MTT assay

We examined the cytotoxic potential of nine N-[2-substituted-2-(2-thienyl) ethyl] piperazinyl quinolone derivatives on human oral epithelial mouth carcinoma (KB) and human squamous carcinoma (A431) cell lines. Phototoxic properties of these compounds were ...

Three haloderivatives of noscapine 2-4 were synthesized chemoselectively and their in vitro cytotoxicity was assessed by MTT assay on U-87 human glioblastoma cell lines. At 50 lM concentration after 72 h, 9-chloronoscapine 2, 9-bromonoscapine 3 (EM011), and 9-iodonoscapine 4 killed 87.8%, 51.2%, and 56.8% cells, respectively, however noscapine kills only 40% of the cells; revealing 9-chloronoscapine as a potential cytotoxic agent than noscapine and 9-bromonoscapine (EM011). At low concentration (1 lM) 9-bromonoscapine (46.7%) and 9-chloronoscapine (45.7%) did not show any significant difference.

The occurrence of the two new cis-fused A/B rings furostanol saponins (25S)-26-O-β-Dglucopyranosyl-5β-furostan-1β,3β,22α,26-tetraol-1-O-β-D-glucopyranoside and (25S)-26-Oβ-D-glucopyranosyl-5β-furostan-1β,2β,3β,5β,22α,26-hexaol and the known compounds

Pseudomonas aeruginosa, a common agent of septicemia, enters into human endothelial cellsin vitro but the effects of bacterial infection have not been addressed properly. In this study, human umbilical vein endothelial cells (HUVEC) were infected by the noninvasive PA103 and ...

Gallic acid-based indanone derivatives have been synthesised. Some of the indanones showed very good anticancer activity in MTT assay. Compounds 10, 11, 12 and 14 possessed potent anticancer activity against various human cancer cell lines. The most potent indanone (10, IC 50 = 2.2 lM), against MCF-7, that is, hormone-dependent breast cancer cell line, showed no toxicity to human erythrocytes even at higher concentrations (100 lg/ml, 258 lM). While, indanones 11, 12 and 14 showed toxicities to erythrocytes at higher concentrations.

The surface of polyethersulfone (PES) membrane was modified by blending triblock copolymers of methoxyl poly(ethylene glycol)-polyurethane-methoxyl poly(ethylene glycol) (mPEG-PU-mPEG), which were synthesized through solution polymerization with mPEG Mns of 500 and 2000, respectively. The PES and PES/mPEG-PU-mPEG blended membranes were prepared through spin coating coupled with liquid-liquid phase separation. FTIR and 1 H NMR analysis confirmed that the triblock copolymers were successfully synthesized. The functional groups and morphologies of the membranes were studied by ATR-FTIR and SEM, respectively. It was found that the triblock copolymers were blended into PES membranes successfully, and the morphologies of the blended membranes were somewhat different from PES membrane. The water contact angles and platelet adhesion were decreased after blending mPEG-PU-mPEG into PES membranes. Meanwhile, the activated partial thromboplastin time (APTT) for the blended membranes increased. The anti-protein-fouling property and permeation property of the blended membranes improved obviously. SEM observation and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5diphenyl tetrazolium bromide (MTT) assay proved the surfaces of the blended membranes promoted human hepatocytes adhesion and proliferation better than PES membrane.

The organotin(IV) compounds used in chemotherapy due to its lipophilicity, affected by the number of carbon atoms and the cytotoxicity. These are affected by the obtainability of Sn coordination bond and bond stabilization between ligand and tin. Two novel organotin(IV) complexes were synthesized, characterized, and tested against human lung cancer cells (A549). The cytotoxic effect of the prepared organotin(IV) complexes against human lung cancer cells (A549) was investigated using the MTT colorimetric assay. Apoptosis was investigated by flow cytometry. The cytotoxicity assay reveals that the Bu 2 SnL 2 complex is more active to inhibit the growth of A549 cells compared to the Ph 2 SnL 2 , and doxorubicin, nevertheless at high concentration (50 and 100) µg/mL the doxorubicin was more affective to inhibit the viability of A549 cells.

Honokiol and magnolol, two major phenolic constituents of Magnolia sp., have been known to exhibit antibacterial activities. However, until now, their antibacterial activity against Propionibacterium sp. has not been reported. To this end, the antibacterial activities of honokiol and magnolol were detected using the disk diffusion method and a two-fold serial dilution assay. Honokiol and magnolol showed strong antibacterial activities against both Propionibacterium acnes and Propionibacterium granulosum, which are acne-causing bacteria. The minimum inhibitory concentrations (MIC) of honokiol and magnolol was 3 -4 Ag/ml (11.3 -15 AM) and 9 Ag/ml (33.8 AM), respectively. In addition, the killing curve analysis showed that magnolol and honokiol killed P. acnes rapidly, with 10 5 organisms/ml eliminated within 10 min of treatment with either 45 Ag (169.2 AM) of magnolol or 20 Ag (75.2 AM) of honokiol per ml. The cytotoxic effect of honokiol and magnolol was determined by a colorimetric (3-(4,5-dimetyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT) assay using two animal cell lines, human normal fibroblasts and HaCaT. In this experiment, magnolol exhibited lower cytotoxic effects than honokiol at the same concentration, but they showed similar cytotoxicity when triclosan was employed as an acne-mitigating agent. In addition, they reduced secretion of interleukin-8 and tumor necrosis factor a (TNF-a) induced by P. acnes in THP-1 cells indicating the anti-inflammatory effects of them. When applied topically, neither phenolic compound induced any adverse reactions in a human skin primary irritation test. Therefore, based on these results, we suggest the possibility that magnolol and honokiol may be considered as attractive acne-mitigating candidates for topical application. D

