DETC Induces Leishmania Parasite Killing in Human In Vitro and Murine In Vivo Models: A Promising Therapeutic Alternative in Leishmaniasis

Background: Chemotherapy remains the primary tool for treatment and control of human leishmaniasis. However, currently available drugs present serious problems regarding side-effects, variable efficacy, and cost. Affordable and less toxic drugs are urgently needed for leishmaniasis. Methodology/Principal Findings: We demonstrate, by microscopy and viability assays, that superoxide dismutase inhibitor diethyldithiocarbamate (DETC) dose-dependently induces parasite killing (p,0.001) and is able to ??????sterilize?????? Leishmania amazonensis infection at 2 mM in human macrophages in vitro. We also show that DETC-induced superoxide production (p,0.001) and parasite destruction (p,0.05) were reverted by the addition of the antioxidant N-acetylcysteine, indicating that DETC-induced killing occurs through oxidative damage. Furthermore, ultrastructural analysis by electron microscopy demonstrates a rapid and highly selective destruction of amastigotes in the phagosome upon DETC treatment, without any apparent damage to the host cell, including its mitochondria. In addition, DETC significantly induced parasite killing in Leishmania promastigotes in axenic culture. In murine macrophages infected with Leishmania braziliensis, DETC significantly induced in vitro superoxide production (p = 0.0049) and parasite killing (p = 0.0043). In vivo treatment with DETC in BALB/C mice infected with Leishmania braziliensis caused a significant decrease in lesion size (p,0.0001), paralleled by a 100-fold decrease (p = 0.0087) in parasite burden. Conclusions/Significance: Due to its strong leishmanicidal effect in human macrophages in vitro, its in vivo effectiveness in a murine model, and its previously demonstrated in vivo safety profile in HIV treatment, DETC treatment might be considered as a valuable therapeutic option in human leishmaniasis, including HIV/Leishmania co-infection

Interference with Hemozoin Formation Represents an Important Mechanism of Schistosomicidal Action of Antimalarial Quinoline Methanols

Heme is an essential molecule to most living organisms, but once in a free state it exerts toxic effects. Blood-feeding organisms evolved efficient ways to detoxify free heme derived from hemoglobin digestion. A key mechanism present in some hematophagous organisms consists of the crystallization of heme into a pigment named hemozoin. Schistosoma mansoni is one of the etiologic agents of human schistosomiasis, a parasitic disease that affects over 200 million people in tropical and subtropical areas. Hemozoin formation represents the main heme detoxification pathway in S. mansoni. Here, we report that the antimalarial quinoline methanols quinine and quinidine exert schistosomicidal effects notably due to their capacity to interfere with hemozoin formation. When quinine or quinidine were administered intraperitoneally during seven days to S. mansoni-infected mice (75 mg/kg/day), both worm and eggs burden were significantly reduced. Interestingly, hemozoin content in female worms was drastically affected after treatment with either compound. We also found that quinine caused important changes in the cellular organization of worm gastrodermis and increased expression of genes related to musculature, protein synthesis and repair mechanisms. Together, our results indicate that interference with hemozoin formation is a valid chemotherapeutic target for development of new schistosomicidal agents

HD 46375: seismic and spectropolarimetric analysis of a young Sun hosting a Saturn-like planet

HD 46375 is known to host a Saturn-like exoplanet orbiting at 0.04 AU from its host star. Stellar light reflected by the planet was tentatively identified in the 34-day CoRoT run acquired in October-November 2008. We constrain the properties of the magnetic field of HD 46375 based on spectropolarimetric observations with the NARVAL spectrograph at the Pic du Midi observatory. In addition, we use a high-resolution NARVAL flux spectrum to contrain the atmospheric parameters. With these constraints, we perform an asteroseismic analysis and modelling of HD 46375 using the frequencies extracted from the CoRoT light curve. We used Zeeman Doppler imaging to reconstruct the magnetic map of the stellar surface. In the spectroscopic analysis we fitted isolated lines using 1D LTE atmosphere models. This analysis was used to constrain the effective temperature, surface gravity, and chemical composition of the star. To extract information about the p-mode oscillations, we used a technique based on the envelope autocorrelation function (EACF). From the Zeeman Doppler imaging observations, we observe a magnetic field of ~5 gauss. From the spectral analysis, HD 46375 is inferred to be an unevolved K0 type star with high metallicity [Fe/H]=+0.39. Owing to the relative faintness of the star (m_hip=8.05), the signal-to-noise ratio is too low to identify individual modes. However, we measure the p-mode excess power and large separation Delta nu_0=153.0 +/- 0.7 muHz. We are able do constrain the fundamental parameters of the star thanks to spectrometric and seismic analyses. We conclude that HD 46375 is similar to a young version of Alpha-CenB. This work is of special interest because of its combination of exoplanetary science and asteroseismology, which are the subjects of the current Kepler mission and the proposed PLATO mission.Comment: Accepted in Astronomy & Astrophysics. 8 pages, 9 figure

