI = 0.60) (p = 0.0038). IFN-y level during co-cultivation of CAR-T anti-CD19 with HeLa_CD19 at the ratio of Е/Т = 1:1 was 64,852 pcg/mL, which was 3.5 times higher than IFN-y level during co-cultivation of CAR-T NKG2D with HeLa_CD19 (18,635 pcg/mL) (p = 0.0360). The degree of tumor infiltration by CAR-T anti-CD19 cells was higher than that by CAR-T NKG2D. The absence of NKG2D proliferating CAR-T cells in mice peripheral blood confirms their low persistence. IFN-y concentration in mice auto-plasma was 11.89 pcg/mL after CAR-T anti-CD19 infusion and 0.57 pcg/mL after CAR-T NKG2D infusion (p = 0.0079). The mean weight of tumor xenografts in experimental groups 10 days after CAR-T anti-CD19 injection was 0.72 g (p = 0.0142), after Т-lymphocyte and NKG2D CAR-T cell infusions it was 2.12 g and 1.2 g, respectively. Conclusion. CAR-T anti-CD19 cells are characterized by more pronounced cytotoxic effect under both in vitro and in vivo experimental conditions compared with CAR-T NKG2D cells. The degree of CAR-T anti-CD19 proliferation and their infiltration in mice xenograft models is considerably higher than the levels reached with NKG2D CAR-T cell injections. A single CAR-T NKG2D injection results only in short-term tumor reduction.

POROUS MATERIALS BASED ON BASALT FIBER AS CARRIERS FOR SUBSTRATE-DEPENDENT CELL CULTURE

The influence of samples of 4 innovative materials developed at the Belarusian National Technical University on the basis of basalt fiber with additives of porcelain, oxides of copper, iron, and zirconium on the transplanted culture of cattle kidney cells was evaluated in this work.
 During the experiments, it was determined that the sample containing basalt, 4 % of porcelain and 4 % of FeO does not have a negative effect on the culture of MDBK cells. The material of this sample is promising for further use as a carrier for substrate-dependent cell cultures.
 Sample materials showing a pronounced cytotoxic effect (basalt + 4 % porcelain + 4 % CuO, basalt + 4 % porcelain + 8 % CuO, basalt + 4 % ZrO2 + 4 % CuO) can be considered for the manufacture of bactericidal filters. This action is explained, obviously, by the manifested biocidal properties of copper, which is part of their composition.

Effect of a novel thiazole derivative and its complex with polymeric carriers on the activity of antioxidant enzymes in murine lymphoma cells

Background. Previous studies have shown a pronounced cytotoxic effect of thiazole derivatives in combination with polymeric carriers on tumor cells. At the same time, the derivatives were not cytotoxic against non-cancerous cells in vitro. It was shown that thiazole derivatives at concentrations of 10 and 50 μM affected the prooxidant and antioxidant systems of lymphoma cells in vitro. The aim of this work was to study the effect of the complex of thiazole derivative N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) in combination with polymeric carriers poly(VEP-co-GMA)-graft-mPEG (Th1), poly(PEGMA) (Th3) and poly(PEGMA-co-DMM) (Th5) on the antioxidant defense system of the NK/Ly cell in vitro. Materials and Methods. The experiments were performed on white wild-type male mice with grafted NK/Ly lymphoma. Tumor cells were inoculated into mice intraperitoneally. Ascites was drained from the abdominal cavity of anaesthetized mice with a sterile syringe on the 7th-10th day after inoculation. Investigated compounds BF1, BF1 + Th1 (Th2, Th12), BF1 + Th3 (Th4, Th14), BF1 + Th5 (Th6, Th16) at a final concentration of 10 μM were added to the lymphoma samples and incubated for 10 min; the activity of antioxidant enzymes was determined according to the techniques described previously. Results. It was found that all the studied complexes based on thiazole derivative BF1 and polymeric carriers poly (VEP-co-GMA)-graft-mPEG (Th2, Th12), poly (PEGMA) (Th4, Th14) and poly (PEGMA-co-DMM) (Th6, Th16) at a concentration of 10 μm increased the activity of SOD, while the activity of CAT and GPX were reduced compared to control. Complexes Th2, Th12 and Th4 increased the significance of the BF1 influence on lymphoma cells from P <0.05 to P <0.01. Pure polymeric carriers did not affect the level of the antioxidant defense system enzymes. Conclusions. Thus, it was found that the polymeric carriers in combination with thiazole derivative BF1 increased the significance of thiazole derivative BF1 influence on the activity of the antioxidant defense system of lymphoma cells, while pure polymeric carriers did not affect the activity of SOD, CAT or GPX. The results of this work can be used for further studies of complexes of thiazole derivative and PEG-containing polymeric carriers as potential antitumor drugs.