In the present study, reconstructed human epidermis (RHE) was used as an in vitro model to discriminate 1-chloro-2,4-dinitrobenzene (DNCB), nickel sulfate (NiSO 4 ), oxazolone (OXA), 2,4-dinitrofluorobenzene (DNFB) and 2,4,6-trinitrobenzenesulfonic acid (TNBS) as skin sensitizers from benzalkonium chloride (BC), benzoic acid (BA) and sodium lauryl sulfate (SLS) as skin irritants. Our criteria were (a) the differential IL-1a and IL-8 synthesis and release (b) cytotoxicity assessment by MTT assay. When the RHE are topically treated with the sensitizers, very low levels of extra-and intracellular IL-1a are observed although they induce significant cytotoxicity. In contrast, they exhibit a sharp maximum of IL-8 release.

Ten commercially available denture adhesives, nine soluble formulations (six creams, three powders) and one insoluble product (pad), were analyzed regarding the cytotoxicity profile in direct and indirect assays using L929 fibroblast cells. In the direct assay, fibroblasts were seeded over the surface of a thick adhesive gel (5%, creams; 2.5%, powders and pad). In the indirect assay, cells were cultured in the presence of adhesive extracts prepared in static and dynamic conditions (0.5-2%, creams; 0.25-1%, powders and pad). Cell toxicity was assessed for cell viability/proliferation (MTT assay) and cell morphology (observation of the F-actin cytoskeleton organization by confocal laser scanning microscopy). Direct contact of the L929 fibroblasts with the thick adhesive gels caused no, or only a slight, decrease in cell viability/proliferation. The adhesive extracts (especially those prepared in dynamic conditions) caused significantly higher growth inhibition of fibroblasts and, in addition, caused doseand time-dependent effects, throughout the 6-72 h exposure time. Also, dose-dependent effects on cell morphology, with evident disruption of the F-actin cytoskeleton organization, were seen in the presence of most adhesives. In conclusion, the adhesives possessed different degrees of cytotoxicity, but similar dose-and time-dependent biological profiles.

The semi-automated MTT colorimetric assay has previously been applied on Leishmania promastigotes based on the ability of viable parasites to reduce the tetrazolium salt to an insoluble formazan product. As promastigotes are non-adherent, application of the MTT assay in its original form has a major drawback of a high and variable background absorbance due to incomplete removal of phenol red, a component of most media. We have accordingly optimised a modified MTT assay wherein the absorbance linearity was maintained for cells ranging from 1 Â10 4 to 1 Â10 7 being 0.04 F 0.003-2.38 F 0.04. In contrast, the original MTT assay had a narrower linearity range of 1 Â10 6 -1 Â10 7 cells, absorbances being 0.05 F 0.005-1.54 F 0.005. The modified MTT assay was effectively applied to study growth kinetics and identification of antimonial resistant field isolates. Considering the growing problem of antimonial unresponsiveness in the Indian subcontinent, this modified MTT assay is a useful tool for Leishmania research. D

Artemisinin regarded as one of the most promising anticancer drugs can bind to DNA with a binding constant of 1.04 9 10 4 M -1 . The electrochemical experiments indicated that for longer incubation time periods, the reduction peak current of artemisinin on carbon nanotube modified electrode increases. Therefore, the uptake of drug molecules from a solution into CNTs will be achieved automatically by adsorption of 88.7% of artemisinin onto carbon nanotubes surface without alteration in drug properties. Hence, capability of carbon nanotubes to have synergistic effect on the bioavailability of artemisinin was investigated. Experimental tests on K562 cancer cell lines growth by MTT assay proved that multi-walled carbon nanotubes can enhance the cytotoxity of artemisinin to the targeted cancer cells with unprecedented accuracy and efficiency. The IC 50 values were 65 and 35 lM for artemisinin and artemisinin loaded on multi-walled carbon nanotubes, respectively; demonstrating that artemisinin loaded on multi-walled carbon nanotubes is more effective in inhibition of cancer cell lines growth.

The aim of the work was early identification of preventable risk factors connected with the consumers usage of products of everyday use, such as cosmetics, toys and children products, and other materials intended for contact with human skin. The risk factor is represented by substances with irritation potential and subsequent possible sensitisation, resulting in negative impact on human physical and psychical health with social and societal consequences. The legislation for cosmetics, chemical substances and other products requires for hazard identification the application of alternative toxicological methods in vitro without the use of animals. For this reason we used a battery of alternative assays in vitro, based on cell cultures. Progressive methods of molecular biology, based on fluorimetry and fluorescence, were employed for identification of early morphological and functional changes on cellular level. Four colorants frequently used in cosmetics (P-WS Caramel, Chlorophyllin, Unicert Red K 7054-J and Unicert Red K 7008-J) were tested on cell line NIH3T3 (mouse fibroblast cell) and 3T3 Balb/c with/without UV irradiation (dose 5 J cm À2 ). Fluorescence methods for the study of cell damage using fluorescence probes offer results for the evaluation of cytotoxicity and cell viability of adherent cells. We detected intracellular production of ROS investigated by molecular probe CM-H 2 DCFDA, which is primarily sensitive to the increased production of hydrogen peroxide or its downstream products. Toxic effects on the cellular level were identified by viability tests using Neutral Red uptake and MTT assay, where the live cells reduce yellow soluble 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to insoluble formazan crystals. The reaction was investigated on mitochondrial membrane of living cells and the type of cell death was determined using Apoptosis detection kit. Cytotoxicity tests revealed health risks of using Chlorophyllin and Unicert Red K 7054-J.

