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.

Background: Various MTT assay methods are proposed to obtain the cell survival parameters.Objective: Determining the survival curve characteristics of two cancerous cells of interest based on a common and a novel MTT assay method after exposing them to ionizing radiation.Method: A common and a novel MTT assay method were used and compared for obtaining the F10B16 melanoma and 4T1 breast adenocarcinoma survivals after exposing them to ionizing radiation from a Co-60 machine. To obtain the survival parameters of the cells based on the common method, the cells were inoculated in 96-well plates. After irradiating the plates, the MTT assay was performed over the following days for a period of 8 days. Thereafter, the survival fraction was calculated from a simple equation for every day from which the best day was selected. To acquire the cells’ survival parameters based on the novel method, extensive experiments were performed on a large number of samples. Then, the MTT assay was done in every day following various experimental treatments to acquire the exponential growth. Finally, the cells’ survivals were determined by measuring the space between relevant growing curves.Results: At low doses (<4Gy) the two MTT assay methods indicated the same results. However, at higher doses there were significant differences among the findings.Conclusion: Both of the MTT methods indicated that the cells’ responses are dependent on the dose levels used. Although the implementation of the common MTT assay method is simpler, the novel method seems to show more precise and reliable results at all levels of radiation doses.

Background and purpose: Penicilliums have high diversity among fungi species and some of them are found to be very useful. Some studies have evaluated their antimicrobial and cytotoxic effects.Penicillium citrinum is a genus of the penicilliums that produces mycotoxin citrinin. Therefore, it is worthy to assess its cytotoxic effect.Materials and methods: The DNA of the fungus obtained from the soil samples from the campus of Mazandaran University of Medical sciences was extracted. Then DNA sequencing was done and the ethanolic extract including metabolites was taken out. The effect of different concentrations of test solution were evaluated on cancer cell lines of human liver (HepG2), lung (A549), ovary (SKOV3), and breast (MCF7) and also on kidney (LLCPK1) and ovary of Hamster (CHO) normal cell lines using MTT method. Cisplatin was considered as positive control. The data was analyzed using Prism Ver.3, ANOVA and t-test.Results: The findings revealed significant differences between the levels of IC50 of fungus metabolites and cisplatin in all cell lines (P<0.005). Also, the level of IC50 of fungus metabolites on normal cell lines was significantly different from that of the cancer cell lines (P<0.05).Conclusion: This study showed that ethanolic extract of P. citrinum metabolites did not have a considerable toxicity effect on cancer and normal cell lines. However, it increaesd the inhibitory effect of cancer cell proliferation.

Background & objectives: Pentavalent antimonials are the first-line drugs for treatment of leishmaniasis, which have multiple side effects such as drug toxicity. Moreover, parasite resistance to these drugs is rising around the world. Second-line drugs, including Amphotericin B and pantamidine have also side effects and expensive for patients. According to the cytotoxic effects of paraquat, this study was conducted to evaluate the effect of paraquat on Leishmania major promastigotes and HUVECs viability. Methods: A number of 2. 5×10 of Leishmania major promastigotes were treated in each well of 96 well plates with different concentrations of paraquat. Cells were incubated for 48 hours in 24 ° C. MTT test was performed for evaluating paraquat impact on promastigotes. The absorbance was measured using a microplate reader at 570 nm. The trypan blue staining assay was performed to evaluate the number of viable Leishmania major promastigotes following paraquat treatment. Furthermore, the effect of paraquat concentrations on HUVECs viability was evaluated under the cell culture condition. Results: The results of the MTT test showed that increasing concentrations of paraquat could significantly reduce the viability and the number of Leishmania major promastigotes in comparison to control group 6 (p<0. 05). In this study, the IC50 for Leishmania major promastigotes was calculated as 272. 46 µ g/ml. Trypan blue results were in line with the finding of MTT assay. Moreover, we found that HUVECs were susceptible to paraquat (IC50=188. 99 µ g/ml ). Conclusion: Paraquat has a strong inhibitory effect on Leishmania major promastigotes and human endothelial cells. Although more comprehensive studies on the effects of the topical use of paraquat on Leishmania major lesions in animal model and its side effects are necessary.

Background: It is a continuing effort to develop new anticancer drugs. It has been reported that albendazole has antiproliferative effects on several tumor cell lines. The aim of this study was to evaluate the killing effect of albendazole on HT-29 human colorectal cancer cells. Materials and methods: Cultured HT-29 cells were treated with albendazole in 0. 1, 1 and 10 µ g/ml concentrations for 1, 3 and 5 days. Cellular viability and proliferation were evaluated by MTT and Trypan blue assays at determined days after treatment. Results: The comparison of cells count and viability in control group and treated groups with albendazole at different concentrations showed significant differences in days 1, 3 and 5. Conclusion: Albendazole can suppress HT-29 cells proliferation and induces cell death in a dose and time dependent manner.

