Background and objective: In addition to the use of chemicals in food, pharmaceutical, cosmetic and hygiene industries, use of herbal and natural compounds have also increased. It seems natural ingredients compared with the chemical components show less side effects, such as mutagenicity and carcinogenesis; however, their use can also be associated with side effects and toxicity. Some significant medicinal effects of saffron have been proven, which reveals the necessity to investigate the toxicity of the plant. This study aimed to evaluates the cytotoxic effects of saffron on cell lines Vero, Hela, and Hep2 using MTT assay method.Material and Method: Firstly the stigma of saffron was extracted by methanol: water 80:20 using maceration method. Cytotoxic effects of different dilutions of saffron extract on Vero normal cell lines and Hela and Hep2 cancerous cell lines was investigated using MTT assay method in a 96 well microplate. Reading optical density (OD) was done by microplate reader apparatus and IC50 (Inhibition Concentration 50%) was calculated for each cell line. Results: Aqueous - methanolic saffron extract on cell lines Vero, Hela, and Hep2, showed IC50 equal 162,653 and 208 mg/ml respectively.Conclusion: Saffron has a toxic effect on normal Vero cell line. Therefore, must be cautious to not to use it in food and as herbal medicine in high concentrations. Also the extract of this plant revealed toxic effects on two cancerous cell lines, Hepa2 and Hela. However, its toxicity, especially on Hela cancerous cell line was lower than the Vero normal cell line.

Purpose: Lipopolysaccharide (LPS) is a major constituent of gram negative bacteria cell wall. LPS consistes of three covalently linked components of (i) lipid A (endotoxine A), (ii) core (oligosaccharide) and (iii) the a-antigen polymer (polysaccharide). LPS preparations are used extensively for research in the elucidation of the LPS structure, metabolisme, toxicity and biosynthesis. The LPS is also used as a mitogen for B- lymphocytes proliferation. The purpose of this study was to evaluate the extracted LPS as a mitogen by the MTT assay.Materials & methods: Escherichia coli was grown in the nutrient broth (NB) and after the harvesting, the bacteria were washed with ethannol, aceton or chloroform- methanol. The LPS was extracted by two different methods of Phenol- chloroform- petroleum ether (PCP) and Methanol-Chloroform (MC). In the MC method, the bacteria was sonicated and then the LPS was extracted. In the PCP method, Petroleum ether and chloroform were removed on rotary evaporator or in a high vacuum at a degree bellow 0°C and the precipitated LPS was washed two to three times with ether to remove any remaining phenol. We measured the B cell proliferation by using the MTT assay. We also used the cell viability assay with the trypan blue dye as a qualitative assay. In the MTT9 assay different concentrations of extracted LPS and standard LPS (Sigma) were added to 96 well plates containing 2.5x104 cells/well. These cells were separated from the whole blood by ficoll and suspendd in a complete RPMI-1640 medium. We measured the 0.0 of the MTT dye at 570 nm and 620 nm respectively.Results: Our result showed that the cellular proliferation by the PCP-LPS was more than the MC-LPS, compared to the control (cells without LPS) and was similar to the standard LPS.Conclusion: In conclusion, the PCP-LPS has more mitogeneic activity compared to the MC-LPS.

BACKGROUND: Cancer is the fifth leading cause of death worldwide. There are considerable efforts to identify naturally occurring substances for use as new drugs in cancer therapy. Some components of animal venoms have been identified that possess substantial anticancer properties. In our previous studies, the cytotoxic effects of ICD-85 (venom-derived peptides) have been reported on HL-60 and MDA-MB231 cell lines. This has prompted us to investigate the comparative cytotoxic effects of ICD-85 on the HeLa cell line and normal lamb kidney (LK) cells.METHODS: Cells were exposed to various concentrations (8´10-4 to 5.6´10 mg/ml) of ICD-85 at various incubation times (24, 48 and 72 hours). Cell viability was measured by the MTT assay. A morphological study was also carried out using an inverted microscope. Caspase-8 activity was assayed by the Caspase-8 Colorimetric Assay Kit in HeLa cells that were exposed to ICD-85 for 48 hours.RESULTS: Data analysis showed that ICD-85 has a dose-dependent cytotoxic effect on HeLa cells with an inhibitory concentration 50% (IC50) of 26.62±2.13 mg/ml at 24 hours, 27.33±2.35 mg/ml at 48 hours, and 28.13±2.52 mg/ml at 72 hours. Results also indicated that the cytotoxic effect of ICD-85, at 48 and 72 hours incubation times did not show significant alteration compared to 24 hours of exposure. Interestingly, the minimum concentration of ICD-85 which showed a cytotoxic effect on LK cells was found to be 3500-fold less than the minimum concentration that showed a cytotoxic effect on the HeLa cancer cells. While morphological analysis revealed a significant difference that included the characteristic rounding of dying cells by treatment with ICD-85 compared with untreated HeLa cells, this difference was not observed in normal cells. ICD-85 increased caspase-8 activity in HeLa cells after 48 hours of exposure.