Lead is a non-essential element that exhibits a high degree of toxicity, especially in children. Most research on lead has focused on its effects on organ systems such as the nervous system, the red blood cells, and the kidneys which are considered to be the primary targets of lead toxicity. However, the molecular mechanisms by which it induces toxicity, and carcinogenesis remain to be elucidated. In this research, we performed the MTT assay to assess the cytotoxicity, and the CAT-Tox assay to assess the transcriptional responses associated with lead exposure to thirteen different recombinant cell lines generated from human liver carcinoma cells (HepG2), by creating stable transfectants of mammalian promoter chloramphenicol (CAT) gene fusions. Study results indicated that lead nitrate is cytotoxic to HepG2 cells, showing LD50 values of 49.0 ± 18.0 mgr;g/mL, 37.5 ± 9.2 μg/mL, and 3.5 ± 0.7 μg/mL for cell mortality upon 24, 48 and 72 h of exposure, respectively; indicating a dose- and time-dependent response with regard to the cytotoxic effect of lead nitrate. A dose-response relationship was also recorded with respect to the induction of stress genes in HepG2 cells exposed to lead nitrate. Overall, six out of the thirteen recombinant cell lines tested showed inductions to statistically significant levels (p < 0.05). At 50 μg/mL of lead nitrate, the average fold inductions were: 2.1 ± 1.0, 5.4 ± 0.4, 12.1 ± 6.2, 5.0 ± 1.7, 2.5 ± 1.3, and 4.8 ± 4.5 for XRE, HSP70, CRE, GADD153, and GRP78, respectively. These results indicate the potential for lead nitrate to undergo biotransformation in the liver (XRE), to cause cell proliferation (c-fos), protein damage (HSP70, GRP78), metabolic perturbation (CRE), and growth arrest and DNA damage (GADD153). Marginal but not significant inductions were also obtained with the GSTYa (1.5 ± 0.8), and GADD45 (5.7 ± 8.1) promoters, and the NF-κB (2.0 ± 1.7) response element, indicating the potential for oxidative stress. No significant inductions (p > 0.05) were recorded for CYP1A1, HMTIIA, p53RE, and RARE.

Homolytic aroylation of pyrazine nucleus with various substituted aromatic carbaldehydes afforded a series of 5-aroylpyrazine-2carboxylic acid derivatives. The synthetic approach, analytical and spectroscopic data of all compounds synthesized, their preliminary in vitro evaluation of antituberculotic and antifungal activities, cytotoxicity data and subsequent SAR studies are presented. Among all derivatives prepared, only 5-(4-chlorobenzoyl)-pyrazine-2-carbothioamide (3d) showed promising activity (90% inhibition) against Mycobacterium tuberculosis . The highest antifungal effect (MIC B/1.95 mmol ml (1 ) against Trichophyton mentagrophytes , the most susceptible fungal strain tested, was found for 5-benzoylpyrazine-2-carbothioamide (3a). Thioamides exhibited higher in vitro antimicrobial activity than the corresponding amides.

Cell encapsulation represents an alternative nonviral technique to treat multiple diseases, leading to a reduction or even absence of administration of immunosuppressants. Hydrogels have been introduced as novel materials suitable for cell encapsulation. This study involves agarose–gelatin blend hydrogels with four different weight percentage ratios (100:0, 75:25, 50:50, 25:75) of agarose to gelatin. Prepared blend hydrogels were assessed in terms of rheological behavior (gel point by using complex viscosity), cell attachment (hemocytometer), cell viability and cytotoxicity (3-(3,4-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliium bromide, MTT assay), and mechanical and integral stability (Bradford test and shear force rupture assay, respectively). Based on the obtained rheological experimental results, the sol-gel transition point for 50:50 was in the physiological condition range (35°C–37°C). The percent of nonattached cells on the surface of the hydrogel decreased from 92% for the 100:0 sample to 46.3% for the 50:50 sample, and the cell viability was more than 95%. A good structural integrity was achieved for samples with weight ratio of 50:50; 20.195% gelatin was released during the 24 h in phosphate buffer solution at 25°C and the mechanical stability of agarose–gelatin microcapsules under shear force were improved about 14% rather than pure agarose microcapsule.

Bioactive glass ceramic nanoparticles (nBGC) were synthesized by sol-gel process and characterized using FTIR, TEM and XRD. Composite scaffolds of chitosan (CS)-gelatin (CG) with nBGC were prepared by blending of chitosan and gelatin with nBGC. The prepared CG/nBGC nano-composite scaffolds were characterized using FTIR, SEM and XRD. The effect of nBGC in the scaffold matrix was evaluated in terms of scaffold properties and biocompatibility. Our results showed macroporous internal morphology in the scaffold with pore size ranging from 150 to 300 m. Degradation and swelling behavior of the nanocomposite scaffolds were decreased, while protein adsorption was increased with the addition of nBGC. Biomineralization studies showed higher amount of mineral deposits on the nano-composite scaffold, which increases with increasing time of incubation. MTT assay, direct contact test, and cell attachment studies indicated that, the nano-composite scaffolds are better in scaffold properties and it provides a healthier environment for cell attachment and spreading. So, the developed nano-composite scaffolds are a potential candidate for alveolar bone regeneration applications.

