The effect of Lonicera caerulea L. (blue honeysuckle) phenolic fraction (18.5% anthocyanins) on cell viability and against oxidative damage in low density lipoproteins (oxLDL), in rat microsomes and in primary cultures of rat hepatocytes and human umbilical vein endothelial cells (HUVEC), was tested. The phenolic fraction was nontoxic to rat hepatocytes and HUVEC at tested concentrations (1-1000 microg/mL) and time intervals up to 24 h inclusive. Phenolic fraction inhibited rat liver microsome peroxidation, induced by tert-butyl hydroperoxide (tBH), with IC(50) values of 160 +/- 20 microg/mL. The fraction at 0.5, 1.0, and 2.0 microg/mL delayed LDL oxidation, induced by Cu(2+), by 130 +/- 20%, 200 +/- 30%, and 400 +/- 10%, respectively. The treatment of HUVEC with oxidatively modified LDL induced an increase in lactate dehydrogenase (LDH) leakage and thiobarbituric acid reactive substances (TBARS) formation, and resulted in lower formazan formation from 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) uptake, most pronounced for 200 microg/mL (24 h oxidation) after 2 h of incubation. The protective effect of the phenolic fraction against cell damage caused by oxLDL was noted at 0.1 microg/mL for HUVEC and against tBH at 1000 microg/mL for both HUVEC and hepatocytes. The observed protective effects were probably due to the antioxidant properties of L. caerulea constituents, mainly anthocyanins. Microsome peroxidation and LDL oxidation inhibition results provide promising perspectives into the prevention of some oxidative stress-associated diseases. Other data are important in in vitro systems but seem to be accidental in vivo.

• MeSH
• antioxidancia farmakologie   MeSH
• Caprifoliaceae chemie   MeSH
• cévní endotel účinky léků   metabolismus   MeSH
• dospělí MeSH
• hepatocyty účinky léků   metabolismus   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• oxidační stres účinky léků   MeSH
• potkani Wistar MeSH
• rostlinné extrakty farmakologie   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antioxidancia MeSH
• rostlinné extrakty MeSH

Lithium accumulates in the thyroid gland and can cause goiter or thyroid dysfunction. The aims of our work were: 1) to verify whether lithium stimulates proliferation of thyroid cells; as methods, the 3H-thymidine incorporation assay and the MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) were used; as a model system the FRTL-5 (Fischer rat thyroid cells in low serum) cell line was selected, 2) to test whether lithium can have a cytotoxic effect on FRTL-5 cells, using the cytotoxicity assay with 51Cr release and the trypan blue exclusion method. Without TSH stimulation, lithium at 0.35-2 mM concentrations significantly increased the 3H-thymidine incorporation. A similar effect was observed in the case of the MTT assay: without TSH stimulation, lithium at 0.4-2 mM concentrations showed a significant stimulation of proliferation. Surprisingly, under TSH stimulation, lithium at the 2 mM concentration significantly inhibited proliferation of FRTL-5 cells. With the cytotoxicity assay, lithium was found to increase 51Cr release at 1.4-2 mM concentrations. Additionaly, the percentage of viable FRTL-5 cells at 0.35-2 mM concentrations of lithium was lower than in the controls without lithium. In conclusion, lithium was found to stimulate proliferation of FRTL-5 cells in conditions without TSH and, surprisingly, lithium in higher concentrations diminished proliferation of FRTL-5 cells under TSH stimulation. A cytotoxic effect of higher lithium concentrations was observed.