INTRODUCTION: Sperm MTT viability assay is a new method for assessment of percentage of viable sperms in semen samples. The objective of this study was to evaluate the capability of sperm MTT viability assay, in comparison with eosin-nigrosin and hypo-osmotic swelling tests, in severe and moderate asthenospermia and normospermia.METHODS: MTT, E&N and HOST were carried out simultaneously on 37 human semen samples. Nine semen samples had less than 5% motility, eleven semen samples had 10-25% motility and seventeen semen samples had more than 50% motility. Two paired t-tests were conducted using SPSS, statistical software.RESULTS: No significant differences were observed between the above 3 sperm viability tests between the aforementioned patients.DISCUSSION: Sperm MTT viability assay can be used as a diagnostic test for differentiating viable from non-viable sperms in all of these three groups.

Background: Toxoplasma gondii is intracellular parasites that cause many symptoms such as encephalitis and congenital disorders. Marrubium vulgare, Salvia officinalis and lippia citriodora are herbal drugs that used to kill this organism. Our objective were to evaluate the therapeutic effect of Marrubium vulgare, Salvia officinalis and lippia citriodora in killing of toxoplasma gondii tachyzoite and evaluation by MTT assay Methods: In this survey we used Marrubium vulgare, Salvia officinalis and lippia citriodora suspension plus RPMI medium containing peritoneal tachyzoite after incubation at 37 C0. The measurement done by MTT assay and statistial assesment done by spss19 software. Our investigation based on two mediam (invitro and invivo) to approve our results. Results: In RPMI medium the killing with three herbal drugs were seen and the decrease of Absorbance were seen by MTT assay. The means of OD were 0. 05 to 0. 16 in presence of herbal drugs but the means of OD were 0. 9 to 2. 3 in absence of herbal drugs. Conclusion: Marrubium vulgare, Salvia officinalis and lippia citriodora are herbal drugs that are very useful to kill toxoplasma tachyzoite in invitro condition.

Background: Leishmaniasis is a parasitic disease that occurs in subtropical and tropical regions with approximately 350 million people worldwide and 2 million new cases annually. The annual increase in cutaneous leishmaniasis (CL) is observed, especially in endemic areas such as Iran. Since there is no effective vaccine, the detection of natural anti-leishmanial products is essential. Objective: The purpose of this study was to evaluate the in vitro anti-leishmanial activity of two herbal medicine including Artemisia dracunculus L. and Heracleum persicum Desf. (Golpar). Materials and Methods: The extracts of selected plants were obtained by maceration, and in vitro anti-leishmanial activity was assayed on Leishmania major and Leishmania infantum promastigotes using colorimetric MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] assay in comparison with glucantime as a reference. Results: Based on the results, 50% inhibitory concentration (IC50) values of selected plants and glucantime solutions were determined at 24, 48, and 72 hours incubation. Further, the anti-leishmanial activity of the leaf extract of A. dracunculus with IC50 values of 1. 85 and 3. 5 μ, g/mL and the fruit extract of H. persicum with values of 31. 32 and 11. 7 μ, g/mL were evaluated against L. major and L. infantum promastigotes, respectively. Conclusion: These results revealed anti-leishmania properties of the above-mentioned plants and the need to study the effects of these extracts on the Leishmania genus in animal models and in vivo assay in the future.

Background: The importance of cellular dosimetry in both diagnostic and radiation therapy is becoming increasingly recognized. Objective: This study aims to compare surviving fractions, which were predicted using Geant4 and contained three types of cancer cell lines exposed to 188Re with the experimentally surviving fraction determined by MTT assay. Material and Methods: In this comparative study, Geant4 was used to simulate the transport of electrons emitted by 188Re from the cell surface, cytoplasm, nucleus or medium around the cells. The nucleus dose per decay (S-value) was computed for models of single cell and random monolayer cell. Geant4-computed survival fraction (SF) of cancer cells exposed to 188Re was compared with the experimental SF values of MTT assay. Results: For single cell model, Geant4 S-values of nucleus-to-nucleus were consistent with values reported by Goddu et al. (ratio of S-values by analytical techniques vs. Geant4 = 0. 811– 0. 975). Geant4 S-values of cytoplasm and cell surface to nucleus were relatively comparable to the reported values (ratio =0. 914– 1. 21). For monolayer model, the values of SCy→ N and SCS→ N, were greater compared to those for model of single cell (2%– 25% and 4%– 38% were larger than single cell, respectively). The Geant4 predicted SF for monolayer MCF7, HeLa and A549 cells was in agreement with the experimental data in 10μ Ci activity (relative error of 2. 29%, 2. 69% and 2. 99%, respectively). Conclusion: Geant4 simulation with monolayer cell model showed the highest accuracy in predicting the SF of cancer cells exposed to homogeneous distribution of 188Re in the medium.