This study aims at the formulation of curcumin with biodegradable thermoresponsive chitosan-g-poly (N-vinylcaprolactam) nanoparticles (TRC-NPs) for cancer drug delivery. The spherical curcumin-loaded nanoparticles of size 220 nm were characterized, and the biological properties were studied using flow cytometry and cytotoxicity by MTT assay. The in vitro drug release was higher at above LCST compared to that at below LCST. TRC-NPs in the concentration range of 100-1000 lg/mL were non-toxic to an array of cell lines. The cellular localization of the curcumin-loaded TRC-NPs was confirmed from green fluorescence inside the cells. The time-dependent curcumin uptake by the cells was quantified by UV spectrophotometer. Curcumin-loaded TRC-NPs showed specific toxicity to cancer cells at above their LCST. Flow cytometric analysis showed increased apoptosis on PC3 compared to L929 by curcumin-loaded TRC-NPs. These results indicate that novel curcumin-loaded TRC-NPs could be a promising candidate for cancer drug delivery.

Tattooing is an ancient art and is still widely practiced all over the world. Since the biocompatibility of tattoo dyes has not been well researched, we studied the toxicity of a commercial tattoo ink, commonly used in tattoo lab and esthetic centers, on human Wbroblasts. To test cell viability, MTT assays were carried out and scanning electron microscopy to visualize changes in the cell surface after the dye exposure was performed. A possible inXuence of the pigment on the expression of procollagen 1 type I protein was visualized by western blotting analysis. The results showed a reduction in cell viability, and electron microscopy demonstrated an unmodiWed cell surface completely covered by pigment particles. Western blotting analysis demonstrated a clear interference of the pigment on the expression of procollagen 1 type I protein. These data demonstrated that the commercial tattoo dye has a time-dependent eVect on protein expression. A possible connection of the inXuence of the tattoo ink with clinical eVects is discussed.

Thymoquinone (TQ), the active constituent of Nigella sativa or black cumin exhibited cytotoxic effects in several cancer cell lines. In this study, the cytotoxicity of TQ in human cervical squamous carcinoma cells (SiHa) was investigated. TQ was cytotoxic towards SiHa cells with IC50 values of 10.67±0.12 and 9.33±0.19μg/mL as determined by MTT assay and trypan blue dye exclusion test, respectively,

Due to their properties and common occurrence, fungi constitute the most frequent cause of biodeterioration of building materials and a serious health hazard to occupants. The authors present the methods of mycological analysis used to assess contamination caused by moulds and their metabolites in buildings.

Introduction Several different synthetic and allograft bone graft substitutes are used clinically to treat large bone defects. In contrast to the ''gold standard'' of autologous bone grafts, these do not contain bone-forming (MSC) or vessel-forming (EPC) cells. In order to achieve the same level of success enjoyed by autologous bone grafts, they must be compatible with mesenchymal stem cells (MSC) and endothelial progenitor cells (EPC). In a previous study, we seeded MSC onto six different bone graft substitutes and then measured the cell adhesion, viability, differentiation, and morphology. In the present study, we seeded both MSC and EPC onto the same six bone graft substitutes and measured the same parameters. Methods In vitro, 125,000 MSC and 125,000 EPC were seeded onto Chronos Ò , Vitoss Ò , Actifuse Ò , Biobase Ò , Cerabone Ò , and Tutoplast Ò . Cell adhesion (fluorescence microscopy) and viability (MTT assay) were measured on days 2, 6, and 10. Osteogenic (cbfa-1, alkaline phosphatase [ALP], osteocalcin, collagen-1 alpha [Col1A]) and endothelial (von Willebrand factor [vWF], vascular endothelial growth factor [VEGF], kinase domain receptor [KDR]) gene expression were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR). Morphology was described by scanning electron microscopy (SEM) at day 2. Results MSC adhered significantly better to Tutoplast Ò , Chronos Ò , Actifuse Ò , and Biobase Ò . EPC adhered better to Actifuse Ò , Chronos Ò , Biobase Ò , and Tutoplast Ò . Viability increased over time when seeded on Tutoplast Ò and Chronos Ò . Osteogenic and endothelial gene expression were detectable at day 10 in cells seeded on Chronos Ò , Actifuse Ò , and Tutoplast Ò . The best morphology of MSC and EPC was found on Tutoplast Ò , Chronos Ò , Actifuse Ò , and Biobase Ò . Conclusion When bone graft substitutes are used to help fill large defects, it is important that their interaction with these cells be supportive of bone healing.

