The Effect of Alkanna tincturia and Peganum harmala Extracts on Leishmania major (MRHO/IR/75/ER) in vitro

"nBackground: Cutaneous leishmaniasis is an important health problem caused by Leishmania spp. As there is no vaccine, drug treatment is the only way to tackle leishmaniasis. In the present study, inhibitory and kill­ing effects of Peganum harmala and Alkana tinctoria extracts on amastigotes and promastigotes forms of Leishmania were evaluated in-vitro."nMethods: The seeds of Peganum harmala, Stems and roots of Alkanna tictoria were collected and crude extrac­tion carried out. In this experimental study,  Leishmania major promastigotes were cultured in RPMI-1640 with 10% FBS at 22-26°C, and infected macrophages with amastigotes   were cultured in RPMI-1640 with 10% FBS at 37°C in 5% CO2. Then the extracts of each plant were added to cultivated parasites and incubated for 3 days. Promastigote and amastigote assay was carried out using   counting assay based on growth inhibition."nResults: The results indicated that both extractions can inhibit the growth of promastigotes, and in concentra­tions of 40µg/ml of P. harmala , 200µg/ml of A. tincturia, and 20 µg/ml of equal combination of P. hamala and A. tincturia are Inhibitory Concentration (IC50) for parasites growth. By adding these concentra­tions of the extracts to the infected macrophages in the culture, their effects were separately eva­lu­ated. The mean of amastigotes number in macrophages in the culture with P. harmala, A. ticturia, combina­tion and control groups were 0.7, 0.7, 0.6, 2.3 amastigotes per macrophage, respectively."nConclusion: By this method, inhibition of intracellular and extracellular growth of L. major was demon­strated suggesting that, plant drugs with efficacy and safe products can be applied as new treatment for cutane­ous leishmaniasis

Evaluation of Anti-amoebic Activity of Peganum harmala Ethanolic Extract on Acanthamoeba In vitro

Abstract Background: Acanthamoeba is an opportunistic protozoan pathogen that is known to infect the cornea to produce eye keratitis and the central nervous system to produce lethal granulomatous encephalitis. The overall aim of the present study was to determine the anti-amoebic potential of natural compound Peganum harmala against the trophozoites and cysts of Acanthamoeba in vitro. Materials and Methods: In this experimental study, a clinical isolate of Acanthamoeba was cultured and genotyped. The ethanolic extract of Peganum harmala was prepared. The trophozoites and cysts were collected by washing in page's saline. Various concentrations (1.25, 2.5, 5, and 10 mg/ml) of the ethanolic extract and polyhexanide 0.02% drop as positive control were tested at three different times (24, 48 and 72 h) on trophozoites and cysts of Acanthamoeba in vitro. The viability of trophozoites or cysts was tested by eozin method, MTT, and flowcytometry analysis. Results: The results revealed that alcoholic extract had remarkable inhibitory effect on the proliferation of Acanthamoeba cysts as compared to non-treated control, and the inhibition was time and dose dependent. In the presence of 10 mg/ml ethanolic extract in medium culture after 72 h, no viable trophozoites were determined and 21.10% cysts of Acanthamoeba were viable. Percentage of trophozoites and cysts viability after adding polyhexanide 0.02% drop in medium culture after 72 hours was 0% and 23.71%, respectively. Conclusion: Ethanolic extracts of Peganum harmala could be considered a new natural compound against the Acanthamoeba trophozoites and cysts. Further works are required to evaluate the exact effect of this extract on Acanthamoeba agents in animal models

Evaluation of Immunogenicity of Cocktail DNA Vaccine Containing Plasmids Encoding Complete GRA5, SAG1, and ROP2 Antigens of Toxoplasma gondii in BALB/C Mice

Background: Severe and fatal complications of toxoplasmosis urge development of effective vaccines against the disease. The current study was performed to evalu­ate cocktail DNA vaccine containing plasmids encoding GRA5, SAG1, and ROP2 genes of Toxoplasma gondii in BALB/c mice in Tarbiat Modares University in 2012. Methods: The plasmids containing complete GRA5, SAG1, and ROP2 genes were mass extracted and then the recombinant plasmids were administered via intramuscu­lar injections according to immunized mice three times with three-week intervals. Then splenocytes were cultured, and proliferation as well as cytokine as­says were carried out. The other mice in each group were inoculated by the parasite and mortality of the mice was evaluated on a daily basis. Results: The results of cytokine assay for INF-γ were higher in the mice that re­ceived the cocktail DNA containing recombinant plasmids. Evaluation of prolifera­tion of splenocytes using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazo­lium bromide) assay indicated induction of cellular response. Measurement of total IgG and the isotypes of IgG1 and IgG2a showed that the cocktail DNA stimulated IgG and IgG2a production in comparison with the control groups (P<0.05). Furthermore, the survival rate of mice in the groups that received the cocktail DNA was significantly higher than that in the control groups (P<0.05). Conclusion: Administration of the cocktail DNA vaccine led to production of higher levels of IFN-γ, confirmed by secretion of IgG2a, and the immune response was shifted toward Th1. Thus, the cocktail DNA containing the recombinant plas­mids can be an appropriate candidate for immunization against toxoplasmosis