Phenolic Compound Composition and Biological Activities of Fractionated Soybean Pod Extract

The objective of this study was to determine the potential of dry extracts from soybean pods as a source of bioactive compounds. The phenolic compound composition of the extract (E) and fraction (F1-F3) as well as their respective lipophilicity were analyzed by UPLC-ESI-QTOF-MS. The biological activity as measured by antioxidant and cytotoxic activity was also evaluated. The in vitro anticancer potential of the ethanolic extract and fraction of soybean pods was measured through MTT assay using human colorectal carcinoma (HTC-116) and prostate cancer (PC-3) cell lines. The F2 fraction, with medium lipophilicity, produced the most pronounced cytotoxic effect in PC-3 cells. Analysis of Pearson’s correlation demonstrated a high relationship between the content of phenolic compounds, the antioxidant activity of the extract and fractions, and cytotoxic activity towards PC-3 cells. UPLC-ESI-QTOF-MS analysis of soybean pod extract resulted in putative identification of 50 polyphenols belonging to three different chemical families (phenolic acids, flavonoids, and other polyphenols), the first time such an analysis has been performed. Among the fractions with variable lipophilicity, fraction F2 was the most abundant in phenolic compounds. This was confirmed in the quantitative analysis, as a result of which the highest total content of phenolic compounds was found in the extract (E) and fraction F2. Soybean pods might be useful material for obtaining preparations with anti-radical and anti-cancer properties. They can also be an active food additive or a component in dietary supplements.

Toxic Effect of Streptozotocin on Cultured Mouse Hippocampal Neurons.

In primary dissociated hippocampal cell cultures from 18-day-old mouse embryos, streptozotocin in concentrations of 2-5 mM produced a dose-dependent cytotoxic effect on day 3 in vitro, whereas on day 11 of culturing, the neurons were resistant to streptozotocin. The neurons in the 3-day cultures were functionally immature, which was seen from their weak spontaneous bioelectric activity in the form of rare single action potentials; by day 11 of culturing, the neurons reached a high level of differentiation and their functional properties acquired a character of network burst activity. Thus, streptozotocin had the most pronounced cytotoxic effect on immature hippocampal neurons in vitro.

Evaluation of the effect of enzymatic hydrolyzate of the bursa of broiler chickens on cytotoxic activity and nonspecific immunity

The use of biologically active substances of tissue origin for the creation of biologically active additives and immunomodulatory drugs is an important area of scientific research in the field of nutrition and pharmacology. The aim of the work is to evaluate the effect of enzymatic hydrolysate from the fabricium bag of broiler chickens on the cytotoxicity and nonspecific immunity of mice against the background of experimental salmonella infection using biotechnological methods. For the experiment, three equal experimental groups of white mongrel mice were formed. All laboratory animals were fed daily for seven days with enzymatic hydrolysate at a dose of 750 mg/kg (therapeutic dose), 150 mg/kg (0.2 therapeutic dose) and 3,750 mg/kg (five therapeutic doses). At the same time, the control group of animals received intragastric water in the same volume. 24 hours after the last intake of the enzymatic hydrolysate, the animals were intraperitoneally infected with a culture of Salmonella enteritidis 92. The study reveals the absence of cytotoxic properties and impaired cell viability in cultures L929, J774.1A, HeLa S3, K562, and HST116 against the background of exposure to various concentrations of enzymatic hydrolysate from 0.02 to 10 mg/ml. It should be noted that there is a pronounced cytotoxic effect on MCF-7 tumor cells of the concentration of enzymatic hydrolysate in culture of 5 and 10 mg/ml, which suggests the possibility of using enzymatic hydrolysate for the prevention of tumor diseases. As a result of the experiment, it has been proved that the administration of enzymatic hydrolysate to mice increased the LD50 index and the survival rate of mice infected with the culture of the causative agent of salmonellosis by 1.5 times. Based on the conducted studies, it has been shown that the enzymatic hydrolysate of the lymphoid tissue of broiler chickens does not violate the viability of mammalian cells and does not exhibit cytotoxic properties on the metabolism of healthy mammalian cells, which indicates its safety. In an experiment on mice, the effect of enzymatic hydrolysate of the fabricium bag of broiler chickens on the activation of nonspecific immunity against salmonella infection has been revealed.