• MeSH
• buněčné dělení účinky léků   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lithium farmakologie   MeSH
• radioizotopy chromu MeSH
• štítná žláza cytologie   účinky léků   metabolismus   MeSH
• tetrazoliové soli metabolismus   MeSH
• thiazoly metabolismus   MeSH
• thymidin metabolismus   MeSH
• thyreotropin farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• lithium MeSH
• radioizotopy chromu MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč
• thymidin MeSH
• thyreotropin MeSH

The production of cytotoxic molecules interfering with mammalian cells is extensively reported in cyanobacteria. These compounds may have a use in pharmacological applications; however, their potential toxicity needs to be considered. We performed cytotoxicity tests of crude cyanobacterial extracts in six cell models in order to address the frequency of cyanobacterial cytotoxicity to human cells and the level of specificity to a particular cell line. A set of more than 100 cyanobacterial crude extracts isolated from soil habitats (mainly genera Nostoc and Tolypothrix) was tested by MTT test for in vitro toxicity on the hepatic and non-hepatic human cell lines HepG2 and HeLa, and three cell systems of rodent origin: Yac-1, Sp-2 and Balb/c 3T3 fibroblasts. Furthermore, a subset of the extracts was assessed for cytotoxicity against primary cultures of human hepatocytes as a model for evaluating potential hepatotoxicity. Roughly one third of cyanobacterial extracts caused cytotoxic effects (i.e. viability<75%) on human cell lines. Despite the sensitivity differences, high correlation coefficients among the inhibition values were obtained for particular cell systems. This suggests a prevailing general cytotoxic effect of extracts and their constituents. The non-transformed immortalized fibroblasts (Balb/c 3T3) and hepatic cancer line HepG2 exhibited good correlations with primary cultures of human hepatocytes. The presence of cytotoxic fractions in strongly cytotoxic extracts was confirmed by an activity-guided HPLC fractionation, and it was demonstrated that cyanobacterial cytotoxicity is caused by a mixture of components with similar hydrophobic/hydrophilic properties. The data presented here could be used in further research into in vitro testing based on human models for the toxicological monitoring of complex cyanobacterial samples.

• Klíčová slova
• Cell lines, Cyanobacteria, Cytotoxicity, Hepatocytes, MTT test, Primary cultures,
• MeSH
• buněčné linie MeSH
• buňky BALB 3T3 MeSH
• buňky Hep G2 MeSH
• cytotoxiny analýza   MeSH
• fibroblasty MeSH
• HeLa buňky MeSH
• inhibiční koncentrace 50 MeSH
• komplexní směsi toxicita   MeSH
• lidé MeSH
• myši MeSH
• sinice chemie   MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytotoxiny MeSH
• komplexní směsi MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč

Cryopreservation offers the potential to maximize the use and availability of biological materials that have a limited supply. This study demonstrates an enhanced technique for the parallel cryopreservation of a series of liver tissue slices using a tray modeled from aluminium foil and low concentrations of a cryoprotectant. Cooling and warming rates of approximately 2000 and 3900 degrees C min(-1), respectively, were achieved as the thermal capacity of the foil-tray was significantly reduced compared to the aluminium sandwich device introduced by Day et al. [S.H. Day, D.A. Nicoll-Griffith, J.M. Silva, Cryopreservation of rat and human liver slices by rapid freezing, Cryobiology 38 (1999) 154-159]. Additionally, the two critical steps involved in the sandwich approach, i.e., clamping the plates and complete filling of the entire space between the plates with liquid, can be omitted using the foil tray. The viability of the slices was verified by measuring tetrazolium salt reduction capacity, cytosolic enzyme lactate dehydrogenase leakage, and ethoxycoumarin metabolism.

• MeSH
• časové faktory MeSH
• játra * MeSH
• kryoprezervace metody   MeSH
• L-laktátdehydrogenasa metabolismus   MeSH
• myši MeSH
• tetrazoliové soli metabolismus   MeSH
• thiazoly metabolismus   MeSH
• uchovávání orgánů * MeSH
• umbeliferony metabolismus   MeSH
• viabilita buněk MeSH
• zmrazování * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• L-laktátdehydrogenasa MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč
• umbeliferony MeSH

