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Three arylcarboxamides issued from 2-amino- 4,6-dimethylpyridine, benzamide 1, furan-2-carboxamide 2 and 5-bromofuran-2-carboxamide 3 were evaluated against Leishmania promastigotes, at three doses, 0.5, 5, and 50 µM. Although benzamide 1 exerted but a moderate inhibition against cultured extracellular promastigotes at a concentration of 50 µM, furan-2-carboxamides 2 and 3 had potent activity at the same concentration. The IC₅₀ of 2 against L. donovani and L. braziliensis were 29.9 and 28.3 µM and those of 3 were 25.9 and 36.6 µM, respectively. In a BALB/c mice model of visceral leishmaniosis, an intraperitoneal administration of 10 mg/kg of 2, for 5 days, led to a consistent parasite burden reduction in the spleen smears (75.8 ± 5.7%) and in the microdilution spleen cultures (72.1 ± 0.6%). A very significant correlation (r = 0.96) was found between the two methods of spleen parasite burden quantification. These amides also exhibit potent antiinflammatory activity and it was previously stated that they inhibit arachidonic acid production by interfering in the PLA₂ activation. This suggests fact that they could disrupt Leishmania membrane lipid integrity by protein kinase C inhibition.
Dysfunction of T-helper 1 mediated immune responses is a hallmark of the progression of visceral leishmaniosis (VL). Several factors such as altered antigen presentation, and abnormalities in MHC/HLA, antigen processing, and T cell receptor recognition regulate the onset of immunosuppression. Recent investigations on VL patients suggest that susceptibility to visceral leishmaniosis is genetically determined and varies between populations in different geographical locations. Emerging evidence also indicates the importance of the role played by myeloid derived suppressor cells in progressive VL. This study provides a mechanistic view of means to target the signaling mechanisms of immunosuppression to determine potential therapeutic interventions.
The therapeutic armentarium against leishmaniasis is very restricted, using mainly up to now drugs showing several side effects and resistance. Consequently, there is a real need to find new compounds as alternatives for the treatment of leishmaniasis. However, most of the classic antileishmanial primary screening assays are not suitable for measuring the cytotoxicity of large number of drug candidates because of the manipulations required. We have established a new assay that incorporate a fluorometric growth indicator based on the detection of a metabolic activity in culture medium after the chemical reduction of alamar Blue® by cells. This antileishmanial test was evaluated on amphotericin B, meglumine antimoniate, allopurinol, ketoconazole and edelfosine. The results reported show the advantages oj this fluorochrome on the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) for Leishmania cytotoxicity measurement. Moreover, this study is the first report of the use of alamar Blue® fluorogenic assay for activity assessment of potential antileishmanial drugs against promastigotes.
The use of toll-like receptor agonists in immunotherapy is a new approach in the prevention of immunosuppression during fatal Leishmania parasite infection. The objective of such immunotherapy is to activate specific cell-mediated immune responses, macrophage activation and antigen-responsive inflammation, to kill intracellular amastigotes. Toll-like receptor agonist-based treatment in immunocompetent hosts can be effective either by selective use of the agonists alone or in combination with the anti-leishmanial drug stibanate. Recent investigations suggest that toll-like receptor signal pathways constitute a possible new mode of anti-leishmanial treatment. This article describes the prospect of toll-like receptor – mediated signal pathways in the immunotherapy of cutaneous and visceral leishmaniosis, as well as post kala-azar dermal leishmaniosis (PKADL), a skin-sequel of visceral infection. Suitable synthetic agonists need to be developed for toll-like receptors to overcome immunosuppression.
Leishmania parasites are the etiological agents of leishmaniosis, with severe course and often fatal prognosis, and the global number of cases has increased in recent decades. The gold standards for the diagnosis of leishmaniosis are microscopic examinations and culture in vitro of the different clinical specimens. The sensitivity of these methods is insufficient. Recent development in specific and sensitive molecular methods (PCR) allows for detection as well as identification of the parasite species (subspecies). The aim of the study was to estimate the usefulness of molecular methods (PCR) for detection of Leishmania species and consequently for the implementation of such methods in routine diagnostics of leishmaniosis in Polish patients returning from endemic areas of the disease. In our investigations we used 54 known Leishmania positive DNA templates (from culture and clinical specimens) received from the CDC (Atlanta, GA, USA). Moreover, 25 samples of bone marrow, blood or other tissues obtained from 18 Polish individuals suspected of leishmaniosis were also examined. In PCR we used two pairs of primers specific to the conserved region of Leishmania kinetoplast DNA (kDNA) minicircle (13A/13B and F/R). Using these primers we obtained amplicons in all DNA templates from the CDC and in three Polish patients suspected for Leishmania infection. In one sample from among these cases we also obtained positive results with DNA isolated from a blood specimen which was previously negative in microscopic examinations.
Cutaneous leishmaniasis is a protozoan infectious disease and widespread in Mediterranean basin including Turkey. Lesions usually start with erythematous papules, gradually enlarges and afterwards it ulcerates. We present a 12-year-old boy with diffuse persistent lip swelling mimicking granulomatous cheilitis. Systemic glucantime was started. However, severe hypotension and bradycardia was developed after injection. Oral fluconazole was started and the lesion resolved completely. Cutaneous leishmaniasis can have varied clinical manifestations and should be suspected especially in endemic areas. Oral fluconazole seems to be safe and effective treatment modality in paediatric cases.
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