In vitro influence of photodynamic antimicrobial chemotherapy on staphylococcus aureus by using phenothiazines derivatives associated with laser/LED Light

ABSTRACT The aim of this study was to evaluate the effect of photodynamic antimicrobial chemotherapy (PACT) using phenothiazinium dyes -PTZ irradiated with red laser (ʎ660nm) or red-orange LED (ʎ632±2nm) on Staphylococcus aureus in vitro. triplicate tests were performed in 10 groups: control, Laser (L1 + P -and L2 + P -) bacterial suspensions were irradiated only with laser energy 2.4 and 4.8 J/cm 2 respectively, (Led1 + P -and Led2 + P -) irradiated only with LED energy 2.4 and 4.8 J/cm 2 respectively, (L1 + P + and L2 + P + ) irradiated with laser in the presence of 1μg/ml of photosensitizer, (Led1 + P + and Led2 + P + ) irradiated with LED in the presence of 1μg/ml of photosensitizer and finally (L -P + ) only in the presence of PTZ dye. Bactericidal effect of the PACT was assessed by counting colony-forming units. The results showed no significant difference on regards different energy densities on group PACT for both lights. PACT groups (L2 + P + and Led2 + P + ) compared to the Control showed significant reduction of CFUs. LED/Laser groups (L2 + P -and Led2 + P -) compared to control and PTZ groups showed also significant differences as groups LED/Laser (4.8J/cm 2 ) increased the average of CFUs. Although the results of this study have shown a reduction in average number of colonyforming units by the appropriate Laser or LED-dye treatment combination, it this topic requires further investigation

Histamine Derived from Probiotic Lactobacillus reuteri Suppresses TNF via Modulation of PKA and ERK Signaling

Beneficial microbes and probiotic species, such as Lactobacillus reuteri, produce biologically active compounds that can modulate host mucosal immunity. Previously, immunomodulatory factors secreted by L. reuteri ATCC PTA 6475 were unknown. A combined metabolomics and bacterial genetics strategy was utilized to identify small compound(s) produced by L. reuteri that were TNF-inhibitory. Hydrophilic interaction liquid chromatography-high performance liquid chromatography (HILIC-HPLC) separation isolated TNF-inhibitory compounds, and HILIC-HPLC fraction composition was determined by NMR and mass spectrometry analyses. Histamine was identified and quantified in TNF-inhibitory HILIC-HPLC fractions. Histamine is produced from L-histidine via histidine decarboxylase by some fermentative bacteria including lactobacilli. Targeted mutagenesis of each gene present in the histidine decarboxylase gene cluster in L. reuteri 6475 demonstrated the involvement of histidine decarboxylase pyruvoyl type A (hdcA), histidine/histamine antiporter (hdcP), and hdcB in production of the TNF-inhibitory factor. The mechanism of TNF inhibition by L. reuteri-derived histamine was investigated using Toll-like receptor 2 (TLR2)-activated human monocytoid cells. Bacterial histamine suppressed TNF production via activation of the H2 receptor. Histamine from L. reuteri 6475 stimulated increased levels of cAMP, which inhibited downstream MEK/ERK MAPK signaling via protein kinase A (PKA) and resulted in suppression of TNF production by transcriptional regulation. In summary, a component of the gut microbiome, L. reuteri, is able to convert a dietary component, L-histidine, into an immunoregulatory signal, histamine, which suppresses pro-inflammatory TNF production. The identification of bacterial bioactive metabolites and their corresponding mechanisms of action with respect to immunomodulation may lead to improved anti-inflammatory strategies for chronic immune-mediated diseases

Blood-Feeding Induces Reversible Functional Changes in Flight Muscle Mitochondria of Aedes aegypti Mosquito

Background: Hematophagy poses a challenge to blood-feeding organisms since products of blood digestion can exert cellular deleterious effects. Mitochondria perform multiple roles in cell biology acting as the site of aerobic energytransducing pathways, and also an important source of reactive oxygen species (ROS), modulating redox metabolism. Therefore, regulation of mitochondrial function should be relevant for hematophagous arthropods. Here, we investigated the effects of blood-feeding on flight muscle (FM) mitochondria from the mosquito Aedes aegypti, a vector of dengue and yellow fever. Methodology/Principal Findings: Blood-feeding caused a reversible reduction in mitochondrial oxygen consumption, an event that was parallel to blood digestion. These changes were most intense at 24 h after blood meal (ABM), the peak of blood digestion, when oxygen consumption was inhibited by 68%. Cytochromes c and a+a3 levels and cytochrome c oxidase activity of the electron transport chain were all reduced at 24 h ABM. Ultrastructural and molecular analyses of FM revealed that mitochondria fuse upon blood meal, a condition related to reduced ROS generation. Consistently, BF induced a reversible decrease in mitochondrial H2O2 formation during blood digestion, reaching their lowest values at 24 h ABM where a reduction of 51% was observed. Conclusion: Blood-feeding triggers functional and structural changes in hematophagous insect mitochondria, which may represent an important adaptation to blood feedin