We studied the antioxidant activities of the essential oils of Zataria multiflora Boiss. (ZEO) and Bunium persicum (Boiss.) B. Fedtsch (BEO) in mixed form on linseed oil using (DPPH.), (ABTS.+), H2O2 scavenging, and reducing power assays. After calculation of IC50 for ZEO and BEO separately, interactions of the essential oils were investigated at the form of mixture and the results were given in isobologram. The interactions between antioxidant effects of ZEO, BEO, TBHQ, andα-tocopherol; and isobologram results showed synergistic effect for DPPH. except for BEO with TBHQ and a-tocopherol (1: 1) and for ABTS+ in ZEO with TBHQ (1: 1). However, we could not find any synergistic effect for H2O2 scavenging and reducing power assays in any of the interactions. Statistical results showed that the best antioxidant levels of reductive oxidation were 600 ppm for ZEO and BEO, and 20 ppm for TBHQ in mixed form in linseed oil.

Background and objectives: The leishmaniases are considered among the major infectious diseases affecting public health in several regions. There are many chemical agents which are effective in treatment of visceral leishmaniasis. But, overall treatment of visceral leishmaniasis is often difficult. Thus, identification of new chemotherapeutic agents is important for treatment of disease. Since targeting of the ergosterol synthesis pathway of Leishmania may be useful therapeutically, the aim of this study was to investigate the effect of alone or in combination of amiodarone and ketoconazole on Leishmania infantum.Methods: To obtain logarithmic promastigotes of L. infantum, the parasites were cultured in BHI medium with FCS 10% together with antibiotics of penicillin and streptomycin and incubated at 24oC. Amastigote forms were obtained in BHI medium supplemented with 20% FCS at pH of 5.5 which incubated in 37oC. L.infantum susceptibility to amiodarone and ketoconazole was evaluated by proliferation of parasites in the absence or presence of these drugs with MTT assay. For evaluation of antiproliferative synergism against promastigotes and axenic amastigotes, fractional inhibitory concentrations (FIC) were calculated. An isobologram curve was constructed too.Results: Amiodarone produced a marked reduction in the viability of L.infantum promastigotes and axenic amastigotes. On the other hand ketoconazole induced a dose dependent effect on the parasites proliferation for promastigotes and axenic amastigotes. When the drugs were used in combination, the results indicated clear synergistic as shown by a concave isobologram and FIC value.Conclusion: The present study represents the evidence that the combination of amiodarone plus ketoconazole acts synergistically in controlling L. infantume in vitro. It is possible that amiodarone could be used in combination with ketoconazole to combat infection at low doses, thus reducing its side effects such as cardiotoxicity, thyroid dysfunction and pulmonary fibrosis.

Purpose: Currently, there is a crucial need for alternative strategies to control leishmaniasis, which threatens more than 1 billion people worldwide. The simultaneous use of combination therapy and nanostructured lipid carriers aimed to assess the leishmanicidal activity of silver and dapsone niosomes co-administration in vitro and in silico. Methods: After preparing the niosomal formulations of dapsone and silver using the film hydration method, Span 40 and Tween 40/cholesterol with a 7/3 molar ratio was selected as the optimal formulation. Consequently, the arrays of experimental approaches were conducted to compare the anti-leishmaniasis efficacy of ready niosomes with amphotericin B and obtain a deeper understanding of their possible mechanisms of action. Results: Our findings showed higher potency of silver-loaded and dapsone-loaded niosomes co-administration compared to amphotericin B as a positive control group. The results of isobologram and combination index (CI) analyses confirmed the synergic potential of this mixture. This combination triggered anti-leishmanial pathways of macrophages, which promoted the expression level of Th1 cell-related genes, and the downregulated expression of the phenotypes related to Th2 cells. Furthermore, a high level of antioxidant and apoptotic profiles against the parasite provides a rational basis and potential drug combination for cutaneous leishmaniasis. Moreover, the outcomes of the molecular docking showed a high binding affinity between dapsone and iNOS, stimulating the immune response in Th1 direction. Conclusion: In general, the multifunctional leishmanicidal activity of dapsone and silver-niosomes co-administration should be considered for further in vivo and clinical studies.

Purpose: The phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin (PI3K/AKT/ mTOR) pathway is a complex intracellular metabolic pathway that leads to cell growth and tumor proliferation and plays a key role in drug resistance in breast cancer. Therefore, the anti-cancer effects of oleanolic acid (OA), maslinic acid (MA), and their combination were investigated to improve the performance of the treatment strategy. Methods: We investigated the effect of OA and MA on cell viability using the WST-1 method. The synergistic effect of the combination was analyzed by isobologram analysis. In addition, the effects of the two compounds, individually and in combination, on apoptosis, autophagy, and the cell cycle were investigated in MCF7 cells. In addition, changes in the expression of PI3K/AKT/mTOR genes involved in apoptosis, cell cycle and metabolism were determined by quantitative RT-PCR. Results: MA, OA, and a combination of both caused G0/G1 arrest. Apoptosis also increased in all treated groups. The autophagosomal LC3-II formation was induced 1. 74-fold in the MAtreated group and 3. 25-fold in the MA-OA-treated group. The combination treatment resulted in increased expression of genes such as GSK3B, PTEN, CDKN1B and FOXO3 and decreased expression of IGF1, PRKCB and AKT3 genes. Conclusion: The results showed that the combination of these two substances showed the highest synergistic effect at the lowest dose and using MA-OA caused cancer cells to undergo apoptosis. The use of combination drugs may reduce the resistance of cancer cells to treatment.

Background and Purpose: Fluconazole resistance in Candida species is on the rise, posing a significant clinical challenge. There is a growing interest in using complementary therapies, especially those from natural sources. This study aimed to evaluate the synergistic and apoptotic effects of Bunium persicum essential oil (BPEO) and its two pure components, cuminaldehyde (CA) and γ-terpinene (γ-TPN), combined with fluconazole (FLC) on susceptible and resistant C. albicans isolates. Moreover, molecular docking was used to study the interactions between lanosterol 14-alpha-demethylase and each agent.Materials and Methods: The BPEO was prepared using the Clevenger apparatus and the hydro-distillation method. The in vitro antifungal activity was evaluated according to the Clinical and Laboratory Standards Institute guideline (M60). The checkerboard and isobologram assays assessed the interaction between BPEO, CA, γ-TPN, and FLC. The necrotic and apoptotic effects of different agents were analyzed using a flow cytometry assay. An in-silico study was performed to examine the receptor-ligand interaction.Results: The CA showed the lowest minimum inhibitory concentrations and minimum fungicidal concentrations, compared to BPEO and γ-TPN.  Statistical analyses indicated significant differences between resistant and sensitive C. albicans isolates regarding minimum inhibitory concentration values of BPEO, CA, and γ-TPN. The most synergistic effect was obtained for FLC combined with CA (n=7, 63.6%), followed by BPEO (n=6, 54.5%), and γ-TPN (n=3, 27.2%). Statistical analyses indicated the synergistic effect of FLC in combination with CA was more than γ-TPN (p=0.023). Apoptotic indicators confirmed that the tested compounds could cause cell death in yeast cells. Combination of each natural component with FLC resulted in a greater apoptosis effect than each tested agent alone. The docking study indicated that both pure compounds have interactivity with the protein residue of 14α-demethylase.Conclusion: The results indicated that the synergistic properties of natural products combined with synthetic antifungal agents available in the market could contribute to developing effective therapeutic strategies, particularly in resistant fungal species.