Aim & Background: Frankincense is a herbal product that has diverse therapeutic effects. Beta-boswellic acid, the main ingredient of frankincense, is poorly soluble in water and its bioavailability is very low. The aim of present study was to evaluate the effect of loading of beta-boswellic acid in dendrosomal nanoparticles on its bioavailability and cell uptake by MTT assay.Materials and Methods: For MTT assay, B65 cells were treated with different doses of nave beta-boswellic acid or dendrosomal beta-boswelic acid for 24, 48 and 72 hours. After adding MTT, colorimetery was done using ELISA reader and IC50 was then calculated. The data was analyzed with One Way Anova program using SPSS v.16 software.Results: The IC50 for nave beta-boswellic acid and dendrosomal beta-boswellic acid was 88.09 and 58.42 mM in 24h, 58.37 and 44.87 mM in 48h and 21.09 and 16.69 mM in 72h respectively.Conclusion: The results showed that loading of beta-boswellic acid in dendrosomal nanoparticles increases the cell toxicity effect of beta-boswellic acid. This observation might be due to the increased cell uptake of beta-boswellic acid in the presence of dendrosomal nanoparticles.

Background and purpose: Today medicinal plants have received much attention in treatment of many cancers.Juglans regia has phenolic compounds and is used in cytotoxicity studies. This study investigated the effects of hydroalcoholic extract of Juglans regia on the growth of human cervical and liver cancer cells.Materials and methods: The hydroalcoholic extract of the fruit of Juglans regia was prepared by percolation method. The HeLa and HepG2 cancer cells were cultivated and further incubated with different concentrations of the extract. After 72 hours, cell growth inhibition was examined using MTT assay. The results were statistically analyzed in Prism ver.3 applying ANOVA and post-test.Results: The inhibitory concentration 50 (IC50) of the plant extract on human HepG2 and HeLa cancer cells were 449.1 ± 18.26 mg/ml and 222.4 ± 2.66 mg/ml, respectively. The IC50of Cisplatin on these cancer cells were found to be 15.96±1.117and 15.79±1.147 mg/ml, respectively.Conclusion: The IC50of Juglans regia on HeLa and HepG2 was more than that in Cisplatin.Accordingly, further investigations are needed on other cancer cell lines. Moreover, due to the fact that the extract was total, future studies should aim at identifying its active substances.

Objective: The rate determination of cellular proliferation and cell viability is critical to the assessment of the effects of drugs on cells and there are different standard methods in evaluation of cell proliferation. Calprotectin is a cytosol metaloprotein in the neutrophil, macrophase, lymphocyte and monocyte cytosol that induce growth-inhibitory against various cell types including tumor cells. These finding indicate it can be as a therapeutic agtent in cancer diseases. Biological characteristics of calprotectin are contained antibacterial, antifungal, immonoregulation, chemotactic activity and inhibition of cell proliferation. In the present study, two staining assays; trypan blue and MTT in vitro evaluation of human calprotectin proliferation inhibition on human gastric cancer cells was compared. Materials and Methods: Calprotectin was purified from human neutrophil by chromatography methods. The human gastric adenocarcinoma cell line (AGS) was used. These cells were incubated in RPMI 1640 medium supplemented with 10% FBS in a humidified incubator (37oC & 5% CO2). AGS cells (10000 cell per well) were incubated to different concentration of calprotectin (25, 50, 100, 200, 400 mg/ml) at 24, 48, 72 h. In this study trypan blue dye exclusion and MTT assay were used for evaluation of cell proliferation inhibition.Results: Results of cytotoxicity effect of calprotectin on AGS cells show that there are significant correlation between two methods (P<0.01) and LC50 value of calprotectin calculated by MTT assay is lower than trypan blue in all time intervals. The LC50 of calprotectin for AGS cell at 24, 48 and 72 h was determined 33.29, 71 and 141.8 mg/ml by trypan blue and 96.78, 38.66 and 9.86 mg/ml by MTT assay, respectively. There are significant correlation between both methods at different time intervals and it is positive linear (P<0.01–P<0.001). Conclusion: Result of cell cyto toxicity of calprotectin by both methods indicated that correlation is positive and significant. It can be said that MTT assay is more sensitive, easy to handle, a large number of probes can be assayed in a relatively short time.