Leber's hereditary optic neuropathy (LHON) is a maternally inherited blinding disease due to mitochondrial DNA (mtDNA) point mutations in complex I subunit genes, whose incomplete penetrance has been attributed to both genetic and environmental factors. Indeed, the mtDNA background defined as haplogroup J is known to increase the penetrance of the 11778/ND4 and 14484/ND6 mutations. Recently it was also documented that the professional exposure to n-hexane might act as an exogenous trigger for LHON. Therefore, we here investigate the effect of the n-hexane neurotoxic metabolite 2,5hexanedione (2,5-HD) on cell viability and mitochondrial function of different cell models (cybrids and fibroblasts) carrying the LHON mutations on different mtDNA haplogroups. The viability of control and LHON cybrids and fibroblasts, whose mtDNAs were completely sequenced, was assessed using the MTT assay. Mitochondrial ATP synthesis rate driven by complex I substrates was determined with the luciferine/luciferase method. Incubation with 2,5-HD caused the maximal loss of viability in control and LHON cells. The toxic effect of this compound was similar in control cells irrespective of the mtDNA background. On the contrary, sensitivity to 2,5-HD induced cell death was greatly increased in LHON cells carrying the 11778/ND4 or the 14484/ND6 mutation on haplogroup J, whereas the 11778/ND4 mutation in association with haplogroups U and H significantly improved cell survival. The 11778/ND4 mutation on haplogroup U was also more resistant to inhibition of complex I dependent ATP synthesis by 2,5-HD. In conclusion, this study shows that mtDNA haplogroups modulate the response of LHON cells to 2,5-HD. In particular, haplogroup J makes cells more sensitive to its toxic effect. This is the first evidence that an mtDNA background plays a role by interacting with an environmental factor and that 2,5-HD may be a risk element for visual loss in LHON. This proof of principle has broad implications for other neurodegenerative disorders such as Parkinson's disease.

Background Phosphatidylcholine formulation has been used to dissolve local fat deposits. This study aimed to evaluate and compare the effects of phosphatidylcholine formulation and its vehicle sodium deoxycholate alone on different cell lines to understand better its mechanism of action. Methods Cells and media including 3T3-L1 preadipocytes, normal foreskin fibroblasts, neonatal human dermal microvascular endothelial cells (CADMEC), and fetal human skeletal muscle cells (HSkMC) were used. After 24 h, cells were exposed in 3-4, 5-dimethylthiazol-2yl-2, 3-diphenyl tetrazolium bromide reagent (MTT assays) to increasing dosages of phosphatidylcholine formulation (0.0156-0.5 mg/ml) or an equivalent vehicle, sodium deoxycholate solution, pH 9.0 (0.0066-0.210 mg/ml). Viability was assessed after 1, 2, and 3 days of treatment. Fat tissue (4 9 4 cm) obtained ex vivo from the dorsal fat pads of five rabbits was injected with 2 ml of phosphatidylcholine formulation (50 mg/ml), sodium deoxycholate (21 mg/ml), or normal saline and incubated for 24 h. These were examined histologically to identify cell lysis and morphologic changes. Results At 0.125-and 0.25-mg/ml doses of phosphatidylcholine solution, CADMEC and HSkMC were more sensitive (P \ 0.001, one-way ANOVA) than adipocytes at all time points examined. Phosphatidylcholine formulation at a dose of 0.5 mg/ml and the equivalent vehicle, sodium deoxycholate, at a dose of 0.21-mg/ml both induced nearly

Recently, the environmental residues of polybrominated diphenyl ethers (PBDEs) have markedly increased. In particular, the levels of certain PBDE congeners in fish have raised concern regarding potential risks associated with dietary PBDEs exposures. However, little is known regarding PBDE-mediated cell injury in relevant in vitro fish cell models. In this study, the cytotoxic effects of 2,2 0 ,4,4 0 -tetrabromodiphenyl ether (BDE-47) and decabrominated diphenyl ether (BDE-209) on RTG-2 cells were investigated.

Phytochemical and bioactivity studies of the flowers of Melastoma malabathricum L. (Melastomataceae) have been carried out. The ethyl acetate extract yielded three compounds, identified as naringenin, kaempferol and kaempferol-3-O-D-glucoside, and methanol extract gave kaempferol-3-O-(2 00 ,6 00 -di-O-p-trans-coumaroyl)glucoside and kaempferol-3-O-D-glucoside. The crude extracts and isolated compounds were screened for their antioxidant and cytotoxic activities. The antioxidant assay was carried out by the DPPH radical-scavenging electron spin resonance (ESR) spectroscopic method. The cytotoxicity was measured by the MTT assay against a MCF7 cell line. Naringenin, kaempferol, kaempferol-3-O-D-glucoside, kaempferol-3-O-(2 00 ,6 00 -di-O-p-trans-coumaroyl) glucoside, ethyl acetate and methanol extracts were found to be active as radical-scavengers with IC 50 values of 0.52 mM, 81.5 lM, 1.07 mM, 35.8 lM, 7.21 lg/ ml and 6.59 lg/ml, respectively. Naringenin and kaempferol-3-O-(2 00 ,6 00 -di-O-p-trans-coumaroyl)glucoside were also found to be active in inhibiting cell proliferation of MCF7 with IC 50 values of 0.28 lM and 1.3 lM, respectively.