With aim to develop effective proof-of-concept approach which can be used in a development of new preparations for the inhalation therapy, we designed a new screening method for simple and rapid simultaneous determination of antibacterial potential of plant volatiles in the liquid and the vapour phase at different concentrations. In addition, EVA (ethylene vinyl acetate) capmat™ as vapour barrier cover was used as reliable modification of thiazolyl blue tetrazolium bromide (MTT) assay for cytotoxicity testing of volatiles on microtiter plates. Antibacterial activity of carvacrol, cinnamaldehyde, eugenol, 8-hydroxyquinoline, thymol and thymoquinone was determined against Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae using new broth microdilution volatilization method. The cytotoxicity of these compounds was evaluated using MTT test in lung fibroblast cells MRC-5. The most effective antibacterial agents were 8-hydroxyquinoline and thymoquinone with the lowest minimum inhibitory concentrations (MICs) ranging from 2 to 128μg/mL, but they also possessed the highest toxicity in lung cell lines with half maximal inhibitory concentration (IC50) values 0.86-2.95μg/mL. The lowest cytotoxicity effect was identified for eugenol with IC50 295.71μg/mL, however this compound produced only weak antibacterial potency with MICs 512-1024μg/mL. The results demonstrate validity of our novel broth microdilution volatilization method, which allows cost and labour effective high-throughput antimicrobial screening of volatile agents without need of special apparatus. In our opinion, this assay can also potentially be used for development of various medicinal, agricultural, and food applications that are based on volatile antimicrobials.

• Klíčová slova
• Antibacterial activity, Colorimetric assay, Cytotoxicity, Essential oil, Vapour, Volatilization method,
• MeSH
• akrolein analogy a deriváty   chemie   MeSH
• antibakteriální látky chemie   MeSH
• benzochinony chemie   MeSH
• buněčné linie MeSH
• cymeny MeSH
• eugenol chemie   MeSH
• fytonutrienty chemie   MeSH
• Haemophilus influenzae účinky léků   MeSH
• lidé MeSH
• mikrobiální testy citlivosti metody   MeSH
• monoterpeny chemie   MeSH
• oxychinolin chemie   MeSH
• Staphylococcus aureus účinky léků   MeSH
• Streptococcus pneumoniae účinky léků   MeSH
• těkavé organické sloučeniny chemie   MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thymol chemie   MeSH
• volatilizace * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrolein MeSH
• antibakteriální látky MeSH
• benzochinony MeSH
• carvacrol MeSH Prohlížeč
• cinnamaldehyde MeSH Prohlížeč
• cymeny MeSH
• eugenol MeSH
• fytonutrienty MeSH
• monoterpeny MeSH
• oxychinolin MeSH
• těkavé organické sloučeniny MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč
• thymol MeSH
• thymoquinone MeSH Prohlížeč

The quantification of cellular metabolic activity via MTT assay has become a widespread practice in eukaryotic cell studies and is progressively extending to bacterial cell investigations. This study pioneers the application of MTT assay to evaluate the metabolic activity of biofilm-forming cells within bacterial biofilms on nanofibrous materials. The biofilm formation of Staphylococcus aureus and Escherichia coli on nanomaterials electrospun from polycaprolactone (PCL), polylactic acid (PLA), and polyamide (PA) was examined. Various parameters of the MTT assay were systematically investigated, including (i) the dissolution time of the formed formazan, (ii) the addition of glucose, and (iii) the optimal wavelength for spectrophotometric determination. Based on interim findings, a refined protocol suitable for application to nanofibrous materials was devised. We recommend 2 h of the dissolution, the application of glucose, and spectrophotometric measurement at 595 nm to obtain reliable data. Comparative analysis with the reference CFU counting protocol revealed similar trends for both tested bacteria and all tested nanomaterials. The proposed MTT protocol emerges as a suitable method for assessing the metabolic activity of bacterial biofilms on PCL, PLA, and PA nanofibrous materials.