Background: Leishmania major is transmitted by sandflies of the genus Phobotomus and causes cutaneous lesions in humans. Curcumin is made from turmeric and various derivatives is derived from it. In the present study the effect of curcumin and it’ s derivatives on Leishmania major in vitro was investigated. Materials and Methods: Curcumin 70% and 90% purity and it’ derivatives such as s, BDMC base methoxy curcumin, diacetyl curcumin DAC, vanadyl curcumin VO (CUR) 2, vanadyl diacetyl curcumin VO (DAC) 2, indium curcumin In(CUR)3 and Ga (CUR) 3 preparation. Curcumin derivatives were synthesized and different concentrations of 500 to 800 μ g/ ml were prepared in glycerin. 106 Leishmania major promastigotes added to each well of 96 well plate and 100 μ L of each curcumin derivative added and stored at 25 ° C for 24 hours. Assay of the lethality of the compounds against Leishmania major and evaluation of their toxic effects on (vero) cells were evaluated using MTT assay. Results: Lethal effect of curcumin and it’ s derivatives against Leishmania, increased with increasing concentration. Anti-leishmanial index (IC50) of 70% and 90% curcumin compounds, (BDMC) bis methoxy curcumin, Diacetyl curcumin (DAC), vanadyl curcumin VO (CUR)2, vanadyl diacetyl curcumin VO (DAC)2, indium curcumin In (CUR)3, Gallium Curcumin Ga (CUR)3 compounds were 110, 93, 101, 103, 98, 103, 51 and 58 μ g / ml and the lethal effect of these compounds against (vero) cells were 33. 1%, 19%, 21%, 20. 3%, 17%, 21%, 25. 3%, 16%, respectively. Conclusion: Gallium curcumin and indium curcumin, compared to curcumin and other derivatives, exhibited the highest anti-leishmanial effect and were the safest derivatives of curcumin for mammalian cells.

Background: All natural anticancer agents are cytotoxic basically and act mainly by the inhibition cell proliferation; but they have different mechanisms. Two assays, thiazolyl blue [3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyl-terazoliumbromide or MTT] and sulforhodamine B (SRB), are used to assess cell growth. This study aimed to compare measurements between MTT and SRB on the cancer cell lines.Methods: Different concentrations of the bromelain were added to cultured cells including mouse breast cancer (4T1), human gastric carcinoma (AGS), and human prostate carcinoma (PC3) cell lines and incubated at 24 and 48 hours. The growth and proliferation rates of the studied cells were investigated using both MTT and SRB assays after treatment with bromelain. The differences between cells were determined using Kruskal-Wallis and Dunns tests.Findings: Bromelain significantly decreased growth and proliferation rate of 4T1, AGS and PC3 cancer cells, in a concentration-dependent manner at different times, in both MTT and SRB assays.Conclusion: Findings showed that both MTT and SRB assays gained similar data regardless of the cell types.

Background: We aimed to investigate different biological properties of aerial parts essential oil of Ferulago trifida Boiss and larvicidal activity of its volatile oils from all parts of plant. Methods: Essential oil was prepared by steam distillation and analyzed by Gas chromatography and GC/Mass. Anti-oxidant, antimicrobial, cytotoxic effects and AChE inhibitory of the oil were investigated using DPPH, disk diffusion method, MTT assay and Ellman methods. Larvicidal activity of F. trifida essential oil against malaria vector Anoph-eles stephensi was carried out according to the method described by WHO. Results: In GC and GC/MS analysis, 58 compounds were identified in the aerial parts essential oil, of which E-ver-benol (9. 66%), isobutyl acetate (25. 73%) and E-β-caryophyllene (8. 68%) were main compounds. The oil showed (IC50= 111. 2μ g/ml) in DPPH and IC50= 21. 5 mg/ml in the investigation of AChE inhibitory. Furthermore, the oil demonstrated toxicity with (LD50= 1. 1μ g/ml) in brine shrimp lethality test and with (IC50= 22. 0, 25. 0 and 42. 55 μ g/ml) on three cancerous cell lines (MCF-7, A-549 and HT-29) respectively. LC50 of stem, root, aerial parts, fruits, and flowers essential oils against larvae of An. stephensi were equal with 10. 46, 22. 27, 20. 50, 31. 93 and 79. 87ppm respectively. In antimicrobial activities, essential oil was effective on all specimens except Escherichia coli, Asper-gillus niger and Candida albicans. Conclusion: The essential oil showed moderate antioxidant activity, strong antimicrobial properties and good toxic effect in brine shrimp test and MTT assay on three cancerous cell lines.