Twenty-nine air samples of total suspended particles (TSP, particles less than 30–60 μm) and thirty samples of particles with aerodynamic diameter smaller than 2.5 μm (PM2.5) were collected at Guiyu, an electronic waste (e-waste) recycling site in southeast China from 16 August 2004 to 17 September 2004. The results showed that mass concentrations contained in TSP and PM2.5 were 124±44.1 and 62.12±20.5 μg m−3, respectively. The total sum of 16 USEPA priority polycyclic aromatic hydrocarbons (PAHs) associated with TSP and PM2.5 ranged from 40.0 to 347 and 22.7 to 263 ng m−3, respectively. Five-ring and six-ring PAHs accounted for 73% of total PAHs. The average concentration of benzo(a) pyrene was 2–6 times higher than in other Asian cities. Concentrations of Cr, Cu and Zn in PM2.5 of Guiyu were 4–33 times higher than in other Asian countries. In general, there were significant correlations between concentrations of individual contaminants in TSP with PM2.5 (i.e. PAHs, Cd, Cr, Cu, Pb, Zn, Mn except Ni and As). The high concentrations of both PAHs and heavy metals in air of Guiyu may impose a serious environmental and health concern. Cytotoxicity of the extract of TSP and PM2.5 of ten 24 h samples collected against human promonocytic leukemia cell line U937 (ATCC 1593.2) was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxicity assay. The results showed that under the same concentrations of extract, PM2.5 cytotoxicity was 2–4 times higher than TSP.

This study reports for the first time the biological properties of Portuguese propolis. The antioxidant potential of propolis samples from Bornes (Northeast) and Fundão (Centre) regions of Portugal was evaluated by their ability to inhibit the 2,2 0 -azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis and lipid peroxidation in human erythrocytes. Bornes and Fundão propolis strongly protected the erythrocyte membrane from hemolysis (IC 50 of 6.3 ± 0.7 and 10.4 ± 2.7 lg/ml, respectively), in a time-and concentration-dependent manner. This effect was found to be significantly higher than that presented by ascorbic acid (IC 50 of 31.0 ± 5.6 lg/ml). In addition, human erythrocytes treated with propolis extracts showed concentration-dependent decrease in levels of malondialdehyde, a breakdown product of lipid peroxidation. Propolis extracts were also assayed for their anticancer properties on human renal cell carcinoma (RCC). Primary cultures of normal and cancerous renal cells derived from RCC patients, in addition to A-498 cell line, were treated with propolis extracts (0-100 lg/ml). Cytotoxic and antiproliferative effects were determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Propolis extracts exhibited selective toxicity against malignant cells compared to normal cells. In vitro RCC growth was strongly inhibited by Bornes and Fundão propolis in a concentration-dependent manner. Our results indicate that Portuguese propolis constitutes an excellent source of effective natural antioxidant and chemopreventive agents.

Background: Blechnum orientale Linn. (Blechnaceae) is used ethnomedicinally for the treatment of various skin diseases, stomach pain, urinary bladder complaints and sterilization of women. The aim of the study was to evaluate antioxidant, anticancer and antibacterial activity of five solvent fractions obtained from the methanol extract of the leaves of Blechnum orientale Linn.

Background: Cancer is a leading cause of death worldwide , with approximately 17.5 million new cases and 8.7 million cancer related deaths in 2015. The problems of poor selectivity and severe side effects of the available anticancer drugs, have demanded the need for the development of safer and more effective chemotherapeutic agents. The present study was aimed at determining the cytotoxicities of 31 medicinal plants extracts, used in Nigerian ethnomedicine for the treatment of cancer. Methods: The plant extracts were screened for cytotoxicity, using the brine shrimp lethality assay (BSLA) and MTT cytotoxicity assay. Rhabdomyosarcoma (RD) cell line, normal Vero cell line and the normal prostate (PNT2) cell line were used for the MTT assay, while Artemia salina nauplii was used for the BSLA. The phytochemical composition of the active plant extracts was determined by high performance liquid chromatography (HPLC) analysis.

This study was undertaken to evaluate the effect of the G93A mutation in the human Cu/Zn-superoxide dismutase gene (hSOD1) on the phosphatidylinositol-3-kinase (PI3K)/Akt and glycogen synthase kinase-3 (GSK-3) pathway in motoneuron, and to determine the role of epigallocatechin gallate (EGCG) on oxidative stress-injured motoneurons. The viability of G93A mutant cells was less than that of wild-type cells, and the activation of PI3K and the phosphorylation of Akt and GSK-3 in G93A mutant cells decreased compared with wild-type hSOD1 4.1 cells. In the experiment to evaluate the effect of oxidative stress and/or EGCG on these motoneurons, after exposure to 400 M H 2 O 2 , the MTT assay revealed greatly reduced viability of G93A mutant cells compared with wild-type cells, and pre-treatment of these cells with EGCG before H 2 O 2 exposure increased the viability of both cell lines. Western blot analysis showed that the G93A mutation and oxidative stress decreased survival signals including PI3K/Akt but increased death signals including GSK-3; however, pre-treatment with EGCG increased survival signals but decreased death signals. These results suggest that PI3K/Akt and GSK-3 activities are altered in G93A mutant cells and EGCG-induced activation of PI3K/Akt and inhibition of GSK-3 could be a new potential therapeutic strategy for ALS associated with oxidative injury.

Context: Some flavonoids have been described as neuroprotectors. Gossypitrin (Gos) is a natural occurring flavonoid and the main bioactive substance from the flowers of Talipariti elatum Sw. (Majagua azul), traditionally used in Cuba as expectorant and antiasthmatic. Only few reports have documented its antioxidant properties.

Aims: To evaluate the antioxidant and cytoprotective effects of Gos against cyanide-induced oxidative stress and cell death in PC12 cells.