• Klíčová slova
• Biofilm, MTT, Metabolic activity, Nanofibers, Nanomaterials,
• MeSH
• biofilmy * růst a vývoj   MeSH
• Escherichia coli * fyziologie   MeSH
• glukosa metabolismus   MeSH
• nanovlákna * chemie   MeSH
• nylony chemie   MeSH
• polyestery * chemie   MeSH
• spektrofotometrie metody   MeSH
• Staphylococcus aureus * fyziologie   MeSH
• tetrazoliové soli * metabolismus   chemie   MeSH
• thiazoly metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glukosa MeSH
• nylony MeSH
• poly(lactide) MeSH Prohlížeč
• polycaprolactone MeSH Prohlížeč
• polyestery * MeSH
• tetrazoliové soli * MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč

BACKGROUND/AIMS: Podocytes are typically cultured on collagen I; however, collagen I is absent from healthy glomerular basement membranes. Erythropoietin (EPO) is thought to protect podocytes in vivo. Here, we studied how various types of extracellular matrix (ECM) proteins and EPO affect podocytes in culture. METHODS: Primary rat podocytes were replated on collagen I, collagen IV, whole ECM extract, laminin, or bare plastic. Cellular adhesion (8 hours after plating), proliferation (5 days, 10 % serum), and resistance to serum deprivation (3 days, 0.5 % serum) were assessed. BrdU incorporation and expression of podocyte-specific markers were employed as measures of cellular proliferation and differentiation, respectively. qPCR was used to verify expression of EPO receptor in cultured podocytes. RESULTS: Cellular adhesion was similar on all ECM proteins and unaffected by EPO. Proliferation was accelerated by laminin and the ECM extract, but the final cell density was similar on all ECM surfaces. Collagen IV supported the serum-deprived cells better than the other ECM proteins. EPO (2-20 ng/ml) improved viability of serum-deprived podocytes on collagen I, collagen IV, and ECM, but not on laminin or bare plastic. The cells expressed mRNA for EPO receptor. CONCLUSION: The physiological ECM proteins are more supportive of primary podocytic cultures compared with collagen I. The protective effects of EPO during serum deprivation are modulated by the cultivation surface.

• MeSH
• barvicí látky MeSH
• erythropoetin farmakologie   MeSH
• extracelulární matrix - proteiny fyziologie   MeSH
• extracelulární matrix účinky léků   metabolismus   MeSH
• glomerulus účinky léků   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• podocyty účinky léků   MeSH
• primární buněčná kultura MeSH
• receptory erythropoetinu biosyntéza   účinky léků   MeSH
• rekombinantní proteiny farmakologie   MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• barvicí látky MeSH
• erythropoetin MeSH
• extracelulární matrix - proteiny MeSH
• receptory erythropoetinu MeSH
• rekombinantní proteiny MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč

The aim of this study was to evaluate the dynamics of the cytotoxicity of gingival margin retraction astringents based on aluminium chloride, aluminium sulphate, and ferric sulphate (solutions and gels) in human fibroblasts isolated from gingiva. The cytocompatibility of ten astringent-based chemical retraction agents: Gingiva Liquid, Alustin, Racestypine, Orbat sensitive, Astringedent®, Alustat, Hemostat, Racécord, Gel cord and ViscoStat®, in dilutions of 1 : 10 and 1 : 20, with human gingival fibroblasts was investigated. The MTT assay was performed to determine oxidoreductive mitochondrial function after 3, 5, 10 min and 24 h of incubation. Cell viability was determined according to the chemical group, concentration, exposure time, and the clinical form of the gingival retraction agents. Ferric sulphate- based agents were the most cytotoxic, followed by aluminium chloride and aluminium sulphate. The form of the astrigents influenced cell viability. The evaluated astringents may have cytotoxic potential for gingival margin tissues under clinical conditions.

