Serine Palmitoyltransferase SPT is the key enzyme in the de novo sphingolipids biosynthesis pathway in eukaryotes, including the intracellular parasite Leishmania. Previous studies showed that this enzyme SPT is expressed only in divided promastigote forms and it is non-essential in the amastigotes form of Leishmania major, which is known as the old world leishmaniasis. In this study we have studied the viability of new world lesihamniasis, Leishmania mexicana. Cytotoxicity test used to determine the effect of the SPT inhibitor myriocin which did not significantly affect the viability of the two forms of the in vitro cultures of the parasite p<0.05, procyclic promastigotes and amastigotes, in which cell viability for miltefosine treatment 100, 50, 25, 12.5, 6.25µM against procyclic promastigotes form was 133.7, 115.8, 103.7, 94.98, 94.78 respectively for 24 hours incubation, while for amastigotes, cell viability for miltefosine treatment was 114.77, 114.34, 104.21, 132.95, 102.74 respectively for the 24 hours incubation and was 81.46, 81.25, 71.40, 68.38, 66.51 respectively for the 48 hours incubation and was 78.99, 90.22, 85.59, 95.18, 84.43 respectively for the 72 hours incubation. While in the old world leishmaniasis, Leishmania major, it has been found that ceramide production is absent and nonessential for the proliferation of intra-amastigotes.
Leishmania are protozoan parasites belonging to the family Trypanosomatidae that cause high morbidity and mortality levels with a wide spectrum of clinical syndrome. This study aimed to investigate the effect of liposomal amphotericin B (AmBisome) drug on promastigote and axenic amastigote stages of Leishmania tropica. From the 20 isolates of cutaneous leishmaniasis collected from patients attended to the AL-Karama Teaching Hospital in Baghdad during the period from October 2013 until February 2014, only three isolates successfully transformed to motile promastigote stage in the culture media. The most active one is included in this study. Different concentrations of liposomal amphotericin B (AmBisome) and pentostam Sb (V) drugs were inv
... Show MoreLeishmaniasis is a worldwide disease still treated with expensive compounds that present severe side effects, and are frequently ineffective emphasizing the importance to search effective compounds against this disease. Miltefosine drug (HePC) that used as antitumor agent has been used against Leishmania tropica in two forms promastigote and axenic amastigote in vitro conditions. Different concentrations (5, 10, 15, 20, 25 and 30 μM) of HePC were performed and exposed to both parasite forms in comparison to sodium stibogluconate (Sb) drug. Parasites viability then was determined using MTT assay after 12, 24, and 48hr of exposure. DNA was extracted from treated and untreated parasites after 48hr of exposure and qualitative analysis of th
... Show MoreLeishmania are protozoan parasites belonging to the family Trypanosomatidae that cause high morbidity and mortality levels with a wide spectrum of clinical syndrome. This study aimed to investigate the effect of liposomal amphotericine B (AmBisome) drug on promastigote and axenic amastigote stages of Leishmania donovani isolate (MHOM/IQ/2005/MRU15)in comparison with pentostam SbV drug. Different concentrations of AmBisome and SbV drugs were investigated against Leishmania donovani promastigote and axenic amastigote. The ICR50R values of SbV and AmBisome drugs on promastigote were10.12 mg/ml and 2.21μg/ml, respectively, while they were 0.77μg/ml for axenic amastigote for both drugs. The present study concluded that axenic amastigote was
... Show MoreSphingolipids (SLs) are major structural constituents of eukaryotes, including the kinetoplastid parasite Leishmania. SLs are important for cellular trafficking and signaling and participate in different cell functions, such as, differentiation and cell death (apoptosis). In this study we have investigated the viability of Leishmania major wild type (W.T) and L. major knockout LmLCB2, one of two subunits of serine palmitoyl transferase (SPT) after treatment with myriocin (potent inhibitor of SPT) in order to detect the survival and proliferation of the parasites in vitro. This is to focus on the de novo sphingolipids biosynthesis pathway in both Leishmania wild type which can synthesize SPT and knockout Leishmania which genetically ablated
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Leishmania species are the causative agent of a tropical disease known as leishmaniasis. Previous studies on the old world species Leishmania major, showed that the amastigotes form which resides inside the macrophage of the vertebrate host, utilize host’s sphingolipids for survival and proliferation. In this study, gene expression of serine palmitoyltransferase (SPT) subunit two (MmLCB2) of the mouse macrophage cell line (RAW264.7), which is the first enzyme in the de novo sphingolipid biosynthesis, was detected in both infected and non-infected macrophages. This was detected under condition where available sphingolipid was reduced, with the new world species Leishmania mexicana. Results of qPCR analysis showed that there was no differen
... Show MoreLeishmania causes disease ranging from self-healing cutaneous to fatal visceral leishmaniasis (VL). Leishmaniasis is reported endemic in 88 countries, including Iraq, in which 82% in low-income countries. The diseases develop following the bite of sand flies injecting Leishmania promastigotes into skin. Promastigotes transform into amastigotes in vivo multiplying within macrophages. In this study we have investigated the ability of axenic procyclic promastigotes of cutaneous Leishmania tropica and visceral Leishmania donovani survive in M199 media with or without Fetal Bovine Serum (FBS) added. Three time incubation periods were adopted (24, 48 and 72) hours and the results showed that L. tropica was able to survive and multiply in both
... Show MoreLeishmaniasis is one of the neglected parasitic diseases, which belongs to the family Trypanosomatidae. Cutaneous leishmaniasis is endemic in Iraq and the available drugs are of side effect or resistant by the parasite. In this study, cytotoxicity of methotrexate was investigated on the promastigotes proliferation of the Iraqi strain ofL.tropica.The results showed a significant (p ≥ 0.05) difference in growth of treated groupsat all concentration (1000, 500, 250, 125.5, 62.5, 31.25, 15.6) μM, after 24 and 48 hours of follow up, while after 72 hours, significant difference was observed at concentration(1000, 125, 62.5) μM.The IC50 measured after 24and 48 hours and it was 40.366 and 44.452 μM, respectively.The present study showed
... Show MoreCutaneous leishmaniasis (CL) is the most form of leishmaniasis disease prevalent in Iraq. CL remains a public health problem in numerous endemic countries because of the absence of safe, effective, and high-cost drugs for treatment. Macrophages are the main inhabitant cell for Leishmania; they phagocyte and allow parasite multiplying. Phagocytosis and anti-leishmanial activity of macrophage are the main factors in the elimination of Leishmania parasites. Phagosome-resident amastigotes also evade innate host defense mechanisms. Silver nanoparticles (Ag NPs) have an important effect in stimulating the production of oxygen species. The objective of this study was to examine macrophages cytotoxicity upon exposure to L. tropica and Ag NPs. Se
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