Methods: Gos effects on DPPH/ ABTS radical scavenging, ferric reducing power and lipid peroxidation were examined. The ischemia/reperfusion neuronal damage was produced by exposing PC12 cells to KCN in glucose-free medium. Gos was added to the incubation medium 30 min before chemical hypoxia induction. The neuroprotective potential of Gos was assessed by measuring cell viability by the MTT assay, the antioxidant enzymes SOD and CAT, and GSH and lipid peroxidation levels. Rutin (Rut), the well-known antioxidant, was used as a reference compound.

Results: Gos showed a potent intrinsic antioxidant capacity evidenced by low IC50 and EC50 values for DPPH/ABTS/malondialdehyde and ferric reducing power, respectively. Pre-treatment of PC12 cells with Gos, significantly increased their survival against KCN, restored the levels of GSH and the SOD and CAT enzymes activities, as well as reduced the level of lipid peroxidation. Its antioxidant effects were higher than those elicited by Rut.

Conclusions: The results show for the first time the neuroprotective potential of Gos against hypoxic cell damage, probably associated to its antioxidant effects.

The search for new anti-cancer drugs is one of the most prominent research areas of natural products. Numerous active compounds isolated from Brazilian Cerrado plant species have been studied with promising results. Aim of the study: To investigate the cytotoxic potential of 412 extracts from Brazilian Cerrado plants used in traditional medicine belonging to 21 families against tumor cell lines in culture. Material and method: Maceration of 50 plant species resulted in 412 hexane, dichloromethane, ethanol and hydroalcohol extracts. The cytotoxicity of the extracts was tested against human colon carcinoma (HCT-8), melanoma (MDA-MB-435), and brain (SF-295) tumor cell lines, using the thiazolyl blue test (MTT) assay. Bioassay-guided fractionation was performed for one active extract. Results and Conclusions: Twenty-eight of the 412 tested extracts demonstrated a substantial antiproliferative effect, at least 85% inhibition of cell proliferation at 50 g/mL against one or more cell lines. Those extracts are obtained from different parts of Anacardiaceae, Annonaceae, Apocynaceae, Clusiaceae, Flacourtiaceae, Sapindaceae, Sapotaceae, Simaroubaceae and Zingiberaceae. Complete dose-response curves were generated and IC 50 values were calculated for these active extracts against four cell lines HCT-8, MDA-MB-435, SF-295 and HL-60 (leukemia), and their direct cytotoxic effects were determined. In summary, 14 extracts of 13 species showed toxicity in all tested tumor cell lines, with IC 50 values ranging from 0.1 to 19.1 g/mL. The strongest cytotoxic activity was found for the hexane extract of Casearia sylvestris var. lingua stem bark, with an IC 50 of 0.1 g/mL for HCT-8, 0.9 g/mL for SF-295, 1.2 g/mL for MDA-MB-435, and 1.3 g/mL for HL-60, and Simarouba versicolor root bark, with an IC 50 of 0.5 g/mL for HCT-8, 0.7 g/mL for SF-295, 1.5 g/mL for MDA-MB-435, 1.1 g/mL for HL-60. Bioassay-guided fractionation of the last extract led to the isolation of glaucarubinone, which showed pronounced activity against the four cell lines studied. Further studies of the active extracts are necessary for chemical characterization of the active compounds and more extensive biological evaluations.

Background: The incidence rate of cervical cancer is increasing and its existing drugs are becoming more and more resistant. Therefore, we extracted the fruiting body of Calocybe indica edible mushroom in 90% ethyl acetate extract (EAE) and evaluated it as an anticancer property against HeLa and CaSki. Method: We performed cytotoxicity assay by MTT, cell morphological study by phase contrast microscope, and apoptosis study by nuclear morphology via DAPI staining under inverted microscopy; the expressions of pro-apoptotic and anti-apoptotic genes and p53 were examined by Western blotting, cell cycle analysis, and cologenic and cell migration assay. Antioxidant content and activity assays were performed for mycochemistry analysis of EAE thin layer chromatography (TLC). Result: EAE-treated HeLa and CaSki cells became round and showed condensed and fragmented nuclei. They inhibited the cell proliferation of both cancer cell lines in a dose-dependent manner. At maximum dose (1250 µg/mL) ...

The purpose of this study was to investigate the potential effects of epothilones (EPOs), a new class of microtubule stabilizing cytotoxic drugs, on glioma cells in vitro. The effects of 1, 10 and 100 nM concentrations of EPO D in four malignant human glioma cell lines were measured using a microtiter-tetrazolium assay. Besides the cell lines U87MG, U138MG and LN405, one cell line was used, which had been derived from a recurrent and therapy-resistant glioblastoma in our laboratory. In addition, changes of the cell morphology were followed by light microscopy and changes in the microtubule and actin cytoskeleton were visualized by a confocal laser microscope. In all four human glioma cell lines, 10 and 100 nM concentrations of the drug, applied for 96 h, lead to a highly significant decrease in the viable cell number (p < 0.001). A mean reduction of the viable cell number between 30% and 40% (60% and 90%) was observed for a drug concentration of 10 nM (100 nM). A round cell morphology occured in most EPO treated cells and the organized network of microtubules was shrunk in these round cells. The tubulin immunostaining now appeared amorphous and was restricted to small perinuclear regions. Large actin filaments also disappeared, but actin staining was present in the whole cytosplasm. These results prove that EPOs have antiproliferative effects in glioma cells and affect their tubulin cytoskeleton, as it was previously observed in several types of carcinoma cells.