FORMULATION AND IN VITRO EVALUATION OF POLY-(D, L-LACTIDE-CO-GLYCOLIDE) (PLGA) NANOPARTICLES OF ELLAGIC ACID AND ITS EFFECT ON HUMAN BREAST CANCER, MCF-7 CELL LINE.

Objective: The aim of this experiment was to develop ellagic acid (EA) loaded poly (D,L-lactide-co-glycolide) (PLGA) nanoparticles for tumour-specific drug delivery. The phytochemical EA is a potential antioxidant, anticarcinogenic and antimutagenic. Due to its low solubility and permeability, it falls under class IV of the BCS classification.
 Methods: PLGA nanoparticles were prepared by a method established on the concept of single emulsification–solvent evaporation by using TWEEN®20 as a cosolvent for solubilizing the drug. While developing this method, polyvinyl alcohol (PVA), was implemented. 
 Results: The stabilized formulation was with a particle size of 174.2 nm, which is ideal for tumour accumulation. The SEM images confirmed that the NPs have spherical shape. The resulting NPs were predominantly spherical and of uniform size and shape. Initial release of EA from nanoparticles in pH 7.4 phosphate buffer was quick, followed by a steady sustained release. The in vitro cytotoxicity study using MTT was also performed on the human breast cancer, MCF-7 cell line and EA-NPs were found to successively reduce the IC50 values which thereby revealed the pronounced cytotoxic effect of the formulation. 
 Conclusion: After performing this experiment, we can conclude that the polymeric nanoparticles are efficient and suitable form of drug delivery of ellagic acid exhibiting potential anti-tumour activity.

PDT with genetically encoded photosensitizer miniSOG on a tumor spheroid model: A comparative study of continuous-wave and pulsed irradiation

BackgroundTherapeutic effects of PDT depend on many factors, including the amount of singlet oxygen, localization of photosensitizer and irradiation protocol. The present study was aimed to compare the cytotoxic mechanisms of PDT under continuous-wave (CW) and pulsed irradiation using a tumor spheroid model and a genetically encoded photosensitizer miniSOG. Methods1O2 detection in miniSOG and flavin mononucleotide (FMN) solutions was performed. Photobleaching of miniSOG in solution and in HeLa tumor spheroids was analyzed. Tumor spheroid morphology and growth and the cell death mechanisms after PDT in CW and pulsed modes were assessed. ResultsWe found a more rapid 1O2 generation and a higher photobleaching rate in miniSOG solution upon irradiation in pulsed mode compared to CW mode. Photobleaching of miniSOG in tumor spheroids was also higher after irradiation in the pulsed mode. PDT of spheroids in CW mode resulted in a moderate expansion of the necrotic core of tumor spheroids and a slight inhibition of spheroid growth. The pulsed mode was more effective in induction of cell death, including apoptosis, and suppression of spheroid growth. ConclusionsComparison of CW and pulsed irradiation modes in PDT with miniSOG showed more pronounced cytotoxic effects of the pulsed mode. Our results suggest that the pulsed irradiation regimen enables enhanced 1O2 production by photosensitizer and stimulates apoptosis. General significanceOur results provide more insights into the cellular mechanisms of anti-cancer PDT and open the way to improvement of light irradiation protocols.