• MeSH
• adstringencia toxicita   MeSH
• barvicí látky MeSH
• chlorid hlinitý MeSH
• chloridy toxicita   MeSH
• fibroblasty účinky léků   MeSH
• gingiva cytologie   účinky léků   MeSH
• kamencové sloučeniny toxicita   MeSH
• kultivované buňky MeSH
• lidé MeSH
• oxidace-redukce účinky léků   MeSH
• sloučeniny hliníku toxicita   MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• viabilita buněk účinky léků   MeSH
• železité sloučeniny toxicita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adstringencia MeSH
• aluminum sulfate MeSH Prohlížeč
• barvicí látky MeSH
• chlorid hlinitý MeSH
• chloridy MeSH
• ferric sulfate MeSH Prohlížeč
• kamencové sloučeniny MeSH
• sloučeniny hliníku MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč
• železité sloučeniny MeSH

The activity in vitro of four types of colicins (A, E1, E3, U) against one human standard fibroblast line and against 11 human tumor-cell lines carrying defined mutations of the p53 gene was quantified by MTT (tetrazolium bromide) assay. Flow cytometry showed that the pore-forming colicins A, E1 and U affected the cell cycle of 5 of these cell lines. Colicins E3 and U did not show any distinct inhibitory effects on the cell lines, while colicins E1 and especially A inhibited the growth of all of them (with one exception concerning colicin E1). Colicin E1 inhibited the growth of the tumor lines by 17-40% and standard fibroblasts MRC5 by 11%. Colicin A exhibited a differentiated 16-56% inhibition, the growth of standard fibroblasts being inhibited by 36%. In three of the lines, colicins A and E1 increased the number of cells in the G1 phase (by 12-58%) and in apoptosis (by 7-58%). These results correlated with the data from sensitivity assays. Hence, the inhibitory effect of colicins on eukaryotic cells in cell-selective, colicin-specific and can be considered to be cytotoxic.

• MeSH
• buněčný cyklus účinky léků   MeSH
• eukaryotické buňky účinky léků   MeSH
• fibroblasty účinky léků   MeSH
• G1 fáze účinky léků   MeSH
• koliciny farmakologie   MeSH
• lidé MeSH
• mutace MeSH
• nádorové buňky kultivované účinky léků   MeSH
• nádorový supresorový protein p53 genetika   MeSH
• tetrazoliové soli metabolismus   MeSH
• thiazoly metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• koliciny MeSH
• nádorový supresorový protein p53 MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč

BACKGROUND: Therapy with 5-fluorouracil (5-FU) and capecitabine is often complicated by skin toxicity (hand-foot syndrome, HFS). Topical application of uridine ointment is beneficial for alleviating HFS and other pyrimidine nucleosides have been described as 5-FU toxicity modulators. We tested pyrimidine nucleosides and their combinations to find the best combination for topical therapy of HFS. MATERIALS AND METHODS: Cellular viability was measured by the real-time cell analyser and methyl thiazol tetrazolium (MTT) assay in order to evaluate the effect of pyrimidine nucleosides on HaCaT keratinocytes treated with 5-FU. The results were confirmed by evaluation of the cellular colonization by microphotography. RESULTS: Cytidine and uridine protected keratinocytes to the same extent. Thymidine enhanced the protective effect when added to cytidine or uridine. Deoxycytidine did not have any protective effect. CONCLUSION: Our findings support the rationale for using uridine or cytidine in combination with thymidine in ointment for HFS treatment.

• Klíčová slova
• 5-FU, Hand-foot syndrome, cytidine, deoxycytidine, keratinocytes, thymidine, uridine,
• MeSH
• antimetabolity antitumorózní škodlivé účinky   MeSH
• dihydrouracildehydrogenasa (NADP) fyziologie   MeSH
• fluorouracil škodlivé účinky   MeSH
• keratinocyty účinky léků   MeSH
• kultivované buňky MeSH
• lidé MeSH
• pyrimidinové nukleosidy farmakologie   MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thymidylátsynthasa antagonisté a inhibitory   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antimetabolity antitumorózní MeSH
• dihydrouracildehydrogenasa (NADP) MeSH
• fluorouracil MeSH
• pyrimidinové nukleosidy MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• thiazolyl blue MeSH Prohlížeč
• thymidylátsynthasa MeSH