A novel series of optically active 2-aminobenzothiazole derivatives were synthesized by reaction of optically active amine (I) with thiophosgene to obtain optically active isothiocyanates (IIa–h) which on condensation with 4-fluoro-3-chloro aniline (III) yielded various optically active thioureas (IVa–h). Further oxidative cyclisation in the presence of bromine and chloroform yielded title compounds (Va–h). The structures of these compounds were established by IR, 1H NMR, 13C NMR, Mass and HRMS. The compounds (IVa–h and Va–h) were evaluated for in vitro cytotoxicity against mouse Ehrlich Ascites Carcinoma (EAC) and two human cancer cell lines (MCF-7 and HeLa). In preliminary MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cytotoxicity studies the optically active thiourea derivatives (IVe, IVf and IVh) were found most effective. In EAC cells the IC50 values for IVe, IVf, IVh and Vg were found in the range of 10–24 μM, whereas in MCF-7 and HeLa cells the IC50 values were observed in the range of 15–30 μM and 33–48 μM, respectively. In alkaline comet assay the compounds (IVe and IVf) showed dose-dependent DNA damaging activity.A series of novel optically active thiourea and their 2-aminobenzothiazole derivatives had been synthesised and evaluated for in vitro cytotoxicity against EAC, MCF-7, HeLa cells.

Nitric oxide is a free radical involved in the pathogenesis of cancer by increasing tumour vascularization and metastasis. Studies using nitric oxide inhibitors have shown decrease in tumour growth and a role in cancer therapy. To analyse the effect of fruits on nitric oxide, we carried out experiments using a nitric oxide donor on the breast cancer cell line, MCF-7. Proliferating MCF-7 cells were treated with the methanolic extract of the fruits. The inhibitory activity of fruit extracts on cell proliferation was measured using the MTT assay. Chiku and dragon fruit showed high inhibitory activity when compared to the other fruits tested. The total polyphenol and flavonoid content and nitric oxide scavenging activity were found to be high in pomegranate, chiku, litchi, durian, grape and apple. This study shows that phytochemicals present in fruits scavenge nitric oxide and inhibit MCF-7 cell proliferation.► We analysed the effects of 13 tropical fruits on nitric oxide. ► Nitric oxide inhibitors are known to decrease tumour growth. ► The fruits inhibited nitric oxide-induced proliferation of MCF-7. ► The fruits also had high polyphenol and flavonoid content. ► Phytochemicals in fruits scavenge nitric oxide and inhibit MCF-7 cell proliferation.

Neuroblastoma is a common childhood tumor derived from the periph- eral nervous system. Favorable neuroblastomas usually express TrkA, the receptor for nerve growth factor (NGF), whereas unfavorable, MYCN- amplified neuroblastomas usually express TrkB and its ligand, brain- derived neurotrophic factor (BDNF). Here, we provide evidence that the TrkB-BDNF pathway is associated with enhanced survival and resistance to chemotherapy in neuroblastoma.

The present study describes the phytochemical profile and the protective effects of Ceratonia siliqua pods essential oil (CsEO), a food and medicinal plant widely distributed in Tunisia. Twenty five different components were identified in the CsEO. Among them, the major detected components were: Nonadecane, Heneicosane , Naphthalene, 1,2-Benzenedicarboxylic acid dibutylester, Heptadecane, Hexadecanoic acid, Octadecanoic acid, 1,2-Benzenedicarboxylic acid, Phenyl ethyl tiglate, Eicosene, Farnesol 3, Camphor, Nerolidol and n-Eicosane. The antimicrobial activity of CsEO was evaluated against a panel of 13 bacteria and 8 fungal strains using agar diffusion and broth microdilution methods. Results have shown that CsEO exhibited moderate to strong antimicrobial activity against the tested species. In addition, the inhibitory effect of this CsEO was evaluated in vivo against a foodborne pathogens Listeria monocytogenes, experimentally inoculated in minced beef meat (2 × 10 2 CFU/g of meat) amended with different concentrations of the CsEO and stored at 7°C for 10 days. The antibacterial activity of CsEO in minced beef meat was clearly evident and its presence led to a strong inhibitory effect against the pathogens at 7°C. On the other hand, the cytotoxic effects of the essential oil against two tumoral human cell lines HeLa and MCF-7 were examined by MTT assay. The CsEO showed an inhibition of both cell lines with significantly stronger activity against HeLa cells. The IC 50 values were 210 and 800 μg/ml for HeLa and MCF-7 cells, respectively. Overall, results presented here suggest that the EO of C. siliqua possesses antimicrobial and cytotoxic properties, and is therefore a potential source of active ingredients for food and pharmaceutical industry.

Background: Considerable interest has been aroused in recent years by the well-known notion that biological systems are sensitive to visible light. With clinical applications of visible radiation in the far-red to near-infrared region of the spectrum in mind, we explored the effect of coherent red light irradiation with extremely low energy transfer on a neural cell line derived from rat pheochromocytoma. We focused on the effect of pulsed light laser irradiation vis-à-vis two distinct biological effects: neurite elongation under NGF stimulus on laminin-collagen substrate and cell viability during oxidative stress.

1National Centre for Alternative Methods (ZEBET), Berlin, Germany; 2ECVAM, Institute for Health & Consumer Protection, European Commission Joint Research Centre, Ispra, Italy; 3Syngenta, Macclesfield, UK; 4Unilever, Sharnbrook, UK; 5L&#x27;Oréal, Clichy, France; 6ECB, ...